Dear Flowers, I have been using Winlist to compensate PeCy5 versus Pe labelled antibodies (CD34-PECy5/CD38-PE) using listmode files and have found it rather difficult to set up the system correctly (using cells labelled with single fluorochromes). The position of the regresion line used to compensate will influence the proportion of double (CD34+CD38hi) and single positive cells (CD34+CD38lo). Similarly we have had problems using a Moflo sorter to compensate (in software) before setting sort gates. This is particularly annoying as we would like to take full advantage of the multiparameter machine. Is this most likely a software or fluorochrome spectral overlap problem? Any advice? Many thanks in advance. Robert Nordon Post Doctoral Fellow Graduate School of Biomedical Engineering University of New South Wales Sydney, 2052 Tel. no. 61-2-9385-1148 Fax 61-2-9663-2108
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