[Histonet] Cryosectioning cryoprotected (30% sucrose) brain
bjrenquist <@t> ucdavis.edu
Tue Apr 5 15:29:38 CDT 2005
I am attempting to slice sheep brainstem and hypothalamus on a cryostat at
-20 C. The tissue was treated as such:
Perfusion through carotid artery (6L of 4% paraformaldehyde)
Postfixation for 24 hours at 4 C
Cryoprotected in a 20% sucrose solution for 5 days at 4 C
Cryoprotected in a 30% sucrose 30% ethylene glycol solution and stored at
I am having some major problems with slicing this tissue. It appears to
never freeze. Does anyone know at what temperature I should be slicing?
Any opinions on use of a vibratome instead of a cryostat? I'm trying to
section tissue at 30-40 um. Thanks in advance for your help.
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