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PRINCIPLE OF THE TEST.
To stain paraffinized tissue sections for cytoplasmic granules, 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.
Fontana Masson Stain.
Silver nitrate solution (Fontana).
Ammonium hydroxide.
Gold chloride solution 1%.
Sodium thiosulfate, 5%.
Sodium thiosulfate.
Nuclear fast red solution.
Nuclear fast red powder.
Aluminum sulfate solution, 5%.
Thymol.
Xylene substitute.
Absolute Flex.
95% Flex.
Permount.
STEP-BY-STEP DESCRIPTION.
Fontana Masson Stain for Argentaffin Granules.
SOLUTIONS:
SILVER NITRATE SOLUTION (FONTANA).
Dissolve 10 gms of silver nitrate in 100 cc water. To 95 cc of this
solution add ammonium hydroxide until a clear solution with no precipitate
is obtained. Add drop by drop enough of the remaining 5 cc of silver
nitrate solution to cause the clear solution to become slightly
cloudy. Let stand overnight before using. When ready to use, dilute
each 25 cc of the silver solution with 75 cc of distilled water and
filter.
GOLD CHLORIDE SOLUTION.
Gold Chloride Solution 1% ... 10 cc.
Distilled Water ............. 40 cc.
SODIUM THIOSULFATE.
Sodium thiosulfate ........ 5 gm.
Distilled water ........... 100 cc.
NUCLEAR FAST RED.
Dissolve 1 gm of nuclear fast red
in a 5% solution of aluminum sulfate,
with the aid of heat.
Cool, filter, and add a grain of thymol,
as a preservative.
STAINING PROCEDURE:
1. Xylene substitute, absolute Flex, 95% Flex, to distilled water.
2. Immerse slides in silver nitrate soluLion for 1 hr. at 56øC.
3. Rinse in distilled water.
4. Immerse in gold chloride for 10 minutes.
5. Rinse in distilled water.
6. Place in sodium thiosulfate solution for 5 minule6.
7. Rinse in distilled water.
8. Counterstain with eosin or nuclear fast red, if desired. Rinse
in distilled water.
9. Flex 95, absolute, xylene substitute.
10. Mount in Permount.
REFERENCES.
1. Bancroft JD.
Cytoplasmic Granules, Organelles, and Neuroendocrine.
Chapter 17, pp. 345-370.
In: Bancroft JD, Gamble M.
Theory and Practice of Histological Techniques. Fifth Edition.
Edinburgh: Churchill Livingstone. 2002;:345-370.
ISBN 0-443-06435-0, 796 pages.
2. Altmann R.
Die Elementarorganismen und ihre Beziehungen zu den Zellen.
[German: Elementary organisms and their relationships to cells].
Leipzig: Veit. 1894;:.
3. Grimelius L.
A silver nitrate stain for alpha 2 cells in human pancreatic islets.
Acta Societatis Medica Uppsala. 1968;73:243.
4. Ham HW.
Histology. Sixth Edition.
Philadelphia: J.B. Lippincott. 1969;:.
5. Hotchkiss RD.
A microchemical reaction resulting in the staining of polysaccharide
structures in fixed tissue preparations.
Arch Biochem. 1948;16:131.
6. Singh I.
A modification of the Masson-Hamperl method for staining
argentaffin cells.
Anat Anz. 1964;115:.
7. Zollinger RM, Ellison EH.
Primary peptic ulceration of the jejunum: Association with islet
cell tumors of the pancreas.
Ann Surg. 1955;12:709-723.
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.