FACS: excluding dead cells?

Derek Davies derek.davies at icrf.icnet.uk
Wed Jan 5 04:35:00 EST 2000


In article 
<Pine.A41.4.05.9912210913490.47780-100000 at node9.unix.Virginia.EDU>, 
Lionel  Berthoux <lb8x at node9.unix.Virginia.EDU> wrote:

> I am FACscanning cells using GFP + a PE-labeled Ab. I would like to
> exclude dead cells. Do you know of any reagent that would be useful in
> that case? (I think 7AAD is not compatible with the use of PE)

Hi Lionel,

7-AAD will actually do the job as it emits much further out in the red 
than PI although you will still need to compensate it out of the PE 
channel and vice versa. If you have access to a dual laser FACSCalibur 
though you can use TO-PRO-3 from Molecular Probes which is 633nm 
excitable and emits at about 660nm and then there are no compensation 
problems.

It may be worth searching the Cytometry mailing list archives at Purdue 
University at:
http://www.cyto.purdue.edu/hmarchiv/index.htm

Hope this helps!

Derek




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