Setting up a simple light sheet microscope for in toto imaging of C. elegans development.
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.
Institut de Biologie du Développement de Marseille, UMR7288 CNRS, Aix-Marseille Université
Claire
C
Chardès
Institut de Biologie du Développement de Marseille, UMR7288 CNRS, Aix-Marseille Université
Pauline
P
Mélénec
Institut de Biologie du Développement de Marseille, UMR7288 CNRS, Aix-Marseille Université; vincent.bertrand@univ-amu.fr.
Vincent
V
Bertrand
Institut de Biologie du Développement de Marseille, UMR7288 CNRS, Aix-Marseille Université; pierre-francois.lenne@univ-amu.fr.
Pierre-François
PF
Lenne
D000818:Animals; D017173:Caenorhabditis elegans / Q000196:embryology*; D004625:Embryo, Nonmammalian; D021621:Imaging, Three-Dimensional / Q000295:instrumentation / Q000379:methods; D008853:Microscopy / Q000295:instrumentation* / Q000379:methods*; D059008:Time-Lapse Imaging / Q000295:instrumentation / Q000379:methods