Figure 7.
Live imaging captures temporal dynamics of morphological states. (a) Live imaging of TC32 cells in the HBV expressing either C shRNA or EF1 shRNA. Sequences were acquired over ∼3 h with 10-min time intervals. XY maximum intensity projections of segmented image pseudo-colored by timepoint demonstrates dynamics of shape change over time. Four representative images of each experimental group are shown. (b) Projection of C shRNA (red, n = 10 cells) or EF1 shRNA (blue, n = 9 cells) timelapse data into the PC space of the fixed cells (gray). The boundaries of C shRNA (dashed line) or EF1 shRNA (solid line)–expressing fixed cells within the HBV, excluding outliers, are indicated for comparison. Slight differences between the fixed and live cell data sets may be due to morphological shifts during fixation and uncontrolled variation in experimental conditions. (c) Relative contribution of PCs explaining the morphological variation of 376 live cell images acquired over all 19 cells. PC3 shows an increased contribution compared to PC3 of the fixed cell analysis (inset, reproduced from Fig. S1 d). (d) PCA of shape features using three PCs in four representative C shRNA (red) or EF1 shRNA (blue) cells shows distinct distributions between the two experimental groups (permutation P value based on the shift in center of mass between 10 C shRNA cells and 9 EF1 shRNA cells <0.001). For each cell, consecutive time points are connected to create a shape change trajectory. (e) Median step between consecutive time points (left) and persistence (right), defined as the net endpoint-to-endpoint displacement divided by the total trajectory length in shape space, for individual cells over time.

Live imaging captures temporal dynamics of morphological states. (a) Live imaging of TC32 cells in the HBV expressing either C shRNA or EF1 shRNA. Sequences were acquired over ∼3 h with 10-min time intervals. XY maximum intensity projections of segmented image pseudo-colored by timepoint demonstrates dynamics of shape change over time. Four representative images of each experimental group are shown. (b) Projection of C shRNA (red, n = 10 cells) or EF1 shRNA (blue, n = 9 cells) timelapse data into the PC space of the fixed cells (gray). The boundaries of C shRNA (dashed line) or EF1 shRNA (solid line)–expressing fixed cells within the HBV, excluding outliers, are indicated for comparison. Slight differences between the fixed and live cell data sets may be due to morphological shifts during fixation and uncontrolled variation in experimental conditions. (c) Relative contribution of PCs explaining the morphological variation of 376 live cell images acquired over all 19 cells. PC3 shows an increased contribution compared to PC3 of the fixed cell analysis (inset, reproduced from Fig. S1 d). (d) PCA of shape features using three PCs in four representative C shRNA (red) or EF1 shRNA (blue) cells shows distinct distributions between the two experimental groups (permutation P value based on the shift in center of mass between 10 C shRNA cells and 9 EF1 shRNA cells <0.001). For each cell, consecutive time points are connected to create a shape change trajectory. (e) Median step between consecutive time points (left) and persistence (right), defined as the net endpoint-to-endpoint displacement divided by the total trajectory length in shape space, for individual cells over time.

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