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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 | | aberrations |
| 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. | | http://creativecommons.org/licenses/by/3.0/ |
| Type: | article |
| URI: | http://hdl.handle.net/2115/60512 |
| 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)
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Submitter: 根本 知己
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