sectioning thick frozens

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From:Gayle Callis <> (by way of histonet)
To:histonet <>
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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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