CCAST: A Model-Based Gating Strategy to Isolate Homogeneous Subpopulations in a Heterogeneous Population of Single Cells
Figure 3
CCAST applied to single cell analysis of T-cells.
A The CCAST gating strategy based on the unlabeled T-cell data in Figure 2, post filtering, showing that 4 cell types can be derived using only Marker 5 and Marker 2 with Marker 5 as the root node. Split points along with the minimum-maximum range for each split point are provided at each node. B Histogram plots for sample split point for each node is obtained via bootstrapping. The multi modal nature of the distributions makes it difficult to calculate a true confidence intervals on the split point estimates. C CCAST result without filtering represented as a 2D scatter plot of the 4 cell types, which each cell type color coded; note that gating the yellow-colored cells will likely result in contamination of green-colored cells. D CCAST result with filtering represented as a 2D scatter plot of the 4 pure cell types, with each cell type color coded. Note all contaminating cells mixed with various clusters have been removed. For manual gating purposes, comparing the two schemes C and D provides a visual evaluation of the expected contamination levels from sorting subpopulations. E CCAST gating strategy for all Tcell types with labels reveals that the key gating markers are CD4 and CD45RA markers. F 2D scatter plot for the four, labeled T-cell types based on CD4 and CD45RA.