Department of Integrative Biology, University of Wisconsin-Madison, Madison, WI, USA.
Program in Genetics, University of Wisconsin-Madison, Madison, WI, USA.
Methods Mol Biol. 2022;2438:345-376. doi: 10.1007/978-1-0716-2035-9_22.
The Caenorhabditis elegans embryo is well suited for analysis of directed cell rearrangement via modern microscopy, due to its simple organization, short generation time, transparency, invariant lineage, and the ability to generate engineered embryos expressing various fluorescent proteins. This chapter provides an overview of routine microscopy techniques for imaging dorsal intercalation, a convergent extension-like morphogenetic movement in the embryonic epidermis of C. elegans, including making agar mounts, low-cost four-dimensional (4D) Nomarski microscopy, laser microsurgery, and 4D fluorescence microscopy using actin and junctional fusion proteins, as well as tissue-specific promoters useful for studying dorsal intercalation.
秀丽隐杆线虫胚胎非常适合通过现代显微镜分析定向细胞重排,因为它的组织简单、世代时间短、透明、谱系不变,并且能够生成表达各种荧光蛋白的工程胚胎。本章提供了用于成像背侧插入(dorsal intercalation)的常规显微镜技术概述,背侧插入是秀丽隐杆线虫胚胎表皮中的一种趋同延伸样形态发生运动,包括制作琼脂载片、低成本的四维(4D)诺玛斯基显微镜、激光显微手术以及使用肌动蛋白和连接融合蛋白的 4D 荧光显微镜,以及用于研究背侧插入的组织特异性启动子。