frozen sections revisited

From:"pruegg@mail.viawest.net"

Histonetters,
I know this subject has been done on HISTONET before, but can we open this
up again.  how do you process frozen sections to get the best slide
adhesion?
do you fix immediately in cold fix in the cryostat or airdry and then fix? 
what do you fix in, ie cold acetone,etc.?  do you always try to stain
slides soon after preparing?  do you store FS for staining later?  how are
FS stored, ?wrapped individually in foil and plastic bags, etc.?  do you
store them unfixed or fixed?  how are stored FS retrieved from the freezer?
Thanks for responding to this mini survey.
Patsy Ruegg 

James,
are you talking about cells in suspension or tissue sections lifting off
silane coated slides?
for keeping FS on plus slides, i pick them up and let them airdry well for
at least 10 min. at RT.  there are some different methods for what to do
after drying to fix and stain right away or store frozen unfixed until
ready to stain, but i think the initial drying at RT goes a long way to
keep them on the slides.  some people have a jar of cold fixative in the
cryostat and fix immediately after sectioning in the cryostat and then air
dry, but i have had trouble with preservation of morphology doing it this
way.  people have their favorite methods that work the best in their hands.
my WS partner said she had some antigenicity loss from storing dryed
unfixed sections in the freezer, but i have always done it that way with no
apparent problems.

Original Message:
-----------------
From:  james.zimmerman@pharma.novartis.com
Date: Fri, 25 Oct 2002 11:52:45 -0400
To: Histonet@pathology.swmed.edu
Subject: 


Has anyone had any problems with lifting off cells for LCM that have been
picked up on SuperFrost Plus Slides?  How

did you resolve the problem?

Also does anyone have any suggestions for keeping fresh frozen cyrostat
sections on any kind of slide.



                         Thanks,

                             JPZ



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