Re: sectioning thick frozens

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From:"M.A. Jennings" <> (by way of histonet)
To:histonet <>
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I have had to compromise with the investigators here. They asked for 20u
sections, then 16u sections, now they settle for 10u. We did a comparitive
on frozen samples with no fixation, where we looked at all of the
parameters on 5-20um sections  (i.e. no wrinkles, no "chatter", stay on the
slide, good immuno staining, high power viewing, etc) kind of like a
checkerboard. In my hands and with a wonderful research grade Zeiss
cryostat the overall best "thick" sections were 10um. Ok now I have to do
serial sections on a cryostat but it is more plausible. I would rather do
40 sections at 10, than 80 at 5 or 20 at 20. We are looking at purchasing a
confocal microscope (50-60um sections and localization of specific FL cells
is routine with "laser sectioning") so I will indeed need to go thicker. I
will also be looking for Help when this day comes. So any help for Lilith
and Gayle would be greatly appreciated by me also. anita

PS Gayle sometimes when I read your questions/answers I think we work for
the same people. At least they seem to have a lot in common. aj

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