Figure 9.
Figure 9. Detection of significant transitions in GC morphology along a neurite outgrowth trajectory. (A) Time series modeling of a DMSO- (left and Video 9) and CK666 (right and Video 10)-treated GC movie in reduced morphology space. Colors: state ID inferred from the HMM/Bayesian model selection. Rings: 1× and 2× the SD of the state distribution centered on the mean. Bottom insets: Plot of detected transition within a 90-s window after respective treatment. (B) CK666 treatment induces reproducible GC morphology state transitions. (i) Plots of detected morphology transitions for six movies (as described in A, bottom inset) 90 s after treatment with DMSO (gray/black shaded vectors) or 25 µM CK666 (blue shaded vectors). (ii) Magnitude of the transition vectors in i. *, P < 0.05, two-tailed t test for equal means. (C) Median of feature distribution for each detected HMM morphology state detected in A (DMSO, left; CK666, right). *, P < 0.05; **, P < 0.01; ***, P <0.001: Kruskal–Wallis test. Insets: Boxplots of the distribution of filopodia lengths in each HMM state. *, P < 0.05; **, P < 0.01; ***, P < 0.001: Tukey–Kramer test. (D) Identification of morphology/motility switches in a heterogeneous population of unperturbed trajectories. (i) Three unperturbed GC movies overlaid on the morphological landscape, each frame colored by neurite outgrowth state as in Fig. 8 E, i. Identification of morphology/motility switch (starred black arrow) similar to that induced upon acute treatment with CK666 (A–C). Black rings: HMM states as defined in A. Colors as in D, i. (iii) “Bag” contours as defined in Fig. 8 D, iii, for GC movies in i. Arrows: Detected morphology state transitions from B and D, ii. Each movie: 10 min total, 5-s intervals. (E) Zoom (left) and neurite outgrowth velocity (right) of unperturbed GC movie identified in D, ii, frame colored by neurite outgrowth state (top) or HMM state ID as defined in A (bottom). (F) Feature/HMM state heat maps (as defined in C) for unperturbed movie identified in D, ii (top). Example GCA segmentations for each HMM morphology state (bottom).

Detection of significant transitions in GC morphology along a neurite outgrowth trajectory. (A) Time series modeling of a DMSO- (left and Video 9) and CK666 (right and Video 10)-treated GC movie in reduced morphology space. Colors: state ID inferred from the HMM/Bayesian model selection. Rings: 1× and 2× the SD of the state distribution centered on the mean. Bottom insets: Plot of detected transition within a 90-s window after respective treatment. (B) CK666 treatment induces reproducible GC morphology state transitions. (i) Plots of detected morphology transitions for six movies (as described in A, bottom inset) 90 s after treatment with DMSO (gray/black shaded vectors) or 25 µM CK666 (blue shaded vectors). (ii) Magnitude of the transition vectors in i. *, P < 0.05, two-tailed t test for equal means. (C) Median of feature distribution for each detected HMM morphology state detected in A (DMSO, left; CK666, right). *, P < 0.05; **, P < 0.01; ***, P <0.001: Kruskal–Wallis test. Insets: Boxplots of the distribution of filopodia lengths in each HMM state. *, P < 0.05; **, P < 0.01; ***, P < 0.001: Tukey–Kramer test. (D) Identification of morphology/motility switches in a heterogeneous population of unperturbed trajectories. (i) Three unperturbed GC movies overlaid on the morphological landscape, each frame colored by neurite outgrowth state as in Fig. 8 E, i. Identification of morphology/motility switch (starred black arrow) similar to that induced upon acute treatment with CK666 (A–C). Black rings: HMM states as defined in A. Colors as in D, i. (iii) “Bag” contours as defined in Fig. 8 D, iii, for GC movies in i. Arrows: Detected morphology state transitions from B and D, ii. Each movie: 10 min total, 5-s intervals. (E) Zoom (left) and neurite outgrowth velocity (right) of unperturbed GC movie identified in D, ii, frame colored by neurite outgrowth state (top) or HMM state ID as defined in A (bottom). (F) Feature/HMM state heat maps (as defined in C) for unperturbed movie identified in D, ii (top). Example GCA segmentations for each HMM morphology state (bottom).

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