Re: Rat Brain

From:Gilmor Keshet <>

Fix the brain by perfusion of cold 4% PFA in PB, take out the brain and 
post fix it overnight in the same solution 4deg, then section the fresh 
brain by the vibratom (50-400um), or let sink in Sucrose 30%/PB and then 
section by sliding microtom (20-40 um).
Brain sections is stained better in floating sections, instead of slide 
mounted, if antibodies are to be used.
What does you friend has to stain with?
Stanford, CA

At 05:53 PM 6/5/2001 , you wrote:
>Dear Histonetters:
>A friend brought me a block of rat brain to cut.  (I have never cut
>brain tissue before).  The tissue had been formalin fixed and paraffin
>embedded.  When I tried to cut it, the tissue just crumbled out of the
>block.  If I used moist heat and then chilled the block, I could get a
>small ribbon,(six or seven sections) but then the tissue  would
>crumble.  I'm not sure it was processed properly.  Does anyone have a
>good protocol for fixing and processing rat brain?  Also, my friend
>would like to cut thick sections (from 40 to 100 microns thick)  What do
>I need to do that?  Any help would be appreciated.
>Louise Strande
>UMDNJ, Camden NJ

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