Nathan, It sounds like the blade holder is not secure enough or even in the tightness on each side.
Hope this helps. Robyn > Date: Thu, 27 Aug 2009 16:44:11 -0400 > From: [email protected] > To: [email protected] > Subject: [Histonet] uneven alternating sections on cryostat > > 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 _______________________________________________ Histonet mailing list [email protected] http://lists.utsouthwestern.edu/mailman/listinfo/histonet
