Thanks, Robyn... I'll make sure the holder is clamped down. (sometimes its the little things...) Best Nate R J VAZQUEZ wrote:
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: [1]natec...@gmail.com > To: [2]histo...@lists.utsouthwestern.edu > 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 > [3]histo...@lists.utsouthwestern.edu > [4]http://lists.utsouthwestern.edu/mailman/listinfo/histonet References 1. mailto:natec...@gmail.com 2. mailto:histonet@lists.utsouthwestern.edu 3. mailto:Histonet@lists.utsouthwestern.edu 4. http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet