[Histonet] Karnovsky and Roots stain

Setlak, Lisa LSetlak <@t> childrensmemorial.org
Fri Feb 4 15:20:19 CST 2011

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


-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of John Kiernan
Sent: Friday, February 04, 2011 3:15 PM
To: Nicole Cosenza
Cc: histonet <@t> 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 <ncosenza <@t> siumed.edu>
Date: Thursday, February 3, 2011 18:45
Subject: [Histonet] Karnovsky and Roots stain
To: histonet <@t> 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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