[Histonet] viability question

[Monfils, Paul]

A reagent we often use in flow cytometry for viability assessment of cells
in suspension is fluorescein diacetate. It quickly permeates cell membranes,
and living cells rapidly convert it into fluorescein through the action of
cytoplasmic esterases.  Fluorescein diacetate is not fluorescent, but
fluorescein is. Therefore if the cells become fluorescent in the presence of
FD, they are alive. I have not tried this with whole tissues, but it's a
quick and easy method to try (assuming of course that you have a
fluorescence microscope available).  Fluorescence is detectable by flow
cytometry within a few seconds of adding the reagent to the cells. It might
take a little longer before fluorescence is directly visible to the eye
because the cytometer is much more sensitive than the human eye is.

> ----------
> From: 	histonet-bounces <@t> lists.utsouthwestern.edu on behalf of Fawn
> Jones
> Sent: 	Monday, June 12, 2006 8:32 AM
> To: 	histonet <@t> lists.utsouthwestern.edu
> Subject: 	[Histonet] viability question
> 
> Dear histonetters,
> 
> I am looking for some information for a friend.  She wants to find out 
> if her tissues (aortas) are live or not.  Does anyone have any 
> experience performing viability stains on tissue samples?  She tried the 
> Molecular Probes Live/Dead kit but found it insufficient due to 
> permeabilty issues. She also suggested MTT, but we're not sure how well 
> that stain works with tissues.  Any help would be greatly appreciated.
> 
> Thank you
> Fawn Jones HT(ASCP)  
> 
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