Biomaterial Database

Rapid volumetric imaging with Bessel-Beam three-photon microscopy. 2018

Bingying Chen, and Xiaoshuai Huang, and Dongzhou Gou, and Jianzhi Zeng, and Guoqing Chen, and Meijun Pang, and Yanhui Hu, and Zhe Zhao, and Yunfeng Zhang, and Zhuan Zhou, and Haitao Wu, and Heping Cheng, and Zhigang Zhang, and Chris Xu, and Yulong Li, and Liangyi Chen, and Aimin Wang

State Key Laboratory of Advanced Optical Communication System and Networks, School of Electronics Engineering and Computer Science, Peking University, Beijing 100871, China.

Owing to its tissue-penetration ability, multi-photon fluorescence microscopy allows for the high-resolution, non-invasive imaging of deep tissue in vivo; the recently developed three-photon microscopy (3PM) has extended the depth of high-resolution, non-invasive functional imaging of mouse brains to beyond 1.0 mm. However, the low repetition rate of femtosecond lasers that are normally used in 3PM limits the temporal resolution of point-scanning three-photon microscopy. To increase the volumetric imaging speed of 3PM, we propose a combination of an axially elongated needle-like Bessel-beam with three-photon excitation (3PE) to image biological samples with an extended depth of focus. We demonstrate the higher signal-to-background ratio (SBR) of the Bessel-beam 3PM compared to the two-photon version both theoretically and experimentally. Finally, we perform simultaneous calcium imaging of brain regions at different axial locations in live fruit flies and rapid volumetric imaging of neuronal structures in live mouse brains. These results highlight the unique advantage of conducting rapid volumetric imaging with a high SBR in the deep brain in vivo using scanning Bessel-3PM.