We describe a fluorescence optical microscope with both single-photon and two-photon non-diffracting light-sheet excitations for large volume imaging. With a special design to accommodate two different wavelength ranges (visible: 400-700 nm and near infrared: 800-1200 nm), we combine the line-Bessel sheet (LBS, for single-photon excitation) and the scanning Bessel beam (SBB, for two-photon excitation) light sheet together in a single microscope setup. For a transparent thin sample where the scattering can be ignored, the LBS single-photon excitation is the optimal imaging solution. When the light scattering becomes significant for a deep-cell or deep-tissue imaging, we use SBB light-sheet two-photon excitation with a longer wavelength. We achieved nearly identical lateral/axial resolution of about 350/270 nm for both imagings. This integrated light-sheet microscope may have a wide application for live-cell and live-tissue three-dimensional high-speed imaging.
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April 2018
Research Article|
April 02 2018
An integrated single- and two-photon non-diffracting light-sheet microscope
Sze Cheung Lau;
Sze Cheung Lau
1
Department of Physics, The Hong Kong University of Science and Technology
, Clear Water Bay, Kowloon, Hong Kong, China
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Hoi Chun Chiu;
Hoi Chun Chiu
1
Department of Physics, The Hong Kong University of Science and Technology
, Clear Water Bay, Kowloon, Hong Kong, China
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Luwei Zhao;
Luwei Zhao
2
Light Innovation Technology Ltd.
, Tseung Kwan O, Hong Kong, China
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Teng Zhao;
Teng Zhao
a)
2
Light Innovation Technology Ltd.
, Tseung Kwan O, Hong Kong, China
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M. M. T. Loy;
M. M. T. Loy
1
Department of Physics, The Hong Kong University of Science and Technology
, Clear Water Bay, Kowloon, Hong Kong, China
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Shengwang Du
Shengwang Du
b)
1
Department of Physics, The Hong Kong University of Science and Technology
, Clear Water Bay, Kowloon, Hong Kong, China
3
Department of Chemical and Biological Engineering, The Hong Kong University of Science and Technology
, Clear Water Bay, Kowloon, Hong Kong, China
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Sze Cheung Lau
1
Hoi Chun Chiu
1
Luwei Zhao
2
Teng Zhao
2,a)
M. M. T. Loy
1
Shengwang Du
1,3,b)
1
Department of Physics, The Hong Kong University of Science and Technology
, Clear Water Bay, Kowloon, Hong Kong, China
2
Light Innovation Technology Ltd.
, Tseung Kwan O, Hong Kong, China
3
Department of Chemical and Biological Engineering, The Hong Kong University of Science and Technology
, Clear Water Bay, Kowloon, Hong Kong, China
a)
Electronic mail: [email protected]
b)
Electronic mail: [email protected]
Rev. Sci. Instrum. 89, 043701 (2018)
Article history
Received:
December 20 2017
Accepted:
March 12 2018
Citation
Sze Cheung Lau, Hoi Chun Chiu, Luwei Zhao, Teng Zhao, M. M. T. Loy, Shengwang Du; An integrated single- and two-photon non-diffracting light-sheet microscope. Rev. Sci. Instrum. 1 April 2018; 89 (4): 043701. https://doi.org/10.1063/1.5020154
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