I am analyzing cell cycle data from flow cytometry using PI as a single dye, and I encountered several questions:
(1) Which signals should be used for gating the cells? Some say FSC-SSC, others say PI Height-PI Width, which one is better? The two different gating strategies give quite different results in terms of proportion of cell cycle stages.
(2) Which method should be used to fit the data? Should I use Watson method or Dean-Jett-Fox? If I use the latter one, should I apply S phase synchronous method?
(3) Except for FlowJo, are there any other software/program that are commonly used for analyze flow cytometry data?
Many thanks
Dear Zepeng
FCS Express 6 is the premier flow and image cytometry data analysis software. I also suggest looking at the link below.
http://www.abcam.com/protocols/flow-cytometric-analysis-of-cell-cycle-with-propidium-iodide-dna-staining
Hi. First I use FSC-A vs FSC-H and PI-W vs PI-A to eliminate doblets, then, FSC-A vs SSC-A and the PI histogram (remember that PI should be in linear scale).
I recommend ModFit software for cell cycle analysis. This link is for trial version http://www.vsh.com/products/mflt/mfTrialVersions.asp
The software is very easy and friendly to use
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