[Histonet] IHC for secreted proteins and cytokines in frozen sections - Pre-fix or post-fix?

[Morken, Timothy]

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 at lists.utsouthwestern.edu] 
Sent: Friday, June 02, 2017 1:01 AM
To: histonet at 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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