sectioning thick frozens
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From: | Gayle Callis <uvsgc@msu.oscs.montana.edu> (by way of histonet) |
To: | histonet <histonet@magicnet.net> |
Reply-To: | |
Content-Type: | text/plain; charset="us-ascii" |
Even though I have the tape transfer system, cryosectioning over
10 micrometers is not fun. I would think the section at 50um would
tend to shatter, since it is so thick.
I have a protocol where people did 40um sections of perfused fixed
tissues, cut it with a vibratome instead of a cryostat. They
had brilliant immunostaining with this technic, to see GFAP in
transgenic mouse brain. (astrocyte specific glial fibrillary acidic
protein). They used both a confocal and ordinary epifluorescent
microscopy to view the sections.
I would be curious to find out if anyone has had success doing thick
cryosections, free floated variety or otherwise?? I tend to avoid
anything over 10 um, and prefer a range of 4 - 7 um in order to maintain
a flat, easy to handle section.
Gayle Callis
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