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3 August 2015 Visible-wavelength two-photon excitation microscopy for fluorescent protein imaging
Masahito Yamanaka, Kenta Saito, Nicholas I. Smith, Yoshiyuki Arai, Kumiko Uegaki, Yasuo Yonemaru, Kentaro Mochizuki, Satoshi Kawata, Takeharu Nagai, Katsumasa Fujita
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Journal of Biomedical Optics, Vol. 20, Issue 10, 101202 (August 2015). https://doi.org/10.1117/1.JBO.20.10.101202
Abstract
The simultaneous observation of multiple fluorescent proteins (FPs) by optical microscopy is revealing mechanisms by which proteins and organelles control a variety of cellular functions. Here we show the use of visible-light based two-photon excitation for simultaneously imaging multiple FPs. We demonstrated that multiple fluorescent targets can be concurrently excited by the absorption of two photons from the visible wavelength range and can be applied in multicolor fluorescence imaging. The technique also allows simultaneous single-photon excitation to offer simultaneous excitation of FPs across the entire range of visible wavelengths from a single excitation source. The calculation of point spread functions shows that the visible-wavelength two-photon excitation provides the fundamental improvement of spatial resolution compared to conventional confocal microscopy.
CC BY: © The Authors. Published by SPIE under a Creative Commons Attribution 4.0 Unported License. Distribution or reproduction of this work in whole or in part requires full attribution of the original publication, including its DOI.
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Masahito Yamanaka, Kenta Saito, Nicholas I. Smith, Yoshiyuki Arai, Kumiko Uegaki, Yasuo Yonemaru, Kentaro Mochizuki, Satoshi Kawata, Takeharu Nagai, and Katsumasa Fujita "Visible-wavelength two-photon excitation microscopy for fluorescent protein imaging," Journal of Biomedical Optics 20(10), 101202 (3 August 2015). https://doi.org/10.1117/1.JBO.20.10.101202
Published: 3 August 2015
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Cited by 20 scholarly publications.
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KEYWORDS
Luminescence
Deep ultraviolet
Absorption
Spatial resolution
Confocal microscopy
Two photon excitation microscopy
Fluorescent proteins
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