Frozen Section
SurSuu
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Suresh Kumar Mahaseth
MLT ( Lecturer)
[email protected]
Introduction:-
The frozen section is a pathological laboratory procedure to perform rapid microscopic
analysis of a specimen .
It is used for rapid examination , technically that is known as cryosection .
Frozen section technique is a valuable tool used to rapidly prepare slides from tissue for
microscopic interpretation.
A frozen section examination is a specific type of biopsy procedure that allows a surgeon to
establish a rapid diagnosis of a suspicious mass during surgery. The technical name for this
procedure is cryosection.
Frozen section is a specimen of tissue that has been quick-frozen, cut by microtome, and
stained immediately for rapid diagnosis of possible malignant lesions.
Procedure :
For the frozen section cryostat microtome is used in this microtome fit a freezer
in side microtome is present .
The microtome capable of slicing section as thin as 1 micron .
The surgical specimen frozen rapidly about -20 to -300 c & the specimen is
embedded in gel like polyethylene glycol .
At this temp. , most tissue become rock hard .
Preparation of sections:-
Place a drop of water in the centre of a precooled block holder.
Place the tissue to be sectioned in the drop of water.
Rapidly freeze the tissue – this is done by either standing the block
holder in a bath of alcohol or acetone containing dry ice or by
placing the block holder in the special freezing attachment &
exposing the tissue to carbon dioxide gas.
when the tissue is frozen , position the holder in the microtome .
Cutting of section :-
Adjust the tissue correctly to the microtome .
Trim the tissue with the aid of the remote control outside the chamber .
About 15-30 min. before cutting starts, place the knife in position in order
to attain the correct cutting temp.
Set the section thickness control ( Usually at 5- 10 micron ) & the
automatic advance mechanism .
Allow the temp. in the chamber to equilibrate by closing the cabinet for
about 2-3 min.
Mounting of frozen section :-
The frozen section are not rigid like paraffin section , so the mounting done
carefully use warm knife for lifted.( Paint brush can be use).
Place the section in D/W.
Dry the section before staining .
For fix tissue ( Section) albumin or glycerol can use as a adhesive .
Staining of frozen section :- They are two method
1. Haematoxylin & Eosin
2. Polychrone methylene blue
1. Haematoxylin & Eosin method:-
i. Fix the air – dried section in pure acetone for 20 seconds.
ii. Place the section in absolute alcohol for ½ min. to 1 min.
iii. Place in 95% ethanol for 5 sec.
iv. Place in D/W until no longer greasy or cloudy .
v. Place in Harris's haematoxylin for 1-2 min .
vi. Place in D/W with agitation for 5-10 secs.
VII. Dip in 0.5 % sodium borate until blue.
VIII. Place in 70 % ethanol for 5 seconds .
XI . Place in 1% alcoholic eosin for 5-20 sec.
X . Wash in water 10- 30 sec .
XI . Dehydrate through graded alcohols .
XII. Clear in xylene & mount in DPX .
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