[Histonet] RE: Bubbles on Fresh Frozen tissue after IHC

Nick Dee NickD <@t> alleninstitute.org
Wed Dec 16 17:24:17 CST 2009




I am doing immunohistochemistry on fresh frozen tissue using the
capillary gap method on Tecans.  The preferred fixative for my antigen
is a cold acetone fix.  I am using MeOH/H2O2 to kill peroxidase before
the blocking step and it's resulting in many small bubbles forming on
the tissue in which I get no staining...in the area where there aren't
bubbles, the staining looks great.  I am aware of using something other
than MeOH to block for peroxidase, however when I don't use an alcohol,
the surface tension pulls all the tissue off my slide when I'm taking
apart the capillary gap chambers to coverslip.


Has anyone experienced this before?  Any ideas/suggestions would be


Thank you, 


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