Re: problem staining thick sections

From:Geoff McAuliffe

Hi Sue:

Antibodies are big molecules.
So what permeabilization methods have you tried? Graded ethanols?
Is this fixed tissue?
What kind of tissue is it?
You can't cut thinner sections because ....?

sue garcia wrote:

> 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

Geoff McAuliffe, Ph.D.
Neuroscience and Cell Biology
Robert Wood Johnson Medical School
675 Hoes Lane, Piscataway, NJ 08854
voice: (732)-235-4583; fax: -4029

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