光学学报, 2017, 37 (3): 0318007, 网络出版: 2017-03-08   

光片荧光显微成像 下载: 1786次

Light-Sheet Fluorescence Microscopy
作者单位
北京大学分子医学研究所膜生物学国家重点实验室, 北京 100871
摘要
在过去的20年, 激光扫描共聚焦显微镜一直是在细胞水平和亚细胞水平上观察生命活动的标准工具, 但是基于针孔的共聚焦显微镜的光学层切是以牺牲焦平面以外的被激发的荧光色团和较大的光毒性为代价的。作为一种新型的荧光显微镜, 光片荧光显微镜采用侧向照明的方式, 对样品直接进行面成像。相对于点扫描的成像方式, 光片显微镜成像速度远远高于激光扫描共聚焦显微镜, 使得研究一些高速的精细生命活动过程成为了可能。光片荧光显微镜的另外一个优点是只有光片处的样品才会被激发, 处于光片以外的样品则不会被激发, 因此光毒性较小, 使得人们能够在更长的时间尺度下观察样品。正是由于光片荧光显微镜特殊的照明和成像方式, 才使其在大样本的三维高速成像中起到不可替代的作用。本文简要回顾了光片荧光显微镜发展的历史及研究现状, 旨在为该领域的科研人员对光片荧光显微镜的现状及未来发展方向提供个人理解。
Abstract
In the past two decades, laser scanning confocal microscope has been the standard tool for observing the process of life at cellular and sub-cellular level. The optical sectioning capacity of pinhole-based confocal microscope comes at the price of unwanted excitation of fluorophores out of focal plane and phototoxic damage to biological samples. As a new type of fluorescent microscope, light-sheet fluorescence microscope (LSFM) uses side illumination to conduct surface imaging of the samples directly. As compared to the point-scanning imaging mode, LSFM excels at its imaging speed, which is much higher than that of laser scanning confocal microscope, thus making it possible to study some high-speed fine life activities. Another advantage of the light-sheet fluorescence microscope is that only the sample at the light-sheet is excited and the sample outside the light-sheet is not excited, so there is less phototoxic dosage and we can observe the sample in a longer time scale. The special illumination and imaging mode of the light-sheet fluorescence microscope make it play an irreplaceable role in three-dimensional high-speed imaging of big biological samples. The history and research status of light-sheet fluorescence microscope are reviewed with the purpose of providing a personal perspective of current situation and future direction of LSFM.

杨豫龙, 宗伟建, 吴润龙, 陈良怡. 光片荧光显微成像[J]. 光学学报, 2017, 37(3): 0318007. Yang Yulong, Zong Weijian, Wu Runlong, Chen Liangyi. Light-Sheet Fluorescence Microscopy[J]. Acta Optica Sinica, 2017, 37(3): 0318007.

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