Visible-wavelength two-photon excitation microscopy with multifocus scanning for volumetric live-cell imaging.

Abstract:

:Two-photon excitation microscopy is one of the key techniques used to observe three-dimensional (3-D) structures in biological samples. We utilized a visible-wavelength laser beam for two-photon excitation in a multifocus confocal scanning system to improve the spatial resolution and image contrast in 3-D live-cell imaging. Experimental and numerical analyses revealed that the axial resolution has improved for a wide range of pinhole sizes used for confocal detection. We observed the 3-D movements of the Golgi bodies in living HeLa cells with an imaging speed of 2 s per volume. We also confirmed that the time-lapse observation up to 8 min did not cause significant cell damage in two-photon excitation experiments using wavelengths in the visible light range. These results demonstrate that multifocus, two-photon excitation microscopy with the use of a visible wavelength can constitute a simple technique for 3-D visualization of living cells with high spatial resolution and image contrast.

journal_name

J Biomed Opt

authors

Oketani R,Suda H,Uegaki K,Kubo T,Matsuda T,Yamanaka M,Arai Y,Smith N,Nagai T,Fujita K

doi

10.1117/1.JBO.25.1.014502

subject

Has Abstract

pub_date

2019-11-01 00:00:00

pages

1-5

issue

1

eissn

1083-3668

issn

1560-2281

pii

JBO-190101SSR

journal_volume

25

pub_type

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