[Histonet] RE: Histogel embedding mouse brain for cryotomy or vibratome

[Bob Nienhuis]

Currently doing Cryotomy on quickly frozen Golgi-Cox impregnated mouse
brains
at 100-120 microns. Cryoprotecting with 30% sucrose. Cutting at -15C.
Tried fresh tissue and brief NBF perfusion(5 min).
If I perfuse with NBF for 15 min,  or immersion fix overnight, it seems to
destroy
spines and create artifact- possibly glial cells. Cutting much easier
though!

Might Histogel help?

Bob Nienhuis
UCLA / VA Medical Center


On Tue, Jan 13, 2009 at 2:10 PM, gayle callis <gayle.callis <@t> bresnan.net>wrote:

> Bob,
>
> So which type of sectioning are you doing now? Cryotomy or vibratome? And
> how thick do you want the sections to be?  If frozen sections on snap
> frozen
> mouse brain, simply turning the temperature up to -16C or so will make
> sectioning much easier and get rid of the shredding, friable sections.
> Also, are you fixing then sucrose cryoprotecting after NBF or 4%
> paraformaldehyde fixation of the brain (can be either perfused or immersion
> fixation)?
>
> Gayle Callis
> HTL(ASCP)HT,MT
> Bozeman MT
>
>
>
>
>
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