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

[Nick Dee]

Hi-

 

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
appreciated.

 

Thank you, 

Nick