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1、Multicolor Analysis,2,Multicolor Analysis Strategies,Plot types Hierarchical gating Population hierarchies Back gating,3,Use Different Plot Types,Histogramsusing histograms can help discriminate between two populations.,Dot Plot: hard to discriminate populations,Dot Plot: displaying fewer events imp
2、roves discrimination,Histogram: population boundary is evident,4,Use Different Plot Types,Contour Plotsusing contour plots can: Help locate population means. Indicate a need for biexponential scaling.,Dot Plot: hard to see populations bunched on axis,Contour Plot: shows that biexponential display mi
3、ght be needed,Biexponential Contour Plot: gives best display of populations,5,Hierarchical Gating,6,Hierarchical Gating,What went wrong here?,Correct Hierarchy,7,Back Gating,Researcher wants to discriminate Tregs based on CD25 and CD127 expression. Need to ensure a gate includes the entire FOXP3+ po
4、pulation.,8,Additional Analysis Strategies,Biexponential scaling Events to display Hinged and offset quadrant gates Derived (boolean) gates Snap-to gates Statistics Analysis templates,9,ICS Data Analysis Results,Information that can be retrieved: Number of collected events, number of live CD3+ cells % of main populations: %CD3+, CD4+, CD8+ % of nave (CD45RA+) and memory (CD45RA) CD4+ and CD8+ cells % CD25+CD4+ cells (Tregs) For the cytokines: % of CD4+ and CD8+making 1 or 2 cytokines,Analysis Exercise,10,ICS Panel Analysis,