Ana, you should fix before sectioning. Chris Van der Loos did an excellent
study on loss of cytokines in frozen sections vs whole cells. See his paper:
Immunohistochemical Detection of Interferon-y, Fake or Fact? CM Van der Loos,
et.al., J Histochem Cytochem, 49(6):699-709, 2001. Www.jhc.org
Tim Morken
Pathology Site Manager, Parnassus
Supervisor, Electron Microscopy/Neuromuscular Special Studies
Department of Pathology
UC San Francisco Medical Center
-Original Message-
From: Ana Maluenda via Histonet [mailto:histonet@lists.utsouthwestern.edu]
Sent: Friday, June 02, 2017 1:01 AM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] IHC for secreted proteins and cytokines in frozen sections
- Pre-fix or post-fix?
Hi everyone,
Just wondering what is people's opinion and protocols for IHC in mouse frozen
sections targeting secreted proteins and cytokines. I see lots of places using
fresh frozen sections/snap-freeze and cold acetone or methanol/ethanol
post-fixation. Is this an issue when it comes to diffusion of such proteins in
the tissue, since they are not well localized or linked to cellular structures?
Would it be better to pre-fix (either immersion in 4%PFA or infusion with
fixative) than post-fix?
Any thoughts would be much appreciated.
Kind regards,
Ana
Ana Maluenda
Research Assistant
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