Figure 3

Cellular heterogeneity in 2-D and 3-D environments.

(A) Strategy to analyze cell morphology in 3D collagen matrices. Z-stack images are obtained through sequential imaging at different z positions. A low-pass filter is then applied to individual images followed by maximum z-projection. Cell morphology in projected image is then obtained and subjected to VAMPIRE analysis. (B) Images of PAC01 cells and PAC09 cells in 3-D collagen matrices after z-projection. Nuclei and F-actin are labeled in blue and green. (C,D) Nucleus shapes modes and cell shape modes are identified by VAMPIRE analysis for cells in 3-D matrices. Histograms show nuclear and cellular shape mode distributions for PAC01, PAC03, PAC06 and PAC09 cells. More than 100 cells were analyzed for each sample. (E) Cell morphological heterogeneity properties in both 2D and 3D environments. Strong positive correlation is observed for CV of nucleus size (Spearman’s correlation coefficient ρ=0.8), CV of cell size (ρ=1.0), shape entropy of nucleus (ρ=0.6) and shape entropy of cell (ρ=0.4).