Institut de Génétique et de Biologie Moléculaire et Cellulaire, Illkirch 67404, France.
Centre National de la Recherche Scientifique, UMR7104, Illkirch 67404, France.
Philos Trans R Soc Lond B Biol Sci. 2018 Sep 24;373(1759):20170332. doi: 10.1098/rstb.2017.0332.
Organogenesis involves extensive and dynamic changes of tissue shape during development. It is associated with complex morphogenetic events that require enormous tissue plasticity and generate a large variety of transient three-dimensional geometries that are achieved by global tissue responses. Nevertheless, such global responses are driven by tight spatio-temporal regulation of the behaviours of individual cells composing these tissues. Therefore, the development of image analysis tools that allow for extraction of quantitative data concerning individual cell behaviours is central to study tissue morphogenesis. There are many image analysis tools available that permit extraction of cell parameters. Unfortunately, the majority are developed for tissues with relatively simple geometries such as flat epithelia. Problems arise when the tissue of interest assumes a more complex three-dimensional geometry. Here, we use the endothelium of the developing zebrafish dorsal aorta as an example of a tissue with cylindrical geometry and describe the image analysis routines developed to extract quantitative data on individual cells in such tissues, as well as the image acquisition and sample preparation methodology.This article is part of the Theo Murphy meeting issue 'Mechanics of development'.
器官发生涉及组织形状在发育过程中的广泛和动态变化。它与复杂的形态发生事件相关，这些事件需要巨大的组织可塑性，并产生大量的瞬时三维几何形状，这些形状是通过整体组织反应实现的。然而，这种全局反应是由组成这些组织的单个细胞行为的紧密时空调节驱动的。因此，开发允许提取有关单个细胞行为的定量数据的图像分析工具对于研究组织形态发生至关重要。有许多可用于提取细胞参数的图像分析工具。不幸的是，大多数工具都是为具有相对简单几何形状的组织（例如扁平上皮）开发的。当感兴趣的组织具有更复杂的三维几何形状时，就会出现问题。在这里，我们以发育中的斑马鱼背主动脉的内皮为例，介绍一种具有圆柱几何形状的组织，并描述了为提取此类组织中单个细胞的定量数据而开发的图像分析例程，以及图像采集和样本制备方法。本文是 Theo Murphy 会议主题“发育力学”的一部分。
