Setting Up a Simple Light Sheet Microscope for In Toto Imaging of C. elegans Development

被引：14
|
作者：
Chardes, Claire [1 ]
Melenec, Pauline [1 ]
Bertrand, Vincent [1 ]
Lenne, Pierre-Francois [1 ]
机构：
[1] Aix Marseille Univ, Inst Biol Dev Marseille, CNRS, UMR7288, Marseille, France
来源：
Jove-Journal of Visualized Experiments | 2014年 / 87期
关键词：
Developmental Biology; Issue; 87; Selective Plane Illumination Microscopy; Light Sheet; Caenorhabditis elegans Embryo Development; in vivo imaging; PLANE ILLUMINATION MICROSCOPY; CAENORHABDITIS-ELEGANS; EMBRYOS; MORPHOGENESIS;
D O I：
10.3791/51342
中图分类号：
O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];
学科分类号：
07 ; 0710 ; 09 ;
摘要：
Fast and low phototoxic imaging techniques are pre-requisite to study the development of organisms in toto. Light sheet based microscopy reduces photo-bleaching and phototoxic effects compared to confocal microscopy, while providing 3D images with subcellular resolution. Here we present the setup of a light sheet based microscope, which is composed of an upright microscope and a small set of opto-mechanical elements for the generation of the light sheet. The protocol describes how to build, align the microscope and characterize the light sheet. In addition, it details how to implement the method for in toto imaging of C. elegans embryos using a simple observation chamber. The method allows the capture of 3D two-colors time-lapse movies over few hours of development. This should ease the tracking of cell shape, cell divisions and tagged proteins over long periods of time.
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页数：9
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