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

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

doi:10.1364/BOE.10.003104

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

Hirokazu Ishii, Kohei Otomo, Jui Hung Hung, Motosuke Tsutsumi, Hiroyuki Yokoyama, Tomomi Nemoto

Research output: Contribution to journal › Article › peer-review

4 Citations (Scopus)

Abstract

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.

Original language	English
Article number	360910
Pages (from-to)	3104-3113
Number of pages	10
Journal	Biomedical Optics Express
Volume	10
Issue number	7
DOIs	https://doi.org/10.1364/BOE.10.003104
Publication status	Published - 2019 Jul
Externally published	Yes

ASJC Scopus subject areas

Biotechnology
Atomic and Molecular Physics, and Optics

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

abstract = "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.",

author = "Hirokazu Ishii and Kohei Otomo and Hung, {Jui Hung} and Motosuke Tsutsumi and Hiroyuki Yokoyama and Tomomi Nemoto",

note = "Funding Information: Brain/MINDS (AMED); MEXT/JSPS KAKENHI (JP18K14659, JP18K06591, JP16K15103, JP15H05953 “Resonance Bio”); the Research Program of “Five-star Alliance” in “NJRC Mater. & Dev.” (MEXT). Publisher Copyright: {\textcopyright} 2019 Optical Society of America under the terms of the OSA Open Access Publishing Agreement.",

year = "2019",

month = jul,

doi = "10.1364/BOE.10.003104",

language = "English",

volume = "10",

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AU - Ishii, Hirokazu

AU - Otomo, Kohei

AU - Hung, Jui Hung

AU - Tsutsumi, Motosuke

AU - Yokoyama, Hiroyuki

AU - Nemoto, Tomomi

N1 - Funding Information: Brain/MINDS (AMED); MEXT/JSPS KAKENHI (JP18K14659, JP18K06591, JP16K15103, JP15H05953 “Resonance Bio”); the Research Program of “Five-star Alliance” in “NJRC Mater. & Dev.” (MEXT). Publisher Copyright: © 2019 Optical Society of America under the terms of the OSA Open Access Publishing Agreement.

PY - 2019/7

Y1 - 2019/7

N2 - 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.

AB - 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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ER -

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

Abstract

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