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STUDENT ORGAN BATH
Introduction:
The organ bath is a traditional experimental set-up that is commonly
used to investigate the physiology and pharmacology of in vitro tissue
preparations. Perfused tissues can be maintained for several hours in a
temperature controlled organ bath.
After a suitable equilibration period, the researcher can begin

experiments. Typical experiments involve the addition of drugs to the
organ bath or direct/field stimulation of the tissue. The tissue reacts by
contracting/relaxing and an isometric or isotonic transducer is used to
measure force or displacement, respectively. From the experimental
results dose-response curves are generated (tissue response against
drug dosage or stimulus potency).
ADInstruments supply a range of organ baths and transducers for

pharmacological research. LabChart software records, displays and
analyzes the data which may then be analyzed or exported to graphing
programs, such as GraphPad Prism®.
Some of the tissues that may be studied with an organ bath system
include:
Smooth Muscle:
Vascular, e.g. arterial/aortic rings
Guinea-pig ileum
Vas Deferens (secretory duct of the testicle)
Uterine
Colon
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RAIGARH COLLEGE OF PHARMACY
STUDENT ORGAN BATH
Skeletal Muscle:
Toad Abdominus Rectus
Chick Biventer Cervis
Mammalian Diaphragm
Cardiac Muscle:
Atrium
Ventricle
Tissues are usually prepared in a petri-dish containing physiological

solution (i.e. Kreb’s solution). The ends of the tissue are then attached to
the mounting hook and transducer using silk. It is important to use non-
compliant material such as silk or fine wire to mount the tissue.
For tissue ring preparations (i.e. arterial rings), a custom tissue-holder is

required (can be sourced by ADInstruments). These may also be made
using triangular-shaped wire holders constructed of non-compliant
material and secured to the mounting hook on the Panlab tissue-support.
Figure 1.
Schemat
ic diagram of a custom-made mounting system for tissue rings.
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STUDENT ORGAN BATH
Panlab Organ Baths:
ADInstruments supply compact and robust, yet versatile organ baths

manufactured by Panlab. Each organ bath can take 5, 10, 25 or 50 ml
tissue chambers. The tissue chamber can be easily changed by the user
and any sized tissue chamber can be used in all the Panlab organ baths.
The tissue chambers are automatically filled and emptied using
electrovalves with each tissue chamber also having it own gas diffuser
which can be regulated.
A heating rod is built into each of the organ bath units and a thermostat
controller is included to control the water temperature surrounding the
tissue chambers (Figure 2). Pre-heating reservoir coils are also
positioned within the water bath to heat the perfusate before entering the
tissue chamber. A highly efficient, magnetic water pump is built into the
water bath to circulate the heated water and ensure even heat
distribution throughout the entire system. Manipulators/micropositioners
are supplied for accurate positioning of the transducers and applying
pre-tension to the tissue preparations prior to experimentation.
Figure 2.
Schematic diagram of a 2 chamber Panlab organ bath.
ADInstruments/Panlab Organ Bath Systems

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STUDENT ORGAN BATH
The Panlab organ baths are supplied as complete, ready-to-use
research systems that include all of the necessary hardware and
software to perform isolated tissue experiments. Each research system
includes:
 An 8 or 16 channel PowerLab data acquisition system

 Multi-chamber Panlab organ baths (4, 8 or 16 chamber systems
available)
 Bridge amplifiers (Quad or Octal Bridge Amps)
 Isometric transducers
 LabChart Pro (including Dose Response)
All of these components may be purchased separately, if required.

Single Chamber Organ Bath
ADInstruments supply a single chamber organ bath by Panlab; however,

it is only recommended for teaching applications and does not include a
thermostat controller.
ADInstruments have a Thermoregulated Water Pump (Model #:

MLE2001) that will work with up to four of the single chamber organ
baths in series; however, there should be no more than 2 meters
distance between the first and last organ baths.
If an external heating system is required, then an "immersion

heater/circulator" is suitable (i.e. the TU1 from Thermoline
Scientific,www.thermoline.com.au). When used with the Single Chamber
Organ Bath, the maximum flow rate of the circulator should not exceed 5
L/min (to avoid excessive pressure within the system). The size of the
support clamp, length of the heating rod and intending heating volume
should also be checked to ensure they are compatible with the
dimensions of the water bath.
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STUDENT ORGAN BATH
Stimulating Electrodes
Electrodes are required if using a stimulator. The platinum electrodes are

recommended for most researchers using field stimulation (see table
and diagrams below). All of the following stimulating electrodes are
supplied with 2 mm plug connections.
Type & Description Platinum Stainless Steel Single Ring (6.5, 8 or 10

mm inner diameter) 20 mm between ring and tissue holder MLA0301/ID
MLA0305/ID Double Ring (6.5, 8 or 10 mm inner diameter) 20 mm
between rings, 6mm above tissue holder MLA0302/ID MLA0306/ID 3
mm Length Pole, 20 mm from tissue holder MLA0303 MLA0307 Double
Ring (6 mm inner diameter) 15 mm between rings, 12 mm from tissue
holder MLA0304 MLA0308
Specify: /ID = 6.5, 8 or 10 mm
Key Benefits
 Available in 2, 4, 8 chamber configuration

 Single water bath provides controlled temperature regulation
 Automated chamber filling and emptying using electrovalves
 All-in-one design provides a compact portable unit
Radnoti Organ Baths:

ADInstruments provide complete 4, 8 and 16 chamber Radnoti tissue-
organ bath systems that include a PowerLab, bridge amplifiers, thermo
bath/circulator and transducers for the recording of data from isolated
tissues.
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STUDENT ORGAN BATH
Radnoti organ baths include a single water-jacketed 2 L reservoir for the

physiological solution. The solution enters the tissue chamber via a
preheating coil that is positioned within the water-jacket of the High-Tech
Tissue Organ Bath. The solution in the chamber can be emptied via a
drain plug (stopcock) at the bottom of the tissue chamber for a single-
pass mode or may be recirculated in a constant flow mode. In the
constant flow mode, the solution exits via an overflow outlet positioned at
the top of the chamber. Aeration of the physiological solution is provided
via needle valves in each bath as well as a frit in the reservoir. Each
chamber has its own 5 mg to 25 g isometric transducer (MLT0201/RAD)
and transducer positioner. The modular construction of Radnoti systems
permits the researcher to easily interchange tissue chambers and use
different sized baths (5, 10, 25, 50, 100, 200, 300 ml). Each organ bath
is individually mounted via adjustable ring clamps and is connected to a
sturdy steel base that has a resistive coating to avoid corrosion.
Radnoti Thermal Circulator Pump (170051B-V)
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STUDENT ORGAN BATH
This unit incorporates a high wattage heater for rapid warming of the
water bath and a two-stage force and suction pump that provides water
circulation. Circulation of the water is necessary to maintain a constant
temperature in the reservoir and tissue chamber jackets.
Specifications
 Reservoir Volume: 17 Liters (4.5 Gallons)

 Temperature range: +12 °C to +200 °C
 Temperature Stability: ±0.01 °C
 Maximum Pump Flow Rate: 15 L/min
 Bright, LED digital display
 Stainless steel reservoir with drainage
 Low water level safety shutoff
Each ADInstruments/Radnoti Tissue-Organ Bath System Includes:
 2L water-jacketed reservoir
 Transducer Positioners
 Oxygenating Bubblers
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STUDENT ORGAN BATH
 Support Stand & Hardware Kit
 Force Transducers (Isometric)
 Cable & Connector Package
 High-Tech Tissue Organ Baths (5-300 ml; User must specify size
when ordering)
 Bridge Amplifiers
 Thermo Bath/Heater
 PowerLab Data Acquisition System
 LabChart Pro (including Dose Response)
These systems require manual refilling and emptying of tissue chamber.

Stimulating Electrodes
Radnoti also provides several combination tissue support

rods/stimulating electrodes that are suitable for use in organ baths
larger than 10 ml. These allow both vertical and horizontal
adjustment of the platinum electrodes in relation to the tissue
sample to ensure maximal stimulation. A general guide for the use
of these electrodes is as follows:
 Tissue strips/spirals: Straight (160152-11), zig-zag (160152-12) or

coil (160152-13) electrodes
 Small samples: Coil (160152-13) or flat (160152-15) electrodes
Page 5/10 Tech Note
 Blood vessel rings: L-type electrode (160152-14). The tissue ring
is slid on the lower support and attached to a tri-shape tissue
support (158817 and 158819) and placed under tension between
the two horizontal platinum wire electrodes
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STUDENT ORGAN BATH
The above product codes are for combination universal support
rods/stimulating electrodes; however, the electrodes may be purchased
separately to be interchanged on a universal support rod.
Key Benefits
Modular components enable tissue chambers to be easily removed and

permit investigations with different sized chambers
Each chamber is individually water-jacketed ensuring uniform circulation

and heating
Reservoir is water-jacketed to maintain solution temperature
Overflow port allows constant recirculation of perfusate to limit the

amount of solution require
Incorporates Quick Disconnect fittings
Large range: of tissue/organ baths volumes (5,10, 25, 50, 100, 200, 300
ml)
Transducers:
Isometric Transducers
Isometric transducers measure the change in force (in grams [g] or

milliNewtons [mN]) produced by muscle relaxation/contraction while the
muscle length remains constant.
Isotonic Transducers
Stimulation and Electrode Selection
Stimulation of tissues may be achieved by a number of methods
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STUDENT ORGAN BATH
Field stimulation may be performed using ring or plate electrodes that
are placed near (but do not touch) the tissue and have a current passed
between them. This method is effective provided that the tissue remains
within the field of stimulation and is the most common technique used in
organ bath preparations.
A field stimulating electrode is available from Radnoti. It is constructed of

platinum wire sealed in glass and designed to insert into Radnoti organ
baths. This electrode features a dual vertical spiral coil design with a 1
cm gap that provides a large, uniform electrical field that surrounds the
tissue. Isotonic transducers measure the change in muscle length (in
millimeters [mm] or degrees [°] traveled) as it contracts/relaxes against a
constant force.
Parts of Student organ bath

Student organ bath or simply organ bath is an apparatus widely
used in pharmacology laboratory and various educational Institutes,
research and development sector.
Organ bath is not a single apparatus but it is a combination of
many small units which include-
1. Bath chamber
2. Organ tube
3. Glass coil
4. PSS reservoir
5. Heater
6. Thermostat
7. Stirrer or mixer
8. Oxygen tube
9. Aerator
10. Leaver
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STUDENT ORGAN BATH
11. Load
12. Sherrington rotating drum (Kymograph Drum)
13. Kymograph Paper
Use- When we have to perform the invitro- studies of any given drug, we
need to isolate
The organ from suitable experimental animal. Once organ is isolated
it should be immediately transferred into physiological salt solution
(PSS). Organ bath is having several important component and assembly
as mentioned above; all are equally important and have individualized
functions. Once organ is placed into PSS other arrangement are made.
1. Cleaning the organ bath

assembly
2. Attaching or arranging all
small hooks, clamp and water
pipes.
3. Fixing the lever and stick
the kymograph paper on
rotating drum.
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STUDENT ORGAN BATH
4. Kymograph paper should

be either smoked or have to
use colour ink at the end of
lever.
5. Fill the water into 2/3
portion of organ bath or up
to suitable length depending
upon the unit of organ bath.
6. Switch on the mains and
heater
7. Wash the organ tube with
PSS once
8. Fill the PSS into organ
bath and hold it.
9. Start providing oxygen by
aerator machine
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STUDENT ORGAN BATH
10. Attach the aeration tube

into the organ bath
11. Now tie the tissue of
isolated organ which you have
to study (This process may be
modified)
12. Tie the other end of thread
into writing lever
13. Attach the lever with
rotating drum and wait till
resting or constant line appears
14. Inject the drug into
organ bath and carefully
examine the changes in
muscle
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STUDENT ORGAN BATH
strength by observing
kymograph paper.
15. Release the clamp of organ
bath to let the PSS flow
outside the organ tube if you
want to reduce the strength of
dose, if you want to increase
the dose, no need to
change the PSS. (It is for same
drug)
16. For every new drug
injection the old PSS must be
change.
17. Ensure the temperature
of water bath; it should not
increase than optimum
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STUDENT ORGAN BATH
temperature.
1. Bath chamber- Bath

chamber is made of good
quality plastic (PVC) it is of
two types-
Single unit and double unit. In
single unit there is only one
place for organ
tube, thus you can perform one
study at one time, or you need
to wash the
tissue with PSS before
changing drug. Double unit
organ bath is having
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STUDENT ORGAN BATH
two holding place for organ

tube, that’s why we can
perform two studies at
same time. It depend on
you weather you want to
check the effect of
various dose of same drug or
you can check two separate
drug’s effect on
muscle. At the bottom of
bath chamber there is two
outlets, one is for
drainage of water and second
is for the PSS.
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STUDENT ORGAN BATH
2. Organ Tube- Organ tube is

the soul of organ bath because
the king of experiment lies or
rest in this chamber. Organ
tube is a glass tube having two
sides opening.
Out of which one is for
expelling used PSS and second
is for inlet of PSS.
PSS is not filled directly from
the top of organ tube, but it is
filled from
this side inlet unit.
assembly
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STUDENT ORGAN BATH

pipes.
rotating drum.
lever.
heater
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STUDENT ORGAN BATH

PSS once
bath and hold it.
aerator machine
into the organ bath
modified)
into writing lever
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STUDENT ORGAN BATH

muscle
kymograph paper.
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STUDENT ORGAN BATH

drug)
change.
temperature.

two types-
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STUDENT ORGAN BATH

place for organ
to wash the
check the effect of
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STUDENT ORGAN BATH

drug’s effect on
outlets, one is for
is for the PSS.
sides opening.
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STUDENT ORGAN BATH

filled from
assembly
pipes.
rotating drum.
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STUDENT ORGAN BATH

lever.
heater
PSS once
bath and hold it.
aerator machine
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STUDENT ORGAN BATH

into the organ bath
modified)
into writing lever
muscle
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STUDENT ORGAN BATH
kymograph paper.
drug)
change.
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STUDENT ORGAN BATH
temperature.

two types-
place for organ
to wash the
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STUDENT ORGAN BATH

check the effect of
drug’s effect on
outlets, one is for
is for the PSS.
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STUDENT ORGAN BATH

sides opening.
filled from
Introduction- Student organ
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STUDENT ORGAN BATH
bath or simply organ bath is

an apparatus widely used in
pharmacology laboratory and
various educational Institutes,
research and development
sector.
Organ bath is not a single
apparatus but it is a
combination of many small
units which
include-
1. Bath chamber
2. Organ tube
3. Glass coil
4. PSS reservoir
5. Heater
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STUDENT ORGAN BATH
6. Thermostat
7. Stirrer or mixer
8. Oxygen tube
9. Aerator
10. Leaver
11. Load
12. Sherrington rotating drum
(Kymograph Drum)
13. Kymograph Paper
Use- When we have to
perform the invitro studies of
any given drug, we need to
isolate
the organ from suitable
experimental animal. Once
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STUDENT ORGAN BATH
organ is isolated it should

be
immediately transferred into
physiological salt solution
(PSS). Organ bath is having
several important component
and assembly as mentioned
above; all are equally
important
and have individualized
functions. Once organ is
placed into PSS other
arrangement are
made. These include
Page | 1
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STUDENT ORGAN BATH
Introduction- Student organ

bath or simply organ bath is
an apparatus widely used in
pharmacology laboratory and
various educational Institutes,
research and development
sector.
Organ bath is not a single
apparatus but it is a
combination of many small
units which
include-
1. Bath chamber
2. Organ tube
3. Glass coil
4. PSS reservoir
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STUDENT ORGAN BATH
5. Heater
6. Thermostat
7. Stirrer or mixer
8. Oxygen tube
9. Aerator
10. Leaver
11. Load
12. Sherrington rotating drum
(Kymograph Drum)
13. Kymograph Paper
Use- When we have to
perform the invitro studies of
any given drug, we need to
isolate
the organ from suitable
experimental animal. Once
Page | 35
STUDENT ORGAN BATH
organ is isolated it should

be
immediately transferred into
physiological salt solution
(PSS). Organ bath is having
several important component
and assembly as mentioned
above; all are equally
important
and have individualized
functions. Once organ is
placed into PSS other
arrangement are
made. These include-
Selection of physiological buffer solution and manner of aeration
The buffer solution(s) chosen for the dissection and maintenance of the
tissue will have a profound effect on the viability of the preparation.
Whole blood from donors may be used, but since whole blood is
generally in limited supply, it must be recycled, for example with an
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STUDENT ORGAN BATH
appropriate peristaltic pump through a membrane oxygenator, without
denaturing blood proteins or causing erythrocyte damage. Whole blood
may also contain biologically active materials, such as hormones and
various cell types, that may affect the tissue preparation. Because of
these limitations, most isolated tissue preparations use an artificial
plasma-like buffer solution to maintain cell viability. Since almost all of
these solutions are water based, water purity is a primary concern. The
water purity should at least be the equivalent of double distilled water
with organic contaminants removed. Achieving this standard used to be
difficult, but with the advent of modern reverse osmosis systems or glass
stills passing on water to charcoal and ion exchange resin filters,
sources of purified water are now readily available with resistivity
measurements in excess of 10MW. Some recipes for buffer solutions still
add 0.1-5 mM EDTA (see Table 1) for chelation of heavy metals that
may be added from salts or leached from metal tubing or other sources.
After storing the water in clean, non-leaching containers, the next
decision to be made is which formulation of the buffer solution should be
used. Reagents should be of high quality (the equivalent of USP or
Analytical Reagent grade) and kept isolated from the rest of the
chemicals in the laboratory to reduce the possibility of misuse or
contamination. There are a number of widely used physiological salt
solutions, such as Ringer’s (5), Tyrode’s (6) and Krebs-Hanseleit (7,
modified as in 8, see Table 1) and their salt concentrations are often
modified to resemble those in the donor’s plasma (see Radnoti Isolated
Perfused Heart Table 1 ). Since these solutions must be buffered, a
choice of buffering agents must also be made. Bicarbonate buffers such
as Tyrode’s and Krebs-Hanseleit, which are based on those naturally
occurring in mammalian blood, are effective when in equilibrium with
relatively high levels of carbon dioxide, hence the use of 95% oxygen
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STUDENT ORGAN BATH
and 5% carbon dioxide gas mixtures to aerate them. The use of 95-
100% oxygen rather than atmospheric levels of -20% is to increase the
oxygen content of the solution to compensate for the lack of hemoglobin
or other oxygen carriers. In most of these solutions, phosphates (or
sulfates) are also added. Besides adjusting the pH and increasing
buffering capacity, the presence of carbonates, bicarbonates and
phosphates aids in maintaining normal anion homeostasis. The
restrictions imposed by bicarbonate buffers are the requirement for a
mixed gas aeration and the adverse effects on solubility products
created by these anions. As an alternative, buffering capacity can be
created through the use of synthetic buffers like HEPES or MES in place
of bi-carbonates with 100% oxygen used for aeration. It is also possible
to use these synthetic buffers in addition to bicarbonate, and then adjust
the sodium balance appropriately to maintain osmolarity. The most
common metabolic substrate is glucose, with pyruvate, lactate, and fatty
acids also added. To aid in glucose metabolism, insulin may be added.
Often salts other than calcium are kept as a concentrated stock in one
container and calcium and glucose kept as a concentrated stock in
another container and then diluted and combined just prior to use. This
is done to reduce precipitation of calcium phosphate and retard bacterial
growth by having hyperosmotic solutions. To maintain an appropriate
oncotic pressure that will reduce edema, albumin, polyvinylpyrrolidine,
dextran or other plasma expanders can be added. Note that the
presence of albumin or other readily denatured or poorly soluble
compounds will necessitate indirect aeration, such as that provided by a
membrane oxygenator, to prevent foaming and precipitation. Always
adjust the pH of the solution while it is at the bath temperature selected
and when it is normally aerated.
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STUDENT ORGAN BATH
Gas flow to the buffer is normally controlled with a two-stage regulator
designed for the gas mixture utilized. Flow rates of 0.5-2 liters per minute
and/or pressures of 1-2 psi are adequate for most preparations. The
aeration stream can be controlled with a needle valve and should be a
steady line of fine bubbles that do not cause a pronounced “boiling”
effect in the bath. Higher pressures will only serve to increase
evaporation, jostle the preparation and create noise in the force trace
and may perforate glass frits used for aeration. For thin tissues with a
large surface area and low metabolic activity (such as blood vessel strips
or rings), aeration using room air may be sufficient; in these cases
simple aquarium pumps are a cost effective replacement for compressed
air tanks.
Selection of tissue donors

Sources of tissue may be from living or dead donors, depending upon
the availability of the tissue type and the requirements of the
experimenter. For tissues obtained from deceased donors, the type of
tissue, the manner of death or euthanasia, the time elapsed between
death and harvesting the tissue and the temperature that the donor’s
body is maintained at will affect the viability of the preparation. In the
case of living donors, the selection and composition of anesthesia or
euthanasia solutions should be made carefully to reduce the impact
upon retrieved tissues. The presence of high concentrations of
barbiturates, general anesthetics, ethanol, potassium or other
substances used in or with the anesthesia or euthanasia may slow
functional recovery, have unexpected long term effects or possibly cause
irreversible damage. The species, variety, age and sex of the donor
should be taken into account, both for the impact on physiological
responses and for the fact that older animals have fatty deposits that
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STUDENT ORGAN BATH
make dissections more difficult.
Advantage And Disadvantage of Student Organ Bath:

An organ bath is a laboratory technique used to study the physiology of
smooth muscle and cardiac muscle in isolated strips of tissue.
Advantages:
 Allows for direct observation and manipulation of muscle

contractions.
 Can be used to study the effects of drugs and other agents on
muscle physiology.
 Can be used to study the physiology of both smooth muscle and
cardiac muscle.
Disadvantages:
 The tissue must be isolated from the body, which can alter
physiological responses.
 The tissue can only be studied for a limited time before it
deteriorates.
 The results obtained may not be directly applicable to in vivo
physiology.
Standard Operating Procedure (SOP) for a student organ bath

typically includes the following sections:
Purpose: The purpose of this SOP is to provide instructions on the
proper use of the student organ bath instrument for conducting
physiological experiments on isolated organs.
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STUDENT ORGAN BATH
Materials:
 Student organ bath instrument

 Isolated organ of interest
 Physiological salt solution
 Temperature controller
 Power supply
 Data acquisition system
 Necessary accessories (tubing, connectors, etc.)
Procedure:
 Prepare the physiological salt solution according to the desired
composition.
 Fill the organ bath with the prepared solution and adjust the
temperature to the desired level using the temperature controller.
 Connect the organ bath to the power supply and start the data
acquisition system.
 Gently place the isolated organ of interest in the bath and secure it
in place using the necessary accessories.
 Adjust the bath parameters (e.g. flow rate, temperature, etc.) as
needed to maintain the desired conditions for the experiment.
 Perform the experiment according to the specific protocol and
collect data.
 After the experiment, carefully remove the organ from the bath and
dispose of the used solution according to established protocols.
Safety:
 Always wear personal protective equipment (PPE) when handling
the isolated organs.
 Avoid contact with the bath solution to prevent skin irritation.
 Do not exceed the recommended temperature range for the organ
of interest to prevent damage to the tissue.
 Follow proper procedures for disposal of biological materials.
 Data Analysis and Interpretation:
 Analyze the collected data according to the specific protocol and
report the results.
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STUDENT ORGAN BATH
 Compare the results to the expected outcomes and interpret any
discrepancies.
References:
 Provide any relevant references or protocols used in the
development of the SOP.
 Include any guidelines or recommendations from the manufacturer
of the organ bath instrument.
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STUDENT ORGAN BATH
Experiment (1)
Aim: To investigate the effect of epinephrine on the contractile activity of isolated

frog heart.
Requirement:
 Isolated frog heart
 Epinephrine solution
 Power supply
 Microscope
Theory: Epinephrine is a hormone that acts as a neurotransmitter and a vasoactive
agent. It binds to specific receptors on the surface of heart muscle cells and causes
an increase in contractile activity. This increase in contractility is mediated by an
increase in intracellular calcium ions, which leads to an increase in the strength and
frequency of cardiac contractions.
Procedure:
 Prepare the physiological salt solution according to the desired composition.
 Fill the organ bath with the prepared solution and adjust the temperature to
the desired level using the temperature controller.
 Connect the organ bath to the power supply and start the data acquisition
system.
 Gently place the isolated frog heart in the bath and secure it in place using the
necessary accessories.
 Adjust the bath parameters (e.g. flow rate, temperature, etc.) as needed to
maintain the desired conditions for the experiment.
 Begin baseline data collection and record the heart rate and contractility.
 Introduce epinephrine solution to the bath and continue to record the heart
rate and contractility.
 Repeat steps 6-7 for different concentrations of epinephrine.
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STUDENT ORGAN BATH
 After the experiment, carefully remove the heart from the bath and dispose of
the used solution according to established protocols.
Result:
The results of the experiment should show an increase in the heart rate and
contractility in response to increasing concentrations of epinephrine.
The experiment should also show that epinephrine causes an increase in the
strength and frequency of cardiac contractions, as predicted by the theory.
The result can be analyzed using graphs and statistical analysis.
Experiment (2)
Aim: To investigate the effect of different temperature on the contractile
activity of isolated frog skeletal muscle.
Requirement:

 Isolated frog skeletal muscle
 Power supply
 Microscope
Theory: Skeletal muscle contractility is affected by the temperature of
the muscle. The muscle fibers have a specific temperature range in
which they function optimally. The muscle fibers are sensitive to changes
in temperature, and contractile activity decreases as the temperature
deviates from the optimal range.
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STUDENT ORGAN BATH
Procedure:
1. Prepare the physiological salt solution according to the desired

composition.
2. Fill the organ bath with the prepared solution and adjust the
temperature to the desired level using the temperature controller.
3. Connect the organ bath to the power supply and start the data
acquisition system.
4. Gently place the isolated frog skeletal muscle in the bath and
secure it in place using the necessary accessories.
5. Adjust the bath parameters (e.g. flow rate, temperature, etc.) as
needed to maintain the desired conditions for the experiment.
6. Begin baseline data collection and record the muscle contraction
force and rate.
7. Slowly decrease the temperature of the bath and continue to
record the muscle contraction force and rate.
8. Repeat steps 6-7 for different temperatures.
9. After the experiment, carefully remove the muscle from the bath
and dispose of the used solution according to established
protocols.
Result:
 The results of the experiment should show a decrease in the

muscle contraction force and rate as the temperature deviates
from the optimal range.
 The experiment should also show that there is an optimal
temperature range for muscle function and contractile activity
decreases as temperature deviates from this range.
Page | 45
STUDENT ORGAN BATH
 The result can be analyzed using graphs and statistical analysis.
Conclusion:
The student organ bath is a useful tool for studying the physiology
of isolated organs. It allows researchers to conduct experiments on
isolated organs in a controlled environment, which can provide
valuable insights into the physiological mechanisms of the organ in
question. The SOP for a student organ bath provides a clear and
detailed guide on how to set up and conduct an experiment, as
well as how to analyze and interpret the data collected.
The experiments described above are examples of how student

organ bath can be used to study the physiology of frog heart and
skeletal muscle. The first experiment demonstrated how
epinephrine affects the contractile activity of the heart. The second
experiment demonstrated how temperature affects the contractile
activity of skeletal muscle.
In conclusion, the student organ bath is a valuable tool for the

study of physiology of isolated organs. The experiments described
above demonstrate how the student organ bath can be used to
study the physiology of frog heart and skeletal muscle. It is
important to conduct these experiments under the guidance of an
experienced researcher and proper safety precautions.
Page | 46

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STUDENT ORGAN BATH

After a suitable equilibration period, the researcher can begin

ADInstruments supply a range of organ baths and transducers for

Vascular, e.g. arterial/aortic rings

Vas Deferens (secretory duct of the testicle)

Toad Abdominus Rectus

Chick Biventer Cervis

Tissues are usually prepared in a petri-dish containing physiological

For tissue ring preparations (i.e. arterial rings), a custom tissue-holder is

ADInstruments supply compact and robust, yet versatile organ baths

Schematic diagram of a 2 chamber Panlab organ bath.

ADInstruments/Panlab Organ Bath Systems

 An 8 or 16 channel PowerLab data acquisition system

All of these components may be purchased separately, if required.

ADInstruments supply a single chamber organ bath by Panlab; however,

ADInstruments have a Thermoregulated Water Pump (Model #:

If an external heating system is required, then an "immersion

Electrodes are required if using a stimulator. The platinum electrodes are

Type & Description Platinum Stainless Steel Single Ring (6.5, 8 or 10

Specify: /ID = 6.5, 8 or 10 mm

 Available in 2, 4, 8 chamber configuration

Radnoti Organ Baths:

Radnoti organ baths include a single water-jacketed 2 L reservoir for the

Radnoti Thermal Circulator Pump (170051B-V)

 Reservoir Volume: 17 Liters (4.5 Gallons)

Each ADInstruments/Radnoti Tissue-Organ Bath System Includes:

These systems require manual refilling and emptying of tissue chamber.

Radnoti also provides several combination tissue support

 Tissue strips/spirals: Straight (160152-11), zig-zag (160152-12) or

Modular components enable tissue chambers to be easily removed and

Each chamber is individually water-jacketed ensuring uniform circulation

Reservoir is water-jacketed to maintain solution temperature

Overflow port allows constant recirculation of perfusate to limit the

Incorporates Quick Disconnect fittings

Isometric transducers measure the change in force (in grams [g] or

Stimulation of tissues may be achieved by a number of methods

A field stimulating electrode is available from Radnoti. It is constructed of

Parts of Student organ bath

1. Cleaning the organ bath

4. Kymograph paper should

10. Attach the aeration tube

1. Bath chamber- Bath

two holding place for organ

2. Organ Tube- Organ tube is

2. Attaching or arranging all

7. Wash the organ tube with

13. Attach the lever with

change the PSS. (It is for same

1. Bath chamber- Bath

Single unit and double unit. In

various dose of same drug or

Out of which one is for

4. Kymograph paper should

10. Attach the aeration tube

1. Bath chamber- Bath

two holding place for organ

2. Organ Tube- Organ tube is

Introduction- Student organ

bath or simply organ bath is

organ is isolated it should

Introduction- Student organ

organ is isolated it should

Selection of tissue donors