[Histonet] brain fixation and native EGFP fluorescence

From:Caroline Bass

Hello Everyone,

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?


Caroline Bass

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