Stepwise Maturation of Lytic Granules during Differentiation and Activation of Human CD8+ T Lymphocytes
Figure 3
Analysis of CTL vesicle physical parameters by flow cytometry.
(A) Upper panel shows size standard beads of 0.5, 1.0 and 4.0 µm used to set-up the SSC and FSC parameters of the flow cytometer. Lower panel shows the design of 4 size-based (FSC) gates (G1 to G4), which borders were placed at the center of distribution of size standard beads. (B) Upper panel shows crude P2 fraction acquired with identical instruments setting used for the size standard beads. The gate “ALL” encompasses the majority of the vesicles. Lower panel shows size based gating analysis of the crude P2 fraction. (C) Doublets were removed by gating on a plot of side-scatter height (SSC-H) versus side-scatter width (SSC-W). Lower panel shows size-based gating analysis of the P2 fraction after the doublet discrimination. Plots corresponding to one representative vesicular extract are shown.