Ultramicroscopy: light-sheet-based microscopy for imaging centimeter-sized objects with micrometer resolution.
Ultramicroscopy: light-sheet-based microscopy for imaging centimeter-sized objects with micrometer resolution.
复制标题
DOI：
10.1101/pdb.top076539
复制
发表时间：
2013-08-01
影响因子：
--
通讯作者：
Dodt, Hans-Ulrich
中科院分区：
文献类型：
--
作者：
Becker, Klaus;Jahrling, Nina;Dodt, Hans-Ulrich
超显微镜（UM）是一种功能强大的成像技术，可以实现精确和准确的三维（3D）重建完整的宏观标本与微米分辨率。它适用于1-15 mm大小的标本，如整个小鼠大脑、小鼠胚胎、小鼠器官和果蝇。在UM中，样品由两个薄的反向传播的激光片垂直于观察路径照射。UM是密切相关的一个不断增长的家庭可比的显微镜方法的基础上，近年来开发的光片照明。本文概述了基于光片的显微镜，并介绍了光片生成的基本物理。详细描述了用于调查固定的化学清除组织的“超显微镜”的组装，并讨论了基本部件，如机械，照相机和物镜的功能。最后，UM研究小鼠器官，小鼠胚胎，果蝇成虫的实际应用进行了说明。
Ultramicroscopy (UM) is a powerful imaging technique that achieves precise and accurate three-dimensional (3D) reconstructions of intact macroscopic specimens with micrometer resolution. It was developed for specimens in the size range of 1-15 mm, such as whole mouse brains, mouse embryos, mouse organs, and Drosophila melanogaster. In UM, the specimen is illuminated perpendicular to the observation pathway by two thin counterpropagating sheets of laser light. UM is closely related to a growing family of comparable microscopy approaches based on light sheet illumination developed in recent years. This article presents an overview of light-sheet-based microscopy and describes the underlying physics of light sheet generation. The assembly of an "ultramicroscope" for investigating fixed chemically cleared tissue is described in detail, and the functions of the essential components, such as mechanics, camera, and objectives, are discussed. Finally, practical applications of UM for studying mouse organs, mouse embryos, and Drosophila adults are described.