[Histonet] RE: Vibratome sectioning of mouse spinal cord

[Mejia, Maria]

Hello Susan,

Before I can help, I need more information from you.  Are your samples fixed or unfixed?  In any case, make sure that the vibratome
is in working order & that all the movable parts are tight before you section on the vibratome.  In addition, during cutting the holding tray
should have cold 0.1M PB buffer & this tray is surrounded by wet ice.  The temperature should be cold during sectioning.  

I don't know what type of vibratome your using, but check the angle of the blade - this may need to be changed slightly - check what
the manual says because it should be possible to use the knife at different angles.  Also check the knife blade, you most likely have
already used a new blade.  Finally, but important during the cutting as the vibratome begins to section & you actually see the section
starting to come off - use your brush to brush away the tissue section from the knife edge during cutting - this should keep the tissue flat. 
Also, maybe try cutting at 50 microns.  Hope this helps you!

Maria Mejia
Senior Histology Supervisor
Department of Neurosurgery
UCSF
San Francisco, CA 94103
________________________________________
From: histonet-bounces <@t> lists.utsouthwestern.edu [histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Michael, Susan [michaels <@t> janelia.hhmi.org]
Sent: Thursday, November 11, 2010 1:18 PM
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] Vibratome sectioning of mouse spinal cord

Can anyone give me some tips on sectioning mouse spinal cord on the vibratome?  Specifically, once the dura is successfully removed, I have been sectioning 40 um horizontal sections, and some cords have been curling up on themselves, both rostral-caudal and medial-lateral, and are impossible to uncurl without destroying the tissue.  I’ve tried every angle of approach, speed, amplitude, the tissue has been in sucrose, embedded in agarose, and chilled.

Thanks,

Susan
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