MODIFICATION OF MOWRY'S 1958 COLLOIDAL IRON STAIN
FOR ACID MUCOPOLYSACCHARIDES
DRAFT COPY ONLY.
(Procedure 71).
http://www.netautopsy.org/axsop/axsop071.htm
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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 stain paraffinized tissue sections for acid mucopolysaccharides,
required in the diagnosis of certain diseases encountered
in anatomic pathology, especially granuloma annulare of skin.
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.
10% buffered neutral formalin.
95% alcohol.
Paraffin sections at 6 microns.
29% Ferric chloride solution.
29% Ferric chloride powder.
Mueller's colloidal iron oxide solution (stock).
29% ferric chloride solution (USP XI).
Colloidal iron solution (working).
Glacial acetic acid.
12% Acetic acid solution.
5% Potassium ferrocyanide solution.
Potassium ferrocyanide powder.
5% Hydrochloric acid solution.
Hydrochloric acid, concentrated.
STEP-BY-STEP DESCRIPTION.
FIXATION: 10% buffered neutral formalin or 95% alcohol. Bichromate
fixatives should be avoided.
TECHNIQUE: Cut paraffin sections at 6 microns.
SOLUTIONS: 29% FERRIC CHLORIDE SOLUTION
Ferric chloride........................................ 29.0 gm
Distilled Water........................................ 100.0 ml
MUELLER'S COLLOIDAL IRON OXIDE SOLUTION (STOCK)
Bring 250 ml of distillod water to a boil. While the water is
still boiling, pour 4.4 ml of 29% ferric chloride solution (USP XI)
and stir. When the solution has turned dark red, remove from heat
and allow to cool. Label: Stock Mueller's Colloidal Iron. This reagent
is stable for many months. In the staining procedures, the stock solution
of colloidal iron is diluted just before use as follows:
COLLOIDAL IRON SOLUTION (WORKING)
Mueller's colloidal iron (stock)........................ 20.0 ml
Distilled water......................................... 15.0 ml
Glacial acetic acid...................................... 5.0 ml
Mix just before use.
12% ACETIC ACID SOLUTION
Glacial acetic acid..................................... 12.0 ml
Distilled water......................................... 88.0 ml
5% POTASSIUM FERROCYANIDE SOLUTION
Potassium ferrocyanide................................... 5.0 gm
Distilled water.......................................... 100.0 ml
5% HYDROCHLORIC ACID SOLUTION
Hydrochloric acid, concentrated.......................... 5.0 ml
Distilled water......................................... 95.0 ml
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. Mowry RW.
Observations on the use of sulphuric ether for the sulphation
of hydroxyl groups in tissue sections.
J Histochem Cytochem. 1956;4:407.
3. Mowry RW.
Alcian blue techniques for the histochemical study
of acidic carbohydrates.
J Histochem Cytochem. 1958;6:82.
4. McManus JFA, Mowry RW
Staining Methods, Histologic and Histochemical.
London: Harper & Row. 1964;:268.
5. Mowry RW
Aldehyde fuchsin staining, dir4ect or after oxidation: problems
and remedies with special reference to human pancreatic B cells, pituitaries,
and elastic fibres.
Stain Technology. 1978;53:141-154.
6. 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.
7. 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.