frozen sectioning

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From:"L. Gibbs" <>
To:Histonet <>
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

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