Hyperspectral two-photon excitation microscopy using visible wavelength

Kubo, Toshiki; Temma, Kenta; Smith, Nicholas I. et al.

Optics Letters, 2020, 46(1), 37-40

Number of Access:220（2026-02-19 11:59 Counts）

Identifier to cite or link to this item: https://hdl.handle.net/11094/103309

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Title
Hyperspectral two-photon excitation microscopy using visible wavelength
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
NRID
1000010362592 1000010362592
ORCID
0000-0002-7835-199X 0000-0002-7835-199X
researchmap permalink
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Scopus Author ID
55262815400 55262815400
CreatorName
Smith, Nicholas I.
CreatorName
Lu, Kai
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
Description
Kubo T., Temma K., Smith N.I., et al. Hyperspectral two-photon excitation microscopy using visible wavelength. Optics Letters 46, 37 (2021); https://doi.org/10.1364/OL.413526.
Abstract
We demonstrate hyperspectral imaging by visible-wavelength two-photon excitation microscopy using line illumination and slit-confocal detection. A femtosecond pulsed laser light at 530 nm was used for the simultaneous excitation of fluorescent proteins with different emission wavelengths. The use of line illumination enabled efficient detection of hyperspectral images and achieved simultaneous detection of three fluorescence spectra in the observation of living HeLa cells with an exposure time of 1 ms per line, which is equivalent to about 2 µs per pixel in point scanning, with 160 data points per spectrum. On combining linear spectral unmixing techniques, localization of fluorescent probes in the cells was achieved. A theoretical investigation of the imaging property revealed high-depth discrimination property attained through the combination of nonlinear excitation and slit detection.
Publisher
The Optical Society
Source Title
Optics Letters
Volume (Issue)
46(1)
Page
37-40
Date of Issued
2020-12-18
Language
English
Handle URL
https://hdl.handle.net/11094/103309
PISSN
01469592
eISSN
15394794
Relation.isVersionOf
DOI (Publisher Version)
https://doi.org/10.1364/OL.413526
Access Rights
open access
CopyRight
© Optical Society of America 2021 Optica Publishing Group. One print or electronic copy may be made for personal use only. Systematic reproduction and distribution, duplication of any material in this paper for a fee or for commercial purposes, or modifications of the content of this paper are prohibited.
oaire:version
Accepted Manuscript
Category
学術雑誌論文 Journal Article