Freezing Microtome and Cryostat

MEDICAL INSTRUMENTS-II
Ghulam Rasool
Lecturer
Department of Allied Health
Sciences, SMC,UOS
Freezing microtome
 Simple type of freezing microtome is clamped to the
edge of a bench and is connected to a cylinder of
carbon dioxide by means of a specially strengthened
flexible metal tube.
 It consist of a radial arm attached to a central pivot.
On this arm, two clamps hold a wedge profile
microtome knife mounted with a simple block
holder, with a cutting edge inclined in a horizontal
plane.
Working principle
 The object is mounted on a block holder (chuck) known as
freezing stage, with a centrally advancing screw. The block
holder is perforated and attached to a feed pipe carrying
carbon dioxide gas, which is sprayed on to the tissue for
freezing. The knife moves over the block around a horizontal
axis when once the tissue hardens.
 Thermoelectric cooling device units may be used in place of
carbon dioxide gas to freeze the tissue and cool the knife.
The cooling produced by thermoelectric units depends upon
the flow of direct current, which may be regulated by means
of power packs. In this stage, temperature can be reduced
from ambient to -36C in 60 seconds, but the optimum
cutting temperature for the tissue is usually about -20C.
Freezing microtome
Advantages:
 Freezing microtome is used in the demonstration of fat
 Ideal for brain sectioning.
 Simple design with no complex moving parts.
 Can be of diagnostic use when affordability of cryostat is not
possible.
 Better demonstration of soluble and diffusible substances.
 Easy to operate and maintain.
Disadvantages:
 The sliding knife tends to jump on striking hard tissue.
 Difficult to sharpen the long knife.
 No serial sectioning possible.
 Sections less than 8 u cannot be cut under the best of
conditions.
CRYOSTAT
 The first cryostat was introduced in 1959.
 (‘cryo’ meaning cold and ‘state’ meaning stationary)
 Cryostat is a refrigerated cabinet in which a microtome is
housed. The microtome used is usually a rotary type, but may
of sliding type or even rocking type and is rust proof. The
microtome is mounted in a stainless steel cabinet at an angle
of 45 degree. It has an antifogging air circulating system, a
drain for defrosting and a shelf for four to six metal block
holders. Cabinet temperature is -5 to -30 ^C. All microtome
control operation are outside the cabinet.
Harris International Microtome is most commonly used.
This unit operates on the ‘open top-cold box’ principle.
Working principle
 To create a cold atmosphere
around tissue block holder and
microtome by means of a special
refrigerator type compressor
capable of taking temperature
below -30 to -50 C. The reason for
freezing the tissue is to provide a
hardened matrix for sectioning
the tissue and at the same time,
preserving biochemical or
immunological properties of a cell
or tissue. The coolant used is
usually Freon 22.
 Advantages
 Used extensively for rapid diagnosis, fat stains and enzyme
histo-chemistry in neurological applications as well as in
fluorescence microscopy.
 Both the knife and tissue are maintained at same low
temperature.
 Capable of slicing sections as thin as 1 um
 Serial sectioning is possible.
 Automatic defrosting and sterilization
 Antifogging air circulatory system.
 Disadvantages
 Constant supervision and maintenance of temperature is
required.
 The whole instrument should be kept in an air-conditioned
room to prevent excessive cryostat compressor functioning.
 Lubricants of special type with a low congealing point have to
be used. This prevents the lubricants solidifying at a cooler
temperature within the chamber.
 Freeze artifacts seen as holes in the tissues.
 If the temperature is too low, the tissue become hard and
crumbles, and becomes difficult to cut.
 Difficulty in sectioning fixed tissue.
 High cost of the instrument.
 A Microtome is used to cut very thin sections at room
temperature, on the other hand a Cryostat is used to cut
frozen sections at sub zero temperatures (generally -30
deg C).
 A cryostat is used in situations where rapid analysis of

tissues is required. The water rich tissue is frozen on a
quick freezing shelf inside the cryostat, this makes it very
hard and it is then ready to be cut into thin sections in a
microtome, also placed inside the cryostat chamber.
 On the other hand to be cut in a simple microtome, the
tissue needs to be first dehydrated and fixed in
paraffin before it can
be sectioned. It is a long procedure compared to quick
sectioning in a cryostat.
 The quality of sections cut in a cryostat is inferior

compared to those cut in a microtome because
dehydrated and paraffin embedded tissues give better
sections but when it comes to quick sectioning,
Cryostat is the choice.

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MEDICAL INSTRUMENTS-II

 A cryostat is used in situations where rapid analysis of

 The quality of sections cut in a cryostat is inferior

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