title: GMars-Q Enables Long-Term Live-Cell Parallelized Reversible Saturable Optical Fluorescence Transitions Nanoscopy.
abstract: The recent development of reversibly switchable fluorescent proteins (RSFPs) has promoted reversible saturable optical fluorescence transitions (RESOLFT) nanoscopy as a general scheme for live-cell super-resolution imaging. However, continuous, long-term live-cell RESOLFT nanoscopy is still hindered mainly because of the unsatisfactory properties of existing RSFPs. In this work, we report GMars-Q, a monomeric RSFP with low residual off-state fluorescence and strong fatigue resistance attributed to a biphasic photobleaching process. We further demonstrate that GMars-Q is particularly suitable for long-term parallelized RESOLFT nanoscopy as it supports an order of magnitude longer imaging durations than existing RSFPs. The excellent photophysical properties of GMars-Q also suggest that it would be of general interest for other RESOLFT nanoscopic methods.
authors.affiliation: State Key Laboratory of Membrane Biology, Biodynamic Optical Imaging Center (BIOPIC), School of Life Sciences and ‡Department of Biomedical Engineering, College of Engineering, Peking University , Beijing 100871, China.
authors.forename: Sheng
authors.initials: S
authors.lastname: Wang
authors.affiliation: State Key Laboratory of Membrane Biology, Biodynamic Optical Imaging Center (BIOPIC), School of Life Sciences and ‡Department of Biomedical Engineering, College of Engineering, Peking University , Beijing 100871, China.
authors.forename: Xuanze
authors.initials: X
authors.lastname: Chen
authors.affiliation: State Key Laboratory of Membrane Biology, Biodynamic Optical Imaging Center (BIOPIC), School of Life Sciences and ‡Department of Biomedical Engineering, College of Engineering, Peking University , Beijing 100871, China.
authors.forename: Lei
authors.initials: L
authors.lastname: Chang
authors.affiliation: State Key Laboratory of Membrane Biology, Biodynamic Optical Imaging Center (BIOPIC), School of Life Sciences and ‡Department of Biomedical Engineering, College of Engineering, Peking University , Beijing 100871, China.
authors.forename: Ruiying
authors.initials: R
authors.lastname: Xue
authors.affiliation: State Key Laboratory of Membrane Biology, Biodynamic Optical Imaging Center (BIOPIC), School of Life Sciences and ‡Department of Biomedical Engineering, College of Engineering, Peking University , Beijing 100871, China.
authors.forename: Haifeng
authors.initials: H
authors.lastname: Duan
authors.affiliation: State Key Laboratory of Membrane Biology, Biodynamic Optical Imaging Center (BIOPIC), School of Life Sciences and ‡Department of Biomedical Engineering, College of Engineering, Peking University , Beijing 100871, China.
authors.forename: Yujie
authors.initials: Y
authors.lastname: Sun
keywords: GMars-Q; biphasic photobleaching; fluorescent protein; live-cell imaging; parallelized RESOLFT; reversible photoswitching; super-resolution fluorescence microscopy