Fine-grained, nonlinear registration of live cell movies reveals spatiotemporal organization of diffuse molecular processes
Fig 4
Analysis of spatial coherence in diffuse molecular signals.
a) Select frames of a movie visualizing the activation of the GTPase Cdc42 (middle) and its upstream GEF Asef (bottom) using multiplex FRET-based biosensors. Shown are normalized FRET-ratio signal indicating local activation of the molecues. For registration the donor signal of the Cdc42 biosensor is used as the location fiducial (top). b) Remapped, normalized FRET-ratio signals for the same time points as in a). c) Spatial coherence of the Cdc42 donor (left), Cdc42 FRET-ratio (center), and Asef FRET-ratio (right) time-series extracted from the remapped (top) and raw time-lapse sequences. d) Distributions of average spatial coherence of the Cdc42 donor, Cdc42 FRET-ratio, and Asef FRET-ratio time-series. Box plots illustrate 25th, 50th, and 75th percentile of average values in for m = 4 movies. Whiskers indicate the 5th and 95th percentile. P-values are calculated by one-way ANOVA testing. Scale bars 10 μm.