Hi Histonetters,
I have the following problem: I am trying to stain thick (200 microns) 
free-floating frozen sections for IHC using different antibodies. I have 
tried different permeabilization methods, but so far the best I can get is 
to stain 30 microns deep into the tissue, using 0.8% Triton X-100 in PBS 
with overnight incubation. This lack of staining penetration seems to be 
independent of the tissue or antibody I use.
Any protocol out there for the staining/permeabilization of free floating 
thick tissue sections would be greatly appreciatted.
Sue





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