Figure 2 | Scientific Reports

Figure 2

From: Three-dimensional super-resolution protein localization correlated with vitrified cellular context

Figure 2

Photophysical properties of Dronpa at CT.

(a) Photoswitching capability of Dronpa at CT (113 K). Purified Dronpa proteins were first switched off by a 488 nm laser, followed by alternative illumination with 405 nm and 488 nm lasers. Each purple arrow indicates a 5 s 405 nm laser pulse. (b) Fluorescence decay kinetics of Dronpa at RT and CT under the irradiation of a 1.5 kW·cm−2 488 nm laser. The decay curves at RT and CT were both fitted with a bi-exponential function27,28. The time constants/amplitudes at RT are 0.06 s/0.87 (fast phase) and 2.37 s/0.04 (slow phase) and the corresponding values at CT are 0.87 s/0.37 (fast phase) and 8.32 s/0.33 (slow phase). Please note that at RT, the amplitude of the slow phase is very small (4%), indicating fast bleaching and off-switching mainly in the fast phase (c,d). (c) shows the evolution of the emission spectra of Dronpa subjected to a temperature gradient from 293 K to 143 K, (d) is the normalized presentation and the inset is the magnified view of the red area in (d).

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