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Flowjo backgating3/30/2023 ![]() To create a colored dot overlay, navigate to the I llustration E ditor, select a plot type of dot a nd turn on the Overlaid button. Blue events were not captured by manual gates, showing shortcomings in the gating strategy) ( An incomplete collection of manually gated populations are overlaid on the corresponding viSNE map of CD45+ PBMC (“Ungated” after viSNE). Here we used viSNE as an example but the same workflow can be applied to other DR algorithms. Pair colored overlay dot plots with viSNE to inform the quality of a manual gating strategy by comparing the location of colored events from gated populations to their location on top of a viSNE map. (A rare signaling population is visualized in a larger context.) Overlay populations to backgate for i mmunophenotyping or to visualize small populations of cells among a greater overall background. See how timepoints stack up or compare donors or doses. In the Cytobank platform, any dimension can be used to create a colored overlay figure. Separate plots can be easily converted into a colored overlay for a different visualization ) ( pSTAT3 expression is compared between different populations gated from PBMC stimulated with IL6 and IL10. The overlaid plots can be colored according to different gated populations or samples within an experiment. A useful tool in the analysis of cytometry data is the ability to overlay plots that would otherwise be viewed separately.
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