Figure 3.
Quantification of irreversible photobleaching and dark-state isolation. (a) Qfocal normalized to the value at onset of interphase 14 in two embryos, one imaged once per 45 s (bullets) and one imaged once per 90 s (squares). t is in minutes. Each time point represents a dark conversion and reactivation series of 24 scans at 496 nm and three scans at 405 nm. (b) Histogram of difference in changes in Qfocal between the embryos in a. δ = 2 for the more frequently imaged embryo and δ = 1 for the less frequently imaged embryo. The quantity plotted is for all n. The mean value is –0.005±0.006, where 0.006 is the standard error. (c) Sfocal shown for three embryos (bullets) extracted from individual images. Arrows indicate 405-nm photoactivation pulses, occurring between consecutive images. , where is taken immediately preceding the 405-nm pulse, and is taken immediately after. A positive value of would indicate maturation of new Dronpa-Bcd into the dark state. t is in minutes. (d) Distribution of over all embryos.