[Histonet] uneven alternating sections on cryostat

[R J VAZQUEZ]

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: natecrmr <@t> gmail.com
> To: histonet <@t> 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
> 
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