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Analysis of high throughput screen for cell surface markers - (Jan/31/2012 )

Hi,

I'm totally new to the world of flow cytometry data analysis and am looking for some help. I have 4 cell lines that were analyzed by high throughput screen flow cytometry for over 300 CD and other surface molecules. I received an excel file with MFI and %gate+ for each molecule for each cell line. I want to compared the level of expression between the cell lines. My questions are:

1) How do I normalized the data (I do not have a control or anything)
2) Do I use MFI or %gate+ for the anaylsis. For either, what is a good cut-off value (i.e. what is background level?)
3) How do I normalize the data?

Any insight would be greatly appreciated!

-kuby007-

Do all of your cells express the proteins of interest. There might be some negative cell lines and maybe you could use them for comparison.

-scolix-