Re: Acetylcholinesterase (AChE) method request

From:"J. A. Kiernan"

On Tue, 28 Aug 2001, Sharon Cassar wrote:

> I have been asked to stain for acetyl cholinesterase on formalin fixed
> paraffin tissue.  I will appreciate a staining protocol.

If anyone sends you one, treat it with great suspicion.
Ask your boss for the information. AChE activity histochemistry
requires frozen sections and various controls that are not the
same for every species and tissue. The only way to detect AChE
in a paraffin section would be immunohistochemically, with a
species-specific primary antibody. Pretty well all of the
indications for detecting AChE make sense only with thick
(30 to 300 um) sections. These are awkward (to say the least)
from paraffin and rather difficult with a cryostat.

For activity histochemistry of choline esterases, consult one of
the bigger textbooks. Pearse & Stoward Vol 3 is the best. If you
have to do immuno on paraffin sections, you'll need guidance
from papers in peer-reviewed academic journals. It's your
boss's job, not yours, to find and critically evaluate the
research literature. 

If your boss wants to do AChE histochemistry only to detect
motor end plates, a  method can be found in any textbook
of histochem or histotech published since about 1955. Refer
him to a 2nd-hand bookshop.

John A. Kiernan
Department of Anatomy & Cell Biology
The University of Western Ontario
London,  Canada   N6A 5C1

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