Berthet Béatrice, Maizel Alexis
Center for Organismal Studies (COS), University of Heidelberg, Heidelberg, Germany.
J Microsc. 2016 Aug;263(2):158-64. doi: 10.1111/jmi.12393. Epub 2016 Mar 28.
Light sheet fluorescence microscopy (LSFM) is increasingly used to investigate biological processes in animals as well as in plants. LSFM achieves optical sectioning by the selective illumination of a single plane of the sample with a sheet of laser light while simultaneously recording emitted fluorescence orthogonally to the illumination plane. A 3D image of the sample can then be generated with a temporal resolution ranging from seconds to several days, and at scales ranging from subcellular to whole organ. By design, LSFM provides fast imaging, and low phototoxicity, two key criteria for live imaging under physiological conditions. Despite its potential, LSFM remains underutilized in plant biology. This review aims to highlight challenges of live imaging in plants, to describe key steps in using LSFM on live plant samples and finally at providing an overview of published examples of applications of LSFM in plants.
光片荧光显微镜（LSFM）越来越多地用于研究动物和植物中的生物过程。LSFM通过用一束激光选择性照射样品的单个平面来实现光学切片，同时正交于照明平面记录发射的荧光。然后可以生成样品的3D图像，其时间分辨率范围从几秒到几天，尺度范围从亚细胞到整个器官。从设计上来说，LSFM提供快速成像和低光毒性，这是生理条件下活细胞成像的两个关键标准。尽管有其潜力，但LSFM在植物生物学中的应用仍然不足。本综述旨在突出植物活细胞成像的挑战，描述在活植物样品上使用LSFM的关键步骤，并最终概述已发表的LSFM在植物中的应用实例。
