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Correcting spherical aberrations in a biospecimen using a transmissive liquid crystal device in two-photon excitation laser scanning microscopy

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Title: Correcting spherical aberrations in a biospecimen using a transmissive liquid crystal device in two-photon excitation laser scanning microscopy
Authors: Tanabe, Ayano Browse this author
Hibi, Terumasa Browse this author →KAKEN DB
Ipponjima, Sari Browse this author
Matsumoto, Kenji Browse this author
Yokoyama, Masafumi Browse this author
Kurihara, Makoto Browse this author
Hashimoto, Nobuyuki Browse this author
Nemoto, Tomomi Browse this author →KAKEN DB
Keywords: adaptive optics
liquid crystals
phase modulation
scanning microscopy
Issue Date: Oct-2015
Publisher: Society of Photo-Optical Instrumentation Engineers (SPIE)
Journal Title: Journal of biomedical optics
Volume: 20
Issue: 10
Start Page: 101204
Publisher DOI: 10.1117/1.JBO.20.10.101204
PMID: 26244766
Abstract: Two-photon excitation laser scanning microscopy has enabled the visualization of deep regions in a biospecimen. However, refractive-index mismatches in the optical path cause spherical aberrations that degrade spatial resolution and the fluorescence signal, especially during observation at deeper regions. Recently, we developed transmissive liquid-crystal devices for correcting spherical aberration without changing the basic design of the optical path in a conventional laser scanning microscope. In this study, the device was inserted in front of the objective lens and supplied with the appropriate voltage according to the observation depth. First, we evaluated the device by observing fluorescent beads in single-and two-photon excitation laser scanning microscopes. Using a 25x water-immersion objective lens with a numerical aperture of 1.1 and a sample with a refractive index of 1.38, the device recovered the spatial resolution and the fluorescence signal degraded within a depth of +/- 0.6 mm. Finally, we implemented the device for observation of a mouse brain slice in a two-photon excitation laser scanning microscope. An optical clearing reagent with a refractive index of 1.42 rendered the fixed mouse brain transparent. The device improved the spatial resolution and the yellow fluorescent protein signal within a depth of 0-0.54 mm.
Rights: (C) The Authors. Published by SPIE under a Creative Commons Attribution 3.0 Unported License. Distribution or reproduction of this work in whole or in part requires full attribution of the original publication, including its DOI.
Type: article
Appears in Collections:電子科学研究所 (Research Institute for Electronic Science) > 雑誌発表論文等 (Peer-reviewed Journal Articles, etc)

Submitter: 根本 知己

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