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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。
For the past two decades, laser scanning confocal microscopy has been the standard tool for observing life activity at the cellular and subcellular levels, but pinhole-based confocal microscopy is performed at the expense of sacrificial focal plane excitation Fluorescent chromophores and greater light toxicity at the expense of. As a new type of fluorescence microscope, light fluorescence microscope with lateral illumination mode, the sample directly to the surface imaging. Compared to point-scan imaging, light microscopy is much faster than laser scanning confocal microscopy, making it possible to study some high-speed, fine-life activities. Another benefit of a light-based fluorescence microscope is that only the sample at the light plate will be excited, and the sample outside the light plate will not be excited, so less photo-toxicity allows one to observe the sample over a longer time scale . It is precisely because of light fluorescence microscopy special lighting and imaging methods that make it in the large sample of three-dimensional high-speed imaging plays an irreplaceable role. This review briefly reviews the history and status quo of the development of light fluorescence microscopy and aims to provide a personal understanding of the current state and future direction of light fluorescence microscopy in this field.