CRYOTOMY
The History of Freezing Techniques
Frozen section techniques were developed in an attempt to eliminate artifacts in tissue specimens caused by exposure to normal processing procedures. Boyle, who sectioned frozen eyes, accomplished the first recorded frozen section in 1663. The initial freezing techniques were accomplished by outdoor freezing in cold weather or with volatile chemicals such as ether and rigolene. As the techniques advanced, freezing was accomplished by using carbon dioxide, Freon, isopentane, liquid nitrogen slush and other cryogens. Thermoelectric cooling is now also an accepted method for routine cryotomy for light microscopic evaluation.
|
Catalog # |
Product Name |
|
Cryo-STAT |
Freezing Compound, 4 oz
bottle |
|
SL202 |
Statfreeze 3™ Freezespray |
|
SF-1 |
Stat-Fix, Alcoholic Formalin Fixative, 1 gallon
bottle |
|
00960-16 |
Acetone, 16 oz. bottle |
|
SL97-1 |
Gill
1 Hematoxylin, 1 gallon bottle |
|
SL97-16 |
Gill
1 Hematoxylin, 16 oz bottle |
|
SL97-2.5 |
Gill
1 Hematoxylin, 2.5 gallon cubitainer |
|
SL94-1 |
Gill
2 Hematoxylin, 1 gallon bottle |
|
SL94-16 |
Gill
2 Hematoxylin, 16 oz bottle |
|
SL94-2.5 |
Gill
2 Hematoxylin, 2.5 gallon cubitainer |
|
SL95-1 |
Gill
3 Hematoxylin,1 gallon bottle |
|
SL95-16 |
Gill
3 Hematoxylin, 16 oz bottle |
|
SL95-2.5 |
Gill
3 Hematoxylin, 2.5 gallon cubitainer |
|
SL90-1 |
Harris Hematoxylin, 16 oz.
bottle |
|
SL90-16 |
Harris Hematoxylin, 1 gallon
bottle |
|
SL90-2.5 |
Harris Hematoxylin, 2.5 gallon
cubitainer |
|
SL93-1 |
Treosin, 1 gallon bottle |
|
SL93-16 |
Treosin, 16 oz. bottle |
|
SL93-2.5 |
Treosin, 2.5 gallon
cubitainer |
|
SL98-1 |
Eosin, 1 gallon bottle |
|
SL98-16 |
Eosin, 16 oz bottle |
|
SL98-2.5 |
Eosin, 2.5 gallon cubitainer |
|
SL30-1 |
Deionized water, 1 gallon
bottle |
|
SL30-2.5 |
Deionized water, 2.5 gallon
container |
|
00100-16 |
Acetic Acid, Glacial, 16 oz
bottle |
|
33780-16 |
Hydrochloric Acid, 16 oz
bottle |
|
SL99-1 |
Bluing Solution, Scott’s formula, 1 gallon
bottle |
|
SL99-16 |
Bluing Solution, Scott’s formula, 16 oz
bottle |
|
SL99-2.5 |
Bluing Solution, Scott’s formula, 2.5 gallon
cubitainer |
|
05870-16 |
Ammonium Hydroxide, 16 oz
bottle |
|
6900-1 |
Reagent Alcohol, 200 proof, 1 gallon
bottle |
|
9500-1 |
Reagent Alcohol, 190 proof, 1 gallon
bottle |
|
69100-1 |
ProMet™ Alcohol blend, 200 proof, Reagent grade, 1 gallon
bottle |
|
6995-1 |
ProMet™ Alcohol blend, 190 proof, Reagent grade, 1 gallon
bottle |
|
8400-1 |
Xylene, Lab Grade, 1 gallon
bottle |
|
7400-1 |
XS-3™
Xylene substitute, 1 gallon bottle |
|
6400-1 |
Citra
Clear Xylene substitute |
|
SL80 |
Acrymount™ Mounting media |
|
SL180 |
M-1™
Mounting media |
Suggestions for
Frozen Section Staining Procedures
The following procedures are recommended for frozen sections cut at 4µm
to 6µm, mounted on glass slides, fixed using the method of choice and rinsed in
tap water to remove all traces of the
fixative.
1. Stain in Gill 3 Hematoxylin for 15 seconds to 1 minute
2. Rinse in several changes of water
3. Place in Bluing Solution for several seconds
4. Rinse in several changes of water
5. Begin dehydration in 80% alcohol until solution runs smoothly from slide
6. Rinse in 95% alcohol until solution runs smoothly from slide
7. Counterstain in 1% Eosin Y or Treosin for 15 to 20 dips (or to desired intensity)
8. Rinse briefly in 95% alcohol
9. Complete dehydration in 2 or 3 changes of absolute alcohol
10. Clear in 3 changes of xylene or xylene substitute until solution runs smoothly from slide
11. Mount with a synthetic resin
RESULTS
USING 1% EOSIN Y
Nuclei – blue to
blue/black
Cytoplasm, collagen – shades of pink
RESULTS
USING TREOSIN
Nuclei – blue to
blue/black
Collagen and skeletal muscle – bright red
Smooth muscle – pink/red and distinct from collagen
RBC and fibrin – bright red/orange
Rapid Regressive Staining using Harris Hematoxylin (CAT# SL90 ) and T reosin(CAT# SL93) or Eosin (CAT# SL98)
1. Stain in Harris Hematoxylin for 1 minute or until desired intensity is achieved
2. Rinse in several changes of water
3. Differentiate quickly in acid alcohol or a dilute aqueous acid solution
4. Rinse in several changes of water
5. Place in Bluing Solution for several seconds
6. Rinse in several changes of water
7. Begin dehydration in 80% alcohol until solution runs smoothly from slide
8. Rinse in 95% alcohol until solution runs smoothly from slide
9. Counterstain in 1% Eosin Y or Treosin for 15 to 20 dips (or to desired intensity)
10. Rinse briefly in 95% alcohol
11. Complete dehydration in 2 or 3 changes of absolute alcohol
12. Clear in 3 changes of xylene or xylene substitute until solution runs smoothly from slide
13. Mount with a synthetic resin
RESULTS
USING 1% EOSIN Y
Nuclei – blue to
blue/black
Cytoplasm, collagen and erythrocytes – shades of pink
RESULTS
USING TREOSIN
Nuclei – blue to
blue/black
Collagen and skeletal muscle – bright red
Smooth muscle – pink/red and distinct from collagen
RBC and fibrin – bright red/orange