Journal Article

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

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

2015

Overview

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.

• Publisher: Nature Publishing Group UK,Nature Publishing Group
• Subject: 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