Hi! I'm currently doing flow cytometry samples and looking at surface proteins. I'm using FITC for fluorescence, however, the histograms are hard to interpret. The curve seems ''shifted to the right'' rather than a ''clear positive population''. I therefore struggle to draw the line between positive and negative cells (compared to unstained). Does anyone have any ideas?
Picture for reference to the issue.
Thanks a lot in advance!
I encountered the same issue as you. I found a method involving Mean Fluorescence Intensity (MFI) calculation, but I'm unsure about its reliability. Have you managed to resolve this problem?
Linpeng Li I typically use median fluorescence Intensity instead of mean. I'm still searching for ways to better identify ''positive cells'' from unstained/untreated. One way I read is to use the 95th percentile value of FITC from the untreated/unstained and ''draw'' the positive population from there. However, not sure what the ''standard'' is.
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