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United States Government Work, uncopyrighted, public-domain,
DRAFT COPY ONLY. This document does not necessarily represent the views
or policies of any United States Government agency.
This document is provided "as is", without warranty of any kind,
express or implied, including but not limited to the warranties
of merchantability, fitness for a particular purpose and
non-infringement. In no event shall the authors be liable
for any claim, damages or other liability, whether in an
action of contract, tort or otherwise, arising from, out of,
or in connection with the document or the use or other dealings
made with the document.
See also: Frozen sections policies:
24,
25,
27,
41,
86.
PRINCIPLE OF THE TEST.
Frozen sections are available for examination promptly
after being received in the pathology department.
SPECIMEN REQUIRED.
All fresh human tissue excised at surgery, along with a filled-out
Tissue Examination Form (U. S. Standard Form 515, USSF515).
REAGENTS, INSTRUMENTATION.
10% buffered formalin fixative.
70% alcohol.
Lithium carbonate (LiCO3) solution.
Hematoxylin solution.
Eosin solution.
Histosol (Xylene substitute).
OCT Compound.
Cryostat.
Frozen section solutions for hematoxylin and eosin staining.
STEP-BY-STEP DESCRIPTION.
1. Dip slide 10 seconds into 10% buffered formalin fixative.
Allow to drain.
2. Dip five times into 70% alcohol.
3. Rinse in running tap water until water sheets off slide.
4. Place into hematoxylin for one minute.
While waiting for stain, prepare labels for slides.
5. Rinse in water until most of the excess stain is removed.
6. Dip four times in LiC03 to blue the section.
7. Rinse in tap water for 10 seconds.
8. Dip for 30 seconds in eosin, allow to drain.
9. Dehydrate through three absolute alcohols (several dips each,
draining after each).
10. Clear by dipping several times in 2 changes of histosol.
11. Coverslip.
REFERENCE.
1. Prophet EB, Mills B, Arrington JB, Sobin LH.
Laboratory Methods in Histotechnology, pp. 67-70.
1992: Armed Forces Institute of Pathology,
Washington, DC. 20306-6000.
ISBN: 1-881041-00-X 1992.