Cryosectioning and Temp.

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From:"John C. Dennis" <> (by way of histonet)
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Dear Folk

We've been sectioning rat brain (small pieces of rat brain) pretty
well.  But, when we've tried to section embryos (E14-E16) the tissue
crumbles.  These were immersion fixed in 4% paraform. and 0.1%
glutaraldehyde (pH 7.4), cryoprotected in a series of sucrose solutions
(10-30% in PBS), placed in OCT (Sakura) and frozen in liquid
nitrogen-cooled isopentane, and sectioned at the same temperature
(specimen and chamber) as were the brain tissues.

In my limited hands-on experience, I seem to remember that by playing with
the temperatures I could vary the quality of sections.  Or, different
tissues require different temperatures while sectioning.

Is this, or something like this, other people's experience as well?

Or, are there any other helpful hints?

Yours in fragments,

John Carroll Dennis
Anatomy, Physiology, and Pharmacology
109 Greene Hall
Auburn University, AL  36849

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