Fig. 2
Mesoscale heterogeneity detected by the pulse-count analysis. (a–d) Spatiotemporal changes in the cAMP pulse-counts in an area (440 × 440 μm2) containing approximately 200 cells. Mesh size for analysis regions of interest (ROIs), 110 μm. Raw fluorescence images of mRFPmars showing the cell position (a) and schematics ((b–d) left) illustrating the temporal cAMP sequence at the 4th (b), 5th (c), and 10th waves (d) at cell resolution. The number of cAMP pulses experienced at the end of the indicated cAMP wave is color-coded in each cell (pulse-count). For better visibility, cells with a higher pulse-count are shown as a larger dot with a warmer color, whereas non-pulsing cells are shown as gray dots. The gray lines outline the high-pulse cluster in which the collapsing waves propagated. A spatially coarse-grained pulse-count at the ROI level is shown on the right side (b–d). (e) Population-averaged cAMP pulses in green and orange boxes in (a) ROIs with high and low pulse-counts, respectively. (f) Spatial distribution of the pulse-counts at 6:00 in half field of view (FOV). (g) Shapes of the high-pulsing clusters in (f) whose size was 0.39 ± 0.30 mm2 (Mean ± SD). (h) The wave propagation area from 6:00–6:30 roughly corresponds to 1.5 cycles of wave propagation. (i) Superimposition of the high-pulsing cluster at 6:00 (g) and wave propagation area after 6:00 (h). Black: double-negative; light orange: double-positive; purple and dark orange: single-positive for high-pulsing clusters and wave propagation, respectively. Mesh size for analysis ROIs (pixels), 110 μm.
