Hi David, Assuming you have access to a sorter with the required facilities, my approach would be to sort them into 96-well plates, one cell per well, with no DNA staining at all. (A sort of high-tech alternative to limiting dilution cloning :~) Depending on what your cells are, there may be antibodies that can be used to identify potential candidates or eliminate cells of no interest, with no cytotoxic effects? 30% of wells should contain aneuploid cells, which then can be cultured and later screened. Joseph. At 03:06 20/8/2002, David Chambers wrote: >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] -- Joseph Webster, Flow Cytometry Facility Centenary Institute, Sydney, AUSTRALIA. Phone +61-2-9565-6110
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