学術雑誌論文

学術雑誌論文

text

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

Visible-Wavelength Multiphoton Activation Confocal Microscopy

American Chemical Society

英語

text

2021-08-30

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

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.

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.

103307

Visible-Wavelength Multiphoton Activation Confocal Microscopy

American Chemical Society

英語

2021-08-30

ACS Photonics

8

9

2666

2673

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

https://hdl.handle.net/11094/103307