Figure 2
From: Three-dimensional super-resolution protein localization correlated with vitrified cellular context

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).