[Histonet] Questions concerning: Mouse Brain Cryosections

From:Jens Schwamborn

Dear Histonet-people,
I want to image EGFP fluorescence (endogenous) in cryosections of embryonic mouse brains (stages: E12.5 to E18.5). Furthermore I would like to perform antibody-stainings for immunofluorescence on those sections. Now I am searching for the best protocol to do so.
Do you fix the brains before the sections? If yes, how? (I guess perfusion is impossible at E12.5)
Or do you freeze the complete brain, perform the sectioning and fix afterwards? If so, how do you freeze, with dry ice ?

Thanks for the help.

Jens
email: jschwamborn@yahoo.de
 		
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