frozen sectioning

<< Previous Message | Next Message >>
From:"L. Gibbs" <lgibbs@u.washington.edu>
To:Histonet <HistoNet@pathology.swmed.edu>
Reply-To:
Content-Type:TEXT/PLAIN; charset=US-ASCII

Hi Scott-

If the specimen is tissue fixed in paraformaldehyde or formalin
or Bouins/Zambonis, you should cryo-protect (at 4C) by soaking in
25-30% sucrose in phos buff until the tissue sinks. The specimen
can then be sectioned on a sledge surrounded by the sucrose
medium or if no sledge is available, the specimen can be frozen
in OCT and sectioned in a cryostat. I think 80um on a cyostat may
be pushing it but it should work if you good quality knives. You
could thaw mount on a slide or collect as free-floating sections
in buffer depending on your application.
If the specimen will be unfixed, you could try the cryostat with
OCT embedding or a vibratome sans embedding.

Good Luck!


Lorraine Gibbs
Physiology & Biophysics
University of Washington




<< Previous Message | Next Message >>