Figure 3
Morphological changes can be tracked over time to distinguish between populations.
(a) Density plot displaying population distributions of cell area versus cell roundness in H3255 cells in the absence (left; n = 1,014 cells) and presence (right; n = 499 cells) of 1 μM erlotinib (E). Cells were segmented based on CellTracker orange intensity thresholds. (b) Nuclear area distribution was tracked over time in H3255 cells with and without 1 μM erlotinib. (c) Top – Representative images of H3255 (red) and CCD-19Lu (blue) classification as compared with labeled fibroblasts (pseudo-stained blue). Bottom Left – Venn diagram illustrates concordance between fluorescent (assumed truth) and morphologic classification of H3255 and CCD-19Lu, which was determined to be 92.6%. Bottom Right - Classification based off of morphological characteristics was evaluated against fluorescence intensity over time and in the presence of erlotinib (1 μM). (d) Cell morphology heterogeneity was evaluated for primary colorectal cancer associated fibroblasts isolated from an individual patient, CAF12347. Parameter values, including cell area, nuclear area, cell roundness and cell width-to-length ratio, were quantitated (n = 41) and cell segmentation mask colors correspond to parameter values for each cell displayed in boxplots. (e) Heterogeneity of cell area across primary colorectal cancer associated fibroblasts isolated from different patients (CAF12347, CAF12380, CAF12415 and CAF12436) was calculated. Scale bars, 200 μm (c) 100 μm (d).
