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@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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