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1、流式细胞多色分析流式细胞多色分析23-13536-00 Rev.01本讲稿第一页,共十页Multicolor Analysis StrategiesPlot typesHierarchical gatingPopulation hierarchiesBack gating223-13536-00 Rev.01本讲稿第二页,共十页Use Different Plot TypesHistogramsusing histograms can help discriminate between two populations.Dot Plot:hard to discriminate populati
2、onsDot Plot:displaying fewer events improves discriminationHistogram:population boundary is evident 323-13536-00 Rev.01本讲稿第三页,共十页Use Different Plot TypesContour Plotsusing contour plots can:Help locate population means.Indicate a need for biexponential scaling.Dot Plot:hard to see populations bunche
3、d on axisContour Plot:shows that biexponential display might be needed Biexponential Contour Plot:gives best display of populations 423-13536-00 Rev.01本讲稿第四页,共十页Hierarchical Gating523-13536-00 Rev.01本讲稿第五页,共十页Hierarchical GatingWhat went wrong here?Correct Hierarchy623-13536-00 Rev.01本讲稿第六页,共十页Back
4、GatingResearcher wants to discriminate Tregs based on CD25 and CD127 expression.Need to ensure a gate includes the entire FOXP3+population.723-13536-00 Rev.01本讲稿第七页,共十页Additional Analysis StrategiesBiexponential scalingEvents to displayHinged and offset quadrant gatesDerived(boolean)gatesSnap-to gat
5、esStatisticsAnalysis templates823-13536-00 Rev.01本讲稿第八页,共十页ICS Data Analysis ResultsInformation 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 cytokinesAnalysis Exercise923-13536-00 Rev.01本讲稿第九页,共十页ICS Panel Analysis1023-13536-00 Rev.01本讲稿第十页,共十页
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