[Histonet] RE: Histonet Digest, Vol 108, Issue 26

Davis, Cassie CDavis <@t> che-east.org
Tue Nov 20 12:46:18 CST 2012

Hi Christina,
      try post fixing the FS slides in ice cold acetone (in a freezer) for 30 minutes. That is what was reccommend to me when I was trying to do IHC on ThinPrep slides.

Cassandra Davis
CDavis <@t> che-east.org
Saint Francis Hospital
Saint Francis Facebook Page

Hi all,
I am performing IHC on tissue that was formalin fixed for 48 hours and then embedded in OCT.  I have been using Gold Plus slides from Thermo Fisher.    I am staining rat spleen and mandibular lymph nodes.  With a peroxidase method, the tissue completely digests off the slides as soon as I apply the peroxidase block (looks like pouring hydrogen peroxide on a cut - just turns white and bubbles off of the slide).  I have tried various concentrations of peroxidase block with no luck.  With the alkaline phos method, the tissue is staying adhered to the slide, but I am getting significant 'tissue lifting' which makes the pathologists interpretation very difficult.  Is there anyone out there with experience in doing IHC on formalin fixed frozen sections that has experienced similar 'tissue lifting' problems?  I have ran this antibody on this tissue with immunofluorescence successfully - that may be the only choice we have for this particular tissue.  Would really like to be able to perform this staining with either an HRP or AP method for light microscopy if possible.
Christina Thurby
Bristol Myers Squibb
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