Re: TO-PRO-3 or something else.

From: Hazel Davey (hlr@aber.ac.uk)
Date: Wed Sep 15 1999 - 07:14:19 EST


Alessandra Nardin wrote:

> 
> Dear all:
> 
> we are searching for a dead-cell exclusion die to be detected in FL-4 (we
> have a 2 laser FACSCalibur). Has any of you had any experience with TO-PRO-3
> (sold by Molecular Probes), so to either suggest or warmly disencourage its
> use? Alternatively, did you come across to some other possible candidate?
> Thanks.

You didn't mention the cell type that you were using, however we've 
had some good results with this dye with bacteria on an Elite 
(633ex / 675 em) and on a Microcyte (635ex / >650em). The 
important things were to keep incubation times short (~1 min) and 
concentration low (0.1 uM) and of course appropiate controls must 
be run. If you are using animal cells the timings / concentrations 
etc may be very different. If you are using plant cells you probably 
won't get a good topro3 signal because of the autofluorescence. 

Hope that helps,

Haze
----------------------------------------------------------
|                  Hazel Davey    hlr@aber.ac.uk         |
|Sefydliad y Gwyddorau Biolegol*Inst. Biological Sciences|
|Prifysgol Cymru               *      University of Wales|
|     ABERYSTWYTH, Ceredigion, CYMRU / WALES SY23 3DD    |
|            http://pcfcij.dbs.aber.ac.uk/index.htm      |



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