Doublet Correction for cell cycle analysis

317 ("Phil)
Tue, 22 Aug 1995 17:00:24 -0500 (EST)

I'm currently collaborating with scientists in several projects that analyze
propidium iodide staining of tissue culture cells. We generate standard
DNA-type histograms, normally with G0/G1 CV's under 4 %.

Our question has to do with doublet correction. When we observe the
raw data in (dual parameter) Peak versus Integral fluorescence scatter plots,
we sometimes observe a fair percentage of so-called doublets, i.e., cells that
have 4N amounts of integral fluorescence with 2N amounts of "peak"
fluorescence.

Our question is this: what are the pros and cons of excluding these cells
from our analysis using a gating approach versus doublet correction using
software routines? I know that this subject has been approached in other
forums, but I'd like to hear current thoughts on the matter.

Phil Marder
Lilly Research Labs

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