Hello,
It shouldn't be much of an issue for this live/dead application.
The stuff you will be gating out is bright, while the viable cells are
very dim, so you won't have any bright DAPI cells to worry about.
Spectral overlap would be something to be concerned about if
you were using the DAPI/FITC combination for cell cycle/immunofluorescence.
Peter Lopez
The Aaron Diamond AIDS Research Center
212.448.5188 (office)
212.448.5159 (fax)
212.448.5190 or 5110 (lab)
MSchwa3722@AO
L.COM To: Cytometry Mailing List
<cytometry@flowcyt.cyto.purdue.edu>
03/04/2001 cc:
06:36 PM Subject: viability gating
I'm thinking of trying DAPI in the UV channel for live/dead gating instead
of
using 7-AAD in FL3, in order to free FL3 for three color gating and sorting
strategies, on our FACS Vantage. My question is: Is there any spectral
overlap/interference with the standard FL1, FL2 or FL3, ie. FITC, PE and
PE-Cy5 or any other considerations I should be thinking of to do live/dead
gating with DAPI?
Your assistance, as always, is appreciated.
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