23 February 2018 Wide field video-rate two-photon imaging by using spinning disk beam scanner
Author Affiliations +
The microscope technology with wider view field, deeper penetration depth, higher spatial resolution and higher imaging speed are required to investigate the intercellular dynamics or interactions of molecules and organs in cells or a tissue in more detail. The two-photon microscope with a near infrared (NIR) femtosecond laser is one of the technique to improve the penetration depth and spatial resolution. However, the video-rate or high-speed imaging with wide view field is difficult to perform with the conventional two-photon microscope. Because point-to-point scanning method is used in conventional one, so it’s difficult to achieve video-rate imaging. In this study, we developed a two-photon microscope with spinning disk beam scanner and femtosecond NIR fiber laser with around 10 W average power for the microscope system to achieve above requirements. The laser is consisted of an oscillator based on mode-locked Yb fiber laser, a two-stage pre-amplifier, a main amplifier based on a Yb-doped photonic crystal fiber (PCF), and a pulse compressor with a pair of gratings. The laser generates a beam with maximally 10 W average power, 300 fs pulse width and 72 MHz repetition rate. And the beam incident to a spinning beam scanner (Yokogawa Electric) optimized for two-photon imaging. By using this system, we achieved to obtain the 3D images with over 1mm-penetration depth and video-rate image with 350 x 350 um view field from the root of Arabidopsis thaliana.
© (2018) COPYRIGHT Society of Photo-Optical Instrumentation Engineers (SPIE). Downloading of the abstract is permitted for personal use only.
Yasuhiro Maeda, Yasuhiro Maeda, Kazuo Kurokawa, Kazuo Kurokawa, Yoko Ito, Yoko Ito, Satoshi Wada, Satoshi Wada, Akihiko Nakano, Akihiko Nakano, "Wide field video-rate two-photon imaging by using spinning disk beam scanner", Proc. SPIE 10498, Multiphoton Microscopy in the Biomedical Sciences XVIII, 104982K (23 February 2018); doi: 10.1117/12.2288969; https://doi.org/10.1117/12.2288969


A video rate laser scanning confocal microscope
Proceedings of SPIE (February 11 2008)
Improved no-moving-parts video-rate confocal microscope
Proceedings of SPIE (June 25 1992)
Laser Scanning In Microscopy
Proceedings of SPIE (October 25 1983)

Back to Top