Introduction
Verhoef’s Van Gieson staining technique is used for the staining of Eeastic tissue fibers.
There are numerous methods for the staining of elastic fibers.
Verhoeff’s method, orcein method, Weigert’s resorcin-fuchsin method, and the aldehyde fuchsin method are most popular methods for the staining of elastic fibers.
5% acid haematoxylin solution, 10 % aqueous ferric chloride solution and Lugol’s iodine are necessary for this procedure.
Moreover, 2% aqueous ferric chloride solution is also necessary for the differentiation of nuclei and elastic fibers.
Working Verhoeff’s mixture is prepared by mixing 20 ml of 5% Alcoholic Haematoxylin, 8 ml of 10% aqueous Ferric chloride solution and 8 ml of Lugol’s iodine.
This solution should be prepared freshly.
- First deparaffinize and rehydrate the tissue sections.
- Then stain with Verhöeff’s solution for about 15–30 minutes.
- Wash well in water.
- Differentiate with 2% aqueous ferric chloride solution until the nuclei and elastic fibers get black.
- The background is still weakly stained.
- Wash with water. Then rinse by using 95% alcohol to remove any staining due to iodine alone.
- After that counter stain with van Gieson stain for few seconds.
- Blot to remove excess amount of stain.
- Dehydrate the tissue section rapidly through ascending grades of alcohol solutions.
- Clear the slides with xylene and mount with DPX.
Observations
Elastic fibers and nuclei stain in black colour.
Furthermore, collagen stain in red colour and other tissue elements stain in yellow colour.
Special Discussion
This stain can be done after using routine fixatives.
Coarse fibers are stained intensely.
However, fine fibers may be stained less satisfactorily.
Working solution can be stored up to 48 hours.
But some text books state that the working solution is stable for 2-3 hours.
Eosin stain also can be used as counter stain.
The colour of other tissues depend on the counter stain used.
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