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Journal of Biomedical Optics • Open Access

Visible-wavelength two-photon excitation microscopy for fluorescent protein imaging
Author(s): Masahito Yamanaka; Kenta Saito; Nicholas I. Smith; Yoshiyuki Arai; Kumiko Uegaki; Yasuo Yonemaru; Kentaro Mochizuki; Satoshi Kawata; Takeharu Nagai; Katsumasa Fujita

Paper 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.

Paper Details

Date Published: 3 August 2015
PDF: 11 pages
J. Biomed. Opt. 20(10) 101202 doi: 10.1117/1.JBO.20.10.101202
Published in: Journal of Biomedical Optics Volume 20, Issue 10
Show Author Affiliations
Masahito Yamanaka, Osaka Univ. (Japan)
Kenta Saito, Osaka Univ. (Japan)
Nicholas I. Smith, Osaka Univ. (Japan)
Yoshiyuki Arai, Osaka Univ. (Japan)
Kumiko Uegaki, Osaka Univ. (Japan)
Yasuo Yonemaru, Osaka Univ. (Japan)
Kentaro Mochizuki, Osaka Univ. (Japan)
Satoshi Kawata, Osaka Univ. (Japan)
Takeharu Nagai, Osaka Univ. (Japan)
Katsumasa Fujita, Osaka Univ. (Japan)

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