EM IHC questions
Dear EM Netters,
Here I am again with questions. I have been asked to help the EM people with
immuno electron histochemistry, and this is a "Halt leading the Blind"
situation! I did EM work back in the middle ages, but did not do any EM IHC
(there was no such thing), so I am somewhat familiar with EM. The EM tech is
not familiar with any IHC whatsoever.
They are working with human skin.
I have finally convinced them not to fix in gluteraldehyde, although they
insist on going with 1% gluter in paraformaldehyde, and to use low temp
(Lowcryl) processing. I gave them a K15 Ab which I use with great success and
which I figured would stain lots of basal cells at least. I also gave them my
method and suggested they bracket their dilutions and incubate overnight.
They got no reaction.
I am aware that not all Abs work with EM sections, obviously I don't know which
ones do/don't! So the 1st question is: can anyone tell me which don't and/or at
least tell me one that does. Since I don't know whether their method is the
problem or the K15 just won't work, I can't trouble shoot.
The 2nd question is: can one do HIER (pressure cooker) and or enzyme retrieval
on EM sections?
Thanks for any help/advise anyone can offer.
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