Figure 5. SHG anisotropy measurement of cardiomyocytes differentiated from genetic disease patient-derived hiPSCs.

(A) A schematic drawing of the experimental procedure. (B) A typical example of dynamic measurement of the sarcomere movement of parameter γ with 80 ms time resolution in Mybpc3^t/+ (top), Mybpc3^t/t (middle), and Mybpc3^+/+ cells (bottom). Right panels are histograms of γ-value in each trace. Solid lines in the right panels are fitting results with double Gaussian distributions. Magenta lines indicate γ_cont and orange lines indicate γ_rest in each trace obtained by the fitting. Asterisk indicates the occasional change of the beating cycle. (C) Violin plots of γ_rest (left) and γ_cont (right) for Mybpc3^t/+ (red, N = 21 FOVs), Mybpc3^t/t (blue, N = 36), and Mybpc3^+/+ (green, N = 28). (D) Violin plots of dγ_ave = γ_cont - γ_rest in Mybpc3^t/+ (red, N = 21), Mybpc3^t/t (blue, N = 36), and Mybpc3^+/+ (green, N = 28). (E) Bar graphs of SL in the resting state (filled) and in the contracting state (opened) (upper) and contraction yield (lower). Error bars are SD. (F) Relationship between SL_rest or SL_cont and γ_rest or γ_cont for Mybpc3^t/+ (top), Mybpc3^t/t (middle) and Mybpc3^+/+ (bottom). Filled and opened marks are for the resting state and for the contracting state, respectively. Solid line is a line linked between averaged value of data for the resting state and for the contracting state. Overlapped gray marks and line in middle and bottom panels are those for Mybpc3^t/+. (G) Averaged time courses of γ-value (left) and sarcomere motion (right) and average trace of a pulsation in all analyzable data for Mybpc3^t/+ cell (red, N = 8), Mybpc3^t/t cell (blue, N = 13), and Mybpc3^+/+ cell (green, N = 6). Light colors are SD. (C, D, E) Asterisks in (C, D, E) indicate less than 0.01 of the P-value in the t test.