流式细胞FACS多色分析PPT课件

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 even

2、ts improves 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 di

3、splay might 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 en

4、tire FOXP3+ population.,.,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+ an