Thawing frozen tissue in OCT

From:Gayle Callis (by way of Histonet)


Take the frozen block and drop it into room temperature PBS. Be sure to
change the PBS several times to remove all OCT.

We have thawed frozen blocks by dropping them into room temperature
formalin, but you need to avoid further aldehyde fixation is you want to do
whole mount immunostaining.  Hopefully, all will go well since you have
snap frozen(?)to minimize ice crystal damage, and a rapid thaw should be
adequate.  We have done this on NBF fixed and/or unfixed frozen tissues,
and morphology was not bad.  I am sure there may be a bit of residual
damage from freeze/thaw, but what the heck - you may have total success.  I
didn't have to do immunostaining after thawing, just paraffin processing.

Gayle Callis
Research Histopathology Supervisor
Veterinary Molecular Biology - Marsh Lab
Montana State University - Bozeman
S. 19th and Lincoln St
Bozeman MT 59717-3610

406 994-6367 (lab with voice mail)
406 994-4303 (FAX)


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