HISTOPATHOLOGY

AUTOMATION
JMF
 Automation
Automationin
inhistopathology
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Some automated machines
1. Microtome
2. Tissue embeding machines
3. Coversliper
4. Sliding microtome
5. 2000 processor
6. Cryostat
7. Paraffin dispensor
8. Tissue floter waterbath
9. Setup slide warmer
10. Rotatary microtome
11. Gusto high speed centrifuge
A complete modern techniques
view in histopathology lab
 Benefits of automation in histopathology labs
• It reduces the time and increases the speed of work
flow .
• Histopathology workflow. Biopsies and larger
specimens can be processed at the same time.
• “By changing to an automated method of working in
which specimens are continuously loaded for
processing and embedding, the peaks and troughs can
be eliminated and replaced by a steady workflow
which can be handled by fewer people throughout the
day”
Tissue tek an automated machine in histolab that is totally automated perfom all functions in flow with constant speed
 A. AUTOMATED TISSUE
PROCESSING
 The basic principle for tissue processing requires the exchange of
fluids using a series of solutions for a predetermined length of time
in a controlled environment

2 types
1. Tissue Transfer type processor (Linear / Carousal)
2. Self-contained fluid exchange systems
Transports tissue blocks
Carousal Type Processor
(Tissue Transfer)
in baskets

Through a series of reagents housed

in stationary containers

Submerged specimen’s length of time in each reagent

container should be
electronically controlled

Vertical oscillation or the rotatory

movement of the tissue basket provided
the
agitation
1.
A microprocessor used
in this instrumentt
Self-contained fluid
exchange systems
Tissue Cassettes are loaded into a
retort chamber where they remain
stationary
throughout the process

Reagents & melted paraffin wax moved sequentially into & out of
the
retort chamber by using vacuum & pressure

Agitation is achieved by Tidal

action

Each step could be

customized by controlling
time, temperature, or
pressure/vacuum
2.
 B. Microwave ovens or
processors
 Microwave ovens specially designed for tissue
processing are now common in a recent tissue
processing technique.
 This technique is 1st used by Boon and Kok in 1985.
 It shortens the processing time from hours to minutes.
 The processing time depends on the thickness &
density of the specimen.
 Comparison between tissue
processers
Manual TP Automated TP Microwave TP
Reliability Reliability Less reliable More reliable More reliable
as human error may
occur
Time consumption Time consumption Have customised Shortens the
schedules for TP processing time
Cost Inexpensive Costly More costly
Chemicals like Xylene Employs noxious Vacuum & heat can be Not used so eliminates
& formalin chemicals used toxic fumes &
carcinogens
Graded concentration Monitored Monitored Not required
of solution
Shrinkage of tissue Evaluated Less evaluated Lesser evaluated
Rapid Processors with multi-
sectioned retorts
This processor uses :
Microwave technology
Vacuum infiltration
‘Molecular-friendly’ proprietary
reagents
Microwaves & agitation are used to
accelerate the diffusion of solvents in tissue.
The microwave technology utilized, operates
at a continuous low power instead of
pulsing high levels of microwave energy.
The retort chamber is cylindrical.
Microwaves circle around the cavity, taking
advantage of the physical principle of the
‘whispering chamber’ effect that eliminates
hot &cold spots.
 TISSUE MICROARRAY
Tissue microarray (TMA) was developed as a
method to evaluate numerous samples of tissue in
a short period.
 The automated arrayer is easy to use and
includes a specimen tracking software system.
The instrument marks, edits, and saves punch
coordinates using an on-screen display and
software tools.
Automated tissue arrayer ATA27.
 AUTOMATED EMBEDDING
STATION
EMBEDDING - the tissue is
surrounded in a molten medium
by using a mould. Subsequently
this medium is solidified to make a
block for cutting thin section of
tissue.
 CRYOSTAT

The cryostat is the instrument that has the

arrangement to freeze the tissue and also to cut the
frozen tissue for microscopic section.
First cryostat was introduced in 1954.
The cryostat is a machine in which a slicer called a
microtome is placed in a freezer. The temperature
may be regulated between-10ºc to -40ºc.
 PARTS

 Cryocabinet (-5 to -30⁰C) – Most tissues section properly between −15 °C

and −25 °C.
 Microtome (Rotary/ Sliding/Rocking) of any type but preferably
rustproof, which is enclosed and operated within a deep freeze cabinet.
 Knife or blade: low- or high-profile disposable blades /profile C steel
blade. The angle of the knife is kept in between 5° and 7°.
 Antiroll plate : just in front of the knife - prevents the rolling of the cut
tissue.
 Specimen Holder: Small, round, metal CHUCKS
 Fluorescent light lamp
 Electronic control panel
Principle:

When the tissue is frozen, the interstitial

water in the tissue turns to ice, and in this
state the tissue is firm with the ice acting
as the embedding medium.
 Once the
The specimens are specimen is cut
Once frozen(-24⁰
mounted on a to a satisfactory
C) the specimen is
cutting medium on quality, it is
mounted on the
a metal chuck and mounted on a
microtome.
frozen to a cutting warm dry glass
temperature slide, where it
melts and dries.
 INDICATIONS

 Rapid production of sections for intra-operative diagnosis.

 Immunofluorescence study.
 Diagnostic and research enzyme histochemistry for labile
enzymes.
 Diagnostic and research non-enzyme histochemistry, e.g.
Lipids and some carbohydrates
 Silver demonstration methods, particularly in
neuropathology
  
WATER BATH

The temperature of the water bath is

usually controlled automatically by a
thermostat.
The temperature of water in the water bath
should be 10 °c below the melting point of
the embedded paraffin wax and is usually
kept in 40–50 °C.
 DRYING
The temperature should be at the melting
point of the paraffin.
Automated stainers have drying ovens as
part of the instrumentation.
 AUTOSTAINER
HIGH THROUGHPUT STAINER
COMPACT STAINER
 DRYING
Types
1 Dips slides into the stains
2 Applies stain to the slide
 DRYING……
Linear design
 1 slide at a time
 Slides are clipped to slide holders, which are attached to
carrier mechanism.
Batch design
 Linear / Carousal
 Multiple slides
 Slide racks are moved through baths of staining solution
 Capillary Gap Stainer
Force or draw the stain between the
specimen slide and another surface.

Centrifugal Stainer
Spray the stain as the slide rotate past
the spray nozzle in a spinning chamber
 Flat Stainer
Drops the stain onto the
slide, while the slide lies flat
within the stainer
 COVER SLIPPING

Suctioning mechanism for picking up a

coverglass from a stack of coverglasses.
Exert a force onto the coverglass to insure that
it is released from the selecting device and
placed onto the slide.
After placement of the coverglass onto the slide,
capillary action pushes air bubbles out from
underneath the coverglass.
 LABELING
Imprints
Easier to locate.
Alphanumeric characters, barcodes or
logos
THE END.. <3