title: A fast- and positively photoswitchable fluorescent protein for ultralow-laser-power RESOLFT nanoscopy.
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.
authors.affiliation: The Institute of Scientific and Industrial Research, Osaka University, Osaka, Japan.
authors.forename: Dhermendra K
authors.initials: DK
authors.lastname: Tiwari
authors.affiliation: The Institute of Scientific and Industrial Research, Osaka University, Osaka, Japan.
authors.forename: Yoshiyuki
authors.initials: Y
authors.lastname: Arai
authors.affiliation: Department of Applied Physics, Osaka University, Osaka, Japan.
authors.forename: Masahito
authors.initials: M
authors.lastname: Yamanaka
authors.affiliation: The Institute of Scientific and Industrial Research, Osaka University, Osaka, Japan.
authors.forename: Tomoki
authors.initials: T
authors.lastname: Matsuda
authors.affiliation: The Institute of Scientific and Industrial Research, Osaka University, Osaka, Japan.
authors.forename: Masakazu
authors.initials: M
authors.lastname: Agetsuma
authors.affiliation: The Institute of Scientific and Industrial Research, Osaka University, Osaka, Japan.
authors.forename: Masahiro
authors.initials: M
authors.lastname: Nakano
authors.affiliation: Department of Applied Physics, Osaka University, Osaka, Japan.
authors.forename: Katsumasa
authors.initials: K
authors.lastname: Fujita
authors.affiliation: The Institute of Scientific and Industrial Research, Osaka University, Osaka, Japan.
authors.forename: Takeharu
authors.initials: T
authors.lastname: Nagai
mesh_terms: D006367:HeLa Cells; D006801:Humans; D007834:Lasers; D008164:Luminescent Proteins / Q000737:chemistry*; D008969:Molecular Sequence Data; D036103:Nanotechnology / Q000295:instrumentation* / Q000379:methods*
chemical_list: D008164:Luminescent Proteins