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A fast- and positively photoswitchable fluorescent protein for ultralow-laser-power RESOLFT nanoscopy
Dhermendra K Tiwari1, Yoshiyuki Arai1, Masahito Yamanaka2
1The Institute of Scientific and Industrial Research, Osaka University, Osaka, Japan.
Abstract:
Fluorescence nanoscopy has revolutionized our ability to visualize biological structures not resolvable by conventional microscopy. However, photodamage induced by intense light exposure has limited its use in live specimens. Here we describe Kohinoor, a fast-switching, positively photoswitchable fluorescent protein, and show that it has high photostability over many switching repeats. With Kohinoor, we achieved super-resolution imaging of live HeLa cells using biocompatible, ultralow laser intensity (0.004 J/cm(2)) in reversible saturable optical fluorescence transition (RESOLFT) nanoscopy.
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