Figure 3.

Time-lapse imaging of IPS-CM following EB disaggregation. (A,B) Brightfield images of IPS-CM populations taken over a period of 48 h following the disaggregation of 0- and 3-week old EBs [(A,B), respectively]. Images shown correspond to 6 h intervals. The arrow in (A) (0 h) shows an IPS-CM cluster which we would typically select for Ca²⁺ imaging in this study. The full image sequence (with images taken every 20 min) corresponding to (A,B) are given in Supplementary Movies 6, 7, respectively. The white box in the 48 h panel in (A) depicts the area in which we recorded post-disaggregation cell division. The full 48 h time-lapse series of this event is given in Supplementary Movie 8. (C) The number of cells per field of view (FOV, 0.625 mm²) was counted in each frame of the time series. The net migration of cells out of and into a FOV over the course of 48 h was balanced. Data were fitted with non-linear regression lines of best fit and tau and r²-values are given. In this example, the number of adherent IPS-CM from 3-week old EBs is reduced by 35.91 ± 0.10 % (mean ± SE) between 12 and 48 h post-seeding when compared to those disaggregated from 0-week old EBs.