Verhoeffs
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
<< Previous Message | Next Message >>