CFSE staining
Biplab Bose
Tuesday, 21 July 2009 13:20 UTC
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.
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Replies
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FL1-A measures the area of the pulse signal, whereas FL1-H measures the height of the pulse signal. In addition, some flow cytometers can measure the width of the pulse signal (eg FL1-W). These measures can apply to all parameters. As long as you exclude doublets from your analysis, FL1-H and FL1-A should be fine to measure your CFSE intensity.
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Thanks Ben!!!
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