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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 iron, required in the diagnosis
of certain diseases encountered in anatomic pathology,
especially in bone marrow biopsies.
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 ferrocyanide, 5%
Hydrochloric acid, 5%
Van Gieson solution.
Glacial acetic acid solution.
Working colloidal iron solution.
Hydrochloric acid-potassium ferrocyanide solution.
95% alcohol.
Absolute alcohol,
Xylene or xylene-substitute.
Permount or Histoclad.
STEP-BY-STEP DESCRIPTION.
Potassium ferrocyanide, 5%............................. 50.0 ml
Hydrochloric acid, 5%................................. 50.0 ml
Mix just before use.
VAN GIESON SOLUTION
STAINING PROCEDURE. Use control slide.
1. Deparaffinize and hydrate to distilled water.
2. Rinse in glacial acetic acid solution for 3 minutes.
3. Working colloidal iron solution for 1 hour.
4. Rinse in glacial acetic acid solution for four changes,
three minutes each.
5. Hydrochloric acid-potassium ferrocyanide solution for 20 minutes.
6. Wash in tap water for 5 minutes.
7. Rinse in distilled water.
8. Van Gieson's solution for 5 to 7 minutes.
9. Without washing, pass quickly to 95% alcohol, absolute alcohol,
and clear in xylene, two changes each.
10. Mount with Permount or Histoclad.
RESULTS:
Acidic mucopolysaccharides and acidic epithelial mucins
are colored deep (Prussian) blue.
Background depending on counterstain used.
REFERENCES.
1. Churukian CJ.
Pigments and Minerals. Chapter 13, pp. 243-268.
In: Bancroft JD, Gamble M.
Theory and Practice of Histological Techniques. Fifth Edition.
Edinburgh: Churchill Livingstone. 2002;:243-268.
ISBN 0-443-06435-0, 796 pages.
2. Gomori G.
Microchemical demonstration of iron.
Am J Physiol. 1936;13:665.
3. Gomori G.
Silver impregnation of reticulum in paraffin sections.
Am J Physiol. 1937;13:993.
4. Gomori G.
A new histochemical test for glycogen and mucin.
Am J Clin Pathol. 1946;16:177.
5. Gomori G.
Aldehyde-fuchsin: a new stain for elastic tissue.
Am J Clin Pathol. 1950;20:665.
6. Gomori G.
Microscopic Histochemistry.
Chicago: Chicago University Press. 1952;:.
7. Virchow R.
Die pathologischen Pigmente. [German: Pathologic pigments].
Virchows Arch f patholog Anat u Physiol u f klin Medizin. 1847;:379.
8. 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.
9. 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.