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Cryostat help!

nishir at ohsu.edu nishir at ohsu.edu
Wed May 4 17:46:09 EST 1994


In article <pataky-270494154832 at steevlab.generes.ca> pataky at bdc.ubc.ca (Dave
Pataky) writes:
>Anybody out there know why Tissue-Tek, presumably designed for embedding
>samples for cryostat cutting, is so $##@***%* annoying to work with?  The
>problem:  I'm cutting chick embryonic brain tissue, anywhere from 10-40
>microns, at -20 and no matter what I try the sections won't stop curling
>up, rolling into tubes as soon as I lift the antiroll plate.  I've tried
>adjusting the blade angle, the "anti-roll" (that's a joke) plate, the
>temperature, speed of cutting, new blade (disposable), nothing works. 
>Ideally I'd like to see nice flat sections that stay that way, preferably
>"ribboning" off the blade so I can mount several at once.  Know any tricks
>or alternatives to Tissue Tek which may work better?  Any ideas how to deal
>with static electricity? (sometimes the sections stick to the "anti-roll"
>plate).  Feels like I'm battling the antichrist (and losing!),
>
>
>-- 
>Dave Pataky
>Dept of Zoology, UBC
>pataky at bdc.ubc.ca
>
I don't think your problems are with the TissueTek. When I've had problems with
rolling I can usually fix it by adjusting the angle of the anti-roll plate, or
by slightly warming the sample by quickly pressing my thumb against it before
cutting.  It also helps to keep the cover closed as much as possible so that
the knife doesn't warm up too much relative to the sample. One other
suggestion: minimize the amount of Tissue tek around the sample.  Embed in as
small a drop as possible so that most of what you're cutting is tissue and not
big globs of Tissue Tek.

Rae Nishi
Cell Biology & Anatomy
Oregon Health Sciences University
Portland, OR 




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