Large-field high-resolution two-photon digital scanned light-sheet microscopy  

作　　者：Weijian Zong Jia Zhao Xuanyang Chen Yuan Lin Huixia Ren Yunfeng Zhang Ming Fan Zhuan Zhou Heping Cheng Yujie Sun Liangyi Chen 

机构地区：[1]The State Key Laboratory of Biomembrane and Membrane Biotechnology,Institute of Molecular Medicine, Beijing Key Laboratory of Cardiometa-bolic Molecular Medicine, Institute of Molecular Medicine, Peking-Tsin-ghua Center for Life Sciences, Peking University, Beijing 100871, China [2]China Department of Cognitive Sciences, Institute of Basic MedicalSciences, Beijing 100850, China [3]Biodynamic Optical Imaging Center,Peking University, Beijing 100871, China [4]School of Electronics Engi-neering and Computer Science, Peking University, Beijing 100871, China

出　　处：《Cell Research》2015年第2期254-257,共4页细胞研究（英文版）

基　　金：Acknowledgments We thank Fuzeng Niu for his help on the optics, Liwa Shao and Fujian Lu for biological experiments, Yongxiao Li for the Labview programming, and Yi Wu for the help on some of the data analysis. Tg（kdrl：EGFP） fish is a gift from Jingwei Xiong at Peking Uni- versity. The work is supported by grants from the Major State Basic Research Program of China （2013CB531200, 2011 CB809100 and 2012CB518200）, the National Key Technology R＆D Program （SQ2011SF 11 B01041）, the National Natural Science Foundation of China （31327901, 31221002, 81222020 and 21390410）, and the Beijing Natural Science Foundation （7121008）.

摘　　要：Dear Editor, Recent advent of light-sheet fluorescent microscopy （LSFM） has revolutionized three-dimensional biological imaging with high temporal resolution and minimal photodamage, enabling long-term fluorescence imaging of tissues and small organisms [1-2]. By combining two-photon fluorescence excitation with LSFM, Truong et al. [3] have created a two-photon digital scanned lightsheet microscope （2P-DSLM）, allowing for deep-tissue imaging of highly scattering Drosophila embryos and fast beating hearts of zebrafish. Similar to classical LSFM configurations, a 2P-DSLM uses a low numerical aperture （NA 〈 0.1） to achieve a long and homogenous illumination. This leads to a thick light sheet and thus reduces axial resolution and image contrast. On the other hand, Betzig and colleagues have used a Bessel beam to generate thin single-photon light sheet that yields superb axial resolution [4]. However, the field of view and the penetration depth are limited in such system.

关 键 词：荧光显微镜 双光子 大视场 光片 数字扫描 高分辨率 轴向分辨率 高时间分辨率 

分 类 号：TH742.65[机械工程—光学工程] O431[机械工程—仪器科学与技术]