[Histonet] RE: Vibratome sectioning of mouse spinal cord
Maria.Mejia <@t> ucsf.edu
Fri Nov 12 13:06:08 CST 2010
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!
Senior Histology Supervisor
Department of Neurosurgery
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.
Histonet mailing list
Histonet <@t> lists.utsouthwestern.edu
More information about the Histonet