[Histonet] Holzer method for glial fibers

Lee & Peggy Wenk lpwenk <@t> sbcglobal.net
Thu May 9 18:45:21 CDT 2013

Some time ago, someone asked what all the reagents in the Holzer stain were 
for, and John Kiernan made some guess (see in quotes below for his response 
to the PMA). If John KIernan is having to make educated guesses, the rest of 
us probably have NO IDEA. Added to that, I don't know anyone who does this 
stain anymore. If we need to demonstrate glial cells, everyone is doing IHC 
such as GFAP.

John's response: "I don't think there are answers, other than wild 
speculation, to your questions! A few statements can be made about the 
reagents, but they don't necessarily relate to Holzer's method.

When a PMA solution is applied to a section its large anions stick to 
regions of protein that are permeable and rich in basic amino acids. (This 
happens in collagen, in trichrome methods.) PMA also makes insoluble pigment 
precipitates when mixed with cationic triphenylmethane dyes such as crystal 
violet. Non-aqueous PMA may have tissue affinity different from that of an 
aqueous solution. With the chemically related phosphotungstic acid, the 
solvent and other factors influence the tissue components that take up the 
metal when PTA is used as a contrast stain for EM (see Hayat 1993 Stains and 
Cytochemical Methods, various places in book)."

Back to my opinion: I'm really worried about some of the other reagents, and 
anyone who has to use them:
- Chloroform - EXTREMELY flammable
- Aniline oil - EXTREMELY carcinogenic (bladder cancer), and causes liver 
and hematopoetic damage

Is there any way you can convince your instructor to let you do a stain that 
is still in use, clinically relevant, and SAFER!?!?!?!?!

Peggy A. Wenk, HTL(ASCP)SLS
Program Director
Schools of Histotechnology
Beaumont Health System
3601 W. 13 Mile Road
Royal Oak, MI 48073

Opinions expressed are my own, and do not reflect my place of employment.

-----Original Message----- 
From: jesse andrade
Sent: Sunday, May 05, 2013 7:25 PM
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] Holzer method for glial fibers

I have a question about this method and the purpose of the phosphomolybdic 
acid-alcohol solution in this stain? I am astudent and need this for my 
staining procedure assignment.
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