Figure 1. Positive control equine aorta: elastic tissue stains black, collagen stains red.
Verhoeff’s Elastic Staining Procedure
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 500mlVan 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.
Positive
control: aorta
Results:
Elastic Fibers - brilliant blue - black
Nuclei - blue to brownish black
Collagen - red
Other tissue elements - yellow
Figure 1. Positive control equine aorta: elastic tissue stains
black, collagen stains red.
Effective:
July 1, 2004
Cherie Chapman
James R. Turk
References:
Armed Forces Institute of Pathology Laboratory Methods in Histotechnology, eds. Prophet EB, Mills B, Arrington JB, Sobin LH, American Registry of pathology, Washington D.C. , 1992, pp.134.Sheehan DC, Hrapchak BR: Theory and Practice of Histotechnology, 2nd Ed, CV Mosby, St. Louis, 1980, pp. 196-197.