problem staining thick sections

From:sue garcia

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.

The new MSN 8: advanced junk mail protection and 2 months FREE*

<< Previous Message | Next Message >>