RE: Fixation of frozen sections

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From:"Patterson, Noelle" <PattersonN@NMRIPO.NMRI.NNMC.NAVY.MIL>
To:"''" <>, "Coskran, Timothy M" <>
Date:Fri, 28 May 1999 12:40:42 -0400
Content-Type:text/plain; charset="iso-8859-1"


I asked a similar question to histonet not too long ago.  I got enough ideas
to run my own study.  What I can tell you is that monkey and human renal
tissue are best when sectionned, fixed (I use Acetone at -20C), air dried
(about 15-30 min., the minimum to get dry slides), and stored at -70C until
use.  When I remove the slides for IHC, I do a brief fix (10-20 dips in
water then acetone) for reasons based on what our staining system needs.
However, murine renal, pancreas, spleen, and lymph node tissue do just fine
if you fix in acetone, air dry, and store (dust-free) at room temperature.  

There are quite varied protocols to address your question.  Good luck.  Feel
free to ask ant nagging questions you may have about what I've written.

Noelle Patterson
Naval Medical Research Center
Bethesda, Md

	From:  Coskran, Timothy M []
	Sent:  Thursday, May 27, 1999 3:34 PM
	To:  ''
	Subject:  Fixation of frozen sections

	Does anyone have any ideas on whether it is better to cut frozens,
fix and
	store the sections or to cut frozens, store, and then fix when your
ready to
	carry out a stain?  We currently cut frozen sections, dry briefly
and then
	store the slides at -80C until we are ready to do an immuno stain.
Then we
	bring the slides to room temp, fix and proceed with the stain.  I've
	some literature that suggests fixing the slides immediately after
they have
	been sectioned and then storing in PBS until ready to use.

	If anyone has a basic protocol for frozen sections, would they be
willing to


	Tim Coskran

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