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United States Government Work, uncopyrighted, public-domain,
DRAFT COPY ONLY. This document does not necessarily represent the views
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PRINCIPLE OF THE TEST.
To predigest paraffinized tissue sections, with hyaluronidase, 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.
0.1 M Potassium phosphate, monobasic, solution.
Potassium phosphate, monobasic, powder.
0.1 M Sodium phosphate, dibasic, solution.
Sodium phosphate, dibasic, powder.
Hyaluronidase digestion solution.
Testicular hyaluronidase.
Buffer solution.
Controls: Testis or skin slides.
STEP-BY-STEP DESCRIPTION.
0.1M POTASSIUM PHOSPHATE, MONOBASIC
Potassium phosphate, monobasic.......................... 13.61 gm
Distilled water........................................ 1000.0 ml
0.1 M SODIUM PHOSPHATE, DIBASIC
Sodium phosphate, dibasic............................... 14.20 gm
Distilled water........................................ 1000.0 ml
BUFFER SOLUTION
Potassium phosphate, monobasic, 0.1 M.................... 94.0 ml
Sodium phosphate, dibasic, 0.1 M........................... 6.0 ml
HYALURONIDASE DIGESTION SOLUTION
Testicular hyaluronidase*................................ O.05 gm
Buffer solution.......................................... 100 ml
DIGESTION PROCEDURE. Use control slides.
1. Deparaffinize and hydrate duplicate sections to distilled water.
2. Digest slides in hyaluronidase solution
at 37o C for 1 hour.
Incubate undigested slides in buffer solution
at 37o C for 1 hour.
3. Wash both sets of slides in running water for 5 minutes.
4. Stain as desired.
CONTROL: Testis and skin
RESULTS: Staining attributable to hyaluronic acid, chondroitin 4
sulfate or chondroitin 6 sulfate is selectively eliminated.
*Cal Bio Chemical Co., Los Angeles, California
REFERENCES.
1. Totty BA.
Mucins. Chapter 10, pp. 163-200.
In: Bancroft JD, Gamble M.
Theory and Practice of Histological Techniques. Fifth Edition.
Edinburgh: Churchill Livingstone. 2002;:163-200.
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.