[Histonet] Acetylcholinesterase protocol and fixation

[John Kiernan]

The AChE method in Filipe & Lake's book (Histochemistry in Pathology) is the one-step Karnovsky & Roots method, with inhibition of pseudocholinesterase by iso-OMPA and an intensification step using osmium tetroxide. It's intended for diagnosis of Hirschprung's disease - minimally fixed cryosections of rectal biopsies. 

The Karnovsky-Roots method is very reliable and in my experience (with brain and muscle) preferable to the Koelle-type methods. If you don't like using osmium tetroxide, there's an alternative intensification procedure using Nickel-DAB. See Tago G et al 1986 J. Histochem. Cytochem. 34: 1431-1438. The ChE inhibitor iso-OMPA is pretty toxic. Depending on the nature of your investigation, you may not need it.
-- 
-------------------------------
John A. Kiernan
Department of Anatomy and Cell Biology
The University of Western Ontario
London,   Canada   N6A 5C1
   kiernan[AT]uwo.ca
   http://publish.uwo.ca/~jkiernan/
   http://instruct.uwo.ca/anatomy/530/index.htm
_______________________________
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Date: Friday, September 21, 2007 7:31
Subject: [Histonet] Acetylcholinesterase protocol and fixation
To: histonet <@t> lists.utsouthwestern.edu

> Another question...
> 
> Looking for Acetylcholinesterase protocols I´ve found that probably
> Filipe and Lake´s is the most used, is this information correct? 
> 
> And... found using Karnovsky fixation or formaldehyde/calcium
> acetate.. which one is the best?? 
> 
> Tks again
> 
>  
>  Ms. Kátia Catunda
> Produção
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> +55 12 3203-0633 (PABX)
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> 
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