About Flow Cytometry Gating: From Scatter Plots to Spectral Panels

You will learn to read a flow cytometry dot plot the way an immunologist does: what forward and side scatter physically measure, how to exclude debris, doublets, and dead cells before any marker is interpreted, and how to build a defensible hierarchical gating strategy. You will understand why spillover exists, how compensation and modern spectral unmixing correct it, and which control (unstained, single-stain, isotype, FMO) is actually allowed to set a gate boundary. You will finish able to justify every gate on a plot, validate it by backgating, judge whether a rare-event frequency is statistically real, and design a panel that keeps spillover spreading error off your dimmest marker.

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