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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 diastase, 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.
Phosphate buffer solution, pH 6.0.
Sodium Chloride.
Sodium phosphate, dibasic.
Sodium phosphate, monobasic.
Diastase digestion solution.
Diastase of malt.
STEP-BY-STEP DESCRIPTION.
PHOSPHATE BUFFER SOLUTION, pH 6.0.
Sodium Chloride................................ 8.0 gm
Sodium phosphate, dibasic...................... 0.282 gm
Sodium phosphate, monobasic.................... 1.97 gm
Distilled water................................ 100.0 gm
DIASTASE DIGESTION SOLUTION.
Diastase of malt................................. 0.1 gm
Phosphate buffer solution, pH 6.0................ 100.0 ml
Mix just before use.
DIGESTION PROCEDURE. Use control slide.
1. Deparaffinize and hydrate to distilled water.
2. Place slides in preheated diastase solution at 37o C for
1 hour.
3. Wash in running water for 5 minutes.
4. Stain with periodic acid Schiff or other appropriate
procedure.
RESULTS: Staining attributable to glycogen is selectively
eliminated.
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. Saunders AM, Rosan RC.
Histochemical nomenclature of carbohydrate-rich components.
J Histochem Cytochem. 1966 Nov;14(11):869.
PMID: 5971455.
PubMed Entry
3. Rosan RC, Saunders AM.
On the histochemical nomenclature for sialomucins.
J Histochem Cytochem. 1965 Jul-Aug;13(6):518.
PMID: 4221537 [PubMed - indexed for MEDLINE]
PubMed Entry
4. Saunders AM.
Histochemical identification of acid mucopolysaccharides
with acridine orange.
J Histochem Cytochem. 1964 Mar;12:164-170.
PMID: 14187320.
PubMed Entry
5. 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.
6. 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.