Two-photon STED nanoscopy realizing 100-nm spatial resolution utilizing high-peak-power sub-nanosecond 655-nm pulses | Zendy

Hirokazu Ishii | Zendy; Kohei Otomo | Zendy; Jui-Hung Hung | Zendy; Motosuke Tsutsumi | Zendy; Hiroyuki Yokoyama | Zendy; Tomomi Nemoto | Zendy

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Two-photon STED nanoscopy realizing 100-nm spatial resolution utilizing high-peak-power sub-nanosecond 655-nm pulses

Author(s) -

Hirokazu Ishii,

Kohei Otomo,

Jui-Hung Hung,

Motosuke Tsutsumi,

Hiroyuki Yokoyama,

Tomomi Nemoto

Publication year - 2019

Publication title -

biomedical optics express

Language(s) - Uncategorized

Resource type - Journals

SCImago Journal Rank - 1.362

H-Index - 86

ISSN - 2156-7085

DOI - 10.1364/boe.10.003104

Subject(s) - sted microscopy , nanosecond , photobleaching , stimulated emission , optics , materials science , image resolution , temporal resolution , microscopy , optoelectronics , resolution (logic) , picosecond , fluorescence , laser , physics , computer science , artificial intelligence

We developed two-photon excitation stimulated emission depletion (STED) nanoscopy using high-peak-power sub-nanosecond 655-nm pulses. The STED pulse exhibited ideal optical properties and sufficient pulse energy to realize a 70-nm spatial resolution in the compact setup with electrically controllable components. For biological applications, we screened suitable fluorescent dyes or proteins and realized the sub-100 nm spatial resolution imaging of presynaptic protein clusters in fixed primary cultured neurons without severe photobleaching. We expect this method to enable visualization of ultrastructures and the cluster dynamics of biomolecules representing physiological functions in living cells and tissue.

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