流式细胞(FACS)多色分析

Click to edit Master title style,Click to edit Master text styles,Second level,Third level,Fourth level,23-13536-00 Rev. 01,Analysis,*,Multicolor Analysis,23-13536-00 Rev. 01,Multicolor Analysis Strategies,Plot types,Hierarchical gating,Population hierarchies,Back gating,2,23-13536-00 Rev. 01,Use Different Plot Types,Histograms,using histograms can help discriminate between two populations.,Dot Plot: hard to discriminate populations,Dot Plot: displaying,fewer events improves discrimination,Histogram: population boundary is evident,3,23-13536-00 Rev. 01,Use Different Plot Types,Contour Plots,using 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 might be needed,Biexponential Contour Plot: gives best display of populations,4,23-13536-00 Rev. 01,Hierarchical Gating,5,23-13536-00 Rev. 01,Hierarchical Gating,What went wrong here?,Correct Hierarchy,6,23-13536-00 Rev. 01,Back Gating,Researcher wants to discriminate Tregs based on CD25 and CD127 expression.,Need to ensure a gate includes the entire FOXP3,+,population.,7,23-13536-00 Rev. 01,Additional Analysis Strategies,Biexponential scaling,Events to display,Hinged and offset quadrant gates,Derived (boolean) gates,Snap-to gates,Statistics,Analysis templates,8,23-13536-00 Rev. 01,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,9,23-13536-00 Rev. 01,ICS Panel Analysis,10,23-13536-00 Rev. 01,