Ex utero culture and live imaging of mouse embryos.
Ex utero culture and live imaging of mouse embryos.
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DOI：
10.1007/978-1-61779-210-6_9
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发表时间：
2011
期刊：
影响因子：
--
通讯作者：
Hadjantonakis, Anna-Katerina
中科院分区：
文献类型：
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作者：
Piliszek, Anna;Kwon, Gloria S;Hadjantonakis, Anna-Katerina
小鼠遗传方法与实时成像工具相结合，有可能彻底改变我们目前对哺乳动物生物学的理解。各种荧光蛋白的可用性和改进为可视化生物体中的细胞提供了不可或缺的工具。现在有可能产生以组织特异性方式表达荧光蛋白的转基因小鼠菌株。这些表达报告的菌株使得在活胚胎的背景下成像动态细胞行为成为可能。由于小鼠胚胎在子宫内发育，活体成像实验要求培养条件与体内条件非常接近。在过去的几十年里，在培养种植前和种植后阶段胚胎的条件方面取得了重大进展。在本章中，我们将讨论小鼠胚胎着床前和着床后的体外培养方法。特别是，我们将描述在不同阶段收集胚胎的方案，建立成像的培养条件，并使用激光扫描共聚焦显微镜观察表达荧光报告基因的小鼠胚胎的活过程。
Mouse genetic approaches when combined with live imaging tools have the potential to revolutionize our current understanding of mammalian biology. The availability and improvement of a wide variety of fluorescent proteins have provided indispensable tools to visualize cells in living organisms. It is now possible to generate genetically modified mouse strains expressing fluorescent proteins in a tissue-specific manner. These reporter-expressing strains make it possible to image dynamic cell behaviors in the context of a living embryo. Since mouse embryos develop within the uterus, live imaging experiments require culture conditions that closely mimic those in vivo. Over the past few decades, significant advances have been made in developing conditions for culturing both pre- and postimplantation stage embryos. In this chapter, we will discuss methods for ex utero culture of preimplantation and postimplantation stage mouse embryos. In particular, we will describe protocols for collecting embryos at various stages, setting up culture conditions for imaging and using laser scanning confocal microscopy to visualize live processes in mouse embryos expressing fluorescent reporters.