[Histonet] IHC on fresh frozen
Sebree Linda A
LSebree <@t> uwhealth.org
Wed Dec 3 10:12:51 CST 2008
I was taught to get frozen sections into cold acetone immediately after
sectioning, not letting the section air dry. We do C4d staining on
frozen renal bxs and immediately fix the sections in cold acetone for 10
minutes. Then we air dry followed by 2 minutes in Ventana Morphosave
(not a critical step) then keep the slides wet throughout the
immunostain. We get nice clear, crisp staining.
Linda A. Sebree
University of Wisconsin Hospital & Clinics
600 Highland Ave.
Madison, WI 53792
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of
Sent: Wednesday, December 03, 2008 9:17 AM
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] IHC on fresh frozen
Recently I did some IHC(chromagen methods) on mouse fresh frozen
tissues, mainly using insulin antibody on pancreas. The image is
much fuzzier compare to paraffin embedding tissue. And the
staining also smeared to acinar cells which surround the islet.
I airdried slide(>30min) and used a general acetone method(-20C
5min) to fix the tissue before I did IHC.
How can I get a relatively sharp staining on the fresh frozen
tissue?Does anyone here have any experience on it? Any
Many thanks and have a nice day,
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