[Histonet] brain fixation and native EGFP fluorescence
cbass <@t> bidmc.harvard.edu
Tue Jan 3 21:24:34 CST 2006
I have been using a viral vector to express EGFP in the mouse liver
and examining native fluorescence. I generally fix the tissue by
cutting the liver into blocks, immersing in NBF and sectioning on a
cryostat. I have gotten some really good results just looking at
native fluorescence. I am hoping to extend this to the brain. I
have now injected my viral vector into the striatum of a mouse and am
set to collect the tissue. Since I am working with the brain, my
natural tendency is to perfuse, as I have always heard this is the
best method of fixing brain tissue.
Does anyone have advice in terms of picking a good fixative for
perfusion? I have been reading about a lot of different fixatives
for the brain, one which includes sucrose. I am familiar with
sucrose cryopreservation, but I haven't considered adding it to the fix.
Any suggestions would be appreciated. If you have a suggestion
please include the formula. Someone recently suggested a
paraformaldehyde zinc fix to me. What are the advantages of this
paraformaldehyde alone and does anyone have a formulation for it?
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