Hi Alessandra, I have quite a bit of experience using TO-PRO-3 and have found it to be a very versatile dye. On the Calibur we regularly use it as a dead cell dye in experiments where the use of PI is excluded ie when PE or PE/Cy5 is used. I find that it is maybe not quite as bright as PI but does label the same cells. You can also use very low concentrations - something in the order of 500nM (Be careful as the stock is made in DMSO). It can also be used as a quantitative DNA dye, although again not quite as good as PI, it can be used in certain situations where PI cant. It does however have some interaction with BRdU which can be used to advantage (see Tom Frey's paper in Cytometry 17, 310). One caveat I have with it is that when we use it on our sorter when we are using significantly higher laser powers we do see some excitation of TO-PRO at 488nm which means that it comes into the FL3 channel. However, I'd say, give it a go! Best wishes, Derek On Mon, 13 Sep 1999, Alessandra Nardin wrote: > 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. ************************************************************************ Derek Davies Voice: (44) 0171 269 3394 FACS Laboratory, FAX: (44) 0171 269 3100 Imperial Cancer Research Fund, e_mail: derek.davies@icrf.icnet.uk London, UK Web Page: http://www.icnet.uk/axp/facs/davies/index.html In tenebris lux *************************************************************************
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