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Analytical Sciences
Vol. 31 (2015) No. 4 p. 307-313

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http://doi.org/10.2116/analsci.31.307

Original Papers

Original Papers

Multi-point Scanning Two-photon Excitation Microscopy by Utilizing a High-peak-power 1042-nm Laser

Kohei OTOMO1), Terumasa HIBI1) 2), Takashi MURATA3) 4), Hirotaka WATANABE1) 2), Ryosuke KAWAKAMI1) 2), Hiroshi NAKAYAMA5), Mitsuyasu HASEBE3) 4), Tomomi NEMOTO1) 2)

1) Research Institute for Electronic Science, Hokkaido University 2) Graduate School of Information Science and Technology, Hokkaido University 3) Division of Evolutionary Biology, National Institute for Basic Biology 4) Department of Basic Biology, School of Life Science, SOKENDAI (The Graduate University for Advanced Studies) 5) Yokogawa Electric Corp.

Released on J-STAGE April 10, 2015  
Received December 24, 2014   Accepted February 14, 2015

Keywords: Two-photon excitation microscopy, multi-point laser scanning method, confocal microscopy, Yb-based laser


The temporal resolution of a two-photon excitation laser scanning microscopy (TPLSM) system is limited by the excitation laser beam’s scanning speed. To improve the temporal resolution, the TPLSM system is equipped with a spinning-disk confocal scanning unit. However, the insufficient energy of a conventional Ti:sapphire laser source restricts the field of view (FOV) for TPLSM images to a narrow region. Therefore, we introduced a high-peak-power Yb-based laser in order to enlarge the FOV. This system provided three-dimensional imaging of a sufficiently deep and wide region of fixed mouse brain slices, clear four-dimensional imaging of actin dynamics in live mammalian cells and microtubule dynamics during mitosis and cytokinesis in live plant cells.
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