[Histonet] Orcein Variation for EVG to detect Elastosis Perforans Serpiginosum

[RSRICHMOND <@t> aol.com]

A. Kevin Williams asks: >>My pathologist has asked for the Orcein variation 
on the EVG (Elastin Van Gieson) or VVG as some of you know it. I have to get 
the Orcein so that is my first challenge, second is my protocol. There are a few 
out there and I would like to hear from anyone who has a favorite for this 
method [elastosis perforans serpiginosa].<<

Here are my reading notes - never tried it - on the traditional Pinkus orcein 
stain used by dermatopathologists of yore.

See also the short description of the stain in John Kiernan's Histological & 
Histochemical Methods (199), page 162.

Bob Richmond
Samurai Pathologist
Knoxville TN
******************
Mehregan, Amir H. (Wayne State). Pinkus' Guide to Dermatohistopathology. 4th 
ed. Appleton-Century-Crofts, 1986.
Hermann Pinkus, 1905-1985
Acid orcein and Giemsa stain: from
Pinkus H, Hunter R. Simplified acid orcein and Giemsa technique for routine 
staining of skin sections. Arch Dermatol 82:699, 1960
Krobock E, Rahbari H, Mehregan AH. Acid orcein and Giemsa stain. Modification 
of a valuable stain for dermatologic specimens. J Cutan Pathol 5:37, 1978.

Fix in formalin or alcohol, without chromium or mercury.

1. Stain in ORCEIN for 30 minutes. Synthetic orcein stains elastic fibers 
specifically, with very little background staining. The background may be 
decolorized by short immersion in absolute alcohol or 0.1% acid alcohol.
2. Wash in running water 10 minutes.
3. Stain overnight in GIEMSA. Krobock et al. speeded this up by staining 1 
hour in 1% Giemsa solution at 60° C.
4. Wipe slides. Remove excess blue by rinsing in 95% alcohol to which a small 
amount of eosin has been added if necessary. Continue until the collagen of 
the skin looks pink. Then dehydrate.

ORCEIN: dissolve 200 mg of Harleco's synthetic Orcein in 100 mL of 70% 
alcohol. Add 0.6 mL of concentrated HCl. The solution improves on standing and has a 
long shelf life.
GIEMSA: one drop of stock in 20 mL of distilled water or pH 7.0 phosphate 
buffer.

RESULTS: Collagen is rose-pink, while elastic is dark brown to black. Melanin 
is dark green to black. Bacteria and fungi are dark blue.

Looking at the book, I would suppose that Mehregan has just about abandoned 
the use of this stain.