[Histonet] uneven alternating sections on cryostat

[Nathan Cramer]

   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]natecrmr <@t> gmail.com
     > To: [2]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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References

   1. mailto:natecrmr <@t> gmail.com
   2. mailto:histonet <@t> lists.utsouthwestern.edu
   3. mailto:Histonet <@t> lists.utsouthwestern.edu
   4. http://lists.utsouthwestern.edu/mailman/listinfo/histonet