[Histonet] Evans Blue

[Jaume del Valle]

Dear Histomembers
 
Some months ago I've trying to work with Evans blue. My problem is that I inject it (IV or IP) and when I want to see the fluorescence of the dye in the brain there is a lot of... Of red signal here and there. 
I've read that the tissue have to be fixed (which fixative is the best? At which temperature? How much time? Do I have to let the sample to air-dry? Some minutes? Hours? The animal have to be perfused? With PFA? Saline is OK? 
 
When I let to dry at Room temperature the signal spreads all over the sample, when I use Mowiol to Mount the slide it seems that the evans blue gets disolved...
 
Have anybody worked with Evans blue and Brain slides to see extravastion? Please, give me the (whole) details!!!
 
Jaume del Valle
Universitat de Barcelona
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