When cutting PFA fixed, cryoprotected tissue on our cryostat, I
frequently find that every other section gets cut improperly. I'll get
one nicely cut section and then on the next pass I only get half of a
section. This cycle simply repeats over and over and I lose many slices.
It has been a while since our cryostat has been serviced, so I'm
wondering if this an operator error or a machine problem. I thought
maybe the tissue temperature hadn't settled properly, but the uneven
cutting still happens even when I let the tissue sit for 30 minutes in
the chuck holder. (cutting mouse spinal cord at -20C)
On another note, if anyone has any tips for improving white/gray matter
contrast in frozen spinal cord sections stained with luxol fast blue,
I'd be very appreciative. I do defat the slices with chloroform and
differentiate with lithium carbonate but everything is usually either
very dark or very light. I am learning as I go (checking the archives
here often) so any help would be great.
Thanks!
Nathan Cramer
Neurobiology and Behavior
Cornell University
Ithaca, NY 14853
_______________________________________________
Histonet mailing list
[email protected]
http://lists.utsouthwestern.edu/mailman/listinfo/histonet
