[Histonet] re:IHC on FFPE sections of Insects

From:"Carl Hobbs"

I have only carried out IHC on Drosophila embryos. I use a std FFPwE
processing IHC with HIER protocol, before detection of Ag using a Std
stABCpxDAB. IMHO, the fixation/processing times are not a major issue.
However, THE major issue is whether or not the Abs will work on such
sections! Many don’t. So, one has to then  try unfixed/fixed frozen
sections/brief-fixed vibratome sections.
 Adult insects, having chitin “casings” will always be a problem, IMHO.
I cut adult Formalin-fixed Drosophila flies on cryostat and Pwax - embedded
sections for NISH and they are always problematic. Lately I have had better
success using Methyl Benzoate as a clearing agent but……I am not yet
convinced that it is a priori better. Perhaps my techniques have improved ?
You do not mention anything of the insects, tho…….More details would be
http://www.immunoportal.com/index.php   Try here..Image gallery, hit the
“search” box with “fly” to see fly embryo IHC examples.

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