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PRINCIPLE OF THE TEST.
To stain paraffinized tissue sections for reticulum, required in the
diagnosis of certain diseases encountered in anatomic pathology.
SPECIMEN REQUIRED.
All human tissue excised at surgery, outpatient clinics, and postmortems,
fresh or in fixative, along with a filled-out Tissue Examination Form
(U. S. Standard Form 515, USSF515).
REAGENTS, INSTRUMENTATION.
Potassium permanganate solution, 1%.
Metabisulfite solution, 1%.
Ferric ammonium sulfate, 2%.
Hortega's silver solution.
Pyridine solution, 1.5%.
Gelatin solution.
37-40% formalin.
STEP-BY-STEP DESCRIPTION.
Procedure:
1. Deparaffinize and hydrate in distilled water.
2. Potassium permanganate solution 1 minute.
3. Rinse in distilled water.
4. Metabisulfite 1 minute or until color disappears.
5. Rinse in distilled water.
6. Ferric ammonium sulfate for 1 minute.
7. Rinse in distilled water.
8. Hortega's silver solution for 5 to 30 minutes.
9. Rinse in distilled water.
10. Pyridine solution for 6 to 8 dips.
11. Rinse in distilled water.
12. Gelatin solution for 1 to 2 minutes.
13. If slides are not dark enough after your gelatin solution,
rinse in distilled water and place back into your Hortega's solution
a few more minutes. Rinse in distilled water and place in gelatin.
After repeating this process, slides are usually ready.
14. Dehydrate, clear, and mount.
REAGENTS:
POTASSIUM PERMANGANATE
1 gm of potassium permanganate to 100 ml of distilled water.
METABISULFITE.
1 gm of metabisulfite to 100 ml of distilled water.
FERRIC AMMONIUM SULFATE.
2 gm of ferric ammonium sulfate to 100 ml of distilled water.
PYRIDINE.
1.5 ml of pyridine to 100 ml of distilled water.
GELATIN.
.25% of gelatin solution with added 2 cc of 37-40 formalin.
HORTEGA'S SILVER SOLUTION.
To 40 cc of 10% silver nitrate add 120 cc of 5% sodium
carbonate. Then carefully add 28% ammonium hydroxide
drop by drop until the precipitate is dissolved (shake
or stir while adding ammonium). Add 440 cc of
distilled water to the solution you have just made.
Store in refrigerator.
REFERENCES.
1. Jones ML.
Connective Tissues and Stains. Chapter 9, pp. 139-162.
In: Bancroft JD, Gamble M.
Theory and Practice of Histological Techniques. Fifth Edition.
Edinburgh: Churchill Livingstone. 2002;:139-162.
ISBN 0-443-06435-0, 796 pages.
2. Prophet EB, Mills B, Arrington JB, Sobin LH, eds.
Laboratory Methods in Histotechnology.
Washington, DC: Armed Forces Institute of Pathology. 1992;:.
ISBN 1-881041-00-X, 278 pages.
3. Mikel UV, ed.
Advanced Laboratory Methods in Histology and Pathology.
Washington, DC: Armed Forces Institute of Pathology. 1994;:.
ISBN 1-881041-13-1, 254 pages.