[Histonet] embryo cryosection problem

Anna Elisse Beaudin aep10 <@t> cornell.edu
Sun Nov 14 08:15:18 CST 2004


hi all,

   I recently tried cryosectioning embryos taken at dpc13-17, and had a
horrible time because sections were crumbling to pieces, due to what I
believe was some really 'crusty' bone or cartiledge sticking out and
ripping the tissue as the blade passed over it (this is in the head). 
I am new to working with embryos, and I am trying to figure out what I
did wrong!  I fixed embryos 24-48 hours in 4% paraformaldehyde, and
then snap froze in OCT.  should I have decalcified these specimens?  or
cryo-protected them?  I would greatly appreciate anybody's advice on
this!  thanks so much in advance!

Ana Beaudin
Division of Nutritional Sciences
Cornell University
Ithaca, NY





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