Verhoeff - Van Gieson (Elastin)
Solutions
5% hematoxylin solution
Hematoxylin 10g
100% ETOH 200ml
Dissolve hematoxylin in 58-60°C water bath. Filter. Solution is stable for several months
10% aqueous ferric chloride (prepare fresh)
Weigert's iodine solution
Potassieum iodide 2 gm
Iodine 1 gm
ddH2O 100 ml
Verhoeff's staining solution (prepare fresh)
5% alcoholic hematoxylin 20ml
10% ferric chloride 8ml
Weigert ‘s iodine solution 8ml
Mix well , solution should be jet black . Use immediately.
2% aqueous ferric chloride ( prepare fresh ).
5% aqueous sodium thiosulfate.
Van Gieson's counterstain
1% aqueous acid fuchsin 5ml
Saturated aqueous picric acid 100ml
Procedure
- Hydrate slides to distilled water.
- Stain in Verhoff's solution for 1 hour. ( Save solution. Saved solution may be used for restain.
- Rinse in tap water with 2 or 3 changes.
- Differentiate in 2% aqueous ferric chloride and agitate slides gently. Stop differentiation with several changes of tap water and check microscopically for black elastic fiber staining and gray background. Repeat 2% ferric chloride treatment and tap water rinses as necessary. If elastic fiber staining is too pale, restain in the saved verhoeff's solution. It is better to slightly underdifferentiate the tissue. 3 minutes differentiation was used for 10um paraffin sections
- Wash slides in tap water.
- 5% sodium thiosulfate 1 minute. Discard solution.
- Wash in running tap water for 5 minutes.
- Counterstain in Van Gieson's solution for 3-5 minutes .
- Dehyddrate, clear in xylene, and coverslip.
Results
Elastic fibers---blue-black to black
Nuclei---bluie to black
Collagen---red
Other tissue elements---yellow
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