Analytical Sciences

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

Kohei OTOMO,*1 Terumasa HIBI,*1,*2 Takashi MURATA,*3,*4 Hirotaka WATANABE,*1,*2 Ryosuke KAWAKAMI,*1,*2 Hiroshi NAKAYAMA,*5 Mitsuyasu HASEBE,*3,*4 and Tomomi NEMOTO*1,*2

*1 Research Institute for Electronic Science, Hokkaido University, Kita 20 Nishi 10, Kita, Sapporo 001-0020, Japan
*2 Graduate School of Information Science and Technology, Hokkaido University, Kita 14 Nishi 9, Kita, Sapporo 060-0814, Japan
*3 Division of Evolutionary Biology, National Institute for Basic Biology, Nishigonaka 38 Myodaiji-cho, Okazaki 444-8585, Japan
*4 Department of Basic Biology, School of Life Science, SOKENDAI (The Graduate University for Advanced Studies), Nishigonaka 38 Myodaiji-cho, Okazaki 444-8585, Japan
*5 Yokogawa Electric Corp., 2-3 Hokuyodai, Kanazawa 920-0177, Japan

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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