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

• Published in: Cell research Vol. 25; no. 2; pp. 254 - 257
• Main Authors: Zong, Weijian, Zhao, Jia, Chen, Xuanyang, Lin, Yuan, Ren, Huixia, Zhang, Yunfeng, Fan, Ming, Zhou, Zhuan, Cheng, Heping, Sun, Yujie, Chen, Liangyi
• Format: Journal Article
• Language: English
• Published: London Nature Publishing Group UK 01.02.2015; Nature Publishing Group
• Subjects: 631/1647/328/2237; 631/80/2373; Animals; Biomedical and Life Sciences; Cell Biology; Embryo, Nonmammalian - ultrastructure; Imaging, Three-Dimensional - methods; Letter to the Editor; Life Sciences; Microscopy - methods; 光片; 双光子; 大视场; 数字扫描; 荧光显微镜; 轴向分辨率; 高分辨率; 高时间分辨率
• ISSN: 1001-0602; 1748-7838
• DOI: 10.1038/cr.2014.124

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.