CCAST gating strategy on SUM159 breast cancer cell line.

CCAST gating strategy for SUM159 breast cancer cell lines isolates 5 pure cell states (across 9 bins) based on CD24 and EPCAM. Visualization of these 5 subpopulations is clearly not apparent from the biaxial side scatter (SSC) vs. biomarker plots. Split point estimates (dotted red lines) go through density contour plot (orange) on the distributed data providing visual evidence for suitable cut-offs through bimodal contours. Note the split point lines for nodes 3 and 4 concentrate on the zero point mass; this indicates there are several cells with zero expression values for EPCAM or CD24 staining but with higher expression values with respect to CD44.