流式细胞(FACS)多色分析课件.ppt

《流式细胞(FACS)多色分析课件.ppt》由会员上传分享，免费在线阅读，更多相关内容在教育资源-天天文库。

1、MulticolorAnalysis23-13536-00Rev.01MulticolorAnalysisStrategiesPlottypesHierarchicalgatingPopulationhierarchiesBackgatingUseDifferentPlotTypesHistograms—usinghistogramscanhelpdiscriminatebetweentwopopulations.DotPlot:hardtodiscriminatepopulationsDotP

2、lot:displayingfewereventsimprovesdiscriminationHistogram:populationboundaryisevidentUseDifferentPlotTypesContourPlots—usingcontourplotscan:Helplocatepopulationmeans.Indicateaneedforbiexponentialscaling.DotPlot:hardtoseepopulationsbunchedonaxisContourP

3、lot:showsthatbiexponentialdisplaymightbeneededBiexponentialContourPlot:givesbestdisplayofpopulationsHierarchicalGatingHierarchicalGatingWhatwentwronghere?CorrectHierarchyBackGatingResearcherwantstodiscriminateTregsbasedonCD25andCD127expression.Needto

4、ensureagateincludestheentireFOXP3+population.AdditionalAnalysisStrategiesBiexponentialscalingEventstodisplayHingedandoffsetquadrantgatesDerived(boolean)gatesSnap-togatesStatisticsAnalysistemplatesICSDataAnalysisResultsInformationthatcanberetrieved:Num

5、berofcollectedevents,numberofliveCD3+cells%ofmainpopulations:%CD3+,CD4+,CD8+%ofnaïve(CD45RA+)andmemory(CD45RA–)CD4+andCD8+cells%CD25+CD4+cells(Tregs)Forthecytokines:%ofCD4+andCD8+making1or2cytokinesAnalysisExerciseICSPanelAnalysis