i need help for brain cryostat section

June Noble JuneNoble at email.msn.com
Tue Mar 16 12:36:07 EST 1999


Hi,
I regularly cut rodent brain using a cryostat and usually use fresh frozen
tissue, in that I freeze the brain on soft dry ice when taken from the
animal, but i don't see why isopentane shouldn't work just as well - I've
only used this way once.
I let my tissue equilibrate for 30-60mins at cryostat temp of -20 before I
attempt to cut. I cut sections of 10 microns.
Fixed tissue really, in my experience, needs to be embedded in paraffin wax
and can be cut by microtome at room temperature.
Hope this helps.

arnaud2507 at my-dejanews.com wrote in message
<7bivkb$l1i$1 at nnrp1.dejanews.com>...
>hi,
>
>I'm a little problem with brain cryostat section.
>
>I want to cut mice brain with a cryostat in order to stain with thionin and
>see the purkinje cells in the cerebellum (for count), but i don't know the
>better fixation procedure to cut the brain at 10µm max.
>
>For the moment, i froze the brain in isopentane. With this procedure, it's
>really difficult to section it under 25µm, and at this thickness, it's
>pratically impossible to counts the P.Cells, so my question is : is it
>possible to fixed the brain after frozen in isopentane in order to cut it
at
>10µm thickness?
>
>What is the good temperature in cryostat to cut brain tissu?
>
>If i fixed the brain in formalin, what is the procedure, after fixation, to
>cut it in cryostat?
>
>If someone have a good procedure, please help me!!!
>
>bye
>arnaud
>
>ps : sorry for my bad english!!!
>
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