[Histonet] (no subject)

[Anna Elisse Pflaster]

Hello,

  I am going to start processing mouse embryos for in situ and
immunohistochemistry, and since I am a novice, I have a few very basic
questions that I am hoping to find answers to!  First, I was wondering
if it is absolutely necessary to embed embryos in paraffin for
cryosectioning.  I usually work with brain tissue from adult mice, and I
never have to embed in paraffin -- I always freeze in OCT.  In this
case, I will be sectioning embryos at ED14.5 and ED17.5.  Also, I was
wondering about a simple fixation procedure for 'older' embryos.  Is it
feasible for fixation to involve only fixing cryosections in 4%
paraformaldehyde?  Lastly, if my goal is to look at expression patterns
in the embryonic brain, what would be the best approach/angle to
sectioning?  Specifically, I am interested in having a good view of the
ventricular zone and dentate gyrus.

I would greatly appreciate any and all advice on these matters!  Thank you
in advance!

Anna Beaudin, M.Sc.
Division of Nutritional Sciences
Cornell University
Ithaca NY 14850