RE: staining fresh frozen brain sections

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From:"Barry, Lilith" <> (by way of Marvin Hanna)
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Hi Cynthia,

I routinely work on free floating sections. This is on fixed sections
(30-50microns thick). I do not think that it is possible to do
immunohostochemistry on fresh floating sections. For that I cryosection
(10-14 microns), mount them in supperfrost + slides and do the immuno on the
slides. You can cut thick sections, fix them immediately and then proceed
with free floating procedure. The immuno is done in tissue culture well
plares by adding and removing solutions.
As for the processing,  what kind of secondary are you using? If it is
fluorescent, then I mount with wet mount. If it is permanent, then
dehydrate and mount with permount.
Please feel free to ask any question.

Lilith Ohannessian-Barry
National Research Council
Institute of Biological Sciences
e-mail; <>

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