You might be able to separate binucleate cells from two mononucleated cells by looking at the DNA signal, showing peak height against area. This would be best attempted on an instrument that has a narrow laser beam, basically those that use crossed cylindrical lenses to focus the laser beam, such as the Beckam Coulter instruments and some of thte Partec/Dako machines. You would still have difficulty in distinguishing binucleates in G1 from mononucleates in G2, unless you also labelled for cyclin B1. If you have access to a Beckman Coulter Elite, you could also try looking at time-of-flight on peak forward scatter. This will usually separate single cells from doublets. Michael Ormerod 34 Wray Park Road Reigate RH2 ODE Telephone: +44 (0)1737 241726 FAX: +44 (0)1737 226736 Mobile telephone: 07802 293242 Message text written by "Antony Bakke" > I have a PI who wants to sort binucleate from mononucleate myocytes. The problem is that some of the mononucleated cells clump together during preparation. Has anyone sorted binucleate cells from clumps of two mononucleated cells? We only have Hoechst staining as a marker at present, since there are no known surface markers. The binucleates are assumed to stop dividing and to only grow by enlarging. The mononucleates still divide. Thank you, Tony Bakke, PhD Director Oregon Cancer Center Flow Cytometry Core<
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