Re: DNA: Double wides but single brights

From: Derek Davies (daviesd2@icrf.icnet.uk)
Date: Fri Oct 01 1999 - 09:17:46 EST


Hi Ray,

You dont say what type of cells you are looking at. If they are large
epithelial-like cells this could be a bit of non-specific cytplasmic
uptake but that is unlikely to be confined to the G1 population. Or do you
see a general width increase as well?

Alternatively it could be FACSRubbish. Are you triggering on FSC or FL2?
If you trigger on FSC and look at the AvW plot does that wide 'G1"
population look as if it is heading off towards the origin? I have seen
this sort of thing when there is a lot of debris or alternatively when the
culture from which the cells are taken has been contaminated.

In either case, it would probably be productive to have a look down the
microscope and see if you can determine what could be causing it.

Hope this helps etc

Derek


Ray Hetser wrote:
> Several times over the last few months when analyzing cells that have
> been stained with propidium iodide we have seen, using pulse processing,
> cells that have an FL2W signal that would seem to qualify them as
> doublets and yet their FL2A is similar to that of a singlet. E.g., in
> the example below, if xxx represent G0/G1 single cells, yyy are G2/M
> single cells, and zzz doublets, what are the cells represented by ???? ?


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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

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