CFSE based FACS analysis

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biplabbose
biplabbose's picture
CFSE based FACS analysis

We are doing cell proliferation assay using CFSE statining, followed by FACS analysis as per the protocol published in Nature Protocols.
We are measuring the fluorescence intensity of CFSE stained cells in FL1 channel of the FACS. But am bit confused whether to collect FL1-A data or FL1-H data . As I understand one should collect FL1-A data to avoid the effect of change in cell size during cell cycle.
However, I do see many publications where they have collected FL1-H data and have shown that as a histogram. Most of the publications simply put it as “CFSE intensity”, with out saying FL1-A or FL1-H.
It will be nice if you can give some advice on this.

coltifer
coltifer's picture
As far as I know, people used

As far as I know, people used to collect H data as a default before A was available. A is supposed to be more reliable data. Perhaps the people who published with FL1-H just did not have FL1-A available on their instrument. That's at least a simple answer to your query.

sander164
sander164's picture
We always use Fl1-H and have

We always use Fl1-H and have had no problems with it.