| Verhoeff’s
Elastic Staining Procedure
Literature
Purpose:
This standard operating procedure (SOP) establishes the standard
for staining elastic fibers and collagen using the Verhoeff’s
Elastic staining procedure.
Reagents
and Materials
Reagents
Elastic Hematoxylin
Hematoxylin
20gm
95% Alcohol 440ml
10%
Ferric Chloride
Ferric
Chloride 10gm
Distilled water 100ml
Lugol’s
Iodine
Iodine
5gm
Potassium Iodide 10gm
Distilled water 250ml
2%
Ferric Chloride
10%
Ferric Chloride 10ml
Distilled water 40ml
2%
Sodium Thiosulfate
Sodium
Thiosulfate 10gm
Distilled water 500ml
Van
Gieson’s
1%
Acid Fuchsin 12.5ml
Picric Acid 2.5gm
Distilled water 250ml
Materials
Tissue slides and control slides for special stain requested.
Microwave to bake slides.
Staining rack, 20 slide capacity.
Coverslips and synthetic mounting media.
Hazards
and Precautions:
See Material Safety Data Sheets.
Sample Preparations:
Paraffin tissue section slides cut at 4 microns.
Calibration
and Standardization:
A tissue section slide containing elastic fibers and collagen is
stained with each procedure.
Procedure:
1. Deparaffinize and hydrate to distilled water.
2. Elastic Stain - 15 minutes.
Make
fresh: Elastic Hematoxylin 22ml
10% Ferric Chloride 8ml
Lugol’s Iodine 8ml
3.
Distilled water rinse.
4. 2% Ferric Chloride - 1 1/2 minutes.
5. Distilled water rinse.
6. 2% Sodium Thiosulfate - 1 minute.
7. Distilled water rinse.
8. Van Gieson’s - 1 minute.
9. Dehydrate, clear and coverslip.
Results:
Elastic Fibers - brilliant blue - black
Nuclei - blue to brownish black
Collagen - red
Other tissue elements - yellow
Cherie
Chapman
James R. Turk
7/1/04
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