Changes in human embryonic stem cell (hESC) chromatin movement upon differentiation. (A)-(C) Undifferentiated, day-3 differentiated and day-15 differentiated ESC nucleus expressing H2B-GFP, with the chromatin density region tested from each stage of differentiation highlighted by a red box. (D)-(F) Tau phase map of the free NADH content (scale from 2 ns = high free NADH to 5 ns = low free NADH) that surrounds the selected chromatin density region from different stages of differentiation (red box, selected from nuclei (A)-(C)), with the tau phase profile recorded along the actual line scan (black arrow) superimposed. (G)-(I) Intensity carpet of H2B-EGFP fluorescence along the line scan performed across the chromatin density region from the different stages of differentiation. (J)-(L) Auto- and cross correlation analysis of the Gaussian track derived for chromatin movement of the density region centered at two adjacent middle columns in undifferentiated, day-3 differentiated and day-15 differentiated ESC nuclei. (M) Change in amplitude of chromatin vibration in the X-Y plane upon differentiation (N = 5 tracking experiments).