Biostat B: Operating Manual

Operating Manual
BIOSTAT® B
BAE_B Rev. 2.32 - 0400 SW 4.1

Operating Manual BIOSTAT B Introduction
Introduction
This Manual applies to B. Braun Biotech International's bench-top fermentor system BIOSTAT® B,
which is one example of B. Braun Biotech International's product program of sophisticated fermenta-
tion and peripheral laboratory equipment. For further information about this fermentor system and our
complete product program please contact
B. Braun Biotech International GmbH
Schwarzenberger Weg 73-79
D - 34212 Melsungen, Fed. Republic of Germany
phone +49 56 61 - 71 34 00,
fax +49 56 61 - 71 37 02
e-mail: bbi.info@bbraun.com
WebSite http://www.bbraunbiotech.com
or your local sales representative of B. Braun Biotech International GmbH.
Guide Through this Manual
The Operating Manual is structured as shown below. It is not necessary to read this manual from the
beginning. Especially experienced users can go straight on to the section of their interest.
Section : Contents
1 : Overview on design and equipment of the fermentor

2 : Delivery and warranty information, installation and connection
3 : Start-up and operation of the fermentor
4 : Maintenance, assembly and equipment of the culture vessels
5 : Detailed operating information about the digital control unit
6 : Technical data
7 : Supplement with additional documentation like technical drawings, etc.
Documentation Release Notes
The Operating Manual refers to the design and equipment of the fermentor valid for the rev. no. and at
the date of print. However, the characteristics and specifications of the BIOSTAT® B are subject to
change. B. Braun Biotech International GmbH reserves the right to modify the equipment and to
change the operating information without notice. B. Braun Biotech International GmbH assumes no ob-
ligation regarding design, ordering data resp. future manufacture unless otherwise agreed to in writing.
The information given herein has been carefully estimated. However, this document is not necessarily
complete. If you happen to find errors or if you miss specific information concerning special parts and
their setup, please do not hesitate to contact your B. Braun Biotech representative or B. Braun Biotech
International GmbH directly.
BAE_B Rev. 2.31 - 1099 SW 4.1

F190101
Return of Material
Declaration about decontamination and

cleaning of equipment and components
When returning equipment or components, please describe on page 2 of this form the problem(s) or
fault(s) you have found. Please also indicate the remedial actions you require.
To protect our personnel, we require all equipment or components be free of biological, chemical, or
radioisotopic contaminants. We will only accept such equipment or components when:
• the equipment or components have been adequately CLEANED and DECONTAMINATED.
• this declaring document has been completed, signed and returned by an authorized person.
Please help us in assuring a safe, hazard-free work environment.
A. Description of the Equipment or Component(s)
Description / Cat. No.: ........................................................................................................................

...........................................................................................................................................................
Serial no.: ...................No. of invoice/delivery note .................................Date of delivery:.................
B. Contamination / Cleaning
Attention: Please specify exactly the biological, chemical, or radioisotopic contaminant.

The equipment was contaminated with:...........................................................................................
.........................................................................................................................................................
.........................................................................................................................................................
Attention: Please describe the cleaning and decontamination procedure/method.
and it has been cleaned and decontaminated by:............................................................................
.........................................................................................................................................................
.........................................................................................................................................................
C. Legally binding declaration

I (we) certify that all information given in this form is correct and complete.
The equipment and components have been adequately decontaminated and cleaned according to
the legal requirements. No chemical or biological or radioisotopic risks remain that can endanger
exposed persons' safety or health.
Company / Institute: .........................................................................................................................
Address / Country:............................................................................................................................
.........................................................................................................................................................
Tel. / Fax (with area code):........................................................./ ....................................................
Name of the authorized person: .......................................................................................................
Position:............................................................................................................................................
Signature / Date: ................................................................................................. /...........................
F190101E.DOC / Rev. 3 / 11.05.2000 Page 1 of 2 BBI-Dr. Wieneke

F190101
Return of Material
D. Reason for return
wrong delivery exchange repair modification other
E. Please describe the problem(s) or fault(s) you have found (for repair)
and/or indicate the remedial actions you require.
.............................................................................................................................
.............................................................................................................................
.............................................................................................................................
.............................................................................................................................
.............................................................................................................................
.............................................................................................................................
.............................................................................................................................
.............................................................................................................................
.............................................................................................................................
.............................................................................................................................
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F. Vom BBI Service auszufüllen (reserved for BBI service)
Bemerkungen: .....................................................................................................
.............................................................................................................................
.............................................................................................................................
.............................................................................................................................
.............................................................................................................................
Please pack the equipment properly and B.Braun Biotech International

send it to your local service representative Technischer Service
or to B.Braun Biotech International, Germany Schwarzenberger Weg 73-79
(carriage paid to receiver). 34212 Melsungen
Germany
F190101E.DOC / Rev. 3 / 11.05.2000 Page 2 of 2 BBI-Dr. Wieneke

Operating Manual BIOSTAT B
Contents
Contents Page
Introduction
Guide Through this Manual
Documentation Release Notes
Declaration about Decontamination and Cleaning of Equipment
and Components
1 Design and Function

1.1 Control Unit 1-1
1.1.1 Thermostate System 1-2
1.1.2 Drive System 1-2
1.1.3 Air and Gas Supply 1-2
1.2 Culture Vessels 1-3
1.3 Integrated Digital Control System 1-4
2 Delivery and Installation

2.1 Unpacking and Checking Completeness of Delivery 2-1
2.2 Warranty Agreements and Customer Service 2-1
2.3 Customer Service 2-2
2.4 Installing the Fermentor System 2-3
2.4.1 Space Requirements 2-3
2.4.2 P&I - Diagram 2-3
2.5 Connection Specifications 2-5
2.5.1 Connectors, tubings, cables and Fittings 2-5
2.5.2 Mains Connection 2-5
2.5.3 Laboratory Connection for Cooling Water Supply and Return 2-6
2.5.4 Connection of Air or Gas Supply 2-7
3 Starting and Operating the BIOSTAT® B

3.1 Preparing and Connection of the Control Unit 3-1
3.1.1 Preparing the Workplace 3-1
3.1.2 Connecting the Thermostate System 3-1
3.1.3 Connecting the Air/Gas Supply System 3-1
3.1.4 Preparing and Connection of Peripheral Equipment for
Measurement and Control 3-1
3.2 Preparing the Culture Vessels and Accessories 3-2
3.2.1 Checking the Equipment 3-2
3.2.2 Assembling and Equipping the Culture Vessels 3-2
3.2.3 Filling the Jacket of the Culture Vessel 3-3
3.2.4 Filling the Culture Vessel 3-3
3.2.5 Preparing and Connecting Addition Agent Bottles 3-4
3.2.6 Sterilization of the Culture Vessel 3-4
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Contents
3.3 Peparing a Fermentation Run 3-5

3.3.1 Overview 3-5
3.3.2 Final Setting-up and Connection of the Culture Vessel for
Fermentation 3-5
3.3.3 Connecting the Probes and Setting the Measurement and
Control System 3-7
3.4 Carrying-Out a Fermentation Run 3-8
3.4.1 Sterility Test 3-8
3.4.2 Inoculating the Culture Vessel 3-8
3.4.3 Adding Sterile Solutions during the Fermentation Run 3-9
3.4.4 Taking Samples 3-9
3.5 End of a Fermentation Run 3 - 10
3.5.1 Harvesting the Product 3 - 10
3.5.2 Cleaning and Maintenance 3 - 10
4 Equipment of the Culture Vessels

4.1 General Information 4-1
4.1.1 Delivery and Starting up 4-1
4.1.2 Inspecting a Culture Vessel and its Accessories 4-1
4.2.1 Dimensions and Design Features of the Culture Vessels 4-2
4.2.2 Set-up, Equipment and Accessories of the Culture Vessels 4-6
4.2.3 Top plate Design and Placement of the Accessories 4-9
4.3 Equipping and Assembling the Culture Vessels 4 - 10
4.3.1 Overview 4 - 10
4.3.2 Disassembling a Culture Vessel 4 - 11
4.3.3 Assembling a Culture Vessel 4 - 11
4.3.4 Fitting of the Standard Equipment inside a Culture Vessel 4 - 12
4.3.5 Baffle Insert and Baffles for Culture Vessels with Stirring Device 4 - 13
4.3.6 Sparger Pipe 4 - 14
4.3.7 Inoculation Connectors (Septum Port Kits) 4 - 15
4.3.8 1-Channel Inoculation Kit 4 - 16
4.3.9 Universal-Adapter 4 - 18
4.3.10 Quadruple Addition Port B 4 - 18
4.3.11 Reference Thermometer 4 - 19
4.3.12 Harvesting Pipe 4 - 20
4.3.13 Harvest Filter for Removal of Medium at Microcarrier-Cultures 4 - 21
4.3.14 Spinfilter 4 - 22
4.3.15 Silicone Membrane Insert 4 - 24
4.3.16 Blind Closures 4 - 26
4.4 Probes and Electrodes 4 - 27
4.4.1 General Notes 4 - 27
4.4.2 Pt-100 - Sensor for Temperature Measurement 4 - 27
4.4.3 Antifoam and Level Probes 4 - 28
4.4.4 pH - Electrode 4 - 30
4.4.5 pO2 - Electrode 4 - 32
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Contents
4.5 External Equipment for Connection to the Culture Vessels 4 - 34

4.5.1 Exhaust Cooler 4 - 34
4.5.2 Sterile Filters for Air/Gas Supply and Exhaust 4 - 36
4.5.3 Corrective Solution Bottles 4 - 38
4.5.4 By-pass Sampler Kit 4 - 39
4.5.5 Manual Sampling Kit 4 - 41
5 Control Unit
5.1 General Information 5-1
5.1.1 Operating Behaviour 5-1
5.1.2 Switching the BIOSTAT® B „Off“ and „On“ 5-1
5.1.3 Power Failure 5-1
5.1.4 Menu Structure 5-2
5.2 Operation of the Control Unit 5-3
5.2.1 General Operating Information 5-3
5.2.2 Basic Menu Structure 5-4
5.2.3 Operating Information 5-4
5.3 Main Function „PROCESS VALUES“ 5-5
5.4 Main Function „CALIBRATION“ 5-7
5.4.1 Calibration of the pH - Electrode 5-7
5.4.2 Calibration of the pO2- Electrode 5 - 11
5.4.3 Pump Calibration 5 - 14
5.5 Main Function „Control Loops“ 5 - 16
5.5.1 Extent of Controller Functions of the BIOSTAT® B 5 - 16
5.5.2 Controller Operation in General 5 - 17
5.5.3 Parametrization of Controllers in General 5 - 17
5.5.4 PID - Controller Optimization 5 - 18
5.5.5 Temperature Control 5 - 19
5.5.6 Stirrer Speed Control 5 - 21
5.5.7 pH - Control 5 - 22
5.5.8 pO2 - Control 5 - 24
5.5.9 Foam Control 5 - 27
5.5.10 Level Controller 5 - 29
5.5.11 Controller for Substrate Feed SUBS1 und SUBS2 5 - 30
5.5.12 Airflow Controller 5 - 32
5.6 Main function "Maintenance" 5 - 33
5.6.1 Recorders 5 - 33
5.6.2 Printers 5 - 34
5.6.3 Utility Functions 5 - 37
5.6.4 Measurement Ranges 5 - 38
5.6.5 Manual Operation of Digital Outputs, Standard 5 - 39
5.6.6 Manual Operation of Analog Outputs, Standard 5 - 40
5.7 Alarm Messages of System Functions 5 - 41
5.8 Factory Settings 5 - 42
5.8.1 General Information 5 - 42
5.8.2 Factory Settings for Measurement Ranges of Process Values 5 - 42
5.8.3 Factory Settings for Controller Parameters 5 - 42
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Contents
5.8.4 Example: pO2-Controller with Airflow - Massflow Controller and

O2 - Enrichment 5 - 43
5.8.5 Schematic Drawing „Pump Logic of DFC - Software“ 5 - 45
5.8.6 Arrangement Plan Control Unit of BIOSTAT® B 5 - 46
5.8.7 Assignment Plan for Interfaces of the Control Unit BIOSTAT® B 5 - 46
6 Technical Data
6.1 Dimensions 6-1
6.2 Installations 6-1
6.3 Control unit 6-1
6.4.1 Culture Vessel Type B 2 6-2
6.4.4 Culture Vessel B 10, Special Version for Small Working Volumes 6 - 8
6.4.5 Culture Vessel B 2 - Airlift 6-9
6.4.6 Culture Vessel B 5 - Airlift 6 - 11
6.5 Additional Equipment 6 - 13
6.6 Consumables 6 - 13
6.7 Control Unit 6 - 14
7 Optional Equipment and Accessories
This section intentionally left free in this version of the manual.
8 Supplement
8.1 Conversion Table for Degrees of Hardness of Water 8-1
8.2 Characteristic Values for Gases 8-1
8.3 Calculation Methods for Flowrates of the Rotameters 8-2
8.3.1 Sizing of Flowmeters / Rotameters 8-2
8.3.2 Calculation Method 8-2
8.3.3 Manufacturer's Examples for Sizing Calculations 8-3
8.4 Additional Drawings and Flow Charts 8-6
8.4.1 P & I - Diagram of the BIOSTAT® B 8-6
8.4.2 Drawings and Flow-Charts of Fermentor Options or Customized
Fermentors 8-6
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Operating Manual BIOSTAT B 1
Design and Function
1 DESIGN AND FUNCTION
The BIOSTAT® B is a compact, high quality laboratory fermentor system for small scale fermentation
applications. It is especially adapted to the demands of biotechnology research in universities and in
industrial research laboratories. The BIOSTAT® B consists of the following main components :
Compact control unit with integrated digital measurement and control unit.
The digital measurement and control unit for comfortable control of the BIOSTAT® B. Its
functions are specially adapted to the requirements of economic fermentor operation.
Stirrer driven and airlift type culture vessels offering these features:
– Jacketed glass vessels
– Working volumes of 2 l and 5 l (all vessel types), optional of 10 l (stirrer driven type only)
– Internal concave bottom section for optimum mixing even at low stirring speeds
For comfortable handling and transport the vessels are installed in a support which also car-
ries holders for the storage bottles of addition solutions. A culture vessel will be placed be-
sides the control unit, thus minimizing the space requirements of the entire unit.
1.1 Control Unit
The control unit contains the thermostate system and all installations required for power supply, the
supply of cooling water, pressurized air and the waste water removal. All electrical components are
placed in a separate part of the control unit and are protected against spray or splashing water.
The upper part of the control unit contains the operating terminal of the control system. It is
designed in inclined arrangement for easy operation of the terminal.
The lower front panel of the control unit includes the mains control switch, the fill-
thermostate switch and a flowmeter for the control of the air supply.
On left hand of the front panel four peristaltic pumps are built-in, providing short distance to
either the storage vessels and the culture vessel connections for the tubig connections.
On the left side of the control unit, directed towards the side where the culture vessel is usu-
ally placed, all connectors for the supplies of the culture vessels are integrated :
– Thermostate in-/outlet
– Cooling water in-/outlet
– Air inlet with additional O2 – inlet for O2-enrichment operation, air outlet
– Exhaust cooler in-/outlet
– Mains supply and fuse
– Power supply to stirring motor
– Sockets for electrodes (temperature, foam, level, pH and pO2)
– Sockets for peripheral units, such as recorders, printers, Gasmix-unit or host computer.
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Design and Function
1.1.1 Thermostate System
The thermostate system is an open, pressure-free system. It includes a electric heater of 600 or 800 W
heating capacity, which supplies the necessary energy to the system, and a solenoid valve for cooling
water supply. The circulation pump 2.4 (see P&I diagram in section 2 and in the supplement) delivers
water of the preadjusted temperature to the culture vessel.
The operation temperature is set at the control panel. The digital measurement and control system en-
sures a precise and constant temperature control. The minimum temperature in the culture vessel is
about 8 degC above cooling water temperature (for a culture temperature of up to 60 degC).
Cooling water will only be supplied when the temperature needs to be decreased. This design of the
BIOSTAT® B minimizes the cooling water consumption. Under usual operating conditions, i.e. a fer-
mentation temperature below 50 degC, only a minimum amount of water will be spent. If tap water
cannot provide sufficient cooling, external cooling water supplies can be connected.
1.1.2 Drive System
The drive sytem includes a 180 W DC electronic motor. This motor is directly mounted onto the stirrer
shaft in the vessel top-plate via an elastic coupling. Usually two 6-bladed disk impellers are mounted to
the stirrer shaft. However, the standard impellers may be replaced by optional stirrers of other design,
if required, see section 4 for availability and mounting of different types of stirrers.
The default speed range is 50 to 1.200 rpm. For special applications, such as tissue cell cultures,
which are sensitive to shear forces and require gentle stirring, and if the tubing insert for bubble free
gas supply is used, an optional slow speed drive is available (cat.-no. 884 470/4, speed range
20 ... 600 rpm, installation exworks only). The DC motor and speed control system are designed to
guarantee a high speed constancy even in case of large torque variations, as may result from intensive
gassing or by changes of the medium viscosity during the growth of the culture.
1.1.3 Air and Gas Supply
The BIOSTAT® B can be connected to any precontrolled laboratory gas supply installation. Required
flow of air (or inlet gas) is up to 10 l/min at a max. pressure of 0.8 barg (11.6 psig). An additional O2 -
inlet connector allows for connection of a second gas containing oxygen. If no extra oxygen supply
should be connected, the O2-inlet is combined with the air inlet by a corresponding pipe adapter.
Optionally a Gas Mixing System is available for supply of a gas mixture of up to 4 gases (air, oxygen-
enriched gas, nitrogen or CO2, for instance). Please call B. Braun Biotech International GmbH for de-
tailed information, if you require such a device. If your BIOSTAT® B is delivered with a Gas Mixing
System, please note the corresponding Operating Manual.
The flowrate required for aeration of the culture will be adjusted at a rotameter on the the front panel of
the control unit. Membrane filters provide for a sterile gas supply and exhaust of the culture vessel. The
air (or inlet gas) will be introduced into the culture broth via a sparger ring.
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Design and Function
1.2 Culture Vessels
There are different types of culture vessels available for the BIOSTAT® B:
Stirrer driven culture vessels type B 2, B 5 and B 10 offering max. working volumes of 2 l, 5 l
and 10 l, respectively
Airlift type culture vessels B 2 - Airlift and B 5 - Airlift with internal loop system offering max.
working volumes of 2 l and 5 l, respectively.
The culture vessels are made of borosilicate glass and the standard stirrer driven vessels have a
height / diameter ratio of about 2 : 1 (2.5 : 1 with the vessel B 10). All vessels are heated via the jacket.
Connecting of the thermostate system to the culture vessel is made with special quick couplings. The
inlet connector of the jacket (lower connector) has a self-closing male adapter. The upper outlet con-
nector has a female adapter which is open. This design ensures that the jacket is kept pressureless
when the culture vessel is disconnected from the control unit and allows for pressure compensation
with the ambient pressure when the vessel is autoclaved. Thus overpressure cannot occur in the jacket
during autoclaving, which may destroy the glass vessel.
The culture vessels type B 2 and B 5 have 4 integrated side entry ports in the head space for access to
the interior culture vessel. Through these side entry ports, corrective solutions can be supplied to the
culture during the fermentation run, for instance.
The top-plates are mounted to the flanged ring of the glass vessels with knurled screws. Each top-
plate contains ports of 19, 12 and 6 mm for assembly of electrodes and accessories, see section 4 of
this manual for further details. In addition 4 tubing connectors are welded into the top plate.
The vessels have an internal concave

bottom section. This is a vessel design
which is proven in fermentations of mi-
croorganisms and in the cultivation of
animal cells. All parts in contact with
the medium are made of stainless
steel (1.4571) and the O-rings consist
of EPDM (ethylene propylene rubber).
The culture vessels are mounted into a

supporting frame. Two handles allow
easy transport of the vessels. The
holders of the storage bottles for addi-
tion solutions are welded to the bottom
support. This allows easy assembling
and comfortable handling.
When the culture vessel is placed in

the autoclave the max. height of the
culture vessel together with all equip-
ment and the exhaust cooler deter-
mines the required dimensions for the
autoclave. On request the exhaust
cooler is available with a flexible con-
nection piece, which can be bent over.
Fig. 1-1: Culture vessel type B 2 of the BIOSTAT® B
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Design and Function
1.3 Integrated Digital Control System
The BIOSTAT® B has an digital measurement and control system which offers a defined set of func-
tions for a broad range of standard fermentation applications. The measurement and control system is
integrated in the control unit. Operating data are entered at the functional keyboard of the operator
terminal in the upper front panel of the control unit. Process data will be indicated on an easy to read
alphanumerical display.
The hardware consists of a highly integrated microprocessor board which carries the measurement
amplifiers and actuator controls. The microprocessor system includes a 80C188 processor, 128 kByte
RAM and 128 kByte EPROM. The board has 8 analog inputs, 8 analog outputs, 8 digital inputs and 16
digital outputs.
The software offers all functions necessary for fermentor operation, such as measurement and control
of the parameters, calibration of electrodes and the intergrated pumps and for the signal transfer
from/to peripheral equipment, such as recorders, printers and host computer for further data process-
ing. The software version which this manual refers to, is revision 4.1.1-1)
Standard measurement and control functions are for temperature, foam, level, substrate supply, pH-
value, partial pressure of oxygen (pO2) and control of a gasmix unit. The extent of measurement and
control functions implemented in a customized fermentor version can be different, according to the
specifications aggreed to in the order.
After power down the system automatically restarts with the settings before. If the interupt takes longer
than an adjustable interupt time (FAILTIME), the system proceeds as with switching off by the mains
switch, i.e. the system restarts with a definable state of operation.
1-1) If you have a control unit with an elder or more actual software version, please contact B. Braun
Biotech International GmbH for a corresponding version or update of the Operating Manual.
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Delivery and Installation
2 DELIVERY AND INSTALLATION
2.1 Unpacking and Checking Completeness of Delivery
The BIOSTAT® B is delivered after a thorough functional test of all mechanical and electrical compo-
nents. The test includes a sterility test of 100 h for which all the ordered mounting parts are installed.
Packing is done only by qualified personnel. After receipt of the the BIOSTAT® B and in case is inop-
erative due to transport damage and in order not to forfeit possible warranty claims for such damage or
other defects, please pay attention to the following advices :
1. Check whether the delivery is complete according to your order.
The delivery includes all connectors, adapters, cables and tubings required for assembly and
connection of the BIOSTAT® B. You must only these parts.
Never use parts and accessories, which do not fulfill the specifications of B. Braun
Biotech International. In such a case our warranty will lapse. B. Braun Biotech Inter-
national GmbH will not be liable for malfunctions and damages resulting from the
use of parts which were not released for use with the BIOSTAT® B.
2. Check carefully all parts for damages, especially the culture vessels. Never use a damaged
glass culture vessel (even if only hairline cracks occur or damages are suspected).
Warning: A damaged culture vessel might burst during sterilization in the autoclave.
3. If parts are missing or damages occurred in transit, please inform your local B. BRAUN sales
representative or contact B. Braun Biotech International GmbH, Melsungen, directly.
2.2 Warranty Agreements and Customer Service
B. Braun Biotech International GmbH warrants its products according to the „General Terms and Con-
ditions of Business“ and unless other terms were agreed upon in writing. Date of reference is the date
of delivery, which must be evidenced by a corresponding delivery confirmation.
The BIOSTAT® B is designed for use under normal laboratory environmental conditions.
Note: Before a use under specific environmental conditions in the laboratory and/or the
use together with special nutrients or additional solutions, you will have to test the
applicability of all parts.
The warranty is granted against defects of construction, manufacture or in material and any
malfunctioning resulting therefrom.
The warranty is neither granted for parts subject to normal wear and tear (O-rings, seals,
membrane filters), nor for defects or damages caused by improper handling, nor for defects
and malfunctions caused by corrosion (i.e. due to lack of resistance against solvents, etc.,
which are used for the fermentation).
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2.3 Customer Service
Defects and damages can be repaired by your local B. Braun Biotech International GmbH service
center. Defective devices can also be returned to your local sales representative or to B. Braun Biotech
International GmbH. Carriage charges will be payable by the customer. The repairs will be carried out
and charged in accordance with our terms of maintenance.
For return of any equipment please note your local sales representative of B. Braun Biotech
International GmbH or contact directly
phone +49 56 61 - 71 34 00,
fax +49 56 61 - 71 37 02
Note: On return the parts must be clean, in good hygienic condition and carefully packed.
Contagious parts must be desinfected or sterilized, according to applicable chemical,
biological, biotechnogical or genetic safety regulations. The sender has to prove
compliance with releted safety regulations. The sender will be charged for repair of
damages due to transport and for necessary cleaning and desinfection of any parts.
If repairs are to be carried out on the premises or in a customer-owned workshop by own

technical personnel, the necessary spare parts can be bought from B. Braun Biotech Inter-
national GmbH. A detailed list of spare and wearing parts is available on request.
Note: In no event will B. Braun Biotech International GmbH be liable for repairs carried out
by the customer and possible damages resulting therefrom.
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2.4 Installing the Fermentor System
2.4.1 Space Requirements
The BIOSTAT® B is designed for bench-top use. In general the space requirements depend on the
type and number of the components and peripheral equipment which should be connected. Due to the
variety of possible configurations the space requirements can vary considerably.
1. Check the working place, whether it offers sufficient space. It is recommended to consider
some additional space for convenient handling of the equipment.
2. A standard configuration of a BIOSTAT® B with culture vessel B2 requires a bench space of
about W x D = 800 x 600 mm. You can vary the arrangement of the components according
to the lengths of the connecting tubings and cables.
For the dimensions of the control unit see the fig. “Dimensions and connectors of the
BIOSTAT® B. The dimensions of the culture vessels are shown in part 4 of this manual. The
dimensions of additional equipment and peripheral devices are shown in the corresponding
documentation of these devices.
3. Verify that the working place can carry the weight of the fully equipped fermentor system with
the culture vessel and all peripheral devices, ready for operation.
4. Check whether all supplies and installations are available at the operating site, such as for
power supply, air or gas supply, water supply and drain, or prepare them correspondingly.
Tubings, clamps and accessories required for connection are delivered with the fermentor
system. For dimensions of the tubings and connectors see tables below.
2.4.2 P&I - Diagram
Fig. 2-1 P & I - Diagram of the BIOSTAT® B

(example for standard design; a copy of the original drawing or of a P&I
diagram for a fermentor system of customized design will be added to the
supplement of this Operating Manual)
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Fig. 2-2: Dimensions and connections of the BIOSTAT® B
Pos. Connector Use

Air hose connector air inlet from laboratory connection
Air hose connector air supply to culture vessel
O2 õ hose connector or adapter extra inlet for oxygen source (O2-enrichment)
combined with Air õ
Exhaust cooler õ hose connector cooling water backflow from exhaust cooler
Exhaust cooler hose connector cooling water supply to exhaust cooler
Thermostate õ tubing with female connector cooling water backflow from double jacket
Thermostate tubing with male connector thermostate supply to culture vessel
Cooling water hose connector cooling water return to lab or drain
Cooling water õ hose connector cooling water supply from laboratory
pH pH-electrode connection of pH-electrode
Foam Foam-probe connection of antifoam probe
Level Level-probe connection of level probe
pO2 pO2-electrode connection of pO2-electrode
Temp Pt-100 connection of Pt-100
Gasmix Socket, 15 pin connection of a Gasmix-unit
Host Socket, 9 pin, RS485 or RS422 connection of a host computer
Recorder Socket, 15 pin connection of a recorder
Printer Plug, 9 pin connection of a printer
Ext. Signals Socket, 15 pin connection of remote signals
Substrate Plug, 9 pin connection of external substrate pump
Table 2-1: Dimensions and connections of the BIOSTAT® B
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2.5 Connection Specifications
2.5.1 Connectors, tubings, cables and Fittings

The connectors, tubings, cables and fittings required for laboratory connection and assembly
of the BIOSTAT® B are included in the delivery, unless otherwise agreed in the order (i.e. for
customized configurations). Details about the dimensions and specifactions are given below.
However the delivered parts may differ from the specifications below, if necessary for a cu-
stomized configuration or for connection of additional peripheral devices. If questions arise
concerning the applicability of the parts for a specific purpose, please contact your B. Braun
Biotech sales representative or B. Braun Biotech International GmbH directly.
1. You must only use connectors and fittings which B. Braun Biotech International GmbH has
released for use with the BIOSTAT® B and its peripheral devices or for which B. Braun Bio-
tech International GmbH has confirmed the applicability in writing. This is especially true for
replacement of connectors and fittings in case of damages or wear and tear.
2. Allways verify that the supplies of energy, water and air (or gasses) are designed according
to the requirements of the fermentor system. Cooling water and air (or gasses) must be
clean and free of corrosion products from the supply lines. If necessary, install correspond-
ing prefilters in the supplies.
Connection Fittings Dimensions Function, special notes
Mains supply fixed cable / see below at first installation the customer
shock-proof plug or our customer service has to
attach the plug which corre-
sponds with country standard
Cooling water õ hose connector hoses ID = 12 mm cooling water supply to control
PVC fabric hose 12.7 x 18.7 mm unit
Cooling water hose connector tubings, ID = 8 mm connection to cooling water re-
PVC fabric hose 8 x 14 mm turn in the laboratory or to lab.
drain
Air õ hose connector tubings, ID = 8 mm connection to air supply or other
8 x 14 mm gas supply systems;
O2 õ hose connector connection of extra O2 supply;
combined with air inlet, if no ex-
tra gas supply should be used
Table 2-2: Dimensions of the tubings and connectors
2.5.2 Mains Connection
There are 2 different versions available of the BIOSTAT® B which can be connected to the following
voltages. Check on the rating plate of the BIOSTAT® B system delivered to you, whether you received
the correct voltage version :
Cat. No. 884 032/6 with 230 V, 50/60 Hz
Cat. No. 884 033/4 with 115 V, 50/60 Hz
BAE_B Rev. 2.31 - 1099 SW 4.1 2-5

2.5.3 Laboratory Connection for Cooling Water Supply and Return
2.5.3.1 General and Safety Information
The BIOSTAT® B requires a precontrolled laboratory water supply of max. 2 barg, without
pressure deviations, see the above P&I diagram. The cooling water will supplied to the ther-
mostate system and to the exhaust cooler. For details about a thermostate system of cus-
tomized design see the P&I diagram supplied for your fermentor system.
The water must be clean and free of particles. A dirt filter is to be installed either in the labo-
ratory installation or in the feeding line to the fermentor control unit.
The minimum operating temperature in the culture vessel will be about 8 degC above the
cooling water temperature. For operation at a lower temperature you will need an external
cooling device.
The water hardness should be below 12 German Degrees of Hardness in order to prevent
calcareous deposits. Note the conversion table for degrees of hardness in the supplement.
Always verify the correct connection of the cooling water supply and drain/return lines to their
inlet and outlet connections at the control unit. The female and male quick connectors are
especially assigned and labelled correspondingly, so that they cannot be mixed up.
If the BIOSTAT® B is moved to another working place and the utility connections are
rebuilt, take special care to properly connect the supply and the return/drain lines to
their corresponding in- and outlets. Erroneously connection of the cooling water
supply to the “Cooling water outlet “ connector of the control unit can result in
supplying overpressure to the culture vessel, which may cause the vessel to burst.
BIOSTAT® B, cat-no. 8840326 (230 V, 50/60 Hz) of unit no. 3226 and later, as well as
cat.-no. 8840334 (115 V, 50/60 Hz) of unit no. 3216 and later, have a return valve in the
cooling water outlet installation of the control unit. This prevents supplying over-
pressure to the culture vessel, if the cooling water supply is erroneously connected
to the cooling water outlet.
Elder versions of the BIOSTAT® B can be rebuilt with the „Rebuilding kit return valve
heater BIOSTAT® B“, cat.-no. 34108971. Please contact your representative of
B. Braun Biotech International GmbH, if you want to have your BIOSTAT® B rebuilt.
Rebuilding must be done by well trained and qualified service personnel.
The cooling water outlet / drain / return line must not be blocked. The temperature
circuit of control unit and culture vessel must always operate at ambient pressure.
Overpressure must not be supplied to the culture vessel jacket since the glass ves-
sel has a limitted pressure resistance. Overpressure can cause the vessel to burst.
This is also true if the BIOSTAT® B is connected to an external cooling device or a
closed cooling circuit in the laboratory.
2.5.3.2 Connection of the Cooling Water Supply and Drain/Return
1. Connect the laboratory cooling water supply to the „Cooling Water Inlet õ“ located at the left
side of the control unit, with the tubing included in the delivery set.
– connection: 2 bar (29 psig) controlled,
– required flow: max. 5 l/min.
2. Connect the „Cooling Water Outlet “ located at the left side of the control unit to the labo-
ratory drain, waste water line or the return connection of a closed laboratory cooling water
installation. The required tubing is included in the delivery set.
3. When a free drain is applied in the laboratory, place the outlet line in such a way that it de-
scends by a steady gradient, in order to avoid water pockets in the line.
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2.5.4 Connection of Air or Gas Supply

The BIOSTAT® B can be connected to any pre-controlled pressurized air supplies delivering
max. 0.8 barg (approx. 11.6 psig). For O2-enrichment an additional source supplying a gas
enriched with oxygen can be connected. For details see the P&I diagram.
Optionally a Gasmix unit is available for supply of a mixture of 4 gases.
The air or gas (mixture) fed into the system must be dry and free from oil and dust. If neces-
sary, prefilters must be installed on the laboratory side.
Defects and damages caused by contaminated air or gas will not be covered by our
warranty.
In the inlet tubings to the culture vessel and for the exhaust membrane filters provide sterile
gas supply (filter no. 1.2) and exhaust (filter no. 1.4) of the culture vessel.
1. Connect the laboratory air supply to the „Air Inlet“ which is located at the left side of the con-
trol unit, with the tubing included in the delivery set.
2. For O2 - enrichment connect an extra oxygen source to the O2 inlet connector. If no extra
oxygen supply should be connected, the O2 inlet is combined with the air inlet by an adapter.
If the laboratory supply delivers a higher pressure, you must install a pressure reducer to
preadjust the pressure.
3. The air inlet tubing of the culture vessel and the exhaust equipment will be prepared and
connected while preparing the fermentation run. See section 3 and 4 for details.
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Operating Information
3 STARTING AND OPERATING THE BIOSTAT® B
3.1 Preparing and Connection of the Control Unit
3.1.1 Preparing the Workplace

1. Place the control unit, the culture vessel and the peripheral devices together with all acces-
sories at the working place. Verify, that there is enough space for convenient handling.
2. Set up the control unit at the operating site and connect it to the laboratory installations, see
chapter 2 for details. :
– the power supply,
– the cooling water supply and waste water drain,
– if required, an external thermostate system,
– and the air supply (if required, together with the extra oxygen supply)
3.1.2 Connecting the Thermostate System
If the BIOSTAT® B has been moved to another working place and the cooling water
supply and drain/return line has been rebuilt, verify that the inlet and outlet connec-
tors are properly connected. See the installation instructions in section 2 for details.
1. Check the connection of the laboratory cooling water supply to the inlet “Cooling water “ of
the control unit and the connection of the laboratory drain or cooling water return line to the
outlet “Cooling water “. Connect the tubings supplied with the system, if not yet done so.
Verify that the laboratory supplies provide cooling water at 2 barg max. and 5 l/min.
2. Check connection of the cooling water outlet to the laboratory drain or cooling system. When
using a free drain, the hose must descend steadily in order to avoid water pockets.
If an external cooler or a closed cooling system is connected, verify that the thermostate
system can operate at ambient pressure and cooling water is circulating sufficiently in order
to maintain the desired temperature in the culture vessel.
3.1.3 Connecting the Air/Gas Supply System

Check the connection of the laboratory air/gas supply at the air inlet at the left side of the
control unit and connect, if necessary. The laboratory supplies must deliver about 10 l/min of
air (or gas) at 0.8 barg (11.6 psig).
3.1.4 Preparing and Connection of Peripheral Equipment for Measurement and Control
1. For checking and calibration of the pH-electrode, which must be done before it is mounted
into the culture vessel, you can connect the electrode to the corresponding socket at the side
panel of the control unit. For details about calibration see section 4.
2. For checking the pO2-electrode, e.g. after service (replacement of the membrane, etc.) you'll
have to connect the electrode to its amplifier and polarize. For details see section 4.
3. For the start of the fermentation run the electrodes will be connected to the corresponding
sockets at the control unit after autoclaving, when the culture vessel is prepared and con-
nected for the fermentation run.
4. As far as necessary connect the cables of an external recorder, printer, a gas mixing system
or a host computer, respectively. See the corresponding plugs and sockets on the left side of
the control unit.
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3.2 Preparing the Culture Vessels and Accessories
3.2.1 Checking the Equipment

During assembly of the culture vessel and mounting of the probes and accessories, carefully
check all parts for damages. You should examine particularly closely the glass vessel itself,
the seals on the top plate and the O-rings of all mountings. See the instructions in Chapter 4
concerning the testing and maintenance of the vessels.
Warnings:
Damaged glass vessels may no longer have the bursting strength required for auto-
claving or running the fermentor, respectively, and must be replaced immediately.
Damaged seals can be the cause of contaminations of the culture during the proc-
ess. Seals on accessories, which are often refitted, should be replaced regularly.
3.2.2 Assembling and Equipping the Culture Vessels

The following sections include a summary of the required or possible steps. The handling of
the different culture vessels and their equipment is described in details in section 4.
1. Unscrew the top-plate screws and dismount the top-plate (1). Take care not to damage the
O-ring (9) and clean, if dirty or replace, if damaged.
2. Remove, replace or mount, respectively, any internal accessory required for the intended
process. Mount all parts which are to be installed into their top-plate port from below. Any
port where no accessory part is installed, must be fitted with a blind closure.
3. Check all seals carefully and replace any seal which appears to be defective. You may
grease seals slightly with silicone grease to prevent glueing onto their contact surfaces.
5. Mount the top-plate back to the glass vessel. Locate the top-plate with the knurled screws.
Tighten the screws properly handtight. Do not use a spanner, you may damage the threads
and/or the glass vessel. Tighten crosswise to prevent uneven pressure of the top-plate
against the glass flange of the vessel and the sealing.
Uneven pressure applied to the rim of the glass vessel can damage the vessel.
6. Complete equipment of the top-plate with all probes and accessories of the delivery set re-
quired for the fermentation run.
The pH-electrode has to be calibrated before it is installed. The calibration of the pO2 - probe
will be carried out in oxygen-free culture medium after autoclaving.
7. Arrange the electrodes and accessories on the top-plate from the inside to the outside so
that they are easily accessible. For the parts and suggestions of their placement in the top
plate see fig. „Top plate of a BIOSTAT® B culture vessel with recommended placement of
the accessories“ and the corresponding information in section 4.
8. Attach a short piece of silicone tubing (4x7 mm) to one side of the membrane filter for the
air inlet and the other end to the hose connector of the sparger pipe in the top plate. Place
the tubing into the clamping device on the culture vessel so that it is completely clamped off
and no culture solution can enter the air inlet line via the sparger ring.
9. Screw the exhaust cooler directly or with its flexible connector into its port in the top plate.
Connect the membrane filter for the exhaust to the exhaust cooler via a short piece of sili-
cone tubing of 4x7 mm.
10. Adjust the height of the antifoam electrode and the level electrode according to the planned
working volume. The height can be determined by gauging the capacity by liters.
BAE_B Rev. 2.31 - 1099 SW 4.1 3-2

11. Wrap aluminum foil round exposed electrical connectors and clamp off all tubings of those
parts which reach into the culture solution (such as inlet air tubing, harvest pipe, for in-
stance). The air/gas supply line can be clamped off by means of a clamping device („dove-
tail“) and fixed to the vessel.
The exhaust line, must be kept open so that pressure compensation with ambient pressure
can take place, as it is necessary during and after the sterilization in the autoclave.
12. Connect all the necessary peripheral equipment (storage bottles for corrective solutions,
sampling systems etc.). For equipment and preparation of the storage bottles (for acid, alka-
line solution, antifoam agent) with withdrawal pipes, silicone tubes and exhaust filters and
filling the bottles) see corresponding sections below.
3.2.3 Filling the Jacket of the Culture Vessel

Note: The thermostate liquid in the jacket of the culture vessel provides optimum heat transfer
during sterilization and in the process. Therefore, before sterilizing the culture vessel and
prior to any new fermentation run, it is necessary to check the filling of the jacket and refill
with thermostate liquid, if necessary. For this you can proceed as follows:
1. Connect the thermostate tubings to the culture vessel in this way:
– connect outlet at supply unit to lower quick connector at culture vessel jacket (inlet)
– connect upper connector at culture vessel jacket (outlet) to inlet of supply unit.
The connectors are marked correspondingly and cannot be mixed.
2. Switch on the basic unit (MAINS 0 -> I). Press FILL THERMOSTATE switch to fill the ther-
mostate system and the jacket of the culture vessel. After filling the jacket disconnect again
the thermostate hoses and continue preparing the culture vessel for sterilization.
The quick coupling at the lower inlet keeps the jacket inlet closed. The tubing connector at
the upper outlet is open to provide pressure compensation with ambient air.
Caution: Do not close (or clamp off) the upper outlet. The thermostate liquid expands
during heating-up in the autoclave. Excess liquid must be allowed to flow out of the
jacket. Else overpressure will be formed in the jacket which may destroy the vessel.
3.2.4 Filling the Culture Vessel

1. Open any port in the top plate and introduce the culture solution. Please note that the culture
solution partly evaporates during sterilization. The loss amounts to approx. 100 ml per 30
min sterilization time (the exact which loss can only be ascertained empirically).
2. Allow for this by increasing the quantity of culture solution in the vessel before sterilization or
by subsequently introducing additional solution.
3. If the culture medium required for later fermentation is not autoclavable, fill in water instead
to ensure reliable sterilization of the culture vessel (volume to be added should correspond
to the loss through evaporation; recommended volume is approx. 200-300 ml).
BAE_B Rev. 2.31 - 1099 SW 4.1 3-3

3.2.5 Preparing and Connecting Addition Agent Bottles
The delivery set of the culture vessels includes 3 bottles for corrective solutions (acid, alkaline,
antifoam or nutrient solution). These are 250 ml bottles for the vessels B 2 or B°2 - airlift and 500 ml
bottles for the vessels B 5, B°5 - airlift and B 10. These volumes usually will suffice for 1 process.
However, especially for supply of nutrient solutions and during long-term fermentations or continuous
operation we recommend to prepare several bottles in order to have sufficient volumes available.
1. Fill the bottles with acid, alkaline, antifoam or nutrient solutions as required.
Caution: When using acid or alkaline agents take precautions against burns. Use
protective gloves, for instance.
2. Attach a piece of silicone tubing to the hose connector carrying the rising pipe of the bottle.
3. Connect the other end of the silicone tubing to one of the nozzles welded in the culture ves-
sel top-plate. The tubings should have sufficient length to allow for passing through the peri-
staltic pump in the control unit.
4. Place the bottles into the holders on the frame supporting the culture vessel. Clamp-off the
tubings before autoclaving so that no corrective solution can escape from the bottles. For
this you can use the clamping device („multible dovetail“) attached to the culture vessel.
3.2.6 Sterilization of the Culture Vessel

1. Take care that all open ports in the top plate of the culture vessel are closed. Locate all
loose parts so that they cannot get loose during handling and transport.
2. Cover the electrode plugs with some aluminium foil in order to protect them from the impact
of steam. Some electrodes (pH-, pO2-electrode) may have special protective caps for this
included in the delivery.
3. Place the bottles for corrective solutions into their holders at the culture vessel. The holders
will facilitate transport of the vessel and the assemblies and save space in the autoclave.
4. Locate the tubings of the air / gas inlet and of the corrective solution supplies. For this you
can use the clamp devices (dovetail clamps) attached to the culture vessel.
The tubing of the exhaust filter must not be clamped-off. This is necessary to provide
the pressure compensation required during and after the sterilization.
5. The double jacket of the culture vessel must be completely filled with the thermostate liquid
(water). If the vessel is used for the first time or to add missing liquid, connect the in-/outlet
to the control unit and push the FILL THERMOSTATE button to fill the double jacket.
6. Autoclave the culture vessel and all accessories at a temperature of 121 degC.
The temperature in the centre of the culture vessel must be kept at 121 degC for at least
20 min. If necessary, test the reliability of your autoclave in preliminary tests by sterilizing
suitable indicators (commercially available test spores, for instance).
Proper sterilization requires approx. 40 min for the 2 l-culture vessel and approx. 60 min for
the 5 l-vessel. Please check whether these times are sufficient for your individual setup. For
the 10 l vessel the time has to be estimated empirically.
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3.3 Peparing a Fermentation Run
3.3.1 Overview
Note: This overview should help experienced users to check the necessary steps of opera-
tion. For a detailed description of the individual steps see the sections 3.4.2.ff below.
1. After sterilization you should let the culture vessel and the connected equipment in the auto-
clave cool down. Before removing the equipment from the autoclave, check whether
the parts are still hot. Put on protective gloves, if necessary.
2. Now the culture vessel can be brought to the workplace for further set up and connection.
Take special care during transport to prevent loosening of tubings or assemblies connected.
3. Mount the motor onto the stirrer shaft coupling at the culture vessel.
4. Connect the gas/air supply to the inlet gas/air filter.
5. Connect the thermostate system to the culture vessel.
6. Connect the cooling water supply and backflow tubing to the exhaust cooler.
7. Remove protective aluminium foil from the electrode connectors. Connect the cables to the
electrodes and to the corresponding sockets at the control unit.
8. Switch on the control unit. If necessary compensate again for losses of thermostate liquid in
the jacket of the culture vessel.
9. Place the tubings of the additive agent supplies into their corresponding peristaltic pumps
and fill the hose lines manually to compensate for the clearance volume in the tubings.
10. Check display of the operation parameters at the measurement and control system and ad-
just as far as necessary. See below and section 5 or this manual for details.
11. Inoculate thevessel with the seed culture. See below for details about suitable methods.
3.3.2 Final Setting-up and Connection of the Culture Vessel for Fermentation
3.3.2.1 Connecting the Thermostate System
1. Connect tubings of the thermostate system. Check whether the water supplied from the
laboratory has a controlled pressure of 2 bar (29 psi). Readjust before opening the supply.
2. If necessary compensate for possible losses of cooling water in the vessel jacket (see „Fill-
ing the vessel for the first time“). Excessive water will escape from the outlet port.
During the fermentation run cooling water will automatically be introduced into the thermo-
state system when the temperature in the culture vessel exceeds the setpoint temperature.
3.3.2.2 Connecting Air / Gas supplies
Note: The pO2-electrode must be calibrated before starting the air supply. Note the more detailed
information about pO2-calibration in corresponding chapters of section 4 and 5 below.
-> Remove the air supply tubing from its holder on the vessel, then fit the filter into the holder.
Connect the hose to the hose connector marked „AIR out“ on left side of the control unit.
3.3.2.3 Connecting the Exhaust Cooler
-> Connect the cooling water supply and drain to the exhaust cooler. See the fig. in section 4
for the location the connectors.
❏ There will be a constant flow of cooling water through the exhaust cooler according to the
flowrate adjusted in the laboratory connection.
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3.3.2.4 Connection of Corrective Solution Supplies
1. Place the tubings of the bottles con-

taining into the corresponding peristaltic
pumps in the control unit :
2. Lift the lid of the peristaltic pump and
open the inlet/outlet clamps. Place the
tubing into the rotor from the inlet side
(lower clamp). Turn the rotor while in-
serting the tubing. Fasten the tubing
with the clamps.
3. Check the pressure load of pump head
rolls and readjust, if necessary. They
should fully clamp the tubing and pre-
vent any backflow of liquid, but must not
damage the tubings.
4. You may turn the pump head by hand
Fig. 3-1: Pump head of the peristaltic pump; until the adjustment screws are acces-
❏ insertion of tubings and adjustment sible. Turn the screws clockwise to in-
of pressuring load; crease the pressuring load and vice
❏ the pump is switched-off with man- versa to reduce. Finally close the lid.
ual switch in central position !
5. Switch on the pump manually to fill the
tubing with the liquid up to its end. This
will prevent incorrect dosing and calcu-
lation of the delivered volumes.
3.3.2.5 Connecting the Drive Motor
1. The motor is connected to the control

unit by a fixed cable, which is internally
wired in the control unit. It is ready for
use after mounting on to the stirrer shaft
of the culture vessel.
2. Place the motor onto the adapter on the
stirrer shaft of the culture vessel. The
support of the motor fits into the locating
pin at the adapter.
See fig. on left side for correct assembly
of the motor.
Fig. 3-2: Connection of the drive motor
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3.3.3 Connecting the Probes and Setting the Measurement and Control System
3.3.3.1 Overview on Connection and Preadjustment
1. If not done yet, connect the electrode cables to the electrodes and to the corresponding in-
put sockets on upper left side of the control unit.
2. Switch on the control unit and check that the equipment is functioning correctly. For this ob-
serve the display in the operating terminal for error messages.
3. Select required functions and adjust the necessary parameters at the measurement and
control system. See fig. and overview on the necessary steps below.
Note: For detailed information about the measurement and control system of the BIOSTAT® B,
such as display functions, necessary adjustments of measurement and control parameters
or error messages, for instance, see section 5 of this manual.
3.3.3.2 Steps of Operation of the Measurement and Control System
1. Select main function with main function key. Changing to the required main function is pos-
sible from any menu or submenu selected.
2. Select required menu/submenu of the main function by repeatedly pressing the main func-
tion key until the required menu is displayed.
3. For entering data move on to the required position with „Cursor up/down“. Enter numerical
data via the numerical keypad. For entering non-numerical inputs select required input with
„ALTER“nate-key.
3.3.3.3 Preadjustment of Operating Parameters and Actuators
1. Calibrate the pO2 electrode in the culture vessel. See more detailed information about cali-
bration in section 5 or in the documentation delivered with the electrode.
2. Select the required air flow rate on the rotameter on the front panel of the control unit.
3. Enter the required setpoints for the measurement and control parameters, for instance :
– Desired operating temperature
– Stirrer speed
– Minimum and maximum pH values of pH controller
– pO2 limits and mode for controlling oxygen supply
– Response threshold of antifoam probe
– Response threshold of level probe
3.3.3.4 Preadjustment of the Supply of Corrective Solutions
In order to avoid incorrect calculation of dosages or dosing times due to the volume of the
connecting hoses, these must be filled with the necessary corrective solutions before pro-
ceeding to fermentation.
1. If not yet done with the connection of the tubings to the peristaltic pumps, now press again
the MAN - switch on the corresponding pump module in the control unit.
2. Allow the pumps to run until the tubings are filled with the corrective solution right up to the
inlet nozzle at the culture vessel.
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3.4 Carrying-Out a Fermentation Run
3.4.1 Sterility Test
Damaged parts, especially faulty seals, but also incorrect handling while equipping the culture vessels,
can result in contaminations of the culture medium. This can cause the process to fail.
1. In order to detect evidence of incorrect handling or faults, especially concerning seals, you
may carry out a sterility test before starting the fermentation run.
Conditions: all necessary components should be connected to the culture vessel; the nec-
essary operating conditions (temperature, air/gas supply etc.) should be set.
2. You can carry out this sterilization test very simply by starting the test run at the same time
you start the inoculation culture and allowing the fermentor to run until the inoculation culture
is finished.
3. A 24 h test period will detect most infections by common environmental germs. It should
leave you enough time to check, reassemble and prepare the culture vessel again for the
planned fermentation run, if infections occur.
3.4.2 Inoculating the Culture Vessel

Inject the inocculation culture. There are various inoculation techniques among which you
may chose. You will find detailed information on individual procedures below.
3.4.2.1 Inoculating the Using a Syringe
1. Remove the necessary amount of solution from the inoculation culture with a syringe and
transfer it to the culture vessel using suitable measures to avoid contamination.
2. Remove the blind closure from an inoculation port equipped with a septum membrane.
Flame the port with an open flame (e.g. using a bunsen burner) to prevent contamination of
the membran. You may also wet the membrane with disinfectant and allow it to take effect
for approx. 5 min.
Note: Using disinfectants may be less effective. Some of the disinfectant can be introduced into
the vessel while the syringe is pierced through the membrane and then interfere with culture
growth. It may also contain resistant spores which will then contaminate the culture.
3. Fit the syringe with a sterile needle and puncture the membrane in order to transfer the in-
oculation solution to the culture vessel.
3.4.2.2 Inoculating the Culture Vessel with a STT Quick Connector
B. Braun Biotech International GmbH offers a STT quick connector, which provides easy
and convenient sterile access to the culture vessel for supply of additional solutions, seed
cultures, etc..
Refer to section 4 for information about use and connection of the STT-quick connector.
Detailed information are given in the „Operating Information STT Quick Connector“, which is
delivered with this device
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3.4.3 Adding Sterile Solutions during the Fermentation Run
3.4.3.1 General Notes
In certain fermentation processes, it is often necessary to introduce additional nutrient solutions or

special essential nutrients (vitamins, amino acids etc.).
Note: Solutions must be transferred from the sterilized storage bottle and to the culture vessel
while avoiding any contamination of the culture.
Add doses of additional substances under sterile conditions. There are various techniques
among which you may chose. Further information on individual procedures is given below.
3.4.3.2 Transferring Solutions by Means of a Syringe
1. Suck sterile solution into syringe from storage bottle, etc. When using solutions which cannot
be sterilized by heat, use a sterile needle connected with a sterile filter.
2. Proceed as shown above for inoculation using a syringe.
3.4.3.3 Transferring Solutions with a STT Quick Coupling
1. Refer to the „Operating Information STT Quick Connector“, which is delivered with this de-
vice, for information about the use and handling of this device.
2. Transfer the sterile solution to the culture vessel using the force of gravity or by means of a
peristaltic pump.
Note: The female connector can be utilized several times; different male connectors can be in-
serted one by one in order to transfer various solutions.
3.4.4 Taking Samples

Samples will be taken from the vessel when required. There are various sampling tech-
niques among which you may chose. A standard method (use of the harvest pipe) is shown
below. For other suitable methods see section 4.
3.4.4.1 Taking Samples Using the Harvest Pipe
Samples can be removed from the culture vessel via the harvest pipe integrated into the top plate and
to which a sterilized silicone hose clamped off with a hose clamp is connected.
1. Insert the connecting hose into an external receptacle (a bottle or beaker, for instance).
2. Open the hose clamp. The overpressure caused by aeration of the vessel usually suffices to
transfer the sample to the receptacle.
When using high-viscosity culture broths, you can clamp off the exhaust hose for a short
time in order to raise the inner pressure forcing out the sample.
3. Dip the open end of the hose into disinfectant between two samples, in order to avoid a
contamination.
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3.5 End of a Fermentation Run
3.5.1 Harvesting the Product

1. Remove the product and the culture medium containing the cells as required for their further
use. Possible methods are, for instance:
– Removal via the harvest pipe and through a transfer tubing for use as seed culture
(transfer to a larger culture vessel for scaling-up).
– Removal via the harvest pipe under sterile conditions (under a Laminar-Flow - chamber)
and filling into sterile harvesting bottles.
– Sterile or insterile downstream processing of the medium.
Take note of the risks of your application and follow the related safety precautions
and other legal or elsewhere compulsory regulations to avoid personnel hazards.
2. Except for processes where microorganisms are applied, which are „generally regarded as
safe“ we recommend to fill the culture vessel with water and sterilize again in the autoclave
before disassembling for cleaning, maintenance and rebuilding for the next use.
3.5.2 Cleaning and Maintenance
Cleaning or maintenance intervals in general depend on how fast the culture vessel and the equipment
gets dirty (due to sticking of proteins, fouling, etc.).
3.5.2.1 Zwischenreinigung nach Fermentationen
For intermediate cleaning between two fermentation runs it often suffices to rinse the culture
vessel thoroughly with water.
When the system will not be used for a short time it is advisable to fill the vessel with demin-
eralized water. It is not necessary to remove electrodes and accessories for this. The water
protects the electrodes from drying out.
3.5.2.2 Basic Cleaning
The culture vessel and the glass bottles for corrective solution supply can be cleaned in a
laboratory dish washer. For this the glass culture vessel need to be removed from the stain-
less steel supporting frame and the top-plate with all accessories need to be dismounted.
Refer to section 4 for dismounting the culture vessel.
When the culture vessel is contaminated with organic substances you can clean the glass
surface with the special laboratory glass cleaners available commercially (e.g. RBS from
ROTH, NEODISHER etc.) and which are dissolved in warm water.
Anorganic deposits can be dissolved using diluted hydrochlorid acid or a similar agent. Take
care to protect yourself against burns. When using aggressive cleaning liquids carefully
flush the vessel with water afterwards.
The metall parts (top-plate, etc.) can be cleaned mechanically or using alcohol or commonly
used detergents. Also flush the vessel carefully with water afterwards.
O-rings, seals and washers can be cleaned mechanically (chemical detergents may affect
the rubber material, for instance). However you should take special care, not to damage the
seals. You may grease the seals with some silicone grease. Above this, it is recommended
to replace all seals regularly.
BAE_B Rev. 2.31 - 1099 SW 4.1 3 - 10

Equipment of the culture vessels
4 EQUIPMENT OF THE CULTURE VESSELS
4.1 General Information
4.1.1 Delivery and Starting up
All BIOSTAT® B - fermentors undergo a comprehensive final quality check before the delivery. This
includes all components and accessories as ordered for the fermentor system. In addition to this, a
sterility test with a duration of 100 h is carried out on the system.
The culture vessels will usually be delivered premounted with the component parts of the standard
equipment. The electrodes, additional accessories and peripheral equipment included in the order will
be packed separately and can be mounted and connected as required for the intended process.
The section 4 below includes the information about the culture vessels available for the BIOSTAT® B
and their standard equipment. If the BIOSTAT® B is to be used for special cultivation processes, such
as tissue cell cultivation, for instance, it may be necessary to remove or replace existing equipment or
to add additional parts. Section 4 also included the information about commonly used additional
equipment for general use or as applied for specific culture vessels.
If your fermentor is of customized design or has component parts not shown in section 4 you
should refer to section 7 of the manual. Section 7 is reserved for customized vessels, cus-
tomized equipment and optional extensions not commonly used for the BIOSTAT® B.
You will get a corresponding documentation update with the Operating Manual or by extra
mail. Please contact your B. Braun Biotech representative or B. Braun Biotech International
GmbH, if no documentation is delivered for a customized modification of the BIOSTAT® B.
4.1.2 Inspecting a Culture Vessel and its Accessories
For the first setup after delivery and prior to any fermentation run we recommend these measures:
Check whether the equipment is complete, as required for the process
– for internal and external equipment of the culture vessels
– for connection of the culture vessel to peripherals and to the control unit
Handle the culture vessels and component parts with special care and check whether they
are in good condition:
1. Carefully inspect the glass culture vessels and other equipment made of glass. These parts
must be free of damages.
The culture vessel can be subject to pressure during the sterilization. Damages will
reduce its pressure resistance and thus its bursting strength.
2. Inspect all ports in the top plate as well as all electrodes and accessories which should be
fitted into these ports including their seals and O-rings. Make sure that all contact surfaces
are free from adhesive deposits. Clean carefully, if necessary. Replace seals and O-rings at
the slightest suspicion of a damage (when pressure marks, are visible, for instance).
3. We recommend to slightly lubricate the seals and O-rings and their contact surfaces with
silicone grease before fitting. This can usually prevent sticking of the seals due to the impact
of high temperatures during autoclaving, and thus can curb premature wear.
BAE_B Rev. 2.31 - 1099 SW 4.1 4-1

4.2 Culture Vessels
4.2.1 Dimensions and Design Features of the Culture Vessels
Culture vessel B2 B5 B 10 B2 - Airlift B5 - Airlift
total volume [l] 3 6,6 13 3,5 9

working volume [l] 2 5 10 2 5
H/D-ratio (total) 1,6:1 2,2:1 2,5 :1 5,4:1 5:1
H/D-ratio (work. vol.) 4:1 3,7:1
space requirements ca. 587 x 350 692 x 400 828 x 420 692 x 350 850 x 420
(HM,B x D [mm]) incl. motor incl. motor incl. motor
autoklave dimensions
- for standard cooler 550 x 350 700 x 400 820 x 420 740 x 350 900 x 420
- cooler bent over 380 x 350 485 x 400 610 x 420 565 x 350 690 x 420
(H x D) [mm]
height incl. exh. cooler 508 645 767 692 850
(HK, [mm])
vessel height with top 285 390 512 469 595
plate (HG, [mm])
height of glass vessel 240 345 470 430 553
(HGl, [mm])
inner Ø vessel 130 160 190 130 (head room) 190 (headroom)
(Di,Gl, [mm]) 80 (reactor) 110 (reactor)
top plate ports, 2 x Ø 19 2 x Ø 19 4 x Ø 19 4 x Ø 19 3 x Ø 19
number x Ø [mm] 2 x Ø 12 2 x Ø 12 2 x Ø 12 1 x Ø 12 2 x Ø 12
9xØ6 9xØ6 9xØ6 6xØ6 10 x Ø 6
4 hose 4 hose 4 hose 4 hose connec- 4 hose connec-
connectors connectors connectors tors tors
side entry ports 4 4 no no no
top plate locating screws 3 6 6 3 6
You should compare these data with the equipment delivered to you. However, you should
note, that B. Braun Biotech International GmbH reserves the right to change these specifica-
tions without prior notices.
Even for modified culture vessels the possible equipment and required handling will usually
be the same as shown below. For a modified design and equipment, check whether addi-
tional information is added to section 7 of this manual or delivered extra. If you require addi-
tional information about specific component parts, you should contact your representative of
B. Braun Biotech or B. Braun Biotech International GmbH directly.
BAE_B Rev. 2.31 - 1099 SW 4.1 4-2

4 Di,Gl HM,B
HM,M
DR HR HK
5 HG
6
HGl,i
VFl
Fig. 4 - 1: Setup and dimensions of the standard culture vessels type B2, B5 and
B10 with stirrer drive system
(1) motor
(2) exhaust cooler
(3) reference thermometer
(4) side entry hose connector
(5) baffle insert
(6) corrective solution bottles
BAE_B Rev. 2.31 - 1099 SW 4.1 4-3

Fig. 4 - 2: Setup and dimensions of a culture vessel type B2 - Airlift
BAE_B Rev. 2.31 - 1099 SW 4.1 4-4

Fig. 4 - 3: Setup and dimensions of a culture vessel type B5 - Airlift
BAE_B Rev. 2.31 - 1099 SW 4.1 4-5

4.2.2 Set-up, Equipment and Accessories of the Culture Vessels

The tables below give an overview of the accessories available at present and described in
this manual. More detailed information about the individual component parts is shown in the
following chapters of section 4.
You should note, that the equipment of the vessels and the available accessories are subject
to a continuos development and may be modified, added or replaced accordingly. Thus
these information are not necessarily complete. For customized equipment you may refer to
section 7 of this manual or to the documentation delivered extra. If the information given in
this manual and the design and handling of your equipment do not match, please contact
your representative of B. Braun Biotech or B. Braun Biotech International GmbH directly.
4.2.2.1 Basic Equipment of the Culture Vessels
Culture vessel B2 B5 B 10 M2/B2 - Airlift M5/B5 - Airlift
vessel insert baffle insert baffle insert baffles guiding tube guiding tube
with 4 baffles with 4 baffles (loop reactor) (loop reactor)
stirrers 2 x 6-bladed 2 x 6-bladed 3 x 6-bladed -- --
disc impellers; disc impellers; disc impellers;
paddle stirrers paddle stirrers paddle stirrers
optional optional optional
air / gas supply pipe with ring pipe with ring pipe with ring pipe with ring pipe with ring
sparger, sparger, sparger, sparger sparger
silicone mem- silicone mem- silicone mem-
brane insert brane insert brane insert
optional optional optional
exhaust cooler type B2 B5 B5 B2 B5
temperature sensor Pt-100-200/4 Pt-100-300/4 Pt-100-400/4 Pt-100-300/4 Pt-100-300/4
pH-electrode Pa/12 - 200 Pa/12 - 325 Pa/12 - 325; Pa/12 - 200 Pa/12 - 325
or as option:
Pa/12 - 425
pO2-electrode 12/220 12/320 12/320; 12/220 12/320
(12/420 op-
tion)
antifoam probe,
insertion depth 50 mm 50 mm std. 50 mm 50 mm 50 mm
opt. 260 mm
level probe, 122 mm 122 mm 122 mm 122 mm
insertion depth std. 122 mm
opt. 260 mm
3 storage bottles 250 ml 500 ml 500 ml 250 ml 500 ml
blind closures 2 x Ø 19 2 x Ø 19 4 x Ø 19 4 x Ø 19 3 x Ø 19
2 x PG 13,5 2 x PG 13,5 2 x PG 13,5 1 x PG 13,5 2 x PG 13,5
9 x Ø 11 9 x Ø 11 9 x Ø 11 6 x Ø 11 10 x Ø 11
sampling system x x x x x
manual sampler x x x x x
BAE_B Rev. 2.31 - 1099 SW 4.1 4-6

4.2.2.2 Accessories for the Culture Vessels
Cat.-Nr. Accessory Description, use
3833697/9 hose connector hose connector with Øo = 4 mm

- assembly into 6 mm top-plate port
884 431/3 quadruple hose connector B adapter with 4 hose connectors of Øo = 4 mm
884 463/1 inoculation adapter B septum kit with membrane and protective cap,
- to be punctured using a syringe or small needle;
- self-sealing membrane which can be punctured
several times
884 060/1 inoculation adapter B10 septum kit with membrane and blind closure
- for 1-channel inculation kit, cat.-no. 884061/0
884 061/0 1-channel inculation kit inculation kit für inculation port B10,
- hollow needle of Øo = 6 mm
- delivered with sterile sleeve
3833717/7 reduction adapter 19/6 adapter for mounting of parts with Øa = 6 mm into
top-plate ports of Ø = 19 mm
881 007/9 filters for air inlet and exhaust autoclaveable membrane filters in plastic housing
- filter membrane of 0,2 µm
- with hose connectors
3922830/4 filter element for exhaust B 10 plastic filter tube with membrane filter
- filter membrane of 0,2 µm
884 459/3 flexible holder for exhaust cooler piece of tubing for exhaust cooler assembly, can
be bent over for autoclaving, requiring a lower in-
ner height of the autoclave
control thermometer for visual monitoring of temperature,
884 432/1 - for vessels B2 - insertion into pocket in top-plate port of 6 mm;
884 453/4 - for vessels B5 and B10 - removable for autoclave
STT quick connector (female) female connector of STT connector for sterile
880924/0 - for tubings 3.2 x 1.6 mm connection of external supplies, to be installed to
880941/0 - for tubings 1.6 x 1.6 mm the tubing attached to the culture vessel
STT quick connector (male) male connector of STT connector for sterile con-
880920/8 - for tubings 3.2 x 1.6 mm nection of external supplies, to be installed to the
880940/2 - for tubings 1.6 x 1.6 mm tubing attached to the external supplies
884 434/8 Bypass sampler kit M/B for setup of a bypass line for removal of small
- for tubings 1.6 x 1.6 mm samples (or for introducing small volumes of cor-
rective solution);
- membrane holder and septum membrane
- 2-channel connector for top-plate port of 19 mm
- 2 m of silicone tubing 1.6 x 1.6 mm
- 0,2 m PTFE hose 3.5 x 5.5 mm
BAE_B Rev. 2.31 - 1099 SW 4.1 4-7

Cat.-Nr. Accessory Description, use
Harvest filter for removal of cell-free samples out of cultures

884438/0 - for culture vessel B2 with microcarrier-bounded cells
884457/7 - for culture vessel B5 - assembly into 6 mm top-plate port
884050/4 - for culture vessel B10
Silicone membrane insert vessel insert carrying a silicone membrane
884466/6 - for culture vessel B2 wrapped around, which is used for bubble-free
884467/4 - for culture vessel B5 gas supply to the culture mediam (especially ap-
884048/2 - for culture vessel B10 plied for tissue cell cultures); kit delivered with ac-
cesories of gas supply and paddle stirrers
Paddle stirrers for optimum mixing of the medium at low stirrer
3823907/8 - for culture vessel M2/B2 speed
3823954/0 - for culture vessel M5/B5
884054/7 - for culture vessel M10/B10
Spinfilter vessel insert for separation of cell-free medium for
880830/9 - type B 2, 20 µm sampling and harvesting;
880831/7 - type B 2, 40 µm for use together with the short harvest pipe SF,
880832/5 - type B 2, 75 µm cat.-no. 880826/0
880833/3 - type B 5, 20 µm for use together with the short harvest pipe SF,
880834/1 - type B 5, 40 µm cat.-no. 880827/9
880835/0 - type B 5, 75 µm
884055/5 - type B 10, 20 µm for use together with the short harvest pipe,
884056/3 - type B 10, 40 µm cat.-no. 880859/8
884057/1 - type B 10, 75 µm
Harvest pipe SF especially designed harvest pipes for removal
880 826/0 - for spinfilter B 2 media (filtrate) separated by a spinfilter
880 827/9 - for spinfilter B 5 - assembly into 6 mm top-plate port
884 059/8 - for spinfilter B 10
perfusion module modular perfusion kit continuous fermentation
884436/4 - type B 2
884439/9 - type B 5
884049/0 - type B 10
BAE_B Rev. 2.31 - 1099 SW 4.1 4-8

4.2.3 Top plate Design and Placement of the Accessories
Fig. 4 - 4: Top plate Design and Placement of the Accessories

- example for standard culture vessels type B2 and B5
port recommended use port recommended use
N1 exhaust cooler N 10 reserve

N2 Pt-100 temperature sensor N 11 level probe
N3 antifoam probe N 12 harvest pipe
N4 pO2 - electrode N 13 pH - electrode
N5 supply of acid N 14 air / gas inlet (sparger pipe)
N6 supply of alcaline agent N 15 holder of baffle insert
N7 supply of antifoam agent N 16 reserve
N8 supply of substrate N 17 holder of silicone membrane insert
N9 reserve
Note: For the other standard culture vessels the arrangement of the ports is almost the same.
Only their distance to the centre and the distance between the ports will be different.
For special and customized culture vessels the arrangement, type and number of top-plate
ports can be different. Please check your equipment for this.
BAE_B Rev. 2.31 - 1099 SW 4.1 4-9

4.3 Equipping and Assembling the Culture Vessels
4.3.1 Overview
The internal equipment of a vessel and the kind and number of components, which should
be installed in the top-plate, need be planned carefully. Some of the accessory parts can
only be mounted, removed or changed, while the top-plate is disassembled. Some parts are
to be pushed through the top plate from below and screwed tight with a corresponding nut
from above. Other parts will be tightened by corresponding nuts from below.
Parts for assembly while the top-plate is
removed, are in general:
– adapters for antifoam and level probe
– adapters for the harvest pipe,
– the adapter for the inoculation port
– any universal fitting (hose connector)
– the mounting pocket for the optional
reference thermometer (if necessary)
– tubing connectors for the side-entry
ports of culture vessels B2 and B5
For culture vessels with drive shaft and
stirrers such parts are, for instance:
– baffle insert or extra baffles
– the stirrers for the drive shaft
– a spinfilter for media removal
– a silicone membrane insert for buble
free gas supply
For airlift culture vessels without drive
such parts are, for instance:
– guiding tube for loop system
– electrodes, probes, pipes of special
imension considering the height of the
airlift vessels
Blind closures must be fitted to any top-
plate port, which is not required for
equiment. For the blind closures of Ø = 6
mm, this is only possible, while the top-
plate is removed.
Fig. 4 - 5: Culture vessel type B 2; internal After assembly of the top plate to the
setup: glass vessel only these component parts
(1) jacketed glass vessel can be added, removed or changed,
(2) stirrer shaft which are located from outside:
(3) 6-bladed disc impeller – exhaust cooler (or flexible adapter for
(4) baffle insert exhaust cooler
(5) sparger pipe – antifoam and level probe
(6) sparger ring for air inlet – the harvest pipe
(7) harvest pipe – other component parts for assembly
(8) antifoam probe into clamping adapters
(9) O-ring seal of top-plate – 2-way inlet of Bypass sampler
– 4-way inlet adapter
BAE_B Rev. 2.31 - 1099 SW 4.1 4 - 10

4.3.2 Disassembling a Culture Vessel

1. Disconnect the cables and tubings connected to the control unit. Remove the motor and all
electrodes and component parts installed in the top-plate, as far as necessary for cleaning,
maintenance and reequipment of the culture vessels.
2. Turn the top-plate screws counterclockwise. They need only get loose from the flange of the
stand, but need not be fully removed. Lift the top plate. Take care not to damage any seals if
they are sticking. Take care not to damage any electrodes. Remove all parts which are not
required for the next fermentation run.
3. Clean the culture vessel inner and the component parts, if necessary. You need not remove
the glass vessel from the stand for this. You may flush the parts with water. When the cul-
ture vessel is contaminated with organic substances you can clean the glass surface with
the commercially availbale laboratory glass cleaners dissolved in warm water.
You may clean sticking residues of the culture medium mechanically. Make sure that the
glass is not damaged. Anorganic deposits may be removed with a solution of hydrochloric
acid or alike. When using aggressive agents rinse the vessel thoroughly afterwards.
4. If the glass vessel is damaged and must be rebuilt, loosen the screws of the flange of the
stand and remove the vessel. Insert the new glass vessel with special care. Do not hit the
hose connectors of the culture vessel jacket, they can easily break.
5. Check all parts for damages (especially the glass vessel and all seals/O-rings). If necessary
(at sticking residues of the previous culture) clean the parts. Replace any seals and O-rings
which are suspected to be damaged.
Order information for the glass vessels and specifications of the O-rings for the top-plate
for vessel type B2 B5 B10 B2-Airlift B5-Airlift
vessel cat.-no. 3920427/8 3920428/6 3920418/9 on request on request

O-ring cat.-no. 39121992 39120155 3410006 39121992 3410006
specifications 136,12 x 164,69 x 196,44 x 136,12 x 196,44 x 3,53
3.53 EPDM 3,53 EPDM 3,53 EPDM 3.53 EPDM EPDM
4.3.3 Assembling a Culture Vessel

After mounting of all parts, the top-plate has to be assembled in the reverse order.
1. Carefully place the top-plate onto the rim
of the vessel. Turn the top plate in such a
way that the openings for the holding
screws come to rest directly below the
screws. Take care not to warp the seals.
2. Carefully tighten the top-plate screws
handtight in a crosswise manner. This will
suffice to locate the top-plate and ensure
the pressure for sterile connection.
Note: Do not use a spanner or other tools,

since this can apply uneven pressure
to the rim of the glass vessel. This
may damage the vessel.
Fig. 4 - 6: Removal /assembly of the
top-plate, example for ves-
sel type B 2
BAE_B Rev. 2.31 - 1099 SW 4.1 4 - 11

4.3.4 Fitting of the Standard Equipment inside a Culture Vessel
4.3.4.1 Fitting, Arrangement or Rebuilding of Stirrers
The standard equipment includes 6-bladed disc impellers. Since the baffles provide addi-
tional mixing of the medium, they allow sufficient agitation for standard applications.
For special cultures, e.g. requiring low stirring speeds, special stirrers (paddle stirrers, etc.)
can be fitted. These may be fermentation processes using cells, which are sensitive to shear
forces, such as tissue cells, for instance. The standard paddle stirrers are available with two
large blades of fixed angle of adjustment (45°). The vessel B10 and (optional) all other ves-
sels can be equipped with stirrers with adjustable paddles.
Type, for culture vessel stirrer Ø Da Cat. - No.
6-bladed disc impellers B2 53 3821785/6

Paddle stirrers B2 58 3823907/8
Paddle stirrers B5 70 3823954/0
Paddle stirrers B10 110 884 054/7
Mounting or rebuildung of the stirrers:

1. In order to adjust the height of the stirrer
elements, loosen the screws, which lo-
cate the stirrers at their position on the
shaft and move the stirrers to the in-
tended height.
2. In order to fit other stirrers, remove the
old one's and push the new onto the
stirrer shaft to the required extent.
For the paddle stirrers it is recom-
mended to locate them in crosswise ar-
rangement (i.e. to lower stirrer turned by
90° compared to the upper stirrer).
3. Tighten the screws in such a way that
they cannot loosen even when stirring is
carried out at high speeds, with highly
viscous culture media or for long fer-
mentation runs.
Fig. 4 - 7: Mounting or rebuilding of the

stirrers
BAE_B Rev. 2.31 - 1099 SW 4.1 4 - 12

4.3.5 Baffle Insert and Baffles for Culture Vessels with Stirring Device
The baffle insert of the culture vessels B2 and B5 and the two baffles of the vessel B10 serve to in-
crease the turbulance of the culture broth, to improve the mixing and mass transfer even at low stirrer
speeds. The baffle insert consists of a holder with four vertically mounted baffles. For the culture ves-
sel B10 one baffle is attached to the aearation pipe and one is mounted into a 6 mm top-plate port (not
shown in herein, see your vessel).
Specifications of baffle inserts and single baffles
Type, baffle insert B2 baffle insert B5 sparger pipe with baffle,

for culture vessel cat.- no. 38219638 cat.- no. 38219670 cat.- no. 38241080
single baffle B10,
cat.- no. 38241072
Ø Da [mm] 125 154 --

HB [mm] 130 204 280
Htot [mm] 200 300 433
O-ring 7,65 x 1,78 EPDM, cat.- no. 39120945
tightening nut hexagon nut M10x1, cat.- no. 38337185
On delivery the baffle insert for the cul-

ture vessels B 2 and B 5 and the baffles
for the culture vessel B 10 are prein-
stalled in the factory. You will have to
remove them only for cleaning and for
checking and replacing the O-ring. You
can remove them at all if you need to
mount other accessories into the corre-
sponding port.
The baffle insert of the vessels B 2 and
B 5 is usually located in port no. N 15,
see fig. „Top plate with recommended
placement of the accessories“). For the
vessel B 10 one baffle is attached to the
sparger pipe, the others are mounted in
separate top-plate ports.
1. To remove the baffle insert or the baffle,
unscrew the hexagon nut and pull the
adapter out of the port. Check the O-
ring and replace, if necessary.
2. To mount again push the extension rod
with the adapter up through the top-
plate port, see fig. left side. Center the
baffle insert in the glass vessel.
Fig. 4 - 8: Assembly of baffle insert
3. Use the box spanner supplied with the
tool kit to carefully tighten the adapter
with the corresponding hexagon nut.
BAE_B Rev. 2.31 - 1099 SW 4.1 4 - 13

4.3.6 Sparger Pipe
The sparger pipe is made of stainless steel and has a sparger ring with evenly distributed fine holes
through which the air or gas can bubble into the culture broth.
Specifications
for culture vessel B2 B5 B 10 B 2 - Air- B 5 - Airlift

cat.-no. 38217864 38218569 38241080 lift B318270200104
38219395
HE [mm] 229 39 453 411 customized

O-ring (2) 7,65 x 1,78 EPDM, cat.-no. 39120945
tightening nut (3) hexagon nut M10x1, cat.-no. 38337185
threaded pin (4) 39504133 38318334
On delivery the sparger pipe is preinstalled in the factory. You will have to remove it only for
for cleaning and checking and replacing the O-ring. At culture vessel equipped with silicone
membrane insert for bubble-free gassing, the sparger pipe is not installed. See detailed in-
formation about the silicone membrane insert below.
The sparger is usually located in port no. N 14, see fig. „Top plate with recommended
placement of the accessories“). If this is different for your fermentor, you can mark the
placement at the top-plate or in a copy of the above figure.
1. To remove the sparger pipe, unscrew
the hexagon nut (3) and pull the adapter
(1) out of the port. Check the O-ring (2)
and replace, if necessary.
2. For cleaning of the sparger ring un-
screw the threaded pins (4) You can
flush the pipe and ring with water. If the
small holes in the ring are blocked, you
can use a brush or a needle.
3. For assembly push the sparger pipe up
through its top-plate port. The adapter
(1) determines the immersion depth.
Center the sparger ring in relation to the
stirrer shaft. If necessary, readjust the
position of the lowest stirrer in such a
height that the sparger ring is centered
approx. 1 cm below the stirrer.
For an airlift vessel you should center
the sparger ring below the guiding tube.
4. Use the box spanner supplied with the
tool kit to carefully tighten the adapter
Fig. 4 - 9: Assembly of sparger pipe with the corresponding hexagon nut.
After comlete assembly of the culture
vessel and mounting of the top-plate
you can connect the silicone hose for
the air supply. This can be done when
you are finally equipping the culture
vessel and prepare it for autoclaving.
BAE_B Rev. 2.31 - 1099 SW 4.1 4 - 14

4.3.7 Inoculation Connectors (Septum Port Kits)
Inoculation connectors (septum port kits) allow for sterile access to the culture vessel, when the seed
culture or corrective solutions should be supplied. The inoculation connector B/MD is available for all
culture vessels. The septum port kit B10 has been developed for the culture vessel B10 but can be in-
stalled into any top-plate port of Ø 19 mm. External supplies can be connected using a 1-channel in-
oculation kit. The septum membranes are „self-sealing“ and can be punctured serveral times.
Part, accessories Cat.-no. Specifications Notes
Inoculation connector B 884 431/3 assembly in top-plate port

Ø 6 mm
– O-ring 39120954 7,65 x 1,78 (EPDM) seals adapter vs. top-plate port
– O-ring 39121054 12,42 x 1,78 (EPDM) seals and locates cap
– septum membrane 39220672 latex self sealing after being punctured
septum port kit B10 884060/1 assembly in top-plate port for 1-channel inoculation kit cat.-
Ø 19 mm no. 884061/0
– septum membrane 39220486 bromobutyl rubber self sealing after being punctured
Use new septum membranes for assembly of the adapters for a new process. Check all O-
rings of the adapters and clean and replace, if required.
Assembling the septum port B/MD:

1. Push the adapter into the top-plate port from
below and screw tight with the nut.
2. Lay septum membrane onto the adapter.
Tighten carefully handtight with locating
sleeve.
3. Put sterile cap onto the locating sleeve. This
cap will keep the septum membrane sterile
after autoclaving until the port is required for
inoculation.
Fig. 4 - 10: Fitting an inoculation connector

(septum port)
Assembling the septum port B10

1. Lay the septum membrane (2) into the top-
plate port (1). Insert the membrane locating
nut (3) and screw tight to locate the mem-
brane.
2. Insert the blind closure (4) into the adapter
and screw tight. It keeps the membrane
clean until you connect the 1-channel in-
oculation kit for transfer of the seed culture
or the corrective solution.
Fig. 4 - 11: Septum port kit B10 with blind

closure
BAE_B Rev. 2.31 - 1099 SW 4.1 4 - 15

4.3.8 1-Channel Inoculation Kit
Similar to the inoculation kit B/MD the 1-channel inoculation kit allows for sterile access to the culture
vessel, when the seed culture or corrective solutions should be supplied. A special advantage is the
use of larger tubings for an increased flow of medium and the supply of larger volumes. The kit re-
quires a top-plate port of Ø 19 mm equipped with a septum port kit B10.
1-channel inoculation kit 884061/0 for septum port B10 incl. sterile sleeve
– 1-channel inoculation B300940100104 hollow needle Ø 6 x 0,5 mm
adapter of stainless steel 1.4435
– O-ring 39120830 15,6 x 1,78, (EPDM)
– silicone tubing 39971155 Ø 3.2 x 1.6, silicone rubber delivery set of 2 m
Preparing of the 1-channel inoculation kit

1. Check and clean a previously used 1-
channel inoculation kit. No residues of
medium should glue inside the needle
(3) and internal pipe. Check the O-ring
and clean or replace, if necessary.
2. The sterile sleeve is filled with glass silk
(7) which can get moistened and dirty
by time. This can hinder proper dearea-
tion during sterilization. Therefore you
should check the filling regularly. Un-
screw the screw cap (6) and check and
replace the glass silk, if required.
3. Attach the silicone tubing for medium
transfer to the hose connector (4). Cut
as long as required and connect to the
corrective solution bottle or the con-
tainer for the seed culture.
4. Screw on the sterile sleeve. It will keep
the needle (3) of the inoculation kit ster-
ile, until it is connected to the septum
port for supply of the seed culture or of
the corrective solution.
5. Prepare the corrective solution bottle or
the container for the seed culture and
sterilize together with the 1-channel in-
oculation kit and the tubing.
For connection of the 1-channel inocu-
Fig. 4 - 12: 1-channel inoculation kit lation kit to the septum port see the
connection information below.
BAE_B Rev. 2.31 - 1099 SW 4.1 4 - 16

Connection of the 1-channel inoculation kit:

1. Remove the blind closure (6) from the
membrane locating nut (7). Flame the
port with a flame to prevent contamina-
tions of the membrane.
You may also wet the membrane with a
disinfectant and allow to take effect for
some minutes.
Note: The disinfectant must not be in-
troduced into the vessel while you
puncture the membrane with the nee-
dle. It may interfere with the process,
then. It can also contain resistant
spores which will then contaminate the
culture.
2. Unscrew the sterile sleeve (5) from the
inoculation kit (1). Also flame the needle
(3) to prevent contaminations.
3. Puncture the membrane (6), push
through the needle vertically and screw
tight with the screw (2) in the membrane
locating nut (7). Then transfer the seed
culture (or the corrective solution, if ap-
plied) into the culture vessel.
3. You can leeve the inoculation kit
screwed into the membrane locating nut
(7). Then clamp-off the tubing.
Fig. 4 - 13: Connection of the 1-channel inocula- 4. If you want to remove the needle for
tion kit connection of other supplies lateron :
– remove needle from the membrane
– flame needle and port (membrane)
– flame blind closure or the needle of
the new kit and screw tight in the port
BAE_B Rev. 2.31 - 1099 SW 4.1 4 - 17

4.3.9 Universal-Adapter
The universal adapter allows to use single top-plate ports for connection of the supplies of corrective
solution, for instance, if all other port are use for asseblies. In addition you can connect a headspace
dearation, when the culture vessel is gassed and deaerated via the silicone membrane insert for bub-
ble-free gas supply, see below for details.
Part, accessory Cat.-no. Specifications Notes
universal adapter (1) 3833697/9 hose connector 4 mm for free 6 mm top-plate port required
tubings of Ø 1.6 x 1.6
tightening nut (2) 38337185 hexagon nut M10x1
O-ring (3) 3912094/5 7,65 x 1,78 (EPDM)
Assembling information
1. Check the O-ring (3) and clean or replace, as re-
quired.
2. Insert the adapter (1) into a top-plate port Ø = 6 mm
from below and screw tight with the hexagon nut (2).
3. For corrective solution supply connect tubing (1.6 x
1.6) of the required length to the hose connector (2).
4. If to be used for dearation of the headspace connect
a sterile filter via a short piece of tubing to the hose
connector.
Fig. 4 - 14: Universal-Adapter
4.3.10 Quadruple Addition Port B
The quadruple addition port allows for additional connection of up to four corrective solution supplies to
the culture vessel, if the other hose connectors are occupied.
quadruple addition port 884 431/3 hose connectors for top-plate port of Ø 19 mm re-
tubings of Ø 1.6 x 1.6 quired for assembly
O-ring no. 39120830 15,6 x 1,78 (EPDM)
Assembly and connection

1. Check the O-ring (3) and replace, if required. Screw
the adapter (1) into top-plate port Ø = 19 mm
2. Connect the tubings of the corrective solution sup-
plies to the hose connectors (2). If any of the hose
connectors should not be used you can attach a
short piece of silicone tubing and make a knot into
the tubing.
Fig. 4 - 15: quadruple addition port
BAE_B Rev. 2.31 - 1099 SW 4.1 4 - 18

4.3.11 Reference Thermometer
A reference thermometer allows for visual control of the temperature in the culture vessel. The ther-
mometer will be inserted into a pocket, which is to be mounted into the top-plate while the top-plate is
removed. The pocket and the housing protect the glass thermometer against damages. If a ther-
mometer is damaged during the process, the pocket prevents the culture from being contaminated.
The thermometer may be autoclaveable or not. An autoclaveable thermometer can be inserted into the
pocket even before the culture vessel is sterilized. Else you will have to leave the thermometer off the
pocket until you are going to start the fermentation run.
Reference thermometer, cat.-no. 884 4321 884 4534 884 4534 884 4534 884 4534
for culture vessel type B2 B5 B 10 B 2-Airlift B 5-Airlift
HH [mm] 297 297 297 297 297

HE [mm] 200 300 300 300 300
glass thermometer 33045054 33045061 33045061 33045061 33045061
O-ring 2 O-rings 7,65 x 1,78 EPDM, cat.- no. 39120945
Assembling information:
1. Push the insertion (1) pocket
into the top-plate port from be-
low.
2. Screw tight the pocket in the
port with the hexagon nut (5).
3. Insert the thermometer (4) into
the pocket.
If the thermometer is auto-
claveable, you can do this prior
to the sterilization. If not, insert
the thermometer, when visual
temperature monitoring should
be applied for the process.
4. Place the protective housing(6)
over the thermometer and
screw tight.
Fig. 4 - 16: Reference thermometer with insertion pocket

(1) insertion pocket
(2) O-rings (protective housing, adapter of in-
sertion pocket)
(4) glass thermometer
(5) hexagon nut
(6) protective housing
BAE_B Rev. 2.31 - 1099 SW 4.1 4 - 19

4.3.12 Harvesting Pipe
The height-adjustable harvesting pipe is the standard equipment of the culture vessels of the
BIOSTAT® B for harvesting of the product at the end of your process. It can also be used for removal
of samples. Information about other devices, which can be used for sampling, is given further below.
Cat. - no. 38337762 38342731 38345331 38339633 38342731

for culture vessel type B2 B5 B 10 B 2 - Airlift B 5 - Airlift
HE, max. [mm] 180 310 410 380 380

adapter (2) adapter for harvest pipe, cat.- no. 38337630
locking ring (7) 6 x 0,7 DIN 471-A2, cat.- no. 39507521
clamping cone (5) cat.- no. 38337649
screw cap (6) cat.- no. 38336820
O-ring (3) 7,65 x 1,78 EPDM, cat.- no. 39120945
O-ring (8) 6,07 x 1,78 EPDM, cat.-no. 39120821
nut (4) hexagon nut M10x1, cat.- no. 38337185
Assembling the harvest pipe:

1. Check the O-rings (3) and (8). Clean or
replace, if necessary. The O-ring (3)
seals the adapter against the top-plate
port, the O-ring (8) seals the pipe against
the adapter.
2. Insert the adapter for the harvest pipe (2)
into the top-plate port (N12 recom-
mended) and screw tight with the hexa-
gon nut (4). Slide clamping cone (5) and
screw cap (6) onto the harvest pipe.
3. Insert the harvest pipe into the adapter
and push through until its lower end is at
the intended insertion depth. The maxi-
mum insertion depth is achieved if the
locking ring (7) is located at the adapter.
4. Tigthen the harvest pipe carefully hand-
tight with the screw cap (6). The clamping
ring locates the pipe at the intended in-
sertion depth.
5. Connect a piece of silicone tubing to the
hose connector of the pipe. You should
consider the required length for connec-
tion to the container for the samples or
the harvested product and whether the
Fig. 4 - 17: Fitting the harvest pipe
tubing should be connected to a pump.
Caution: Never push the harvest pipe

deeper into the vessel after steriliza-
tion. This may cause infections by
environmental germs.
BAE_B Rev. 2.31 - 1099 SW 4.1 4 - 20

4.3.13 Harvest Filter for Removal of Medium at Microcarrier-Cultures
The harvest filter is an optional device for removal of samples in cultures of suspended cells and with
cells growing on microcarriers. It consists of harvesting pipe combined with a special harvesting filter
(teflon element of pore width of 25 µm). Depending on the size of the cells the filter allows the medium
and cell debris (if occuring) to permeate through the teflon element and be removed through the pipe
while the the suspended cells or the microcarriers with the cells are retained in the vessel.
Specifications of harvest filter
for culture vessel type B2 B5 B10
cat.- no. 884 438/0 884 457/7 884 050/4

adapter 38337630, adapter for harvest pipe
clamping cone 38337649
screw cap 38336820
O-ring 7,65 x 1,78 EPDM, cat.- no. 39120945
O-ring 6,07 x 1,78 EPDM, cat.-no. 39120821
nut hexagon nut M10x1, cat.- no. 38337185
Assembling the harvest filter

1. Insert the harvest pipe into the top-plate port from below, where the corresponding adapter
with clamping cone is installed (port N12, for instance) . Arrange the pipe at the intended
immersion depth, so that the stirrers cannot hit against the filter tube.
2. Locate the harvest pipe with the screw cap carefully handtight.
3. Connect a piece of silicone tubing of the required length for connection to the sampling con-
tainer to the hose connector of the pipe. You should consider, whether the tubing need to be
connected to a pump for sample removal.
Maintenance information
The teflon filter tube can be blocked by cell debris and macromolecules.
1. For cleanig remove the harvest pipe from the top-plate. Connect the inlet of the pipe to a
supply of water and flush by counterflow.
2. If the layer and blocking cannot removed by this, you can unscrew the teflon tube from the
pipe and clean using a brush, for instance.
BAE_B Rev. 2.31 - 1099 SW 4.1 4 - 21

4.3.14 Spinfilter
Spinfilters are used for removal of medium while suspended cells or microcarriers are retained in the
vessel. The sleeve consists of a 4-layer square mesh made of stainless steel 1.4404. There are differ-
ent mesh sizes available. The mesh size determines which particles can permeate and which are re-
tained in the culture. Spinfilters are often used in tissue cell cultures, where microcarrier-bound cells or
suspended cells should be retained, while medium and suspended cell debris should be removed.
Equipment for culture vessel B2 B5 B10
spinfilter pore size 20 µm, cat.-no. 880 830/9 880 833/3 884 055/5
locating pin (5)
O-ring (4), size, cat.-no. 9.25 x 1.78, 39120953 on request 14.0 x 1.78, 39120767
paddle stirrers, number, cat. -no. 2 x 3823907/8 2 x 3823954/0 2 x 884 054/7
harvest pipe SF (7), not adjustable 880 826/0 880 827/9 884 059/8
The spinfilter will be attached to the stirrer shaft. The immersion depth is to be adjusted so that the up-
per edge of the insert is kept above the liquid level (the medium must not flow over the upper egde).
You will mount a special harvest pipe which reaches into the inner compartment of the insert. Before
assemby we recommend to label the filling level for the working volume on the culture vessel.
Assembling of the spinfilter:

1. Remove the top-plate and loosen the stir-
rers. Pull them off the shaft.
2. Mount the harvest pipe SF into one of the
inner top-plate ports, so that it can reach
into the inner compartment of the spinfilter
after assembly (for this the harvest pipes
for the culture vessels B 5 and B 10 have
an angular pipe; the fig. shows the straight
pipe as used for the culture vessel B 2).
3. Slide the spinfilter onto the shaft and up to
the intended height. Consider the filling
level for this. In addition, the lower end of
the harvest pipe should come close to the
bottom of the spinfilter. Locate the spinfilter
with the pin (5).
4. Slide the paddle stirrers onto the shaft. You
should arrange the stirrers crosswise (i.e.
the upper stirrer should be turned by 90°
compared to the lower stirrer. The lower
stirrer should come close to the bottom of
Fig. 4 - 18: Assembly of the spinfilter: the culture vessel.
(2) stirrer shaft 5. After assembly of the culture vessel con-
(3) paddle stirrers nect the tubing for medium removal to the
(4) O-ring harvest pipe SF.
(5) locating screw
(6) Spinfilter
(7) harvest pipe SF
BAE_B Rev. 2.31 - 1099 SW 4.1 4 - 22

Examples for setting-up of a culture vessel with a spinfilter and the accessories:
left hand figure: spinfilter in a standard culture vessel B2, as used for cultures of microcar-
rier-bounded microorganisms and cells, for instance;
right hand figure: spinfilter applied together with a silicone membrane gas supply insert, as
used for tissue cell cultures, for instance.
Fig. 4 - 19: Spinfilter in standard vessel B2: Fig. 4 - 20: Assembly of a spinfilter together
(1) glass vessel with a silicone membrane insert:
(2) stirrer shaft (2) stirrer shaft
(3) paddle stirrers (in crosswise (3) paddle stirrers (crosswise
mounting) mounting)
(5) locating screw (5) silicone membrane insert
(6) spinfilter (6) windings of silicone membrane
(7) harvest pipe SF (7) harvest pipe Sffor vessel B2
(8) level probe (8) level probe
(9) holder for silicone membrane
(10) connection of gas inlet
(11) connection of exhaust
(13) spinfilter
BAE_B Rev. 2.31 - 1099 SW 4.1 4 - 23

4.3.15 Silicone Membrane Insert
A silicone membrane insert is an assembly providing bubble free gas supply to the culture vessel. It is
often used in tissue cell culture or cultures with cells, which are sensitive to shear forces. The mem-
brane insert is installed instead of the sparger pipe. The entire kit includes the following parts:
silicone membrane insert, cat. -no., 884 466/6 884 467/4 884 048/2
for culture vessel type B2 B5 B10
silicone membrane, delivered length 3 x 3.7 / 5,2 m 3 x 3.7 / 10,4 m 3 x 3.7 / 16 m

cat. -no. 3997144/9 3997144/9 3997144/9
paddle stirrers, cat. -no. 2 pcs., 3823907/8 3 pcs., 3823954/0 3 pcs., 884 054/7
membrane filter gas in-/outlet PTFE membrane Ø 50 mm, pore width 0,2 µm, filter area
2
delivery set, cat. -no. 19.6 cm , 2 x 881 007/9
pressure controller setup acc. to drwg. B231.20.08.00.104, order on request
The membrane is optimized for reversible gas diffusion of O2, N2 und CO2. For gas supply you can use
technical gasses or a Gas Mix Unit, cat.-no. 884 171/3, where you can adjust a mixture of gasses op-
timum for the process. O2 and N2 allow a reversible pO2-control. CO2 can be supplied for pH-control
while excess gaseous CO2 of the medium can permeate the membrane and be exhausted via the
membrane outlet. The direction of gas diffusion is determined by the differential partial pressure of the
gasses in the membrane compartment versus the partial pressure of the gas dissolved in the medium.
The pressure inside the membrane is controlled via a pressure controller in the outlet.
Assembly of the silicone membrane kit:

1. Check whether the silicone membrane
is properly wrapped around the holder.
It is wrapped too tight, if the membrane
is depressed at the rods of the holder. It
is wrapped around too loose, if it
sloughes off the indents of the rods.
2. If a membrane is damaged and need to
be replaced, loosen the membrane and
wrap the new membrane around the
rods of the holder. It should be evenly
wrapped around. The inlet side is at the
lower hose connector of the holder, the
outlet is at the upper hose connector.
You should use hose clips to locate the
membranes at the hose connectors.
3. Mount the spinfilter, if it should be ap-
plied. See the section above and the
figure on the next page.
4. Mount the paddles stirrers to the stirrer
shaft and adjust.
Fig. 4 - 21: Silicone membrane insert kit: 5. Mount the holder with ist rod into the
(2) stirrer shaft top-plate and locate with the hexagon
(3) paddle stirrers nut. Recommende top-plate port is N17.
(5) holder for silicone membrane
(6) silicone membrane
(7) harvest pipe
(9) rod for holder assembly
BAE_B Rev. 2.31 - 1099 SW 4.1 4 - 24

6. Mount universal hose adapters for connection of the membranes and of the gas supply and
exhaust into the top-plate. Attach the inlet and outlet tubing of the membrane insert to the
lower hose connectors of these adapters.
7. Connect the membrane filter of the inlet gas to the „inlet“ hose adapter via a short piece of
silicone membrane. Also connect the exhaust filter to the „outlet“ hose adapter.
For optimum gas transfer the internal pressure of the silicone membrane must be precisely
adjusted. For this attach the pressure controller (12) to the vessel and connect to the ex-
haust filter. Gas is supplied to the silicone membrane at a given max. pressure and pread-
justed flow rate. The internal pressure is then adjusted with the exhaust pressure controller.
Note: The internal pressure of the membrane must not exceed 0.8 bar (11.6 psig).
Overpressure can cause the membrane to burst!
8. After assembly, when filling the culture vessel and during operation, you should ensure that
the membrane is allways fully covered with medium. If a spinfilter is installed, the medium
must not flush over the upper edge of this device into the inner compartment.Therefore, after
taking samples, you should refill with medium or use a level control for this.
Fig. 4 - 22:: external connections of the insert: Fig. 4 - 23: complete setup of a culture vessel B2,
(1) glass vessel including the spinfilter in the culture
(2) ... (9) see fig. on previous page vessel (13)
(10) gas inlet filter
(11) exhaust filter (membrane outlet)
(12) pressure controller
BAE_B Rev. 2.31 - 1099 SW 4.1 4 - 25

4.3.16 Blind Closures
Blind closures need to be fitted to any top-plate port, where no other accessory, adapter or culture ves-
sel equipment is installed.
You should note that the blind closures for top-plate ports of Ø 6 mm must be installed be-
fore the top-plate is mounted to the vessel. Those ports cannot be used for assembly of
other culture vessel assemblies lateron. Therefore you must carefully check which assem-
blies are required for the process and need to be mounted into which top-plate port.
Product Information
Blind closures, type DN 11 PG 13,5 DN 19
cat.-no. 38337193 38344416 38316773

for top-plate port Ø = 6 mm Ø = 12 mm Ø = 19 mm
number, for culture vessel:
– B2 7 2 2
– B5 7 2 2
– B 10 7 2 4
– B 2 - Airlift 4 1 5
– B 5 - Airlift 8 2 3
locating screw, type, cat.-no. M10 x 1, 38337185 38344416 38316773

O-ring, type / dimensions 7,65 x 1,78 EPDM, -- 15,6 x 1,78 EPDM,
cat.-no. 39120945 39120830
sealing, cat.- no. -- 38606127 --
Assembling of blind closures

1. Fit the blind closures to those ports
which will not be used for other assem-
blies.
2. Insert a sealing membrane into the
ports of Ø = 12 mm, before fitting the
corresponding blind closure.
Note: It may be advisable to prepare ports,
which are not used by assemblies, as
inoculation ports or with universal
adapters in order to provide sterile ac-
cess to the interior of the culture vessel,
when required lateron during the fer-
mentation run.
Fig. 4 - 24: Blind closures for top-plate ports of

Ø 6 mm, Ø 12 mm and Ø 19 mm
BAE_B Rev. 2.31 - 1099 SW 4.1 4 - 26

4.4 Probes and Electrodes
4.4.1 General Notes

1. Check the requirements for measurement of parameters of your process. Mount all probes
and electrodes as shown below. For recommended placement see the fig. on page 4-9.
The immersion depth is determined by the dimensions of the probes and electrodes and
their adapters. The immersion depth of the level and antifoam probe can be reduced to
adapt it to the working volume of the culture vessel.
2. Before assembling of the probes and electrodes you should check the O-rings and replace,
if necessary. Only O-rings being clean and free of damages will ensure a sterile assembly. It
is recommended to grease the O-rings with some silicone grease. This will prevent the O-
rings from glueing to their contact surfaces of the ports, which can damage them.
3. Carefully tighten the locating screws with their hexagon nuts or screw caps with the span-
ners enclude in the tools kit. Knurled screwes should only be tightened carefully handtight.
4.4.2 Pt-100 - Sensor for Temperature Measurement

The Pt-100 - temperature sensor will usually be mounted into top-plate port N2, see fig. on
page 4-9. If this is not true with your culture vessel you should mark the port at your vessel
correspondingly. The following sensors and attachments are included for the vessels:
for culture vessel type B2, B5 B10 B2-Airlift, B5-Airlift,

cat.- no. 33190887 33190895 00001769 33190895 on request
type 200-4 300-4 400-4 300-4 300-4

HE [mm] 211 316 418 316 316
O-ring (2) 7,65 x 1,78 EPDM, cat.- no. 39120945
locating nut (3) hexagon nut M10x1, cat.- no. 38337185
1. For removal of an installed Pt-100, unscrew

the locating nut (3) and pull the Pt-100 out of
its port.
2. If necessary clean the sensor shaft me-
chanically. The probe can be fouled by cells,
cell debris and floated compenents of the
culture broth, e.g. proteins. Replace the O-
ring (2), if required.
3. Insert the Pt100 into top-plate port of the
culture vessel from top. The immersion
depth is determined by the adapter (1). Slide
the hexagon nut (3) onto the sensor shaft
from below and screw tight.
Before the sterilization of the culture vessel
you should wrap some aluminium foil around
the plug of the cable. This will protect the
plug against the impact of steam. After the
sterilization connect the Pt-100 with the
Fig. 4 - 25: Assembly of Pt-100 multi-conductor plug to the socket on the
control unit.
BAE_B Rev. 2.31 - 1099 SW 4.1 4 - 27

4.4.3 Antifoam and Level Probes
The antifoam probe and the level probe are single conductive electrodes. Their measuring principle is
conductivity measurement; the stainless steel parts of the culture vessel act as the opposing electrode.
Foam or culture medium in contact with the probes causes a current jump by changing the conductivity
of the measuring system.
The probes have a long insulating ceramic coating. This coating prevents short circuit due to fouling.
The following sensors and attachments are included for the vessels:
probe, for culture vessel type B2 B5 B 10 B 2 - Airlift B 5 - Air-

lift
antifoam probe, cat.- no. 884446/1 884446/1 884446/1 884446/1 884446/1

probe type MD 85 MD 85 MD 85 MD 85 MD 85
HE [mm] 50 50 50 50 50
level probe, cat.- no. 884448/8 884448/8 884448/8 884448/8 884448/8
opt. 884469/0
HE [mm] 122 122 122 122 122
opt. 260
O-ring, adapter vs. top-plate port 7,65 x 1,78 EPDM, cat.- no. 39120945
O-ring, probe shaft vs. adapter 6,07 x 1,78 EPDM, cat.-no. 39120821 (not shown)
Maintenance :
1. If necessary clean the probe´s shaft mechanically. The ceramic coating of the probe can
partially be fouled by cells, cell debris and floated compenents of the culture broth, such as
proteins.
2. Check the O-rings of the adapter and inside the adapter (not shown in the fig. below). Clean
or replace if required.
BAE_B Rev. 2.31 - 1099 SW 4.1 4 - 28

Assembling the antifoam and level probe:

1. Insert the adapters for the antifoam and
the level probe into their corresponding
top-plate ports from below (port no. N3
recommended for the antifoam probe,
N11 for the level probe) while the top-
plate is removed. Tighten adapter with
hexagon nut using a spanner.
2. After mounting of the top-plate onto the
glass vessel insert the probes into their
adapters from top down to the required
height and screw tight with their locating
screws.
The possible immersion depths depend
on the probes delivered for the culture
vessel, see above table.
3. For adjustment of the immersion depth
of the level probe you will need to
measure the filling level obtained with
the intended working volume. You
should lable the level on the vessel cor-
respondingly.
The O-ring inside the adapter seals the
probe shaft against the adapter. There-
fore it is possible to pull out the elec-
trode a little even after sterilization for
readjustment, if the immersion depth is
adjusted too deep.
Caution: Never push the probe deeper into

the vessel after sterilization. This
Fig. 4 - 26: Assembling the antifoam and level may cause infections by environ-
probe mental germs.
Other measures
1. Remove the cable before placing the culture vessel with the antifoam and level probe in the
autoclave. The cable must not be sterilized.
2. After autoclaving / before starting fermentor operation connect the probes to their corre-
sponding sockets „Foam“ and „Level“ on the left side of the control unit.
For detailed information about the adjustment of the operation parameters of the antifoam
and level measurement and control see the corresponding chapters in section 5.
BAE_B Rev. 2.31 - 1099 SW 4.1 4 - 29

4.4.4 pH - Electrode
The pH - electrodes are sterilizable combined pH-electrode assemblies, manufactured by INGOLD.

For the BIOSTAT® B electrodes with pasteous bridging electrolyte are used. Such pH - electrodes are
an alternative to electrodes filled with liquid electrolyte, offering several advantages :
pH-electrodes with pasty electrolyte have a pressurized electrolyte compartment and can be
used in environments having an operating pressure of up to 2.5 bar (about 36 psi). The max.
pressure permissible for a culture vessel of the BIOSTAT® B is 1 bar (14.5 psig)
Although a small amount of the electrolyte continuously penetrates the diaphragm, the filling
suffices for the whole lifetime. The electrodes are maintenance-free.
The component parts of the combined glass electrode set up a galvanic chain where the total potential
Eges (in „V“) which is the total sum of all boundary surface potentials Ei, can be measured with a high
impedance voltmeter connected between discharge electrode and reference electrode. The pH value
4-1)
is calculuted from measured potential Eges according to the NERNST equation .
pH-electrode, cat.- no. 33187711 33187703 33187703 08844607 33187703

or 08844640

type Pa 12/200 PA/12/325 PA/12/325 or Pa 12/200 PA/12/325
PA/12/425
HSo [mm] 200 325 325 or 425 200 325
O-ring 10.77 x 2.62 (silicone), included in the delivery of the electrode
4.4.4.1 Reactivation of pH - Electrodes
1. For the first use and after long-term

storage of the electrode in dry state it
need to be reactivated and conditioned
carefully.
Glass membranes, as used for pH
electrodes, built up a thin watery gel
layer, due to a reaction with water. This
layer essentially effects the measuring
quality of the probe (response time,
slope, alkali mistake) and therefore
should regenerate after a long-term
storage.
2. For reactivation you should place the
pH-electrode into a beaker with demin-
eralized water for about 12 ... 14 h. See
also the information in the documents
delivered with the electrode.
Fig. 4 - 27: pH - electrode
4-1) See details about the NERNST equation in the corresponding literature
BAE_B Rev. 2.31 - 1099 SW 4.1 4 - 30

4.4.4.2 Calibration of the pH - electrode
The pH - electrode is calibrated before assembly into the culture vessel. The calibration is
accomplished in two steps, „zero“ and „slope“ calibration. For details about how to proceed
for calibration of the pH-electrode see the corresponding chapter in section 5.
Caution when handing the calibration buffers - they can cause burns.
To consider the possible effects of the heat impact during sterilization and of chemical and
physical reactions of the electrolyte with ingredients of the culture medium, the ph-electrode
can be recalibrated during the process. See also the corresponding chapter in section 5.
4.4.4.3 Assembly at the Culture Vessel
1. Insert the pH-electrode into its top-plate port (port N13 recommended, see fig. „Top plate of
a BIOSTAT® B culture vessel with recommended placement of the accessories“). The im-
mersion depth below the top plate depends on the delivered electrode. The culture vessel
should be filled with water or the culture medium time to prevent drying of the electrode.
2. Tighten the electrode carefully handtight with. Do not use a spanner or similar tools.
3. For autoclaving disconnect the cable. You can cover the plug of the electrode with the pro-
tective cap included in the delivery or wrap some aluminium foil around to protect against the
impact of steam.
4. After autoclaving / before starting fermentor operation connect pH -electrode probe with its
cable to the „pH“ - socket at the left side of the control unit.
4.4.4.4 Maintenance
A pH - electrode is subject to wear and tear. This is caused by the thermal effects during the steriliza-
tion or by chemical reactions of the diaphragm or of the pasteous electrolyte with ingredients of the
culture medium, for instance. Also fouling by cells, cell debris or macromolecules can change the
measurement behavior.
The pH - electrode can be sterilized several times. The number of sterilizations depends on
the process conditions (i.e. the composition of the medium and resulting impacts on the
electrode). If a pH - calibration is malfunctioning, it should be replaced.
Symptoms for wear and tear are a slower response, a reduced slope or a zero drift. If the
measured pH-value is different for the medium at intensive stirring and at stopped stirrer
drive, this may incicate fouling of the diaphragma.
A functional check of the pH - electrode is limitted to checking the electrode´s zero and slope
and to comparing the pH-value measured „online“ with the pH-value measured in a fresh
sample. For details see the section „pH - Calibration“ in section 5 of this manual and the
documentation delivered with the electrode.
4.4.4.5 Storage of the pH - Electrode
1. You should flush the pH - electrode with aqua dest. after any run. Never store an electrode
with dirty diaphragm. The residues will dry and may not be removable.
2. Short-term storage is possible in demineralized water. For a longer storage you can place
the pH - electrode in a beaker filled with 3m-KCL solution. You can also fill a small amount of
this electrolyte into the protective cap delivered with the electrode and attach this cap onto
the electrode´s tip.
BAE_B Rev. 2.31 - 1099 SW 4.1 4 - 31

4.4.5 pO2 - Electrode
The measurement of the dissolved oxygen is made with a sterilizable pO2-electrode (manufactured by
INGOLD). The electrode operates on the polargraphic principle and consists of an Ag-anode and Pt-
cathode, which are seperated by a gas permeable polymer membrane from the solution to be meas-
ured (CLARK-principle). Anode and cathode are connected conductively by an electrolyte; the electro-
lyte builds a small, definable layer between the membrane and the cathode.
At a suitable polarization voltage, the oxygen, which diffuses through the membrane into the cathode,
will be reduced. This chemical reaction produces an electrical current in the nA-range, which is directly
proportional to the oxygen partial pressure (the actual measureable quantity of the measured solution).
Since the permability of the membrane depends on temperature, an exponential rinse (3 %/ degC) in
the electrode current will be observed at rising temperature. This effect will be compensated by auto-
matic temperature compensation by means of an integrated thermistor in the electrode.
pO2 - electrode, cat.- no. 3318241/8 3318240/0 3318240/0 3318241/8 3318240/0

opt. 884468/2

type of electrode 12/220 12/320 12/320; 12/220 12/320
optional 12/420
HSo [mm] 212 312 312, opt. 412 212 312
O-ring 10.77 x 2.62 (silicone), included in the delivery of the electrode
Assembly of the pO2 - Electrode:

1. Insert the pO2 - electrode into the suit-
able port (port N4 recommended, see
fig. on page 4-9). The immersion length
below the top plate depends on the
electrode delivered with the type of cul-
ture vessel.
2. Screw the locating nut carefully hand-
tight, do not use a spanner or similar
tool.
3. For autoclaving disconnect the cable.
You can cover the plug of the electrode
with the protective cap included in the
delivery or wrap some aluminium foil
around to protect against the impact of
steam.
4. After autoclaving / before starting fer-
mentor operation connect the pO2 -
electrode with its cable to the socket
„pO2“ at the left side of the control unit.
The electrode must be polarized for at
least 6 h before calibration or meas-
Fig. 4 - 28: Assembling the pO2 - electrode urement. See detailed information be-
low.
BAE_B Rev. 2.31 - 1099 SW 4.1 4 - 32

4.4.5.1 Calibration of the pO2 - Electrode
Below you will find a summary of important steps of calibration. More detailed information
and the inputs required for the calibration menu are included in section 5.
The calibration of the pO2 - electrode is made in the culture vessel after autoclaving and un-
der the conditions of the fermentation. Before calibration, it must be polarized. You can use a
polarization module available from the manufacturer for this or you can connect the elec-
trode to the control unit and switch-on the system. The polarization must be repeated any
time the electrode is disconnected from the amplifier for more than 10 min, but may require
less time then.
Calibration includes zero and slope calibration. The „zero“ is the electrode´s current, when
no oxygen is present in the culture medium, the „slope“ is usually the pO2 after saturation of
the medium with air at the maximum air supply intended for the process.
1. Adjust the operating temperature in the culture vessel.
2. For „zero“ calibration you can measure the pO2 of the culture medium before starting the air
supply. The medium will be degassed almost completely due to the heat impact during ster-
ilization and thus should not contain dissolved oxygen. Alternatively you can s upply an oxy-
gen-free gas (such as nitrogen of 99.98 % purity) to the culture medium to displace the dis-
solved oxygen until a constant pO2 near „0 %“ can be read at the measurement and control
system. Confirm this value as „zero“ as shown in section 5 of this manual.
3. For slope adjustment activate the air supply and adjust the stirring speed. The medium
should be optimally gassed (max. flow rate intended for your process) and mixed. At a stable
display of the measured value you can calibrate this as „100 % pO2“
4. After calibration the gas supply rate required for the startup of the intended fermentation pro-
cess can be adjusted on the rotameter of the control unit. Note that the rotameter is cali-
brated according to standard conditions (temperature 20 degC, with air at 2 barabs). If it is
important to maintain precise operating air flow-rates for further calculations, this makes it
necessary to recalculate the indicated flowrate with a correction factor. See further informa-
tion the supplement.
4.4.5.2 Function Control of pO2 - Electrode
The electrode should be inspected in regular intervals. This is best carried out outside of fermentor.
The electrode must be polarized at least 6 hours before the inspection. A relative good assessment
can be made of the operating effectiveness of the pO2-electrode by checking the zero current and the
response time, see the short information below. More detailed information will be included in the
documentation of the manufacturer delivered with the electrode.
1. Hold the electrode in the air. Then measure the pO2 and wait for a stable indication of the
measured value. Note this value.
2. Place the electrode in an oxygen-free environment (nitrogen gas of purity >99.98 %, for in-
stance). Wait for a stable indication of the measured pO2.
When changing from ambient air to oxygen-free environment, the measured zero current
must be less than 1.5 % of the current in the air after 5 minutes.
If a malfunctioning is suspected you should service the electrode. Usually the membrane
module and the electrolyt will be changed for this. For detailed information on how to pro-
ceed, see the documentation of the manufacturer.
Caution when handing the electrolyte - it can cause burns.
BAE_B Rev. 2.31 - 1099 SW 4.1 4 - 33

4.5 External Equipment for Connection to the Culture Vessels
4.5.1 Exhaust Cooler
The exhaust cooler can, by condensation, lower the water vapour content in the exhaust. This reduces
the loss of liquid via the exhaust and prevents the sterile filter from being blocked by humidity.
If the exhaust filter is blocked, the air supply to the culture can be reduced or blocked at all.
This can result in inadmissible overpressure.
Wet exhaust filters can be fouled by environment germs introduced via the outlet. The germs
can permeate the filter membrane by time and contaminate the culture.
exhaust cooler, cat.-no. 3824042/4 3824116/1 3824116/1 3824042/4 3824116/1

design features stainless stainless stainless stainless stainless
steel tube steel tube steel tube steel tube steel tube
with angular with straight with straight with angular with straight
pipe pipe pipe pipe pipe
HK [mm] 222 255 255 222 255
O-Ring (1) 15,6 x 1,78 (EPDM), 3912083/0
O-Ring (2) 18,77 x 1,78 29,82 x 2,62 29,82 x 2,62 18,77 x 1,78 29,82 x 2,62
39121763 39121780 39121780 39121763 39121780
extension kit, cat.-no. nein 884 459/3 884 459/3 884 459/3 884 459/3
The exhaust cooler can be mounted directly into ist top-plate port or via the extension kit.
The extension kit includes a tubing and a holder attached to the culture vessel. For the ster-
ilization in the autoclave the exhaust assembly can be removed from this holder and bent
over. This reduces the internal height required for the autoclave.
Assembly without extension kit (standard equipment, see fig. on next page) :
1. Fit the exhaust cooler into its port in the top plate (port no. N1 recommended) and fasten it
there with its cap nut.
2. Attach the exhaust filter to the exhaust cooler via a short piece of silicone tubing as shown in
the corresponding section below. Do not clamp-off this tubing for the sterilization in the
autoclave. The exhaust outlet provides the necessary pressure compensation of the interior
culture vessel with the ambient air.
The hoses for the cooling water supply and drain can be connected when fermentation proc-
ess is initiated.
BAE_B Rev. 2.31 - 1099 SW 4.1 4 - 34

Assembly with extension kit (optional equipment) :

1. Place the tubing adapter of the extension kit into
the top-plate port assigned for the exhaust
cooler (port N1 recommended) and screw tight.
2. Replace the knurled top-plate screw next to the
port for the exhaust cooler by the special screw
of the extension kit. This screw has a locating
pin for the support rod of the extension kit,
where the tubing adapter will be located.
3. Place the support rod onto the pin and screw
tight with the lower fastening screw.
4. Place the rod attached to the upper screw
adapter of the extension tubing into the boring of
the support rod and screw tight with the upper
fastening screw.
5. Screw the exhaust cooler into the upper screw
adapter of the extension tubing.
6. Attach the exhaust filter to the exhaust cooler
via a short piece of silicone tubing as shown in
the corresponding section below. Do not
clamp-off this tubing for the sterilization in
the autoclave. The exhaust outlet provides the
Fig. 4 - 29: Fitting the exhaust cooler necessary pressure compensation of the interior
(version for vessel type B2 culture vessel with the ambient air.
with angular connection pipe)
The hoses for the cooling water supply and
drain can be connected when fermentation pro-
cess is initiated.
Bending-over of the exhaust cooler for autoclaving:

1. Loosen the screw of the support rod, which lo-
cates the rod attached to the upper screw
adapter of the extension tubing. Pull the rod out
of the boring of the support rod.
2. Bent-over the exhaust cooler with the filter to
minimize the height of the entire culture vessel
assembly.
3. After autoclaving place again the rod attached to
the upper screw adapter of the extension tubing
into the boring of the support rod and locate with
the screw.
Fig. 4 - 30: Assembling of the extension

kit
BAE_B Rev. 2.31 - 1099 SW 4.1 4 - 35

4.5.2 Sterile Filters for Air/Gas Supply and Exhaust
The air supply and exhaust filters are autoclaveable membrane filters in plastic housing. They have a
pore size of 0,2 µm and ensure sterile air (or gas) supply to the culture vessel and sterile exhaust.
for culture vessel B2 B5 B10 B2-Airlift B2-Airlift
membrane filter 2 pcs. 2 pcs. air inlet: 1 x 881 007/9; 2 pcs. 2 pcs.
delivered no., cat. -no. 881 007/9 881 007/9 exhaust 1 x filter element 881 007/9 881 007/9
cat.-no. 3922830/4
(not shown)
Filter 881 007/9 : - flat membrane made of PTFE, in plastic housing
- pore width 0,2 µm
- membrane - Ø = 50 mm
2
- filter area 19,6 cm
Filter 3922830/4 : - stareshape folded PTFE - membran cylinder
in plastic cartridge
- pore width 0,2 µm
2
- filter area about 500 cm
4.5.2.1 General Notes
1. The membrane filters can be sterilized several times and therefore be used for several fer-
mentation runs. The number of uses will depend on the application conditions.
2. You should care, that the filters are not applied to vapour or moisture. If moisture conden-
sates at the filter membrane, this may block the filter. This may require to carefully dry the
filter before it can be used again. You should also avoid that foam of the culture enters the
filter. Then the filter will be blocked permanently and must be replaced.
3. Carefully check the pieces of silicone tubing used for connecting the filters. They must not
be damaged, since damages can cause environmental germs to enter the tubing and con-
taminate the culture.
4. Air or inlet gas can be connected and supplied from the control unit or an extra Gas Mix Unit.
An extra Gas Mix Unit can be applied, if the inlet air should be enriched with oxygen.
Above this you can apply a pO2 - control by reversible supply of either O2 and N2. pH-control
is possible by supply of gaseous CO2, for instance. Such control strategies are often applied
for tissue cell cultures using the silicone membrane insert described above in this manual.
For connection of a Gas Mix Unit see the documentation delivered with the device.
BAE_B Rev. 2.31 - 1099 SW 4.1 4 - 36

4.5.2.2 Connecting the Inlet Air Filter
1. Connect a short piece of silicone tubing

(4x7 mm) to the hose connector (2) and to the
sparger pipe (or the inlet of the silicone mem-
1 brane insert, if installed).
2. Attach a piece of silicone tubing of suitable
length for connection to the control unit or to
the Gas Mix System onto the hose connector
(1) of the filter. Secure the tubing against slip-
ping.
3. Connect a piece of silicone tubing of suitable
length for later connection to the external sup-
2 ply at the inlet connector of the filter (1).
4. Clamp-off the tubing prior to autoclaving of the
culture vessel. The tubing can be inserted into
the clamping device assigned for this (the
„dovetail clamp“, see fig. „Top plate design
Fig. 4 - 31: Membrane filter for air inlet and and placement of accessories“ for details).
exhaust, cat.-no. 881 007/9
4.5.2.3 Attaching the Exhaust Filter
1. Connect a short piece of silicone tubing (3.2 x 1.6 mm) to the hose connector (2) of the filter
and to the exhaust cooler. Secure the tubing against slipping. For a standard setup you need
not install a tubing to the „outlet“ connector (1) of the exhaust filter. This provides free ex-
haust into ambient air.
2. If a silicone membrane insert is used for bubble free gas supply to the medium, fit the tubing
to the filter (hose connector (2) in this case) and to the outlet hose connector of the silicone
membrane insert. Connect the outlet of the exhaust filter (hose connector (1)) to the pres-
sure controller. See description of the setup of a silicone membrane insert for details.
3. If you want to dearate from the vessel headspace, you can connect an additional exhaust
filter to a hose connector of an universal adapter installed in the top-plate, for instance.
The exhaust tubing must not be clamped-off during the sterilization, since it provides
the necessary pressure compensation between the culture vessel and the ambient air
during and after the sterilization. The exhaust filter guarantees sterile pressure compen-
sation.
4. Take special care that the tubings are not damaged and do not slip during manipulations of
the culture vessel. If unsterile air enters the air inlet or exhaust tubing, the culture will be
contaminated by environmental germs.
5. Also check the exhaust filter connection and the exhaust filter after the sterilization. If the
culture medium tends to foam, the foam can enter and block the exhaust. Then it may be
necessary to replace the filter and repeat the sterilization.
BAE_B Rev. 2.31 - 1099 SW 4.1 4 - 37

4.5.3 Corrective Solution Bottles
For corrective solutions (acid, alkaline, antifoam or nutrient solution) the delivery set of the culture ves-
sels includes 250 ml bottles, cat.-no. 0882360/0, for the vessels B 2 and B2-airlift and 500 ml bottles,
cat.-no. 0882361/8, for the vessels B 5, B5-airlift and B10. The bottles are autoclaveable glass bottles
made of borosilicate glass.
For supply of solutions during long-term or continuous operation, we recommend to prepare several
bottles to have sufficient volumes available. If „bulk“ volumes of nutrient solutions should be supplied,
storage bottles made of polypropylen may be applied (not shown herein). They offer storage volumes
of 10 l (cat.-no. 882364/2), 20 l (cat.-no. 882365/0) or 50 l (cat.-no. 882366/9).
Equipment of the bottles

Stainless steel cap (3) with 2 integrated hose con-
nectors. A gasket (2) on the rim of the glass bottle
(1) seals the cap. The cap is tightened with the screw
cap (4).
PTFE dispenser hose (7) which is inserted into the
bottle and is resistant to the aggressive action of ac-
ids and bases at high temperatures.
Sterile air filter (5), cat.-no. 3922480/5, for pressure
compensation when solution is removed (1).
Silicone tubing (6) for connecting the bottle to the
culture vessel.
Assembly of a bottle :
1. Attach the PTFE hose (7) to one of the hose con-
nectors from below. Shorten it so that the end is 1-2
mm above the bottom of the bottle.
2. Fill the bottle with acid, alkali or antifoam agent.
Screw the cap (4) onto the bottle.
Warning: When using acid or alkali agents take

precautions against burns.
We recommend not to use hydrochloric acid
(HCL) for pH-control. HCL can cause corrosion
even of stainless steel parts.
3. Cut a length of silicone tubing sufficiently long to

connect the bottle to the control unit (for passing
through the peristaltic pump). Attach it to the hose
connector carrying the PTFE hose.
4. Connect the other end of the silicone tubing to one of
the nozzles for corrective solutions. Locate the
tubings on the hose connectors.
Fig. 4 - 32: Storage bottles of 250 ml 5. Attach the air filter (5) to the other hose connector via
for acid, alkali and a piece of silicone tubing.
antifoam solution
6. Place the bottle into the holders at the culture vessel.
Clamp-off the tubings. No solution should escape
from the bottles during sterilization.
BAE_B Rev. 2.31 - 1099 SW 4.1 4 - 38

4.5.4 By-pass Sampler Kit
The by-pass sampler can be used for the continuous or discontinous removal of very small samples
from the culture for the purpose of off-line-analysis, where almost actual samples are required. It can
also be used for introducing small volumes of addition solutions which should be fed to the culture.
The by-pass sampler consists of a two-channel screw connector which is fitted into the top plate of the
culture vessel, the membrane holder and approx. 2 m of 1.6 x 1.6 mm silicone tubing for the by-pass
hose. The membrane holder includes a septum membrane, which can be punctured using a syringe
(since the membrane is self-sealing, it can be punctured several times, if necessary).
You can use dosing pump such as the FE 411 (B. Braun Biotech International GmbH) to remove the
sample from the culture vessel. The sample will be fed through the bypass sampler and returned to the
culture vessel after it has passed through the by-pass via the second hose connector of the two-
channel screw connector. Thus you will receive representative samples for the state of process, for in-
stance.
Assembly and connection

1. Add the rising pipe (piece of teflon tube)
to one of the lower hose connectors of
the adapter. During equipment of the
culture vessel screw the adapter into a
suitable port in the top-plate.
5. Connect two pieces of silicone tubing to
the upper hose connector:
- one for sample removal from the ves-
sel is to be connected to the hose con-
nector with the rising pipe.
- one ifor return of sample which has
Fig. 4 - 33: Two-channel adapter for connection to passed through the bypass, is to be
the top-plate connected to the free hose connector.
2. Prepare the membrane holder. Insert a
new piercing membrane. Add the
washer and locate with the sleeve nut.
Finally add the protective cap for sterili-
zation.
3. Connect the tubings for sample remo-
mal and return to the membrane holder.
4. Locate all parts for safe handling during
autoclaving. Autoclave together with the
culture vessel.
Fig. 4 - 34: Assembly of membrane holder
BAE_B Rev. 2.31 - 1099 SW 4.1 4 - 39

Taking Samples using the Bypass - Sampler

1. After placement of the entire assembly at
the working place and/or for taking sam-
ples, introduce the removal tubing into a
peristaltic pump. Switch-on the pump to
have a continous flow of culture broth
through the membrane holder.
2. Prepare a sterile syringe for the sample.
3. Remove the cap of the membrane holder.
Burn-off the membrane and the needle of
the syringe with an open flame or moisten
with a desinfectant.
Fig. 4 - 35: Taking a sample via the membrane 4. Pierce-in the syringe into the membrane
holder and pull the plunger to remove the sam-
ple. Afterwards withdraw the syringe from
the membrane and attach again the cap
to keep the membrane sterile until you
pierce-in another syringe.
Notes:
After applying a desinfectant, wait for a sufficient time so that it can take effect. Carefully
check if it could be used with your culture. Residues of the desinfectant entering the medium
must not influence the culture. Alcohol, for instance, may not destroy spores of some envi-
ronmental germs and can cause that such spores to enter and contaminate the medium.
Remove any residues of the desinfectant.
Introducing an additive solution is done in a similar way. Instead of removing the sample
from the bypass, you will have to introduce the addittive solution with the syringe. Prepare a
sterilie additive solution for this.
The membrane of the Bypass-Sampler is „self-sealing“ and can be punctured several times.
If you are manipulating the sterile parts carefully, there is only a minimum risk of contamina-
tion of your culture.
BAE_B Rev. 2.31 - 1099 SW 4.1 4 - 40

4.5.5 Manual Sampling Kit
The manual sampler, cat.-no 884 462/3, can be used for discontinuous removal of samples at a larger
volume. It consists of the following parts, see fig. of the setup:
Sampling tube (pos. 2),
Clamping device for assembly to one of the stands of the culture vessel
Syringe for sample removal with sterile filter assembly (pos. 4)
Silicone tubing 1.6 x 1.6 mm for connection of the syringe and 3.2 x 1.6 mm for connection
to the harvesting pipe in the culture vessel and the sample outlet,
Hose clamp
The sampling tube has a screw cap with three hose connectors. One is used for connection to the
sampling tube of the culture vessel (pos. 1), one for outlet of the sample to external receptacle (pos. 5)
and one for the syringe (pos. 3). The sterile filter assembly of the syringe prevents insterile air to enter
and contaminate the device.
Assembling the manual sampling kit

1. Unscrew the cap of the sampling tube. Attach a
short piece of silicone tubing Ø 3.2 x 1.6to the
rising pipe, it should be reach down to the bot-
tom of the tube. Screw cap onto the tube again.
2. Attach a piece of silicone tubing Ø 3.2 x 1.6 to
the outside hose connector of the rising pipe.
This tubing is used to transfer a sample into an
external receptacle (via outlet tubing, 5).
Attach another piece of silicone tubing

Ø 3.2 x 1.6 to hose connector (1) and con-
nect to the sampling pipe of the culture
vessel. This tubing is used for transfer of
the sample from the harvest pipe into the
sampling tube
3. Mount sterile filter to the syringe and connect to

the third hose connector of the sampling tube
via a short piece of silicone tubing Ø 1.6 x 1.6,
see pos. (3).
4. Attach the clamping device to one of the stand
rods of the culture vessel so that the manual
sampler can easily be handled. Insert the sam-
pling tube and the syringe into the clamping de-
vice.
5. Clamp-off the silicone tubing between the rising
pipe of the culture vessel and the sampling tube
prior to autoclaving.
6. Autoclave the complete assembly together with
Fig. 4 - 36: Manual Sampler the culture vessel.
BAE_B Rev. 2.31 - 1099 SW 4.1 4 - 41

Removal of Samples with the Manual Sampler

1. Prior to taking a sample push down the plunger of the syringe. The outlet tubing (5) should
be open at this time.
2. Remove the hose clamp from the tubing connected to the culture vessel (1) and clamp-off
the outlet tubing (5) now.
3. Draw the plunger of the syringe (4). The sample is sucket through the sampling pipe and the
tubing (1) into the sampling tube. Transfer the required sample volume. Then push the
plunger back a little until the tubing connection to the culture vessel is empty. This avoids
that residues of the sample are kept in the tubing until the next sample is taken.
4. Losen the hose clamp from the outlet tubing and clamp-off again the transfer tubing from the
culture vessel (1).
5. Lead the outlet tubing (5) into a receptacle (flask, beaker, etc.). To transfer the sample
slowly push the plunger of the syringe. The sample is driven through the rising pipe of the
sample tube and the outlet tubing into the receptacle.
Notes:
It is recommended to completely empty the sample tube as shown in step 5. Sample resi-
dues in the tube may contaminate the next sample and change its characteristics.
Between two samples dip the open end of the hose into a disinfectant in order to avoid con-
tamination of the culture broth.
BAE_B Rev. 2.31 - 1099 SW 4.1 4 - 42

Control Unit
5 CONTROL UNIT
5.1 General Information
The control unit of the laboratory scale fermentor BIOSTAT® B includes a digital measurement and
control system. The section below describes all standard functions available with the current software
version 4.15-.1). For a detailed technical information about the hardware design, the interface specifi-
cations and the default settings please refer to the supplement. The protocol for communication with a
host computer is described in details in the manual „DCU-Host Communication“5-2).
5.1.1 Operating Behaviour
The measurement and control system stores all parameters which are adjustable by the operator (set-
points, calibration parameters, etc.) in a battery-buffered memory. These parameters will be backed-up
when the BIOSTAT® B is switched-off and are available again when the fermentor is restarted for an-
other operation sequence and after power failure.
For the starting behavior of certain system functions which have direct access to the fermentor (some
controllers, for instance) and for the outputs of the measurement and control system two different types
of power failure are differentiated :
Switching the BIOSTAT® B „off“ and „on“ via the mains switch on the front panel.
Shut down caused by a power failure (mains cut-off).
5.1.2 Switching the BIOSTAT® B „Off“ and „On“
The BIOSTAT® B will be switched on or off, respectively, via the mains switch on the front panel of the
control unit. After switching-on the measurement and control system is in this state of operation :
All controllers are switched off, actuators are in rest (neutral) position.
5.1.3 Power Failure
In the menu „Maintenance“ a maximum power failure time (FAILTIME) can be set. See section „Utility
Functions“, on page 5-37, for details. In the case of a power failure, after being restarted or when the
mains supply is available again, the system continues all activities as follows:
At a power failure which is shorter than the time set for the FAILTIME all controllers will con-
tinue with the setpoint active before the power failure
If the power failure lasts longer than set by the FAILTIME, the measurement and control
system will be switched over into a defined basic state (default state). It behaves as if being
switched-off by the mains switch.
5-1) If you have a control unit with another software version, please contact B. Braun Biotech Inter-
national GmbH for a corresponding version of the Operating Manual.
5-2) Further information available on request. Please contact your B. Braun representative or
B. Braun Biotech International GmbH, see address in introductionary page.
BAE_B Rev. 2.31 - 1099 SW 4.1 5-1

Control Unit
5.1.4 Menu Structure
The operation display of the BIOSTAT® B is divided into the main function and function level:
Main functions Functions
Process Values Process Values (display of process data in various forms)

Calibration Calibration function for probes and pumps, as used for
– pH - measurement and control
– pO2 - measurement and control
– ACID – pump
– BASE – pump
– foam measurement and control AFOAM
– level measurement and control LEVEL
– substrate control SUBS1 ... 2
Control Loops DDC controller functions:
– temperature control ”TEMP“
– stirrer speed control “STIRR“
– pH-control “pH“- pO2-Control “pO2“
– pump control function “AFOAM“- pump control function “LEVEL“
– pump control function “SUBS1 ... 2“
– gasmix control function “GASMX”
– AIRFLOW - control (only with optional massflow control)
Maintenance Function for system maintenance and trouble shooting; configuration of
in/outputs, such as :
– RECORDER
– PRINTER
– HOST
– UTILITY
– MEASUREMENT RANGES
– manual operation (DIGITAL OUTPUTS)
– manual operation (ANALOG OUTPUTS)
Sterilization Function for culture vessel sterilization
Fig. 5-1: Menu structure of the control system
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Control Unit
5.2 Operation of the Control Unit
5.2.1 General Operating Information

1. You will select the required main function by pressing the corresponding main function key.
The possible main functions are directly labelled on the keys. An active main function will be
displayed in the first row of the LCD and indicated by a lighting LED in the key.
2. Each main function except of „PROCESS VALUES“ has several subfunctions arranged in a
defined order. As an example you'll always find the submenu „pH-Sensor“ at first whenever
you select the main function „CALIBRATION“ or the submenu „Temp“ whenever you select
the main function „CONTROL LOOPS“.
To switch over to subsequent menues of a main function repeatedly press the main
function key. See above figure for the order of the submenues within each main func-
tion. You may change from one main function to another at any time by pressing its
main function key.
3. Non-numerical entries in a submenu are used for selecting the state of operation (such as
switching over „auto“ „man“, „off“ „on“). To switch over to an alternate state of opera-
tion press the „ALTER“-key. Allways confirm the change of an entry by pressing „ENTER“.
Otherwise the change will be ignored.
4. If a menu includes more lines than visible on the actual display, either the top line or the low-
est line of the information entry will include an arrow „ „ or „„. You can switch over to addi-
tional lines using the cursor-keys. Switching over usually takes place line-by-line, except with
the Process Display, which displays a new page, then.
5. The access to the menues for adjustment of system functions, like parametrization of the
controllers, is protected by a password. Note the information about the available passwords
in the corresponding sections below. When you enter a password into the corresponding
entry, the entry will not be displayed.
6. If it is difficult for you to read the LCD, you can change its contrast. This is done with the
main function „MAINTENANCE“, in submenu „UTILITIES“.
Fig. 5-2: Operating terminal of the BIOSTAT® B measurement and control system
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Control Unit
5.2.2 Basic Menu Structure

1st line :
For main functions with several subfunctions (except PROCESS VALUES) the first line is a
headline which identifies the selected function. Data may also be displayed in this line. The
headline will always be displayed, even when you scroll down to following lines of the menu.
Different main functions may show the same headline. Note the LED's in the main function
keys, the LED of the main function selected at this time will illuminate.
Example :
TEMP : 37.5 oC
2nd to 4th line :
Information entry, line - structure. In some cases, if the parameters cannot be displayed
within the three lines available, you can scroll down to following lines. The headline will re-
main unchanged at this time.
Example :
pO2 : 88,5 %
SP : 83,4 %
MODE : auto GASM
PARA :
5.2.3 Operating Information
Symbol/entry meaning, possible input
(in top line) Additional lines available before the actual line.
– You can scroll upward to above lines with the „„-key.
(in lowest line) Additional lines to follow within this menu.
– You can scroll down to following lines with the „ „-key.
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Control Unit
5.3 Main Function „PROCESS VALUES“

The main function PROCESS VALUES only includes one function. The display has no head-
line. If you scroll down to lines beyond the 4th line even the top line will change.
Operating display, page 1:
TEMP : 42.7 oC
STIRR: 800 rpm
pH : 10.3 pH
pO2 : 80.4 %
ACID : 32 ml ñ
BASE : 200 ml
AFOAM: 12 ml
LEVEL: 1300 ml
SU1DC: 25 ml/m ñ
SUBS1: 3.0 %
SU2DC: 25 ml/m
SUBS2: 3.0 %
Optional process value display:
GASMX: ñ
AIRFL: 25.00 l/m
EXT_1: 0.0 %
EXT_2: 0.0 %
additional pages of optional display
EXT_3: 0.0 % ñ
EXT_4: 0.0 %
FOAM : off
LEVEL: off
HEATC: off ñ
MFAIL: off
HIFOM: off
Symbol / entry meaning, possible input
TEMP, etc. display of process value with physical unit, such as

– process values of analog inputs (Temp, Stirr, etc.)
– dosing counters
– process values of digital inputs (Foam/Level)
, Display of additional lines accessible via „ , „-keys
LEVEL Dosing counter of level control pump
SU1DC Dosing counter of substrate pump 1
SUBS1 Display of substrate setpoint of substrate pump 1
SU2DC Dosing counter of substrate pump 2
SUBS2 Display of substrate setpoint of substrate pump 2
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Control Unit
GASMX Display of setpoint of gasmix controller

AIRFL Display of airflow rate
EXT_1 ... 4 Display of up to four input signals of remote devices:
– the entry is in terms of „0 ... 100 %“ for input signals of „0 ... 10 V“
(or of 0/4 ... 20 mA, if inputs are set as current signals).
FOAM, LEVEL Display of „foam“ or „level“ signal
HEATC Display of heater monitoring; in case of malfunctioning the system dis-
plays an additional error message
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Control Unit
5.4 Main Function „CALIBRATION“
5.4.1 Calibration of the pH - Electrode
5.4.1.1 General
The pH-electrode is calibrated by a two-point calibration which determines the electrode parameters
„zero drift“ and „slope“ using buffer solutions. The calibration is to be done before the electrode is in-
stalled in the culture vessel. For calibration you can use technical buffers, but we recommend that you
should prefer original buffer solutions of the manufacturer of the electrode.
For the specifications and the assembly of the pH-electrode see section 4.
Warning: The calibration buffers can cause burns. Be careful when handling the buffers.
You should wear protective gloves during the calibration procedure.
The software of the control system calculates the pH-value from the electrode voltage with regard to
„zero drift“ and „slope“, considering the Nernst equation. To compensate for effects of temperature the
actual temperature can be entered manually during the calibration.
The pH - electrode is calibrated before mounting into the culture vessel, i.e. prior to its sterilization. The
high temperature during the sterilization can cause a shift of the electrodes’ zero. Also chemical com-
pounds of the culture broth may effect the measuring characteristics of the electrode. Therefore the
pH-electrode should be calibrated and checked for proper function regularly.
To compensate for changes of the measurement characteristics of the pH-electrode the software of-
fers a recalibration function. You can measure the pH-value externally in a sample taken from the pro-
cess and enter this pH-value into the calibration menu. The system will then recalculate the calibration
function and apply the new parameters for the pH-control function.
For measuring the pH-value in such a sample, you should consider the following:
Use only fresh samples. Take care that the sampling method and the containers do not in-
fluence the pH-value of the sample
The devices used for external pH - measurement should ensure precise pH-measurement
If the pH - value measured by the BIOSTAT® B control system and the externally measured
pH-value largerly deviate from each other, we recommend to check you remote measure-
ment procedure twice rather than changing the calibration parameters. Entering of wrong re-
calibration parameters into the system can considerably influence the control behaviour and
the supply of corrective solutions, which then will disturb your process.
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Control Unit
5.4.1.2 Operating Display :
pH : 7.30 pH
o
TEMP : 23.4 C man
BUFZ : 7.00 pH ok
BUFS : 4.01 pH ok
pH : 7.30 pH
RECAL: 6.76 pH no
ZERO : 4 mV ok
SLOPE: 58.4 mV ok
auto/man When the menu is selected, the cursor is placed at this entry, showing
the mode which has been selected previously. Press ALTER to change
the temperature compensation mode:
– auto: pH-measurement during the process; with temperature compen-
sation using the measured temperature;
– man: mode for pH-calibration, with temperature compensation by the
temperature which is entered manually
change to „man“ mode for pH-calibration using the temperature labelled
on the buffer bottle for the reference pH-value;
TEMP display of culture vessel temperature, when in the „auto“ – mode input of
the calibration temperature when in the „man“ - mode
BUFZ input of the pH - value of the zero buffer (as labelled on the buffer bottle
for the actual temperature)
ok confirmation of the calculation of the zero point; after confirmation the
cursor jumps to the BUFS - entry
BUFS input of the pH - value of the slope buffer (as labelled on the buffer bottle
for the actual temperature)
ok confirmation of the calculation of the slope. After confirmation the cursor
jumps to the „man/auto“ - entry. The temperature compensation will
automatically switch over to „auto“ mode
RECAL display and input of separately measured pH-value for the recalibration of
the pH-electrode
no / yes ... – no: the electrode will not be recalibrated
– yes: the electrode will be recalibrated
After pressing ENTER the cursor jumps to the „RECAL:“ entry. Here you
can enter the pH-value measured in a sample taken from the process.
After confirmation of the manually entered pH-value with ENTER the
system adapts the „zero“ - calibration to the actual conditions
ZERO display of the electrodes’ zero in [mV]
SLOPE display of the electrodes’ slope in [mV]
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Control Unit
5.4.1.3 pH - Calibration Routine
1. Press the main function key „Calibration“ once to select the menu for pH-calibration. In the
menu the cursor is on the „man/auto“ - entry.
2. To allow for entering the reference temperature labelled on the buffer bottles for the relation-
ship of pH versus temperature of the buffer press „ALTER“ to select „man“ual temperature
compensation. Confirm with „ENTER“. The cursor jumps to the „TEMP“ - entry.
3. Fill „zero buffer“ of pH 7.0 into a beaker. Place the pH-electrode into the beaker. Measure
the temperature of the buffer and enter this as reference temperature into the TEMP entry.
Confirm with „ENTER“. The cursor jumps to the „BUFZ“ - entry.
4. Check the reference pH-value of the „zero“ buffer labelled on the bottle for the actual buffer
temperature. Enter this pH-value in the BUFZ-entry and confirm with „ENTER“. The cursor
then jumps to „ok“ and the system starts an automatic monitoring of the electrode. If the
measured signal is constant within the given limits, the system stores the measured voltage
as the electrode´s zero. Then the cursor automatically jumps to the BUFS entry.
5. Fill some slope buffer of either pH 4.0 or pH 9.0 (depending on the expected pH-range dur-
ing in the process) into a beaker. Remove the electrode from the beaker with zero buffer,
flush with (demineralized) water and place into the beaker with the slope buffer.
Never dry the elektrode tip using a paper cloth. This may disturb the calibration.
6. Check the reference pH-value of the „slope“ buffer labelled on the bottle for the actual buffer
temperature. Enter this pH-value in the BUFS-entry and confirm with „ENTER“. The cursor
then jumps to „ok“ and the system starts an automatic monitoring of the pH-electrode. If the
measured signal is constant within the given limits, the system stores this as electrode slope.
7. The cursor automatically jumps to the „man/auto“-entry and switches over the temperature
compensation to „auto“-mode. For the pH-measurement the system calculates the pH-value
from the electrode voltage using the Nernst equation with regard to the calibrated „zero drift“
and „slope“ and to the temperature measured in the culture vessel.
At step 4 and 6, when the cursor is on the „ok“ entry, you can shorten the automatic moni-
toring of the electrode. Wait about 10 seconds and press ENTER then. The system stores
the actual signal as reference value of „zero“ and „slope“.
5.4.1.4 Notes on Recalibration of the pH-Electrode During the Process
1. Use a fresh sample taken from the process in such a way, that its pH-value cannot be ef-
fected by the sampling procedure.
2. Measure the pH-value using a precise pH measuring system. Enter the values into the cor-
responding entries of the pH-calibration menu, see description of the menu for details.
The „zero“ and „slope“ measured during the first calibration are displayed in the calibration
menu until they were overwritten by manually entered values.
By comparing the pH-value measured in the process and the one measured in the sample
you can evaluate the accuracy of the pH-electrode. This will also indicate whether the sterili-
zation or the process conditions have changed the measuring characteristics of the elec-
trode and whether the pH-electrode should be serviced or replaced.
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Control Unit
5.4.1.5 Notes about the Calibration Routine
The automatic monitoring of the electrode during the calibration routine shown above con-
siders the following references and limits :
– measured signal is stable within 0.2 % of the measurement range for 60 sec.;
– voltage for electrode zero is kept within the range of -30 ... +30 mV;
– electrode slope within the range of 54 ... 60 mV/pH
If the measured signal is outside these limits, the system displays an alarm message, such
as „**ALARM**, Calibr. out of range, date time“. The calibration has failed and the pH cannot
be measured correctly. In this case you should check whether :
– all parts are properly connected;
– you had followed the above calibration procedure accordingly.
If the error message appears again, the electrode must be serviced or replaced.
The pH-value displayed during the calibration procedure is invalid. The system will display
the correct value after completion of the calibration course.
5.4.1.6 Special Notes on Handling of the pH-Electrode
The electrode has a restricted maximum lifetime which depends on the operating conditions
and the application. The electrode should be serviced or replaced, whenever the functional
check and the calibration indicate malfunctioning.
You'll find more detailed information about handling and mounting of the pH-electrode in
section 4 of this manual. Also consider the documentation delivered with the electrode.
Note: Depending on the type of the delivered pH-electrode, the handling, operation and mainte-
nance measures may differ from the information herein. You'll find more specific and de-
tailed information about the electrode, the recommended service periods, the estimated life-
time and proper handling during calibration in the documentation of the manufacturer.
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Control Unit
5.4.2 Calibration of the pO2- Electrode
The pO2 - electrode is calibrated in percentage oxygen saturation, „% pO2“. The calibration is a two-
point calibration. Usually a measuring solution without oxygen gives the „zero“-current and a solution
saturated with oxygen is defined as „100 % saturated“. The resulting electrode parameters „zero cur-
rent“ and „slope“ are used to calculate a calibration function for pO2-measurement of the process.
The pO2-electrode will be calibrated after mounting into the culture vessel and after the sterilization.
For measurement of the electrode´s „zero“ the culture medium can be gassed with nitrogen to replace
any oxygen dissolved in the medium. Supply of air or a specific gas mixture required for the process up
to saturation of the medium with oxygen gives the electrode´s „slope“, 100 % pO2.
The calibration menu for the pO2-electrode displays the actual „pO2“ as percentage saturation „% pO2“,
the zero current and the slope. This allows a simple function control of the pO2-electrode.
5.4.2.1 Calibration
Operating display
pO2 : 89.3 %
TEMP : 23.4 degC auto
NITR : 0.0 % ok
AIR : 100 % ok
pO2 : 89.3 %
AIR : 100 % ok
ZERO : 3.6 nA
SLOPE:160.3 nA
auto/man When the menu is selected, the cursor is placed at this entry, showing
the mode being previously selected; press ALTER to change the tem-
perature compensation mode:
– auto: temp. compensation using the measured temperature;
– man: temp. compensation using the temperature which is entered
manually
TEMP Display of vessel temperature, when in the „auto“ - mode; input of the
calibration temperature when in the „man“ - mode
NITR Display of zero pO2;
ok Confirmation of the calculation of the zero point; after confirmation the
cursor jumps to the AIR - entry;
AIR Display of the pO2 - value at optimum aeration of the culture medium for
slope calculation
ok Confirmation of the calculation of the slope. After confirmation the cursor
jumps to the „auto/man“ - entry. The temperature compensation will
automatically switch over to „auto“ mode
ZERO Display of the electrodes’ zero in [nA]
SLOPE Display of the electrodes’ slope in [nA]
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Control Unit
5.4.2.2 Course of the Calibration Routine
At a first use or at any time the pO2-electrode has been disconnected for more than 5 ... 10
min from the amplifier, it must be polarized. Polarization may take up to 6 h (shorter, only if
the electrode has been disconnected for a few minutes).
In a culture vessel the pO2-electrode will be calibrated after the sterilization. Any interaction
of this environment with the electrode, such as impacts of heat or chemical reactions of
compounds of the culture medium with the membrane or the electrolyte will be considered
for the calibration.
The electrode „zero“ can be calibrated when the sterilization cycle is switched over to
COOL1-phase. Since the heating of the culture medium causes the dissolved gasses to be
exhausted, the pO2 should be near zero, then. This state is identified as „0 % pO2“. Existing
actual currencies measured by the electrode are within the accuracy of the measuring sys-
tem. Therefor this method will be precise enough for most of the applications.
If a precise „zero“ measurement is required, the vessel can be gassed with nitrogen to re-
place any remaining dissolved oxygen. For this you can supply an oxygen-free gas (N2 of
99.8 % purity, for instance) and degas the dissolved oxygen from the culture medium.
The electrode „slope“ will be calibrated while the fermentor is at operation conditions (tem-
perature, etc.) and the culture medium is saturated with oxygen.
Steps of Operation for Calibration :

1. During the sterilization of the culture vessel wait until the BIOSTAT® B has switched to
COOL1-phase. For „zero“ calibration using supply of oxygen-free gasses, wait until the
measured pO2 is constant near „0 %p pO2“
2. Press main function key „Calibration“ twice to enter the menu for pO2-calibration. The cursor
will be in the „auto/man“ entry.
3. If the mode is „man“ press ALTER to change into „auto“-mode for temperature compensa-
tion considering the actual temperature in the culture vessel. Confirm your selection with
ENTER, the cursor jumps to the „NITR“ - entry.
4. For zero calibration confirm „NITR : 0,0 %“ with ENTER. The cursor then jumps to „ok“.
The system starts an automatic monitoring of the electrode. After autoclaving and prior to
supply of any gas containing oxygen the measured NITR-value should be near „0 % pO2“.
and the electrode´s zero current should be about „0 nA“. If the measured signal is constant
within the given limits, the system stores this as electrode zero. The cursor automatically
jumps to the AIR entry.
5. Prior to starting the fermentation run adjust the fermentation conditions and gas the culture
vessel with air or the gas mixture provided for the process at the required flow rate. Confirm
„AIR : 100 pO2“ with ENTER. If the slope reference should not be 100 % pO2, enter the in-
tended value into the AIR entry and confirm a changed input with „ENTER“.
The cursor then jumps to „ok“. The system starts an automatic monitoring of the electrode. If
the measured signal is constant within the given limits, the system stores this as slope. The
cursor automatically jumps to the „auto“-entry.
During the process the system calculates the pO2-value from the electrode current with re-
gard to the „zero“, „slope“ and the temperature. Since the calibration refers to the reference
conditions (flow rate and type of gas) applied for the calibration, the actual pO2 measured
can exceed 100 % O2, depending on the flow rate and kind of gas supplied.
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Control Unit
5.4.2.3 Notes about the Calibration Routine
The pO2 - measuring value displayed during the calibration course is invalid. The correct
process value is only available after running the calibration.
The automatic monitoring of the electrode after confirmation of the NITR and AIR entries
considers the following references and limits:
– measured signal stable within 0.2 % of the measurement range for 60 sec.;
– current for electrode zero is kept within the range of NITR 0 ... +15 nA;
– electrode slope within the range of AIR: 25 ... 200 nA
If the measured signal is beyond these limits, the system displays an alarm message, such
as „**ALARM**, Calibr. out of range, date time“. In this case the calibration routine has failed
and the pO2 cannot be measured correctly. However, the system will accept the calibrated
„zero“ and „slope“. You should repeat the calibration and check whether :
– all parts are properly connected;
– you had followed the above calibration procedure accordingly.
If the error message appears again, the electrode must be serviced or replaced
At step 4 and 6 of the calibration routine, when the cursor is on the „ok“ entry, you can
shorten the automatic monitoring of the electrode. Wait about 10 seconds and press ENTER
then. The system stores the actual signal as reference value of either zero and slope.
5.4.2.4 Special Notes on Handling of the pO2-Electrode
The pO2-electrode is subject to wear and tear, due to the impacts of heat during sterilization
and due to possible chemical reactions of compounds of the medium with the membrane or
the electrolyte. Such effects may alter the membrane structure, ingredients of the media may
bind to the membrane or ions may permeate into the electrolyte. This will change the meas-
urement characteristics more or less by time. Therefore the electrode has a restricted
maximum lifetime which depends on the operating conditions and the application.
The electrode must be serviced whenever the electrode measures currencies exceeding the
limits given above, while no oxygen is present, or the electrode shows a slow response.
The service of the pO2-electrode is restricted to
– cleaning of the anode
– replacement of the membrane module
– refilling of fresh electrolyte
We recommend that you should refill with new electroly after about 5 times of sterilization of
the entire unit or after about 6 weeks of use for measurement. Depending on physi-
cal/chemical properties of the culture solution it may be necessary to do this more often.
If refilling of new electrolyte cannot improve the measurement characteristic, the anode must
be cleanend and/or the membrane module must be replaced. See the information concern-
ing maintenance and service in the documentation of the electrode´s manufacturer.
You will find detailed information about mounting of the pO2-electrode in section 4 of this
manual. However you should note, that the required operation and maintenance measures
may differ from the information herein, depending on the type of the pO2-electrode delivered.
Specific information about proper handling of the electrode, the recommended service peri-
ods, the estimated lifetime is available in the documentation of the manufacturer.
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Control Unit
5.4.3 Pump Calibration
For monitoring of the corrective solution consumption the measurement and control system can total-
ize the dosing times of the pumps and use them as process values. For this it converts the dosing
times into delivered volumes with regard to the specific flowrates of the pumps. This is possible for the
following pumps and operation modes :
st
1 pump: supply of an acid, mode „ACID“
nd
2 pump: supply of alkaline solution, mode „BASE“
rd
3 pump: for supply of antifoam agent, mode „AFOAM“
th
4 pump: substrate supply or harvest pump for level control, mode LEVEL
th st
5 pump: 1 pump for substrate supply, mode „SUBS1“
nd
6th pump: 2 pump for substrate supply, mode „SUBS2“
The flowrates can be calculated automatically from the measured running time of the pumps and the
delivery of the pumps during the calibration. The flowrates can also be entered directly via the operat-
ing terminal. You can zero the dosing counters for the pumps at any time via the operating terminal.
Since the calibration and dosing counter functions are the same for all pumps, only those for
the acid pump will be described in details herein.
5.4.3.1 Operating display
ACID : 1280 ml
MODE : calib strt
TOTAL: 200 ml
FLOW : 1.2 /m
ACID Display of acid volume delivered; the counter will be set to zero :
– at switching on the unit with the mains switch;
– when switching the operation mode over from „calib“ to „total“
for the other pumps instead of ACID the following entries are shown :
BASE Display of delivered volume of alkaline agent („BASE“)
AFOAM Display of delivered volume of antifoam agent „AFOAM“
LEVEL Display of actual value of the LEVEL dosing counter
SU1DC/SU2DC Display of the substrate volume delivered by pump SUBS1 or SUBS2
MODE Enter mode of the dosing counter :
– calib: calibration of flow rate (delivery) of the pump;
– total: dosing counter active;switching from „calib“ to „total“ mode sets
the counter to zero.
strt/stop Enter operation mode „start/stop“ of the pump calibration routine. This
entry is only active in the „calib“-mode.
TOTAL When switched into „calib“ - mode, the delivered volume can be entered
herein
FLOW Display of actual pump delivery or input of delivery rate. It is possible to
enter a delivery rate manually at any time
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Control Unit
5.4.3.2 Calibration of a Pump Installed in the Control unit
1. Place a piece of tubing of suitable length with one end into a beaker filled with water, mount
the tubing into the pump head and place the outlet end into a measuring beaker. The tubing
must have the same size as used for medium transfer (such as 1.6 x 1.6 or 3.2 x 1.6 mm).
2. To avoid that the clearance volume of the tubing is considered for calibration (this will cause
a calibration error) activate the pump manually until the tubing is completely filled.
3. Press the main function key „Calibration“ several times to select the menu of the pump
which should be calibrated (ACID, BASE, AFOAM or LEVEL). Move the cursor to line
„MODE : total“. Press the ALTER-key once to change to „MODE : calib“. Confirm with
„ENTER“. Then the cursor jumps to „strt“.
4. Press „ENTER“ to start the pump for calibration. At the same time the entry at „TOTAL“ will
be deleted. The pump delivers liquid for a certain period. The system automatically switches
over to „stop“ while the cursor remains in this entry. Confirm „stop“ with „ENTER“. Then the
cursor jumps tho „TOTAL“.
5. Measure the volume delivered into the measuring beaker and enter the measured volume in
entry „TOTAL“. Confirm your input with „ENTER“.
Now the system calculates the flowrate, which will then be displayed. Confirm with „ENTER“
as well. The cursor jumps to entry „MODE“. The operation mode automatically changes from
„calib“ to „total“. The existing entry of „TOTAL“ will be deleted.
If you want to calibrate another pump, select the corresponding menu using the main
function key „Calibration“ and repeat the steps shown above.
5.4.3.3 Special Notes
You must use tubings of the same size for calibration and delivering the addition solution
(tubing size Ø 1.6 x 1.6 or 3.2 x 1.6 mm, for instance). Else the system will calculate the
dosage using wrong calibration parameters.
You can do the calibration before start-up and sterilization of the corresponding storage sup-
plies. However, recalibration is possible at any time using tubings of the same size as con-
nected to the storage bottles.
The dosing counter will be set to zero when the control unit is switched-off. When disabling
the controller with „stop“ the dosing counter will not be set to zero but continue with the ac-
tual value after restart.
During a fermentation run you should handle the assemblies with special care to prevent
unintentional loosening of the connections to the culture vessel or damages of the tubings.
Caution: Acid or akaline agent used for pH-control can cause burns, when the tubings get
loose and the agents are released unintentionally.
Damages of the tubings can cause the corrective solutions to be contaminated by
environmental germs. This may result in infections of the culture.
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Control Unit
5.5 Main Function „Control Loops“
All control loops in the measurement and control system system operate as DDC controllers. The con-
trollers are implemented as PID controllers, setpoint controllers or on/off controllers and adapted to the
control loops concerned. Depending on the connected actuators the control output is either continuous
or pulsewidth-modulated. Both single- and split-range control operation are possible. The controller
structure (P-I-D) can be parametrized according to the task, if necessary.
The controller functions possible with the BIOSTAT® B are shown below. The controllers can be
switched over to different modes of operation. Switching between the operating modes is carried out
with bumpless transfer.
OFF : controller switched off with defined output
AUTO : controller in operation
CASC : controller operating as servo controller in cascade mode
In the operating display you can enter the actual value and the operating mode. Controller parametri-
zation is accessible by entering the required password in the „PARAM“ - entry. Then the parametriza-
tion menu appears for adjustment of PID parameters and deadband, for instance.
The BIOSTAT® B can be connected to a host computer and hence operate in remote operation mode.
Setpoints and operating modes of the individual controllers are then determined by the host computer.
The BIOSTAT® B operating terminal will be locked against inputs, then.
5.5.1 Extent of Controller Functions of the BIOSTAT® B

Temp PID cascade controller with pwm-splitrange outputs for heating via steam
supply or electric heater and cooling via cooling water supply
Stirr PID controller with analog output for motor control
pH PID controller with pwm-split-range outputs for the acid & base pump
pO2 PID cascade controller for one or two servo controllers.
Servo controller(s) can be
– STIRR
– AIRFL
– Gasmix
– SUBS1
FOAM On/off (cycle/delay time) controller for antifoam
LEVEL On/off (cycle/delay time) controller for harvest pump
SUBS1/SUBS2 Setpoint controller for external continuous substrate pumps or, as an al-
ternative, for an internal pump, which has no control function
(ACID/BASE or AFOAM/LEVEL)
Gasmix Setpoint controller for pulse-width modulated splitrange outputs for N2
and O2 of a separate Gasmix unit (optional equipment)
Airflow controller Setpoint controller for a massflow controller
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Control Unit
5.5.2 Controller Operation in General
All DDC controllers are operated in almost the same way. The „Control Loops“ menu of each of the
controllers has two different displays:
The settings for the setpoints and the operating modes of the controllers are done in the
„Operating Displays“, which are directly accessible. If more than 3 lines are required for dis-
playing the settings of a controller, you can scroll up and down to following lines showing ad-
ditional parameters using the cursor keys „ , “.
Any settings not required for routine operation, will be made using the parametrization func-
tions.. The „Parametrization Displays“ are only accessible via a two-digit password The sec-
ond page is accessible via a password.
5.5.2.1 Setpoint Adjustment
Setpoints are entered and displayed in the SETP entry. Entered values must be within the
measuring range of the variable. Values exceeding the measuring range will be ignored.
5.5.2.2 Operating Modes
Operating modes are entered and displayed in the MODE - entry. Modes are selected via
the ALTER - key. Some controllers only allow specific operating modes. If a selected mode
is not plausible for the controller, the inputs will be ignored.
– In „off“ - mode the controller is inactive; the output adapts to a defined position (corre-
sponding to the neutral position of the actuator).
– In „auto“ mode the controller is active. Setpoint values can be altered via SETP.
– In „casc“ mode the controller operates as a servo controller in a cascade control loop. The
setpoint value is specified by the master controller; it cannot be adjusted via SETP.
5.5.3 Parametrization of Controllers in General
On delivery the DDC-controllers are configured with such parameters, which ensure a stable operation
of the controls for the individual type of fermentor. The default settings of the BIOSTAT® B are given in
section “Factory Settings for Controller Parameters“ on page 5-42. If the settings are different for cus-
tomized versions of the fermentor system, the corresponding documentation will be attached to this
manual or be delivered extra.
Note: Under normal circumstances is not necessary to change the control parameters. An
exception are those control loops which are strongly influenced by the process, such
as pH and pO2 control.
If necessary for improved adaptation of the DDC controllers to the control loops of the fermentor, you
may change these control parameters via the parametrization display of the corresponding controller :
DEADB : Deadband adjustment (only for PID controllers)
XP, TI, TD : PID-Parameter (only PID controllers)
MIN, MAX : Controller output limits
Within a controller display the parametrization display is only accessible via the required password.
Note: As the passwords are shown in the corresponding sections of this manual below, the
manual should only be available for personnel, which is qualified and authorized to
enter the parametrization displays of the controllers and change the settings.
Inadequate changes of parameters can cause malfunctioning of the control system.
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Control Unit
5.5.3.1 Deadband
A deadband can be adjusted for PID controllers. If actual values vary stochastically a predefined dead-
band allows for more stable control at minimum actuator changes. The controller output remains con-
stant or is set to zero (i.e. a specific feature of pH - controller) if the control deviation is within this
deadband. In the case of splitrange controller outputs, a deadband can avoid oscillating of the output
(which may result in a continuosly alternating acid-base titration by the pH-controller, for instance).
The deadband is adjusted in the entry DEADB, it is the percentage [%] of the measurement
span of the actual value and is symmetrical to the adjusted setpoint.
Example for pH control:
– measurement range pH = 2 - 12 pH, measurement span 10 pH
– adjusted deadband 1% = ± 0.1 pH, adjusted setpoint 6.0 pH
Here the control is inactive at an actual value within 5.9 pH - 6.1 pH. The acid pump will be
activated when the actual value exceeds the upper limit, the alkaline agent pump will be acti-
vated when the actual value falls below the lower limit.
5.5.3.2 PID - Parameters
The PID controller can be optimized via the following PID parameters :
XP Proportional range in % of measurement range (P-section)
TI Reset time in seconds (I-section)
TD Rate time in seconds (D-section)
The PID parameters can be adjusted in the corresponding entries. The digital controller implemented
operates according to the positioning method. Setting individual PID parameters to zero determines the
control principle. For easy optimization of the controllers you can change the parameters during opera-
tion. Set the parameters for the controller functions as follows:
to work as set the PID – parameters as follows
P-controller TI = 0, TD = 0
PI-controller TD = 0
PD-controller TI = 0
PID-controller all PID - parameters have defined set values
5.5.4 PID - Controller Optimization
You will need some detailed knowledge about the control theory for optimum adaptation of a PID con-
troller to the corresponding control loop. For detailed information about proven adjustment rules you
may refer to any standard literature. Therefore the following specifications are only general guidlines:
The D section (TD) should only be activated in the case of rather stable actual values since
the controller output will change considerably and fast if the actual values are varying sto-
chastically. Else this would lead to an unstable control.
As a general rule the TI : TD ratio should be 4 : 1.
In the case of a periodically oscillating control loop you can increase Xp and/or increase
TI/TD to minimize or even prevent oxcillation.
If an actual value adapts to the setpoint to slowly after a setpoint jump or if the actual value
shows a continous change (drift), you can decrease Xp and/or decrease TI/TD
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5.5.5 Temperature Control
5.5.5.1 Functional Principle
The temperature controller is a single-loop PID controller. The vessel temperature is the reference in-
put. The controller output controls the cooling valve and the heater in split-range operation via pulse-
width-modulated outputs.
As a special feature, the master controller can be switched from PD (which is the start-up-mode) to
PID - operation when approaching the setpoint. Thus overshooting is eliminated as far as possible.
When the temperature controller is switched off, an additional digital output also switches off the circu-
lation pump and the heating contactor in the fermentor.
5.5.5.2 Operating Information
Operating display page 1 :
TEMP : 37.5 oC
SETP : 33.4 oC
MODE : auto
PARA : _
TEMP Display of actual value

SETP Display or input of controller setpoint
MODE Display and input of control mode:
– auto: automatic operation according to given setpoint;
– off: controller is switched off, outputs are in operating state
PARAM Enter 2-digit passwort „19“ for access to the controller parameters. When
entering the password it will not be shown in the display.
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Control Unit
Operating display page 2, with cursor within the upper 4 lines :
TEMP : 37.5 oC
MIN : -100 %
MAX : 100 %
XP : 10.0 %
Operating display page 2, with cursor moved down to the lowest line :
TEMP : 37.5 oC
TI : 300 s
TD : 75 s
DEADB: 00.0 %
MIN Display and input of minimum controller output limit. For the temperature
controller this parameter limits the output for „cooling“
MAX Display and input of maximum controller output limit. For the temperature
controller this parameter limits the output for „heating“
Note : The values for „MIN“ and „MAX“ depend on the individual measurement
range. If the measurement ranges are changed, the „min“ and „max“ val-
ues for the outputs are changed correspondingly and cannot be ex-
ceeded
XP Enter proportional range
TI Enter integral portion (reset time)
TD Enter differential portion (delay time)
DEADB Enter dead band
, More lines are available via the „ , “ - keys
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Control Unit
5.5.6 Stirrer Speed Control
The stirrer speed control function works as a PID controller. It sets the stirrer speed, supplies the digital
setpoint signal for the motor control and displays the input signal from the motor control. When the
controller is off, the motor contactor is switched off via a digital output. In conjunction with the pO2-
controller the stirrer speed controller can be servo controller in the pO2 - cascade control loop.
The stirring speed is adjustable in the range of 50 ... 1,200 rpm.
The stirring speed must be limitted to max. 600 rpm for 10 l vessels and to about 300
rpm for vessels with a silicone membrane insert. Higher speeds may cause inadmis-
sible vibrations of the assembly of culture vessel with motor or can result in dam-
ages of devices inside the vessel (i.e. loosening of the silicone membrane).
A system reset will enable the default speed settings. Therefore, after a system reset
the speed limit settings should be checked and the max. speed be reconfigered.
Operating display:
STIRR: 750 rpm

SETP : 750 rpm
MODE : auto
PARA :
STIRR Display of actual value

– auto: automatic operation at given setpoint
– casc: cascade operation. The controller is servo controller and triggers
the output of the master controller;
– off: the controller is switched off, outputs are in operation state
PARAM Enter 2-digit passwort „19“ for access to the controller parameters
Operating display page 2, with cursor within the upper 4 lines :
STIRR: 750 rpm

MIN : 004.2 %
MAX : 100.0 %
XP : 200.0 %
STIRR: 750 rpm

TI : 5 s
TD : 0 s
DEADB: 000.0 %
MIN, MAX For limitting controller output to minimum or to maximum

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Control Unit
5.5.7 pH - Control
The pH - controller of the BIOSTAT® B is a software function which acts as a PID controller. It triggers
the corrective solution pumps for supply of acid and alcaline agent in split-range mode via two pulse-
width-modulated outputs. This allows for a reversible control of both pumps. If gaseous CO2 should be
applied for pH - control instead of acids (as often applied in tissue cell culture), the output of the con-
troller can be connected to external Gas Mixing Unit including a CO2 - control valve.
The acid and base signals will remain switched-off as long as the deviation remains within the
parametrized dead bands. This will prevent unnecessary supply of acid or base to the culture broth and
will limit the consumption of these corrective solutions (this is also true for CO2, if applied).
5.5.7.1 Setting the Setpoint and Controller Mode
Operating Display :
pH : 7.25 pH
SETP : 7.10 pH
MODE : auto
PUMP : ACID/BASE
Operating display page 2:
pH : 7.25 pH
MODE ; auto
PUMP : ACID/BASE
PARAM:
pH Display of actual value in the headline

– auto: automatic operation with the given setpoint;
– off: controller is switched-off; outputs are in the operation state
PUMP Display and selection of the required pump(s)
– ACID/BASE: both acid and base pump are operated
– ACID/-----: the acid pump is operated but not the base pump; thus the
base pump is free for use as the substrate pump
– -----/BASE: the base pump is operated but not the acid pump; thus
the acid pump is free for use as the substrate pump
PARAM Enter 2-digit password „19“ for access to controller parameters; during
entry the password will not be displayed
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Control Unit
5.5.7.2 Parametrization
Parametrization display, with cursor within the upper 4 lines :
pH : 7.25 pH
MIN : -100 %
MAX : 100 %
XP : 30.0 %
Parametrization display page 2, with cursor moved down to the lowest line:
pH : 7.25 pH
TI : 30 s
TD : 0 s
DEADB: 0.5 %
MIN Display and input of minimum controller output limit.

– At the pH controller this parameter limits the output for the acid pump
MAX Display and input of maximum controller output limit.
– At the pH controller this parameter limits the output for the base pump
Note : The „MIN“ and „MAX“ values depend on the individual measurement
range. If the measurement ranges are changed, the „min“ and „max“ val-
ues for the outputs are changed correspondingly and cannot be ex-
ceeded
XP Input of proportional range
TI Input of integral portion (reset time)
TD Input of differential portion (delay time)
DEADB Input of dead band
, More lines are available below; move over using the „ , “ - keys
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Control Unit
5.5.8 pO2 - Control
5.5.8.1 Modes of Operation
The control unit of the BIOSTAT® B includes a pO2-controller for universal use. The pO2 control func-
tion can be applied as required for the intended process. You can adapt the pO2 control to even spe-
cific process requirements. In particular the following control strategies can be performed :
pO2-control by operating a single process value. These servo controllers can be selected:
– stirring speed (STIRR)
– substrate supply (SUBS)
– gas supply by mixing of 2 gasses (GASMIX)
– gas supply using the optional massflow controller (AIRFL)
pO2-control by operating two process values. The same servo controllers can be selected.
5.5.8.2 pO2 - Controller with One Servo Controller
In the pO2 control using one servo controller the pO2 controller acts as the master controller. Its output
directly operates the input of the servo controller. In standard configurations the pO2-controller oper-
ates either the stirrer speed controller, the substrate supply or the Gasmix controller, respectively.
By use of the Gasmix servo controller two gasses can be mixed, such as N2 and O2, for instance. For
this the Gasmix controller operates a valve for O2 supply and a valve for supply of O2 in splitrange
mode via pulse-width modulated outputs. This method is often applied for bubble free gas supply to the
culture medium via a membrane aeration system.
An alternative to the use of the Gasmix controller is the “O2-enrichement” operation. The pO2 controller
then triggers a 3/2-way valve of the air (gas) inlet, where one input is connected to the air supply and
the other input is connected to an O2 – supply. You will find an example for this further below.
5.5.8.3 pO2 - Controller with Two Servo Controllers
For the pO2-cascade control serial operation can be selected. Then the pO2-controller operates two
servo controllers one after another, according to their priority. In the pO2-controller you can predefine a
„min/max“ range for each servo controller. The pO2-controller triggers this servo controller as long as
the related setpoint is within its „min/max“ range. If you set the reverse „min/max“ - settings you will get
a reverse control.
When the pO2-control is switched on, the output of the pO2-controller operates the servo
controller 1 (CASC1) with the given setpoints within the range of the preset „min/max“ set-
tings. If selected, the servo controller 2 receives the „MIN“ setpoint of the pO2-controller.
At increasing O2 consumption, when the setpoint signal reaches the „MAX“ value defined for
the servo controller 1, the output of the pO2-controller will automatically switch over to the
setpoint input of servo controller 2. The delay time for swiching over is 5 min (default setting)
and can be modified. Then the following setpoints will be transmitted:
– predifined MAX output for servo controller 1
– controlled output of pO2-controller for servo controller 2
If the O2-consumption is reduced again, the servo controller 2 receives the setpoint signal
defined in the pO2 controller as “MIN” and the output of the pO2-controller again triggers the
servo controller 1.
This control principle allows for an accurate pO2-control during the process, the pO2 can be kept con-
stant even at larger variations of the O2-consumption without any manual control required. Above this
the PID-parameters of the servo controllers 1 and 2 can be parametrized independently from each
other to provide an optimum adaptation of the controls to the behaviour of the control system.
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Control Unit
Operating display, cursor in the first line:
pO2 : 88.5 %
SETP : 83.4 %
MODE : auto
CASC :STIRR AIRFL
Operating display, cursor in the last line:
pO2 : 88.5 %
MODE : auto
CASC :STIRR AIRFL
PARAM:
Switching over to cascade operation with one servo controller:
pO2 : 88.5 %
MODE : auto
CASC :STIRR STIRR
PARAM:__
pO2 display of actual value in the headline

SETP display or input of controller setpoint
MODE possible modes are:
– off: controller is switched-off; outputs are in the operation state.
CASC display or selection of servo controller 1 and, if necessary, servo con-
troller 2 (the above example shows STIRR selected as servo controller
1 and AIRFL selected as servo controller 2)
STIRR : output of the pO2-controller triggers the setpoint of the stirrer controller;
the operating mode of the stirrer speed controller will automatically be
switched to „casc“ -mode.
SUBS : output of the pO2- controller triggers the setpoint of the substrate 1 con-
troller; the operating mode for the substrate 1 controller will automati-
cally be switched to „casc“ -mode
GASM : output of the pO2-controller triggers the setpoint of the gasmix controller
and the operating mode will automatically be switched to „casc“ -mode.
AIRFL : output of the pO2 controller operates the setpoint of the airflow control-
ler; the operating mode will automatically be switched to „casc“ -mode
Note : For switching over to operation with one servo controller place the cur-
sor at the tag of servo controller 2 and select the same controller as ser-
vo controller 1 via „Alter“;
– confirmation with ENTER deletes the tag of servo controller 2.
PARAM Enter 2-digit passwort „19“ for access to controller parameters
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Control Unit
Parametrization display, page 1, with cursor within the upper 4 lines :
pO2 : 88,4 % ñ
DEADB: 0.5 %
HTIME: 5 min STIR
MIN : 0 %
Parametrization display, page 1, with cursor in the lowest line :
pO2 : 88,4 % ñ
MAX : 100 %
XP : 100 s
TI : 100 s
Parametrization display, page 2, with cursor in the lowest line :
pO2 : 88,4 % ñ
XP : 100 s
TI : 100 s
TD : 0 s

HTIME display or input of the delay time for switching over from cascade 1 to
cascade 2
STIR, etc. selection of servo controller using the ALTER key
STIR : display / setting of the controller parameters for the stirrer controller while
used as servo controller
SUBS : display / setting of the controller parameters for the substrat controller
while used as servo controller
GASM : display / setting of the controller parameters for the gasmix controller
while used as servo controller
AIRFL : Display and adjustment of the controller parameters related to the servo
controller „Airflow“, if this servo controller is available (Option)
, More lines are available below; move over using „ , “ - keys
For the pO2-controller two sets of parameters are internally available. Access is only possible
to the PID - set, which has been activated before within „Mode“.
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Control Unit
5.5.9 Foam Control
5.5.9.1 Function Principles
When foam has contact with the foam sensor for longer than 6 seconds the foam amplifier releases a
limit value signal, which is the input signal for the foam controller. The sensitivity of the foam meas-
urement amplifier can be adjusted in the display of the foam controller. The delay time prevents erro-
neous activation of the foam control and supply of antifoam agent caused by short time contact or
splashes of the liquid against the probe.
The controller output operates the internal „Antifoam“ pump. The pump is switched-on and off periodi-
cally as long as foam is detected (sensor signal = on). Running time and the cycle time (= dosing time
+ delay time) for intermittent operation can be entered via the controller display.
The amount of antifoam agent delivered by the pump depends on the hose dimensions. After pump
calibration via the corresponding calibration function a dosing counter displays the delivered volume.
The internal antifoam pump of the BIOSTAT® B can be used as substrate pump, if the foam control
function is not required.
5.5.9.2 Operation of the Foam Control
Operating Display, page 1:
FOAM : off
MODE : auto chg
CYCLE:01:30 m:s
PULSE:00:05 m:s
Operating Display, page 2, with cursor in the lowest line:
FOAM : off ñ
SENSI: 3
PULSE:00:05 m:s
PUMP : AFOAM
FOAM Display of actual sensor signal FOAM in the headline;

– the tag FOAM will automatically be switched over to SUBS according
to the preselected function of the controller
MODE Enter controller mode :
– auto: controller is active, automatic operation;
– off: controller is switched-off; outputs are in the operation state;
1. Place cursor at „auto“ or „off“, respectively
2. Press ALTER to switch over the mode and confirm with ENTER
CYCLE Enter cycle time in „Minutes:Seconds“ [mm:ss]
PULSE Enter pump running time in „Minutes:Seconds“ [mm:ss]
SENSI Enter probe sensitity; 4 levels of sensitivity are adjustable
PUMP Display and selection of the pump connected to the controller:
– AFOAM: the antifoam controller operates the antifoam pump
– -----: the antifoam controller does not operate the antifoam pump;
the antifoam pump can now be used for substrate supply.
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Control Unit
With the BIOSTAT® B chemical foam control using commercially available antifoam agents
(such as greases, oils, oil-water-emulsions, silicone-oils, paraffines) is applied. You should
note that these agents can affect the growth of the cells and cell metabolism. Therefore the
amount of agent supplied to a culture vessel should be minimized.
Dosing of antifoam agent should only be activated, when foam has contact with the probe.
Adjust sensitivity of the probe in that way, that splashing of bubbles, foam residues or culture
medium against the probe will not activate the controller. Adjust cycle time long enough to
differentiate between splashing or short contact of foam or medium caused by stirring and/or
aeration and longer contact with foam.
The reference potential for the foam electrode is provided via the Pt-100. Thus the foam
measuring system only works, if both the foam electrode and the Pt-100 are mounted into
the top-plate of the culture vessel.
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Control Unit
5.5.10 Level Controller
Taking samples during a fermentation run will reduce the filling volume of the culture vessel. Therefore
you may need to add medium the culture vessel regularly. You will also supply medium or specific sub-
strates for fed-batch or continuous processes. This can cause that the maximum working volume of
the vessel is exceeded and the surplus volume needs to be removed.
The level control function of the BIOSTAT® B provides a monitoring of the filling volume of the culture
vessel. In addition you can automate the removal of surplus medium as well as the supply of medium
and specific substrates, in order to keep the filling volume in the culture vessel constant.
5.5.10.1 Function Principles of Level Control
The insertion depth of the level probe determines the controlled filling volume of the culture vessel.
Therefore you will need to gauge the capacity of the vessel and to determine the insertion depth of the
probe. When the culture medium comes into contact with the probe, the level amplifier generates a
digital signal, which is the input signal for the level control function. You can adjust the sensitivity of the
amplifier to consider the conductivity of the medium and to avoid unintended controller activities due to
contact of foam with the probe or by splashing of liquid against the probe, for instance.
The level amplifier offers the two modes „HARV“ and „FEED“. In the „HARV“ mode the level controller
output operates the „LEVEL“ pump, as long as the level signal is „on“. The pump acts as harvest
pump. In the FEED - mode, it operates the pump, when the level signal is „off“. Then the pump acts as
dosing pump. The pump operates at running and delay times, which can be adjusted in the corre-
sponding operating display. The amount of medium which is removed or supplied, depends on the
running time and the hose dimensions. If the pump is calibrated using the corresponding pump calibra-
tion function, the dosing counter displays the exact delivery volumes.
You will operate the level controller in the same way as the antifoam controller. See the
above section for details.
5.5.10.3 Additional Notes
For a precise level control at certain filling volume you´ll need to adjust the level probe at the
required height. For this you'll have to measure the vessel capacity at a given assembly of
internal vessel component parts, such as inserts, stirrers, electrodes, etc.
You may label the intended filling volumes on the vessel. However, you should note, that
high stirring speeds and/or intentisified gassing of the culture can produce a whirl or an in-
creased level of the culture medium, compared to the level without or at low stirring speed or
gas supply, respectively.
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5.5.11 Controller for Substrate Feed SUBS1 und SUBS2
The two controllers support external continuous pumps intended for substrate feeding. The controllers
does not use feedback of the actual speed of the pump. The display of the process value „SUBS1“ and
“SUBS2” represents the controller´s setpoint. The amount of substrate supplied with the pump de-
pends on hose dimensions and running time. If the pump is calibrated using the pump calibration func-
tion (refer to the pump calibration instructions), the dosing counters SU1DC and SU2DC display the
exact deliveries.
You can switch the substrate controller to an internal peristaltic pump, if no external continuous pump
is available but one of the internal pumps is (if unused). For switching over proceed as follows:
1. Clear the unused pump in the operating display of the controller assigned to it.
2. Assign this pump to the substrate controller.
You can operate the substrate controller using a timer setpoint profile. Such a profile can have step
changes and ramp changes with max. 10 segments and be started and stopped at any time.
5.5.11.1 Operation of the Substrate Supply (Example for SUBS1)
Operating display
SUBS1: 20 %
SETP : 020.0 %
MODE : auto
PUMP : SUBS1
Operating display with cursor in the lowest line:
SUBS1: 20.0 % ñ
MODE : auto
PUMP : SUBS1
PARAM: __
SUBS1 / 2 Display of setpoint of substrate controller SUBS1 (/ 2, if applied)

SETP Display or input of the setpoint for the pump delivery (flow rate)
MODE Enter controller mode :
– off: controller is switched-off; outputs are in the operation state;
– casc: substrate controller is switched to cascade operation for the
pO2-controller; i.e. the pO2-controller delivers the setpoint for the sub-
strate controller
– profil: automatic operation using a defined setpoint profile
PUMP Display and selection of the connected pump :
– SUBS: the controller operates an external continuous pump;
– ACID: the substrate controller operates the ACID pump, i.e. the pump
is not used by the pH-controller;
– BASE: the substrate controller operates the BASE pump, i.e. the
pump is not used by the pH-controller;
– AFOAM: the substrate controller operates the AFOAM pump, i.e.the
pump is not used by the AFOAM-controller;
– LEVEL: the substrate controller operates the LEVEL pump, i.e. the
pump is not used by the LEVEL -controller
PARAM Access to the controller parameters via 2-digit password „19“
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Control Unit
5.5.11.2 Parametrization of the Substrate Dosing Function SUBS1, Input of a Setpoint Profile
Parametrization display page 1
SUBS1: 15.0 %
MIN : 000 %
MAX : 100 %
TIME0: 0:00 SP : 10 %
TIME1: 1:10 SP : 15 % ñ
TIME2: 2:00 SP : 20 %
TIME3: 3:15 SP : 35 %
TIME4: 4:00 SP : 40 %_
TIME5: 4:50 SP : 55 % ñ
TIME6: 5:30 SP : 70 %
TIME7: 6:00 SP : 80 %
TIME8: 6:15 SP : 90 %
TIME9: 7:00 SP : 90 % ñ
MIN Display and input of the minimum controller output limit

MAX Display and input of the maximum controller output limit
TIME0...9 Display and input of the time for max. 10 profile changes
SP Display and input of the setpoints for the profile changes
The above inputs

will create this
setpoint profile
Additional notes
If the time for the first breakpoint is not set to „00:00 h:m“ the system uses the current set-
point for the start of a profile.
In a program for a setpoint jump (step change) both setpoints can have the same time.
When a profile is defined and started for pO2-control, correlating active profiles for „SUBS1“
will be stopped automatically. The controller will be switched over to „cascade“-mode.
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Control Unit
5.5.12 Airflow Controller
5.5.12.1 Modes of Operation and Equipment
Airflow controllers allow an automatic continuous control of the air supply according to the oxygen con-
sumption in the process. The measurement and control system triggers the massflow controller via an
analog setpoint signal. The airflow controllers is a massflow controller module integrated in the inlet air
line behind the flowmeter, see P&I – diagram for details.
Standard equipment is a massflow - controller for air supply rates up to 10 l/min
Depending on the size of the culture vessel and the intended intensity of aeration different
airflow controllers can be applied
Operating display :
AIRFL: 8.00 slpm

SETP : 10.00 slpm
MODE : auto
PARAM:_
Parametrization display :
AIRFL: 8.00 slpm

MIN : 0 %
MAX : 100 %
AIRFL display of actual flowrate

SETP entry of setpoint for the flowrate
MODE enter controller mode :
– off: controller is switched-off; outputs are in the operation state;.
PARAM access to controller parameter, enter 2-digit password „19“ for this
MIN „min“ limit of controller output
MAX „max“ limit of controller output
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Control Unit
5.6 Main function "Maintenance"
5.6.1 Recorders
5.6.1.1 Technical Set-up
Two types of recorders are available as standard equipment of BIOSTAT® B - fermentors:

Arucomp SK 10 recorder with digital display, Hartmann und Braun
Arucomp SK 10 recorder with analog display, Hartmann und Braun
5.6.1.2 Operating the Recorder
The recorder is contained in an extra housing and will be connected to the corresponding
connector „Recorder“ of the control unit.
Below you will find an overview on the most important (default) adjustments for connection to
a BIOSTAT® B. Additional information about individual functions and the adjustment of op-
5-3)
eration parameters can be found in the Operating Manual of the recorder.
5.6.1.3 Recorder Configuration
The control unit includes a recorder output board with plug connection. This board offers 6 analog out-
puts 0 ... 10 V for the connection of a 6-channel dot printer. It is possible to assign up to 6 process val-
ues to the 6 recorder outputs. The signal ranges of the outputs are fixed and correspond with meas-
urement range for the process values.
Operating display page 1:
Recorder
CHAN1:TEMP
CHAN2:STIR
CHAN3:pH
Operating display page 2, with the cursor in the lowest line :
Recorder
CHAN4:pO2
CHAN1 Enter process value for recorder output channel 1.

– Select other process values with the „ALTER“ key.
– Confirm selection with ENTER.
– Moving on with a cursor key without ENTER will ignore your selection
CHAN2..4 see CHAN1
, More lines are available. Move over using „ , “
5-3)
Included in the documentation set if part of the system configuration on delivery. Otherwise
available on request. Please contact B. Braun Biotech International GmbH for this.
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Control Unit
5.6.2 Printers
5.6.2.1 Technical Set-up
Connection of a protocol printer to the BIOSTAT® B allows for easy online - recording of the running
process. The protocol is a „hardcopy“ of the course of the fermentation run. Thus the course of the im-
portant parameters can be monitored and/or unexpected events can easily be detected, for instance.
The printer can be a standard matrix printer with serial interfaces and small fonts. It will print the pa-
rameters numerically into a corresponding table. The protocol will include the data of the PROCESS
DISPLAY and the following parameters (see example of a protocol below) :
Date, time of the print (showing the time of an event during the fermentation run)
All process values
Alarm messages
1. Connect the printer to the RS 232 interface of the control unit (socket „printer“). Note the in-
formation for connecting and presetting the printer in the operating manual of the device.
2. Switch over to the menu for the printer settings, see the menu, the tags and the information
about the entries below.
3. Set the cycle time for the output of data.
4. Start the printout. Starting and stopping is possible at any time during the process.
Operating display :
Printer
CYCLE: 20 min
MODE :start
PARAM: __
CYCLE Enter cycle time for printout of one line of the protocol in the table. Possi-
ble cycle times are in the range of 1 ... 60 minutes.
MODE Enter operation mode of the printer, select via the „ALTER“ - key. :
– stop: no data will be printed
– start: printout of data is active
PARAM Enter 2-digit password „19“ for access to interface parameters.
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Control Unit
Parametrization display for printer:
Printer
SPEED: 9600 bd
DATA : 8 bit
STOP : 1 bit
next page of parametrization display:
Printer
PARTY: no
1 : TEMP 2 : STIRR
3 : PH 4 : PO2
Parametrization display with cursor in the lowest line (see print protocol example below) :
7 : SUBS1 8 : SUBS2
9 : AIRFL 10: EXT_1
11: EXT_2 12: EXT_3
13: EXT_4 14:
SPEED Enter signal transfer rate within the range of 300..19,200 baud; default is
9,600 baud
DATA Enter number of data bits: 7 ... 8; default is 8
STOP Enter number of stop bits: 0 ... 2; default is 1
PARTY Enter kind of parity bit: „even“, „odd“, „no parity“; default is „no“
1: TEMP. etc. Selection of a process value for the column 1 ... 14 in the print
(possible with software version 4.14 and later versions)
Fig. 5-3:
Example for a
printer protocol
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Control Unit
5.6.2.3 Host Computers and Connections
The BIOSTAT® B can be connected to a host computer system via the integrated serial interface RS
422. The 4-wire network allows for „Multidrop“ - operation. The communication protocol is compatible
with the protocol of the DCU measurement and control system. The data contained are measurement
values or setpoints (at fixed order).
Adjustment of interface parameters is possible via protected operation display, entries are
accessible via password. The user can define transfer rates within 300 ... 19.200 Baud.
1. Connect the signal cable to the RS-422 interface on the left side panel of the control unit.
Note the documentation of the host computer for the required settings for data transfer.
2. Switch over the operating terminal to the menu for host communication.
Operating display:
Host
ADR : 2
PARAM: XX
ADR Display of BIOSTAT® B - address

PARA Enter 2-digit password „19“ for access to interface parameters
Operating display „Parameters":
Host
ADR : 2
SPEED: 9600 bd
DATA : 7 bit
Host
DATA : 7 bit
STOP : 1 bit
PARTY: even
ADR Enter BIOSTAT® B - address 1 ... 16

SPEED Enter transfer rate within 300 ... 19,200 baud; default is 9,600 baud
DATA Enter number of data bits: 7 ... 8; default is 7
STOP Enter number of stop bits: 0 ... 2; default is 1
PARTY Enter kind of parity bit: even, odd, no parity; default is „even“
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Control Unit
5.6.3 Utility Functions
Within the „Utility“ - menu you can find and reset, if necessary, some general system settings, such as
the „date“ or „real time clock“. The display also shows the implemented software version and any spe-
cific configuration, if applied.
Utility
DATE :17.04.1999
TIME :14:30 h:m
VERS : 4.1
Utility
CONFG:BA01D 5-4)
FAILT:02:00 h:m
CONTR:alter
Operating display, page 2, with cursor in the lowest line :
Utility
FAILT:02:00 h:m
CONTR:alter
PARAM: ___
DATE Display or input of date

TIME Display or input of time (system clock)
VERS Display of software version
CONFG Display of configuration identification
FAILT Display or input of the failtime for power failures.
– If a power failure is shorter than the preset „failtime“, the process will
be continued with the given settings of all parameters.
– If the power failure takes longer, the system behaves like being
switched off with the mains switch after return of the power supply.
CONTR Select contrast of the LCD - display using the „ALTER“ - key.
PARAM Entry of password for access to the configuration parameters; this is a
special password only available for authorized service personnel.
5-4) Example for configuration code, see „Utility Functions“ menu of your system for your code. You
must refer to this code if you contact B. Braun Biotech International GmbH for questions about
your system and if problems arise with your configuration.
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Control Unit
5.6.4 Measurement Ranges
The menu shows the measurement ranges of the process values. Setting of the parameters is possible
in a submenu, which is also protected by a password.
Only authorized service personnel is allowed to change the measurement ranges!
Meas. Ranges
PARAM:___
Page 2 of the Operating Display, with cursor within the upper 4 lines :
Meas. Ranges
PV :TEMP
MIN : 0.0 %
MAX : 100.0 %
PARAM Enter password „753“ to select the second page of the display.
PV Display or input of a process value. For selecting another process value
use the „ALTER“ - key.
MIN Display or input of lower limit threshold of a measurement range.
MAX Display or input of upper limit threshold of a measurement range.
On delivery the following measurement ranges of process values are adjusted as default
(you should note that this can be different for the delivered version of the BIOSTAT® B):
Process value Limits of measurement range Physical unit
lower limit upper limit

TEMP : 0.0 150,0 degC
5-5
STIRR: 0 1.200 rpm
pH : 2.00 12,00 pH
pO2 : 0.0 100,0 %
ACID : 0 500 ml
BASE : 0 500 ml
AFOAM: 0 500 ml
LEVEL: 0 10.000 ml
SU1DC/SU2DC: 0 10.000 ml
SUBS1/SUBS2: 0 100 %
5-6
AIRFL: 0 10.00 slpm (lS/min)
EXT1....4: 0 100,0 %
5-5
Max. stirring speed for 2 l - vessel; delivery setting depends on size of the culture vessel and / or
is specified according to customized specifications
5 -6
BIOSTAT B with 5 l culture vessel, may vary for other vessels or customized specifications
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Control Unit
5.6.5 Manual Operation of Digital Outputs, Standard
The menu „Manual Operation - Digital Outputs Std.“ is used for service support. It displays the state of
the digital outputs of the system. Inputs in this menu directly trigger the corresponding digital outputs.
Menu display :
MAN OP DIG OUT STD

1: a off 2: a off
3: a off 4: m 20
5: a o 6: a off
Symbol / entry Meaning
1 ... 16 Digital output, channel 1 ... 16

a, etc Display of mode of output:
a : „automatic“: internal function acts on output, i.e. controller
o : „operating state“: no internal function acts on output
s : „sterilization“: sterilization program acts on output (only available for
control systems with sterilization programs, i.e. not valid for
BIOSTAT® B, for instance)
h : „host“: host computer connected can act on output
m : „manual“: manual settings mode of the digital outputs menu enabled;
the displayed state will be transmitted
off/on : Display of state (or manual setting of state in “manual” mode) of:
– off: output statically switched off
– on: output statically switched on
0 ... 100 : Display of state (or manual setting of state in “manual” mode) of pulse-
width modulated output :
– 0: output switched off
– 100: output fully (100%) switched on
– 20: output switched on in pulse-with modulated state.
In this example the output is switched on at a rating of 20%. The cycle
time (pulse - pulse) depends on the controlled system and is in the range
of 5 ... 10 s
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Control Unit
5.6.6 Manual Operation of Analog Outputs, Standard
The menu „Manual Operation - Analog Outputs Std.“ is used for service support. It displays the state of
the analog outputs of the system. Inputs in this menu directly trigger the corresponding analog outputs.
Menu display :
MAN OP ANA OUT STD

1: a o 2: a o
3: m 3 4: a 50
5: a 10 6: a 90
Symbol / entry Meaning
1 ... 8 analog output, channel 1 ... 8

a, etc display of mode of output:
a : „automatic“: internal function acts on output, i.e. controller
o : „operating state“: no internal function acts on output
m : „manual“: manual settings mode of the analog outputs menu enabled;
the displayed state will be transmitted
0 ... 100 Display of state of output (or manual setting of state in “manual” mode):
0 : output switched off (voltage of 0 V)
100 : output switched on (voltage of 10 V)
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Control Unit
5.7 Alarm Messages of System Functions
When they occur, alarms will be displayed within an extra alarm menu, showing date and time of the
alarm. If several alarms occur, they will be stored in an internal buffer, which can hold up to ten alarms.
A single alarm message will dissappear after pressing any main function key. For more than one
alarm, the next alarm will be displayed afterwards.
Example of alarm display :
** ALARM **
Power failure
22.01.91 14:33
another example
** ALARM **
HEATER failtime
22.01.91 14:33
Heater failure Display of heater alarm (malfunction of heater)

Motor failure Display of a motor alarm (malfunction of motor or motor control)
Power failure Display of general power failure of mains supply
BUFRAM read error System error; a software reset has been applied or the battery is mal-
functioning
EEPROM read error System error; the parameters specific for the system cannot be proc-
essed; not all functions are available (system partly out of order)
How to Answer Alarm Messages :

1. If the process continuous although an alarm has occured, check which kind of alarm it is and
whether the process has been interfered. Disturbances of the process may be indicated at
the chart paper of the recorder or the printout of the printer. You may continue the process, if
possible or need to interupt it.
2. If the alarm has stopped the fermentation run, you may try to continue by restarting the fer-
mentor. Check at the display and at the chart paper of the recorder or the printout of the
printer at which state of operation the fermentation run has been stopped, which kind of
alarm it is and whether the process has been interferred.
3. At malfunctioning of the related device of fermentor and if the system cannot be restartet:
– check all adjustments for the related component
– readjust given setpoints, etc., if necessary.
For information about the system parameters adjusted in the factory see section „Factory
Settings“ on next page.
4. If it is impossible for you to restart the fermentor system by this, and in any case one of the
system errors „BUFRAM read error“ or „EEPROM read error“ occurs you should contact
your B. Braun service representative or directly the B. Braun Biotech International GmbH.
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Control Unit
5.8 Factory Settings
5.8.1 General Information
For the laboratory scale fermentor BIOSTAT® B the integrated measurement and control system offers
a defined range of standard configurations. Below you will find a listing of all parameter settings config-
ured in the factory which can be changed by the user according to his specific requirements. In the
case of unintended changes of the parameters you can recreate the original state.
5.8.2 Factory Settings for Measurement Ranges of Process Values

Note section “ Measurement Ranges“ further above for an overview of the default settings.
If your fermentor is of customized design and/or has different factory settings, please note
the corresponding extra documentation delivered with your equipment. It includes the most
actual information on the settings of your BIOSTAT® B.
If your fermentor is of customized design and the corresponding documents are not deliv-
ered, please contact B. Braun Biotech International GmbH for this.
5.8.3 Factory Settings for Controller Parameters
5.8.3.1 PID-Parameters und MIN/MAX-Limits
CL-Tag MIN MAX DEADB XP TI TD RAMP
[%] [%] [%] [%] [s] [s] [%]
TEMP --- --- 0 7 1000 250 ---

STIRR 1.7 100 0 200 5 --- ---
pH -100 100 0.5 30 30 0 ---
pO2/STIRR 0 100 0,5 150 100 0 ---
pO2/AIRFL 0 100 0,5 90 50 0 ---
pO2/GASMIX -100 100 0,5 5 200 0 ---
pO2/SUBS1 -100 100 --- 120 60 0 ---
SUBS1 / 2 0 100 --- --- --- --- ---
GASMIX -100 100 --- --- --- --- ---
AIRFL 0 100 --- --- --- --- ---
If your fermentor is of customized design and/or has different factory settings, please note
the corresponding extra documentation delivered with your equipment. It includes the most
actual information on the settings of your BIOSTAT® B.
If the corresponding documents are not delivered with the unit, please contact B. Braun
Biotech International GmbH for this.
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Control Unit
5.8.4 Example: pO2-Controller with Airflow - Massflow Controller and O2 - Enrichment

After switching on the pO2 - controller first operates the AIRFL - controller. At this time the
GASMIX - controller receives the „MIN“ setting assigned in the pO2 - controller.
If the setpoint setting of the AIRFL - controller reaches its maximum, the output of the pO2 -
controller switches over to the setpoint input of the GASMIX - controller, see fig. on the next
page. The AIRFL - controller receives the „MAX“ setting defined in the pO2 - controller. For
control of the pO2 the GASMIX - controller operates a 3/2-way valve for supply of air and O2.
Operating display:
pO2 : 88.5 %
SETP : 83.4 %
MODE : auto
CASC : AIRFL GASMX
pO2 Display of actual value des Istwertes

SETP Display and input of the controller´s setpoint
MODE Display and input of the controller´s operating mode:
– off: controller is switched off; outputs are in operating state
CASC Display and selection of servo controller 1 and 2:
AIRFL : The pO2 - controller operates the setpoint of the Airflow - controller
GASM : The pO2 - controller operates the setpoint of the Gasmix-controller
PARAM Access to the controller parameters via 2-digit password „19“
5.8.4.1 Parametrization
Parametrization display page 1 :
pO2 : 88,4 % ñ
DEADB: 0.5 %
HTIME: 5 min GASMX
MIN : 0 %
Parametrization display page 2 :
pO2 : 88.4 % ñ
MAX : 100 %
XP : 100 s
TI : 100 s
DEADB Display and input of the Totzone („Dead band“)

HTIME Display and input of delay time for switching from cascade 1 to 2
AIRFL, etc. Selction of the servo controller using the ALTER key
AIRFL : Display and input of the parameters for the Airflow - controller
GASM : Display and input of the Parameter for the Gasmix - controller
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Control Unit
5.8.4.2 Schematic Drawing, Set-up of „Oxygen Enrichment System“
Fig. 5-4: Set-up of „Oxygen Enrichment System“
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Control Unit
5.8.5 Schematic Drawing „Pump Logic of DFC - Software“

The schematic drawing below shows the different modes of pump control:
pH The pH - controller operates the ACID - pump;
– switching over is possible for control by the SUBS1/2 - controllers
pH The pH - controller operates the BASE - pump;
FOAM The antifoam controller operates the AFOAM - pump;
LEVEL The level - controller operates the LEVEL – pump;
– the level controller can be switched over to the modes „Harvest“ or
„FEED“ for operation of the LEVEL - pump;
SUBS1/2 The substrate - controllers operate the external substrate pump;
– the substrate controller can be switched over for cascade operation by
the pO2 - controller (CASC) and profil mode (PROF):
– switching over is possible for operation of the ACID, BASE, AFOAM or
LEVEL - pump
Fig. 5-5:
Pump switching
logic of DFC -
Software
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Control Unit
5.8.6 Arrangement Plan Control Unit of BIOSTAT® B

See drawing(s) on the next page(s). If the arrangement plan(s) do(es) not match with the
BIOSTAT® B delivered to you and if the customized documentation is not delivered to you
with the equipment or by extra mail, please contact B. Braun Biotech International GmbH for
an update of the documentation.
5.8.7 Assignment Plan for Interfaces of the Control Unit BIOSTAT® B

See drawing(s) on the next page(s). If the assignment plans do not match with the
BIOSTAT® B delivered to you and if the customized documentation is not delivered to you
with the equipment or by extra mail, please contact B. Braun Biotech International GmbH for
an update of the documentation.
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Technical Data
6 TECHNICAL DATA
6.1 Dimensions
Basic unit : 365 x 536 x 450 mm (W x H x D)
Culture vessel (W x H x D) : type B2, 305 x 580 x 270 mm (with stand and motor);
type B5, 400 x 685 x 330 mm (with stand and motor);
type B10, 420 x 828 mm (W x H; with stand and motor)
Weight : control unit approx. 30 kg;
type B2: approx. 16 kg, completely equipped;
type B5: approx. 25 kg, completely equipped;
type B10: depending on equipment
6.2 Installations
Mains connection : 230 V / 50 ... 60 Hz
115 V / 50 ... 60 Hz
Power consumption : max. 900 W
Cooling water : 2 barg (about 29 psig), controlled, connector ID 12 mm;
cooling water flow max. 5 l/min
Cooling water outlet : Connector ID 8 mm
Air inlet : 0.8 barg, controlled,
connector ID 8 mm,
air flow max. 10 l/min
6.3 Control unit

Thermostate system : open thermostate system with circulation pump;
heater of 600 or 800 W, with solenoid valve;
temperature range 8 degC above
cooling water temperature in the range up to 60 degC
Drive system : electronic motor 180 W, speed range 50 ... 1,200 rpm
(need be limitted to 50 ... 600 rpm for 10 l culture vessels by
setpoint adjustment)
Aeration system : autoclavable membrane filter, 0,2 µm, for sterile aeration and
exhaust;
rotameter, manual control valve, air flow max. 10 l/min
optional airflow control equipment
Corrective solution pump : 4 built-in peristaltic pumps for acid, alkali, antifoam, har-
vest/feed, substrate 1, substrate 2;
flow rate max. 8 ml/min for tubings size 1.6 x 1.6 mm; max.
30 ml/min for tubings size 3.2 x 1.6 mm
Storage bottle : 3 glass bottles 250 ml for B 2 and B2-airlift;
3 glass bottles 500 ml for B 5, B5-airlift and B 10
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Technical Data
6.4 Culture Vessels
6.4.1 Culture Vessel Type B 2

Cat.-no. : 884 036/9
Set up : jacketed glass vessel with stainless steel-top plate and baffle
insert; total volume 3 l, working volume 2 l
Materials : glass vessel made of borosilicate glass
- steel parts in touch with medium made of stainless
steel 1.4571 (corresponding with US-standard 316TI),
- other steel parts steel 1.4301 (US-standard 304TI)
- seals made of EPDM (ethylen-propylen rubber)
Stirring system : - rotary shaft with viton lip seals (until January 1993);
- actual design: rotary shaft with mechanical seal;
- stirrer speed range 50 ... 1200 rpm;
- two 6-bladed-disk impellers
6-1)
Ports : 4 side ports in the head space of the glass wall ;
2 top plate ports Ø 19 mm;
4 top plate hose connectors Ø 4 mm;
2 top plate ports Ø 12 mm (Pg 13,5)
Autoclave size (min. required) : Height 550 mm, diameter 360 mm
6.4.1.1 Standard Equipment
Cat.-no. Device Description, specification
3821963/8 baffle insert vessel insert with 4 baffles

3821786/4 aeration pipe with sparger ring
881 007/9 plastic filter for air inlet and exhaust membrane filter, by Millipore
3824042/4 exhaust cooler B2
sampling system harvesting pipe with top-plate adapter
3319088/7 temperature sensor Pt 100 type 200-4
3318258/2 pH-electrode with filling of pres- type Pa/12, length 200 mm
surized pasty electrolyte
3409978/6 pO2-electrode type 12/220
0884446/1 antifoam probe immersion depth 50 mm
0884448/8 level probe immersion depth 122 mm
3821785/6 flat bladed stirrers with 6 blades delivery set of 2 pcs.
manual sampler
0882360/0 storage bottles, 250 ml delivery set of 3 pcs.
6-1) Glass vessels are optionally available without side-entry ports, further information on request.
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Technical Data
6.4.1.2 Optional Equipment for Culture Vessel B2
884 432/1 thermometer kit for vessel B2 for visual temperature monitoring; with pocket
3823907/8 paddle stirrer for vessel B2 delivery of 2 paddle stirrers with option „silicone
membrane insert“
3833697/9 universal hose connector adapter for 1 supply to the culture vessel
884 431/3 quadruple hose connector B adapter for 4 supplies to the culture vessel
3833717/7 reduction connector 19/6 adapter for mounting of devices with Øa = 6 mm in
top-plate ports 19 mm
884 463/1 inoculation kit B for inoculation or dosing of substrate via syringe;
septum with blind closure and membrane is self-sealing and can be punctured
septum membrane several times
884 459/3 flexible holder for exhaust cooler with this adapter the exhaust cooler can be bent
over for autoclave sterilization, this reduces the
height required for the autoclave the
884 434/8 Bypass sampler B - membrane holder with septum membrane
set for assembly of a bypass - 2-channel top-plate adapter 19 mm
- 2 m silicone tubing 1.6 x 1.6 mm
- 0,2 m PTFE-Schlauch 3,5 x 5,5 mm
884 438/0 harvest filter B2 for removal of - rising pipe with teflon filter tube,
samples at microcarrier cultures - pore width 25 µm
884 466/6 kit „silicone membrane insert B5 - silicone membrane, L = 5,2 m, Ø = 3 x 3,7 mm
for bubble-free gas supply“ - membrane filter for gas inlet and outlet, 2 pcs.
- pressure controller
- 2 paddle stirrers
880 830/9 spinfilter B2, 20 µm - mounted to stirrer shaft
- 4-layer mesh made of stainless steel 1.4404
- pore width 20 µm
- harvesting pipe 880826/0 required
880 831/7 spinfilter B2, 40 µm - as above, but with pore width 40 µm
880832/5 spinfilter B2, 75 µm - as above, but with pore width 75 µm
880 826/0 harvesting pipe SF B2 for sample removal via spinfilter
884 436/4 perfusion module B2 perfusion system for continuous fermentation, with
microporous membrane
884041/5 guiding tube for B2 culture vessel recommended for assembly with perfusion module
and spinfilter, if no silicone membrane insert is used
6.4.1.3 Spare Parts for Culture Vessel B2
3920427/8 Spare glass vessel B2
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Technical Data

Cat.-no. : 884 037/7
Set up : jacketed-glass vessel with stainless steel-top plate and baffle
insert, total volume 6,6 l, working volume 5 l
Stirring system : - rotary shaft with viton lip seals (until January 1993);
- actual design: rotary shaft with mechanical seal;
- three 6-bladed-disk impellers
6-2)
Ports : 4 side ports in the head space of the glass wall ;
38219670 baffle insert

38218569 aeration pipe with sparger ring
881 007/9 plastic filter for air inlet and exhaust membrane filter, by Millipore
sampling system
33190895 temperature sensor Pt 100 type 300-4
38218550 flat bladed stirrers with 6 blades delivery set of 3 pcs.
manual sampler
6-2) Glass vessels are optionally available without side-entry ports, further information on request.
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Technical Data
3823954/0 paddle stirrer for vessel B5 delivery of 2 paddle stirrers with option „silicone
membrane insert“
884 463/1 inoculation kit B; for inoculation or dosing of substrate via syringe;
septum with blind closure and membrane is self-sealing and can be punctured sev-
septum membrane eral times
884 459/3 flexible holder for exhaust cooler with this adapter the exhaust cooler can be bent over
for autoclave sterilization, this reduces the height re-
quired for the autoclave
884 434/8 Bypass sampler B - membrane holder with septum membrane
884457/7 harvest filter B5 for removal of - rising pipe with teflon filter tube,
samples at microcarrier cultures - pore width 25 µm
884 467/4 kit „silicone membrane insert B5 - silicone membrane, L = 10,4 m, Ø = 3 x 3,7 mm
for bubble-free gas supply“ - membrane filter gas inlet and outlet, 2 pcs.
- 2 paddle stirrers
- pore width 20 µm
884439/9 perfusion module B5 perfusion system for continuous fermentation, with
884 066/0 guiding tube for B5 culture vessel recommended for assembly with perfusion module
and spinfilter, if no silicone membrane insert is used
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Technical Data

Cat.-no. : 884 038/5
Set up : jacketed-glass vessel with stainless steel-top plate and baffle
insert, total volume 13 l, working volume 5-10 l
Stirring system : - rotary shaft with mechanical seal;
- three 6-bladed-disk impellers
Ports : 4 top plate ports Ø 19 mm;
6.4.3.1 Standard Equipment:
38241072 baffles
38241080 aeration pipe with baffle sparger ring at lower end
881 007/9 plastic filter for Zuluft membrane filter, by Millipore
3922830/4 membrane-filter tube for Abluft filter tube Emflon II,, by Pall
sampling system
00001769 temperature sensor Pt 100
38214881 flat bladed stirrers with 6 blades delivery set of 3 pcs.
manual sampler
BAE_B Rev. 2.31 - 1099 SW 4.1 6-6

Technical Data
884 453/4 thermometer kit for visual temperature monitoring; with pocket
884 054/7 paddle stirrers delivery of 2 paddle stirrers with option „silicone
membrane insert“
884 463/1 inoculation kit B for inoculation or dosing of substrate via syringe;
quired for the autoclave the
884 434/8 Bypass sampler B; - membrane holder with septum membrane
884 060/1 inoculation kit B10 septum port assembly for inoculation connector cat.-
no. 884 061/0
884 061/0 1-inlet inoculation connector for - hollow needle Ø 6 x 0,5 mm made of stainless
inoculation kit B10 steel 1.4435, with sterile sleeve
884 050/4 harvest filter B10 for sample re- - rising pipe with filter tube,
moval at microcarrier cultures pore width 25 µm
884 048/2 kit „silicone membrane insert B10 - silicone membrane, L = 16 m, Ø = 3 x 3,7 mm
for bubble-free gas supply“ - membrane filter for gas inlet and outlet
- 2 paddle stirrers
- pore width 20 µm
884 056/3 spinfilter B10, 40 µm as above, but with pore width 40 µm
884057/1 spinfilter B 10, 75 µm as above, but with pore width 75 µm
884 049/0 perfusion module B10 perfusion system for continuous fermentation, with
884058/0 guiding tube for culture vessel recommended for assembly with perfusion module
B10 and spinfilter, if no silicone membrane insert is used
BAE_B Rev. 2.31 - 1099 SW 4.1 6-7

Technical Data
6.4.4 Culture Vessel B 10, Special Version for Small Working Volumes
Cat.-no. : 884 039/3
Setup : jacketed-glass vessel with stainless steel-top plate;
total volume 13 l, working volume 1,5 - 10 l
Equipment:

pO2-electrode type 12/420
level probe immersion depth 300 mm
BAE_B Rev. 2.31 - 1099 SW 4.1 6-8

Technical Data
6.4.5 Culture Vessel B 2 - Airlift

Cat.-no. : 884 044/0
Setup : jacketed-glass vessel with concave bottom
- H:W (w.v.) = 4 : 1, H:W (total) = 5,4 : 1
- total volume 3,5 l, working volume 2 l
- loop reactor with guiding tube instead of stirrer drive
- other steel parts steel 1.4301 (corresponding with US-
standard 304TI)
4 top plate hose connectors Ø 4 mm
38219395 guiding tube with aeration pipe sparger ring at lower end
881 007/9 plastic filter for air inlet and ex- membrane filter, by Millipore
haust
sampling system
manual sampler
BAE_B Rev. 2.31 - 1099 SW 4.1 6-9

Technical Data
6.4.5.2 Optional Equipment for Culture Vessel B2 - Airlift
884 463/1 inoculation kit B/MD, septum with for inoculation or dosing of substrate via syringe;
blind closure and septum mem- membrane is self-sealing and can be punctured sev-
brane eral times
884 434/8 Bypass sampler B; - membrane holder with septum membrane
6.4.5.3 Spare Parts for Culture Vessel B2 - Airlift
3920429/4 Spare vessel B2 - airlift
BAE_B Rev. 2.31 - 1099 SW 4.1 6 - 10

Technical Data
6.4.6 Culture Vessel B 5 - Airlift

Cat.-no. : 884 045/8
Setup : same as culture vessel B2 - airlift, except
- H:W (w.v.) = 3,7 : 1, H:W (total) = 5,0 : 1
- total volume 9 l, working volume 5 l
Materials : same as culture vessel B2 - airlift
6.4.6.1 Standard Equipment:
n.n. guiding tube acc. to drwg. B318.27.04.00.104

n.n. aeration pipe, sparger ring at acc. to drwg. B318.27.02.00.104
lower end
881 007/9 plastic filter for air inlet and ex- membrane filter, by Millipore
haust
sampling system
n.n. temperature sensor Pt 100 on request
manual sampler
BAE_B Rev. 2.31 - 1099 SW 4.1 6 - 11

Technical Data
6.4.6.2 Optional Equipment for Culture Vessel B5 - Airlift
884 453/4 thermometer kit B5 for visual temperature monitoring; for mounting in
thermometer pocket
884 463/1 inoculation kit B / MD for inoculation or dosing of substrate via syringe;
884 434/8 Bypass sampler B; - membrane holder with membrane
6.4.6.3 Spare Parts for Culture Vessel B5 - Airlift
n.n. spare glass vessel B5 - airlift accord. to drwg. B318.27.00.06.103
BAE_B Rev. 2.31 - 1099 SW 4.1 6 - 12

Technical Data
6.5 Additional Equipment
Cat.-no. Device Description, specifications
880 924/0 STT-quick connector, female con- - for sterile tubing connections
nector of stainless steel for tubings - delivery with blind closure
Ø = 3.2 x 1.6 mm - connection at culture vessel
880 920/8 STT-quick connector, male con- counterpiece to female connector 880 924/0
nector for tubings 3.2 x 1.6 mm - delivery with cap
- connection der externen Zuleitung
880 941/0 STT-quick connector, female con- like STT - female connector 880 924/0
nector for tubings Ø = 1.6 x 1.6
880 940/2 STT-quick connector, male con- counterpiece to female connector 880 941/0
nector for tubings Ø = 1.6 x 1.6 like STT-male connector 880 920/8
882364/2 Storage and harvesting bottle - large bottle made of polypropylen, 10 l
10 ltr., autoclaveable - stainless steel headpiece with 3 inlets
- silicone tubing 3.2 x 1.6 mm, 40 cm length
- top-plate screw with washer
- deaeration filter
882365/0 Storage and harvesting bottle large bottle made of polypropylen, 20 l, equipment
20 ltr., autoclaveable as shown above, delivered with 0,7 m of silicone
tubing 3.2 x 1.6 mm
882366/9 Storage and harvesting bottle large bottle made of polypropylen, 50 l, equipment
50 ltr., autoclaveable as shown above, delivered with 2 m of silicone tub-
ing 3.2 x 1.6 mm,
6.6 Consumables
Cat.-no. Device Description, specifications
618201/1 silicone tubing 1.6 x 1.6 mm minimum order 10 m

3997115/5 silicone tubing 3.2 x 1.6 mm minimum order 10 m
3997078/7 PTFE-tube
880 926/7 slotted membrane for connection for STT-female connector 880924/0 or 880941/0;
of STT - quick connector kit with 20 pcs.
3925071/7 pH calibration buffer pH 4 bottle 250 ml
3925072/5 pH calibration buffer pH 7 bottle 250 ml
3925073/3 pH calibration buffer pH 9 bottle of 250 ml
3409947/6 membrane kit for pO2-electrode for electrodes with Ø = 12 mm
882157/7 elektrolyte for pO2-electrode bottle of 50 ml
3922480/5 filter for storage bottle and manual sampler
3880485/9 septum membrane (washer) for Bypass- sampler, set of 10 pcs.
882 165/8 Pepsin-HCl - cleaning agent bottle of 250 ml
882 166/6 Thio-Urea - cleaning agent bottle of 250 ml
BAE_B Rev. 2.31 - 1099 SW 4.1 6 - 13

Technical Data
6.7 Control Unit

System set-up : highly integrated microprocessor-board (DFC-2) with inte-
grated measurement amplifiers and actuator controls;
measurement amplifiers for temperature, pH, pO2, foam and
level;
microprocessor-system with 80C188 processor, offering
- 128 kByte RAM, 128 kByte EPROM,
- 8 analogue inputs, 8 analogue outputs,
- 8 digital inputs and 16 digital outputs;
automatic restart after power down
Operating panel : illuminated LCD-Display with 4 lines x 20 characters;
Film keyboard with 20 keys
Temperature control : sensor Pt 100, measurement range 0 ... 100 degC
digital PID-controller with split range outputs for heating and
cooling
Stirrer-control : RPM-measurement with Hall sensors,
- measurement range 0 ... 1,200 rpm
digital PID-controller for external motor controller
pH-control : Ingold pH-electrode with pasty electrolyte (Ø 12 mm)
digital sensor calibration
measurement range 2 ... 12 pH
digital PID-controller with split range outputs for acid and al-
kali
dosing counter for acid-/alkaline agent - pump
pO2-control : Ingold pO2-Electrode (Ø 12 mm);
digital sensor calibration;
measurement range 0 ... 100 % pO2 (with reference to oxy-
gen saturation of the measurement solution);
digital PID-cascade controller for stirrer speed sequential
controller or for control of a Gas Mix unit (optional)
Antifoam-Control : conductive foam sensor;
digital on/off controller for antifoam agent pump;
dosing counter for antifoam agent pump
Level control : conductive level sensor;
digital liwith controller for harvest or substrate pump
dosing counter for harvest/substrate pump
Substrate pump control : max. 2 pumps via pulse width modulated outputs;
adjustable delivery rate 0 ... 100%;
dosing counters for feed pumps
Recorder connection : 6 analogue outputs 0 ... 10 V;
assignment of parameters to channel configurable
Protocol printer connection : via serial interface RS 232 C; printing intervals and process
variables for protocol configurable
Host connection : via serial interface RS 422; protocol with measured values,
set points and controller mode
BAE_B Rev. 2.31 - 1099 SW 4.1 6 - 14

Optional Equipment and Accessories
7 OPTIONAL EQUIPMENT AND ACCESSORIES
Detailed information about optional equipment and accessories will be included herein by
time oder will be sent to you together with the equipment or by extra mail.
If your fermentor system BIOSTAT® B includes optional equipment or is of special design,
which is not described in the above manual or by the documentation of this section or as ex-
tra delivered, please contact B. Braun Biotech International GmbH for such documentation:
phone +49 56 61 - 71 34 00,
fax +49 56 61 - 71 37 02
BAE_B Rev. 2.31 - 1099 SW 4.1 7-1

Supplement
8 SUPPLEMENT
8.1 Conversion Table for Degrees of Hardness of Water
Note: In this table the german decimal comma (",") is used instead of the english decimal
point (".")
alkaline alkaline german CaCO3 engl. french.

earth ions earth ions degrees degrees degrees
[mmol/l] [mval/l] [°d] [ppm] [°e] [°f]
1 mmol/l alkaline earth ions 1,00 2,00 5,50 100,00 7,02 10,00
1 mval/l alkaline earth ions 0,50 1,00 2,80 50,00 3,51 5
1 german degree [°d] 0,18 0,357 1,00 17,80 1,25 1,78
1 ppm CaCO3 0,01 0,020 0,056 1,00 0,0702 0,10
1 engl. degree [°e] 0,14 0,285 0,798 14,30 1,00 1,43
1 french degr. [°f] 0,10 0,200 0,560 10,00 0,702 1,00
8.2 Characteristic Values for Gases
Gas : Density
3
Carbon dioxide CO2 : 1,977 kg/m
3
Air : 1,293 kg/m
3
Methane : 0,717 kg/m
3
Oxygen : 1,429 kg/m
3
Nitrogen : 1,251 kg/m
BAE_B Rev. 2.31 - 1099 SW 4.1 8-1

Supplement
8.3 Calculation Methods for Flowrates of the Rotameters
8.3.1 Sizing of Flowmeters / Rotameters
Rotameters of the Gas Mixing System or flowmeters are usually calibrated and graduated according to
standard conditions. These calibration conditions are marked on every glass tube or on the support
connector of any rotameter. For instance, such calibration conditions may be
Calibration gas : air
Temperature : 20 °C = 293 K
Pressure : 1 bar overpressure
-> Please check which values are marked on your rotameters / flowmeters and must be con-
sidered for further evaluations.
As far as your operation conditions (type of gas, density, temperature, pressure, etc.) deviate from the
standard conditions and if you need the exact flowrates for the interpretation of the fermentation proc-
ess, you may calculate the reference flowrates using the procedures or the tables / nomographes
shown below. Additional information about calculation of flowrates of a gas under special conditions of
temperature, pressure and specific density (specific heat) can be found in the corresponding literature
or is available from manufacturers of rotameters / flowmeters.
You may also use these formulas, tables or nomographes to evaluate which design of the Gas Mixing
System will meet your specific requirements. For selection of an appropriate Gas Mixing System it is
important, that its design allows the maximum flowrates of the individual gas which are required for the
projected fermentation process.
If the rotameters are inadequately sized delivering to small flowrates, the automatic control
may not function properly.
Especially, when the pO2-Control with the gas supply does not operate accurately, this may
be due to wrong sized rotameters.
8.3.2 Calculation Method
The corrective calculation for the flowrates includes the following formula (1) - (3) :
(1) q1 = q2 * F
q1 = air-equivalent flowrate of the gas (flowrate read on the flowmeter
q2 = actual flowrate of the gas
F = conversion factor
BAE_B Rev. 2.31 - 1099 SW 4.1 8-2

Supplement
According to Brooks, „Sizing calculations“, the conversion factor F can be calculated by equation (2) :
ρ2 · T2 · p1
F=
ρ1· T1· p2
3
ρ1 = density of the calibration gas; e.g. for air the density is ρ1 = 1,293 kg/m
ρ2 = density of the gas to be controlled 8-1)
T1 = temperature in [Kelvin] at calibration conditions
T2 = temperature at measurement conditions, in [Kelvin]
p1 = pressure at calibration conditions, in [bar]
p2 = pressure at measurement conditions, in [bar]
This leads to equation (3):
q1 q1
q2 = =
F ρ2 · T2 · p1
ρ1· T1· p2
Example for calculating:

Evaluated gas : for calibration : air ;
3
density (20 °C) ρ1 = 1,293 kg/m
gas for measurement CO2;
3
density (20 °C) ρ2 = 1,977 kg/m
Standard flow : q1 = 10 l/min
Pressure : for calibration : p1 = 1 barabs
operation pressure : p2 = 1,5 barabs
Temperatur e : for calibration : 20 °C (= 293 K)
operation temperature 25 °C (= 298 K)
These values give the following result for the actual flow of CO2 at operating conditions :
10 l / min 10 l / min
q2(CO2) = =
F 1977
, · 298 · 1
1293
, · 293 · 15,
q2 (CO2) = 9,8 l/min
8.3.3 Manufacturer's Examples for Sizing Calculations

The following tables and nomographes give examples of manufacturer's of metering tubes
and rotameters for calculation of actual flowrates at given operating conditions.
If you are using other brands or types of flowmeters / rotameters, you may find correspond-
ing information in the documentation delivered with your specific equipment or you may
contact the manufacturer of these parts for further support.
8-1) Densities of usual gases applied in fermentation processes can be found in the above table
"Characteristic Values for Gases" as well as in corresp. literature
BAE_B Rev. 2.31 - 1099 SW 4.1 8-3

Supplement
BAE_B Rev. 2.31 - 1099 SW 4.1 8-4

Supplement
BAE_B Rev. 2.31 - 1099 SW 4.1 8-5

Supplement
8.4 Additional Drawings and Flow Charts
8.4.1 P & I - Diagram of the BIOSTAT® B

See copy attached on next page. This is an example of the default design of the
BIOSTAT® B at the time this document was printed.
8.4.2 Drawings and Flow-Charts of Fermentor Options or Customized Fermentors

Drawings will be completed with delivery of the manual, especially when drawings are avail-
able of optionally designed fermentor systems BIOSTAT® B.
BAE_B Rev. 2.31 - 1099 SW 4.1 8-6

Supplement
Declaration of Conformity with EC-Directives, Standards and

Technical Specifications
By the copies of the „Declaration of Conformity“ enclosed in this Operating Manual or delivered extra
with the device the B. Braun Biotech International GmbH confirms, that the fermentor systems
BIOSTAT® B fulfill the requirements of all mentioned EC-directives, harmonized and national stan-
dards and technical specifications. The signatures of the English forms substitutionally confirm the va-
lidity of the declaration for all other language forms.
BAE_B Rev. 2.31 - 1099 SW 4.1 8-7

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or your local sales representative of B. Braun Biotech International GmbH.

Guide Through this Manual

1 : Overview on design and equipment of the fermentor

Documentation Release Notes

B. Braun Biotech International GmbH

BAE_B Rev. 2.31 - 1099 SW 4.1

Declaration about decontamination and

A. Description of the Equipment or Component(s)

Description / Cat. No.: ........................................................................................................................

Attention: Please specify exactly the biological, chemical, or radioisotopic contaminant.

C. Legally binding declaration

Company / Institute: .........................................................................................................................

Tel. / Fax (with area code):........................................................./ ....................................................

Name of the authorized person: .......................................................................................................

Signature / Date: ................................................................................................. /...........................

F190101E.DOC / Rev. 3 / 11.05.2000 Page 1 of 2 BBI-Dr. Wieneke

D. Reason for return

wrong delivery exchange repair modification other

F. Vom BBI Service auszufüllen (reserved for BBI service)

Please pack the equipment properly and B.Braun Biotech International

F190101E.DOC / Rev. 3 / 11.05.2000 Page 2 of 2 BBI-Dr. Wieneke

1 Design and Function

2 Delivery and Installation

3 Starting and Operating the BIOSTAT® B

BAE_B Rev. 2.31 - 1099 SW 4.1 1

3.3 Peparing a Fermentation Run 3-5

4 Equipment of the Culture Vessels

BAE_B Rev. 2.31 - 1099 SW 4.1 2

4.5 External Equipment for Connection to the Culture Vessels 4 - 34

BAE_B Rev. 2.31 - 1099 SW 4.1 3

5.8.4 Example: pO2-Controller with Airflow - Massflow Controller and

7 Optional Equipment and Accessories

This section intentionally left free in this version of the manual.

BAE_B Rev. 2.31 - 1099 SW 4.1 4

1 DESIGN AND FUNCTION

1.1 Control Unit

BAE_B Rev. 2.31 - 1099 SW 4.1 1-1

1.1.1 Thermostate System

1.1.2 Drive System

1.1.3 Air and Gas Supply

BAE_B Rev. 2.31 - 1099 SW 4.1 1-2

1.2 Culture Vessels

The vessels have an internal concave

The culture vessels are mounted into a

When the culture vessel is placed in

BAE_B Rev. 2.31 - 1099 SW 4.1 1-3

1.3 Integrated Digital Control System

BAE_B Rev. 2.31 - 1099 SW 4.1 1-4

2 DELIVERY AND INSTALLATION

2.1 Unpacking and Checking Completeness of Delivery

2.2 Warranty Agreements and Customer Service

BAE_B Rev. 2.31 - 1099 SW 4.1 2-1

2.3 Customer Service

If repairs are to be carried out on the premises or in a customer-owned workshop by own

BAE_B Rev. 2.31 - 1099 SW 4.1 2-2

2.4 Installing the Fermentor System