[Histonet] unfixed tissue

[Sebree Linda A.]

Nathan,
How about 3-5" in acetone?

Linda A. Sebree
University of Wisconsin Hospital & Clinics
IHC/ISH Clinical & Research Laboratory
DM223-VA
600 Highland Ave.
Madison, WI 53792
(608)265-6596
FAX:  (608)262-7174



-----Original Message-----
From: Nathalie Dhomen [mailto:n.dhomen <@t> ich.ucl.ac.uk]
Sent: Thursday, June 17, 2004 4:38 AM
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] unfixed tissue


Hello histonetters,
I believe this subject may have been covered before, but I couldn't quite
find what I wanted in the archives.
I've been doing some immunohistochemistry on some unfixed frozen tissue
(mouse embryos) as my antibody doesn't like the 24 hour 4%
paraformaldehyde fix I usually use before OCT embedding. However, the
tissue is so fragile after the immunohistochemistry, I was wondering what
methods I might use to postfix. I've had suggestions/read about placing
the sections for 20 minutes in 4% paraformaldehyde as well as the use of
methanol. Anyone have good experiences with this? Is it better to fix
right before or after the IHC? All suggestions much appreciated!

Nathalie Dhomen
Institute of Child Health
London


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