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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 collagen fibers,
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.
Cut paraffin sections at 6 microns.
Weigert's Iron Hematoxylin Solution.
Van Gieson's Solution.
Acid fuchsin, 1% aqueous.
Picric acid, saturated aqueous.
95% alcohol.
Absolute alcohol (Flex).
Clearing.
STEP-BY-STEP DESCRIPTION.
FIXATION: Any well-fixed tissue.
TECHNIQUE: Cut paraffin sections at 6 microns.
SOLUTIONS: Weigert's Iron Hematoxylin Solution
Van Gieson's Solution
Acid fuchsin, 1% aqueous.................. 2.5ml
Picric acid, saturated aqueous........... 97.5ml
STAINING PROCEDURE
1. Deparaffinize and hydrate to distilled water.
2. Weigert's hematoxylin solution for 10 minutes.
3. Wash in distilled water.
4. Van Gieson's solution for 1 to 3 minutes.
5. Dehydrate in 95% alcohol, absolute alcohol (Flex) and clearing
(two changes each).
6. Mount.
CONTROL: Skin
RESULTS:
Collagen ................................ red.
Muscle and cornified epithelium ......... yellow.
Nuclei .................................. blue to black.
REFERENCES.
1. Jones ML.
Connective Tissues and Stains. Chapter 9, pp. 139-162.
In: Bancroft JD, Gamble M.
Theory and Practice of Histological Techniques. Fifth Edition.
Edinburgh: Churchill Livingstone. 2002;:139-162.
ISBN 0-443-06435-0, 796 pages.
2. van Gieson I.
Laboratory notes of technical methods for the nervous system.
NY Med J. 1889;50:57.
3. Verhoeff FH.
Some new staining methods of wide applicability.
Including a rapid differential stain for elastic tissue.
JAMA. 1908;50:876.
4. Weigert K.
Über eine Methode zur Färbung elastischer Fasern.
[German: Regarding a method for staining elastic fibers].
Zentralbl f allgem Pathologie u pathol Anat. 1898;9:289.
5. Weigert K.
Eine kleine Verbesserung der Hämatoxylin van-Gieson-methode.
[German: A small improvement in the hematoxylin van-Gieson method].
Z f wissenschaftl Mikroskopie u mikroskop Technik. 1904;21:1.
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.