flow cytometry on formaldehyde fixed tissues (GFP detection)

Hi Flowers,

Screening the archive I have seen that the question of doing flow on 
fixed tissues has been brought up a couple of times,
but it is still an open question for me (especially for doing cell 
surface staining or evern intracellular staining on theses tissues for flow)

But here is the specific question:
Can one run flow on formaldehyde fixed tissues that express GFP ?
What is the best way to prepare the single stain suspension ?

>starting with gfp mice:
>At first the mice were anaesthetized then the thoracic cavity was 
>opened and a needle connected to a 40ml syringe wass inserted into 
>the right ventricle for manual perfusion with phosphate buffered 4% 
>formaldehyde solution. A cut in the heart or vena was made to allow 
>outflow of blod and fixative.
>Afterwards the liver, lungs, spleen, kidney, and brain were dissected 
>and have since been store in the same type of fixative as used for 
>the perfusion.
>So what I would like to know is if it is at all possible to prepare 
>single-cell solutions of these organs and run flow to detect the egfp 
>expressing cells.


But of course I am also still interested in hearing something about the possibility of
antibody staining
of formaldehyde fixed tissues for flow.

Many thanks in advance
Alex




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