Larina Irina V, Dickinson Mary E
Molecular Physiology & Biophysics, Baylor College of Medicine, Houston, TX, USA.
Methods Mol Biol. 2012;872:205-15. doi: 10.1007/978-1-61779-797-2_14.
Live confocal microscopy of vital fluorescent markers, expressed in mouse embryonic tissues, is a powerful and exciting method to study mammalian embryonic development. This chapter discusses imaging approaches to visualize and characterize dynamic changes of the yolk-sac vasculature and blood flow in mouse embryos. We describe static embryo-culture protocols, which allow maintaining early mouse embryos on the imaging stage for over 24 h. We also describe vital fluorescent-reporter lineages, which can be used to image the developing vasculature and characterize hemodynamics by tracking individual blood cells. Imaging approaches described in this chapter can be used to analyze cardiovascular defects in mutant animals and can provide insights into understanding how genetic signaling pathways and physiological inputs regulate development.
对在小鼠胚胎组织中表达的活体荧光标记物进行共聚焦显微镜观察,是研究哺乳动物胚胎发育的一种强大且令人兴奋的方法。本章讨论了用于可视化和表征小鼠胚胎中卵黄囊脉管系统和血流动态变化的成像方法。我们描述了静态胚胎培养方案,该方案可使早期小鼠胚胎在成像阶段维持24小时以上。我们还描述了活体荧光报告谱系,可用于通过追踪单个血细胞对发育中的脉管系统进行成像并表征血流动力学。本章所述的成像方法可用于分析突变动物的心血管缺陷,并有助于深入了解遗传信号通路和生理输入如何调节发育。