Stowers Institute for Medical Research, Kansas City, MO, USA.
Department of Anatomy and Cell Biology, University of Kansas School of Medicine, Kansas City, KS, USA.
Methods Mol Biol. 2021;2179:107-114. doi: 10.1007/978-1-0716-0779-4_10.
Live embryo imaging may provide a wealth of information on intact cell and tissue dynamics, but can be technically challenging to sustain embryo orientation and health for long periods under a microscope. In this protocol, we describe an in vivo method to mount and image cell movements during the epithelial-to-mesenchymal transition (EMT) of neural crest cells within the chick dorsal neural tube. We focus on describing the collection of images and data preparation for image analysis throughout the developmental stages HH15-21 in the chick trunk. Trunk neural crest cell EMT is crucial to development of the peripheral nervous system and pigment cell patterning. The methods we describe may also be applied to other cell and tissue phenomena at various chick developmental stages with some modifications.
活体胚胎成像可以提供大量关于完整细胞和组织动态的信息,但在显微镜下长时间维持胚胎的方向和健康状态在技术上具有挑战性。在本方案中,我们描述了一种在体内方法,用于在鸡背神经管内神经嵴细胞的上皮-间质转化(EMT)过程中对细胞运动进行成像。我们专注于描述在鸡躯干发育阶段 HH15-21 期间收集图像和准备图像分析数据的过程。躯干神经嵴细胞 EMT 对周围神经系统和色素细胞模式形成的发育至关重要。我们描述的方法也可以通过一些修改应用于其他细胞和组织现象在不同的鸡发育阶段。