Dead Cell Marker on 633 nm FL4 ?

Derek Davies davies at icrf.icnet.uk
Fri Nov 8 08:50:19 EST 1996



Hello Matthias,

We have used To-Pro-3 as a dead cell marker but on a FACS Vantage. We have
found that it gives comparable results to using PI. WE get it from
Molecular Probes (Cat No T-3605) and use it at a final concentration of
1uM. Like PI staining of dead cells is rapid. 

Using dual beam excitation (Primary 488, secondary 633 HeNe), we sometimes
see some of the third colour signal in FL4, and although the dead cells
should be brighter than this I can forsee problems. I would also be
interested if anyone has managed this on a 2 laser Calibur (We here in the
UK are still awaiting our upgrades from BD!).

Derek

Derek Davies
FACS Laboratory
Imperial Cancer Research Fund, London, UK.

http://www.icnet.uk/axp/facs/davies/index.html

On Thu, 7 Nov 1996, Matthias Haury wrote:

> 
> Hello me again,
> 
> Those guys out there that have already experience with the FACScalibur 4
> Colors (or equivalent setup on another machine), is there a dye that you
> can use on the FL4 to exclude dead cells, while still using FITC, PE,
> Tricolor for Surface markers ?
> 
> I remember to have heard something about TOPRO etc, but which works the best ?
> 
> Thanks for details (dye, manufacturer, compensations, staining concentrations).
> 
> Merci,
> 
> Matthias
> 
> ______________________________________________________________
> Matthias Haury                      _/_/_/    _/_/_/    _/_/_/
> Flowcytometry - Immunology           _/     _/    _/  _/    _/
> Institut Pasteur Paris              _/     _/_/_/    _/_/_/
> Email: mhaury at pasteur.fr           _/     _/        _/
> Tel:   33 (01) 40 61 31 29      _/_/_/   _/        _/
> Fax:   33 (01) 45 68 86 39      INSTITUT PASTEUR   PARIS
> ______________________________________________________________
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> 
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