流式细胞(FACS)多色分析.ppt

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1、23-13536-00 Rev. 01,Multicolor Analysis,23-13536-00 Rev. 01,2,Multicolor Analysis Strategies,Plot types Hierarchical gating Population hierarchies Back gating,23-13536-00 Rev. 01,3,Use Different Plot Types,Histogramsusing histograms can help discriminate between two populations.,Dot Plot: hard to di

2、scriminate populations,Dot Plot: displaying fewer events improves discrimination,Histogram: population boundary is evident,23-13536-00 Rev. 01,4,Use Different Plot Types,Contour Plotsusing contour plots can: Help locate population means. Indicate a need for biexponential scaling.,Dot Plot: hard to s

3、ee populations bunched on axis,Contour Plot: shows that biexponential display might be needed,Biexponential Contour Plot: gives best display of populations,23-13536-00 Rev. 01,5,Hierarchical Gating,23-13536-00 Rev. 01,6,Hierarchical Gating,What went wrong here?,Correct Hierarchy,23-13536-00 Rev. 01,

4、7,Back Gating,Researcher wants to discriminate Tregs based on CD25 and CD127 expression. Need to ensure a gate includes the entire FOXP3+ population.,23-13536-00 Rev. 01,8,Additional Analysis Strategies,Biexponential scaling Events to display Hinged and offset quadrant gates Derived (boolean) gates

5、Snap-to gates Statistics Analysis templates,23-13536-00 Rev. 01,9,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,23-13536-00 Rev. 01,10,ICS Panel Analysis,