Sorting live cells for DNA content?

From: David Chambers (davidc@ccmi.salk.edu)
Date: Mon Aug 19 2002 - 12:06:33 EST


Hello flowers!

I have an investigator who has a population of mouse cells containing about 30% aneuploid
cells, some with one chromosome missing and some with an extra one.  He wants to sort
(or otherwise process) the cells to get a relatively pure population of these two types.
He wants to keep them alive and culture them afterwards.

How feasible is this, in your opinion?  Apart from using H33342, are there any
other/better stains? - my investigator is concerned about the potential toxicity of
Hoechst; I understood that it wasn't particularly toxic, but maybe I'm wrong?

Yes, I have suggested limiting dilution cloning... this was met with a resounding
silence! (also, a "clonal" population might not be particularly good, in this case).

All advice gratefully received, grovel, grovel :-)

- David

[davidc@ccmi.salk.edu]
[Salk Institute Flow Cytometry lab, La Jolla, CA, USA]



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