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You may be saying right now, what debris? My histogram doesn't have
anything in the channels to the left of the first G1 peak. This could
very well be true as a result of the incorrect setting of the doublet
discrimination gate on the flow cytometry. Many people set their doublet
gate so it starts just below the G1 population and extends just past the
last G2 population. Valuable information is being lost this way. The
doublet gate should be set from just above the origin of the dual
parameter display all the way out to the end of the dual parameter
display.
This way, when you use a sophisticated cell cycle analysis software like
MultiCycle, it looks at the debris information to the left of the first
G1 to calculate how much of the S-phase is debris and how much is real.
Cell fit software is not able to perform this calculation.
MultiCycle was developed at the University of Washington by Dr. Peter
Rabinovitch and is distributed by Phoenix Flow Systems, Inc. It is
available in both PC and MacIntosh versions.
Sincerely yours,
C. Kevin Becker
Phoenix Flow Systems, Inc.
San Diego, CA. USA
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