RE: viability gating

From: Barber, Lesley (PMC) (l.barber@pmci.unimelb.edu.au)
Date: Mon Mar 05 2001 - 18:22:01 EST


I have been using a different compound, Hydroxystilbamidine methanosulfonate
(Fluoro-Gold) from Molecular Probes, for viability staining using the UV
laser. My work was published in Cytometry in 1999 and the research labs at
Peter MacCallum Cancer Institute have been using this compound for more than
two years with good results. Cells sorted using the viable gate from this
procedure have been grown in tissue culture and also  retransplanted with no
adverse results.  See the article:
Fluoro-Gold: An alternative Viability Stain for Multicolor Flow Cytometric
Analysis.
Lesley Barber et al, Cytometry 36:349-354 (1999)
Cheers Lesley
Mrs. Lesley Barber
Cell Manipulation Scientist
Edith Margaret Collie Centre for Blood Cell Therapies
Division of Haematology and Medical Oncology
Peter MacCallum Cancer Institute
St Andrews Place, East Melbourne,
VICTORIA , 3002, AUSTRALIA.
Tel +61 3  9656 1957 or: Page (via switch) +61 3 9656 1111 (pager 1271)
Fax +61 3 9656 1811 Mobile: 0428 977 278


> -----Original Message-----
> From: MSchwa3722@AOL.COM [SMTP:MSchwa3722@AOL.COM]
> Sent: Monday, March 05, 2001 10:37 AM
> To:	Cytometry Mailing List
> 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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