[Histonet] uneven alternating sections on cryostat

[Adam .]

I was having this problem too, but I think I finally figured it out. There
is a screw on our cryostat that attaches the chuck to the rest of the
machine. This screws fits into a small hole in the chuck. Sometimes the
screw isn't well set in the hole or is well set but for some reason comes
loose and causes the chuck to wobble. For some reason, this causes your
problem. Now I make sure that everything is tightly secured, and I rarely
have this problem.

Good luck,
Adam

On Thu, Aug 27, 2009 at 4:47 PM, Nathan Cramer <natecrmr <@t> gmail.com> wrote:

>
>   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
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