posted on 2016-08-19, 00:00authored bySheng Wang, Xuanze Chen, Lei Chang, Ruiying Xue, Haifeng Duan, Yujie Sun
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.