IHC on 50u frozen sections

<< Previous Message | Next Message >>
From:"Instrumedics, Inc." <info@instrumedics.com> (by way of histonet)
To:histonet <histonet@magicnet.net>
Content-Type:text/plain; charset="us-ascii"

Happy Holidays to you all!

Lillith questioned the idea of 50micron sections for immunos.

I believe that 50 micron sections for immunos would present a real problem
for  seeing  where the product is localized. There would so many focal
planes that  you would not know where you are in the cell and  what cell
type is stained. Also it is very likely that the antibody might never  reach
the antigens deep in the section.and  I also would think you would need to
use commensurately more reagent.

I always tried to use the thinnest section I could cut so that the
localization of immunostain could be accurately interpreted. I found that
when we developed the CryoJane Tape-Transfer for frozen sections I was able
to cut 2 micron sections which were  bonded to the slide. Once I found a
"good" monoclonal antibody for the antigen I was studying I could use very
little reagent(10ul) droppered on to the section and because there was no
fear of losing the section I could go through the entire immuno process in
10-15 min. These were truly "fast immunos."


<< Previous Message | Next Message >>