Figure 4

Effects of cell cycle and local cell density on cell heterogeneity.

(A,B) htCIP provides versatile single-cell measurements including cell DNA content (A) and local cell density (ρ_D) (B), for cells on 2D substrates. (C,D) Relation between cell morphology and progression of cell cycle (C) and increase in local cell density (D) for PAC01, PAC03, PAC06 and PAC09. Cells were sorted based on their DNA content and divided into 9 groups with equal sample size. Probability of cells found in different nucleus and cell shape modes (P(NS_k), P(CS_k)) at these groups are shown in heat maps. The averaged DNA contents, normalized nucleus size and normalized cell size for these groups are shown in the plot next to the heat maps. Nucleus and cell size are normalized by dividing the lowest value among all groups. The same procedures are used to show effect of local cell density on cell morphology. (E,F) Nucleus and cell shape modes, filled with colors to show correlation between DNA content and occurrence of individual shape modes (E). Color coding from red to green corresponds to Pearson’s correlation coefficient from −1 to 1. The same procedure is applied to show correlation between local cell density and occurrence of shape modes (F,G) Overall heterogeneity profiles of cell morphology of PAC samples in the G₀/G₁, S and G₂/M phases. (H) Overall heterogeneity profiles of cell morphology of PAC samples under three local density conditions, including singlet (ρ_D = 0) semi-crowded (0 < ρ_D < 4) and crowded (ρ_D ≥ 4). The heterogeneity of these samples is shown in a heat map where colors from red to green indicate an increasing degree of heterogeneity.