Visible-Wavelength Multiphoton Activation Confocal Microscopy

Kubo, Toshiki; Temma, Kenta; Sugiura, Kazunori; Shinoda, Hajime; Lu, Kai; Smith, Nicholas Isaac; Matsuda, Tomoki; Nagai, Takeharu; Fujita, Katsumasa

doi:https://doi.org/10.1021/acsphotonics.1c00685

Visible-Wavelength Multiphoton Activation Confocal Microscopy

Kubo, Toshiki; Temma, Kenta; Sugiura, Kazunori et al.

ACS Photonics, 2021, 8(9), 2666-2673

Number of Access:231（2026-02-20 11:11 Counts）

Identifier to cite or link to this item: https://hdl.handle.net/11094/103307

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Title	Visible-Wavelength Multiphoton Activation Confocal Microscopy
Creator	NRID 1000050975159 1000050975159 ORCID 0000-0002-5696-4111 0000-0002-5696-4111 researchmap permalink kubo_toshiki_8 kubo_toshiki_8 CreatorName Kubo, Toshiki
	NRID 1000070993533 1000070993533 ORCID 0000-0002-9968-3459 0000-0002-9968-3459 researchmap permalink k_temma k_temma CreatorName Temma, Kenta
	CreatorName Sugiura, Kazunori
	NRID 1000090846643 1000090846643 CreatorName Shinoda, Hajime
	CreatorName Lu, Kai
	NRID 1000010362592 1000010362592 ORCID 0000-0002-7835-199X 0000-0002-7835-199X researchmap permalink read0089935 read0089935 Scopus Author ID 55262815400 55262815400 CreatorName Smith, Nicholas Isaac
	NRID 1000050419206 1000050419206 CreatorName Matsuda, Tomoki
	NRID 1000020311350 1000020311350 ORCID 0000-0003-2650-9895 0000-0003-2650-9895 researchmap permalink ng1 ng1 Scopus Author ID 7403433377 7403433377 CreatorName Nagai, Takeharu
	NRID 1000080362664 1000080362664 ORCID 0000-0002-2284-375X 0000-0002-2284-375X researchmap permalink read0089911 read0089911 Scopus Author ID 7404058829 7404058829 CreatorName Fujita, Katsumasa
Subject	cell imaging
	confocal detection
	fluorescence
	nonlinear
	reversibly photoswitchable fluorescent proteins
	super-resolution microscopy
	visible-wavelength two-photon excitation
Description	This document is the Accepted Manuscript version of a Published Work that appeared in final form in ACS Photonics 8, 2666 (2021), © American Chemical Society after peer review and technical editing by the publisher. To access the final edited and published work see https://doi.org/10.1021/acsphotonics.1c00685.
Abstract	Nonlinear optical effects modify the point spread function (PSF) in laser scanning microscopy and have been utilized to enhance the spatial resolution in three dimensions. In this paper, we propose the use of visible-wavelength two-photon excitation to activate negatively switching reversibly photoswitchable fluorescent proteins (RSFPs) to introduce nonlinear relationships between the distribution of excitation light and the fluorescence emission. Single-photon excitation following the two-photon induced photoactivation provides a PSF corresponding to the cube of the excitation intensity distribution and achieves imaging properties equivalent to that using third-order nonlinear optical effects. We experimentally confirmed several species of negatively switching RSFPs can be activated by two-photon excitation in the wavelength range of 530-560 nm. We applied the cubic PSF imaging to the observation of HeLa cells stained by Skylan-NS to confirm the improvement of the spatial resolution and the image contrast in confocal scanning microscopy.
Publisher	American Chemical Society
Source Title	ACS Photonics
Volume (Issue)	8(9)
Page	2666-2673
Date of Issued	2021-08-30
Language	English
Handle URL	https://hdl.handle.net/11094/103307
eISSN	23304022
Relation.isVersionOf	DOI (Publisher Version) https://doi.org/10.1021/acsphotonics.1c00685
Access Rights	open access
oaire:version	Accepted Manuscript
Category	学術雑誌論文 Journal Article

Resource Type	journal article
Local Type	Journal Article
DCMI Type	Text
DCTERMS.bibliographicCitation	ACS Photonics. 2021, 8(9), p. 2666-2673
DC.title	Visible-Wavelength Multiphoton Activation Confocal Microscopy
DC.creator	Kubo, Toshiki
	Temma, Kenta
	Sugiura, Kazunori
	Shinoda, Hajime
	Lu, Kai
	Smith, Nicholas Isaac
	Matsuda, Tomoki
	Nagai, Takeharu
	Fujita, Katsumasa
DC.publisher	American Chemical Society
DC.language' scheme='DCTERMS.RFC1766	English
DC.type' scheme='DCTERMS.DCMIType	Text
DCTERMS.issued' scheme='DCTERMS.W3CDTF	2021-08-30
DC.identifier	https://doi.org/10.1021/acsphotonics.1c00685
DC.subject	cell imaging
	confocal detection
	fluorescence
	nonlinear
	reversibly photoswitchable fluorescent proteins
	super-resolution microscopy
	visible-wavelength two-photon excitation
DC.description	This document is the Accepted Manuscript version of a Published Work that appeared in final form in ACS Photonics 8, 2666 (2021), © American Chemical Society after peer review and technical editing by the publisher. To access the final edited and published work see https://doi.org/10.1021/acsphotonics.1c00685.
DCTERMS.abstract	Nonlinear optical effects modify the point spread function (PSF) in laser scanning microscopy and have been utilized to enhance the spatial resolution in three dimensions. In this paper, we propose the use of visible-wavelength two-photon excitation to activate negatively switching reversibly photoswitchable fluorescent proteins (RSFPs) to introduce nonlinear relationships between the distribution of excitation light and the fluorescence emission. Single-photon excitation following the two-photon induced photoactivation provides a PSF corresponding to the cube of the excitation intensity distribution and achieves imaging properties equivalent to that using third-order nonlinear optical effects. We experimentally confirmed several species of negatively switching RSFPs can be activated by two-photon excitation in the wavelength range of 530-560 nm. We applied the cubic PSF imaging to the observation of HeLa cells stained by Skylan-NS to confirm the improvement of the spatial resolution and the image contrast in confocal scanning microscopy.
DC.format	103307
citation_title	Visible-Wavelength Multiphoton Activation Confocal Microscopy
citation_author	Kubo, Toshiki
	Temma, Kenta
	Sugiura, Kazunori
	Shinoda, Hajime
	Lu, Kai
	Smith, Nicholas Isaac
	Matsuda, Tomoki
	Nagai, Takeharu
	Fujita, Katsumasa
citation_publisher	American Chemical Society
citation_language	English
citation_date	2021-08-30
citation_journal_title	ACS Photonics
citation_volume	8
citation_issue	9
citation_firstpage	2666
citation_lastpage	2673
citation_doi	https://doi.org/10.1021/acsphotonics.1c00685
citation_public_url	https://hdl.handle.net/11094/103307

Visible-Wavelength Multiphoton Activation Confocal Microscopy

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