Verhoeffs

From:valerie hannen

Angela,
 
Here is the info. as promised:  All of he solutions needed for staining with the exception of Distilled water and Xylene are included in the kit mentioned below.
 
VERHOEFF'S STAIN FOR ELASTIC TISSUE
 
Vendor: Polyscientific
 
FIXATION: Any may be used
TECHNIQUE: Parrafin. Cut at 6 microns
 
Catalog # K 059
 
STAINING PROCEDURE:
 
1) Deparaffinize and hydrate to water.
2) Rinse in running tap water- 3 min.
3) Stain in Verhoeff's Working Solution-- 15 min.
4) Wash in distilled water- 10 dips
5) Differentiate in Ferric Chloride 2% Aqeous, few dips at a time, until tissue begins to appear gray. ( we do for about 1 min.)
6) Rinse in tap water to remove excess Ferric chloride. Check under microscope for correct differentiation. Elastic fibers should be clear, sharp black, and surrounding elements colorless. The differentiation is very important. The section should be differentitated to the fine fibers. The slides should be slightly under differentitated to offset the VanGiesons reaction which follows. The Picric Acid in Van Giesons will remove some of the stain from the elastic fibers. If fine fibers are not visible, the slide has been over-
differentiated. Repeat process from step 2.
7) Treat in Sodium Thiosulfate 5% aqeuous for 1 min.
8) Wash well in tap water- 5 min. Recheck under microscope.
9) Counterstain in Van Gieson Solution. Timing differs with type of specimen and size. Consult pathologist before staining. Do not dehydrate in alcohol, because this will remove the Basic Fuchsin part  of the Van Gieson Solution and slides will take on a purple cast.
10) Blot slide dry before placing in Xylene. Clear.
11) Mount with mounting media.
 
RESULTS:
 
Elastic Fibers: Black
Collagen: Red
Muscle, Cornified Epithelium: Yellow
Nuclei: Blue to black
 
 
Sorry this was so long.....   hope this will help you.
 
 
Valerie Hannen
Parrish Medical Center
Titusville,Florida

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