problem staining thick sections
Hi Histolanders,
I have the following problem: I am trying to stain free-floating thick
frozen sections for IHC using different antibodies. Independently of the
tissue or antibody of choice, I can only get stain about 30 microns deep
into the section, while the rest (about 200 microns total) remains
unstained. I have tried different permeabilization methods, but nothing
seems to solve this problem. Lately, I have been doing 0.8% Triton X-100 in
PBS overnight at room temperature, and sometimes I can get up to 40 microns
deep...Any protocol out there for staining of free-floating tissue sections
would be greatly appreciatted.
Sue
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