We do ACH staining on rectal biopsies to evaluate for ganglion cells for
Hirschsprung's disease. Our stain is performed on fresh frozen tissue and I
believe it's the Karnovsky method, but I'm not sure. Feel free to email if you
are interested in out procedure.
Lisa M. Van Valkenberg, B.S., HT- ASCP
Histology Manager
2300 Children's Plaza
Chicago, IL 60614
773-868-8949
-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of John Kiernan
Sent: Friday, February 04, 2011 3:15 PM
To: Nicole Cosenza
Cc: histonet@lists.utsouthwestern.edu
Subject: Re: [Histonet] Karnovsky and Roots stain
Karnovsky Roots is (IMHO) the best histochemical method for choline esterase
activity. In muscles, acetylcholinesterase (AChE) is the only such esterase
shown by this method, and it is in the subneural apparatus of the motor
endplate. Some counterstains (notably silver methods for the innervating axons)
can remove the brown copper ferrocyanide product.
Another way to show motor endplates is with a method that picks up all
esterases. In muscle, the endplate AChE shows up sooner than the enzymes
present in all cells. Indigogenic esterase methods can be followed by silver
staining of axons.
Why do you need or want to use fresh frozen, unfixed tissue sections? This
makes no sense in the world of esterase activity histochemistry. There haven't
been any developments in this field since the 1960s other than labelled
alpha-bungarotoxin and immunohistochemistry.
An inexpensive book is Van Noorden CJF Frederiks WM 1992. Enzyme
Histochemistry. Oxford Univ Press and Royal Microscopical Soc. ISBN0198564341.
Another one is Lojda, Gossrau Schiebler 1979. Enzyme histochemistry. Berlin:
Springer. ISBN 0387092692.
A quick web search indicates that both are available and cost less than $10
second-hand.
John Kiernan
Anatomy, UWO
London, Canada
= = =
- Original Message -
From: Nicole Cosenza ncose...@siumed.edu
Date: Thursday, February 3, 2011 18:45
Subject: [Histonet] Karnovsky and Roots stain
To: histonet@lists.utsouthwestern.edu
I am looking into a project involving motor end plate staining.
Literature that I've found continually references Karnovsky and
Roots from the 60s. However the papers are not
supplying all the details.
Does anyone do AchE staining by this method on fresh frozen,
unfixed tissue sections? If so, can I get a more detailed
protocol (fixation steps, washes, etc)?
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