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