Imaging mouse embryonic cardiovascular development.
Imaging mouse embryonic cardiovascular development.
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DOI：
10.1101/pdb.top071498
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发表时间：
2012-10
影响因子：
--
通讯作者：
I. Larina;Monica D. Garcia;T. Vadakkan;K. Larin;M. Dickinson
中科院分区：
文献类型：
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作者：
I. Larina;Monica D. Garcia;T. Vadakkan;K. Larin;M. Dickinson
哺乳动物心血管系统的早期发育是一个高度动态的过程。实时成像是分析正常和异常心血管发育和动力学的重要工具。本文介绍了两种用于小鼠胚胎心血管发育实时动态成像的光学方法：共聚焦显微镜和光学相干断层扫描（OCT）。与荧光蛋白报告线结合使用的共聚焦显微镜能够以亚微米分辨率可视化发育和重塑心血管系统，甚至可以可视化标记结构的亚细胞细节。我们描述了小鼠转基因系，可用于成像发展中的血管系统和表征血液动力学通过跟踪单个血细胞。重要荧光标记物的共聚焦显微镜揭示了关于细胞形态发生和运动的独特细节;然而，这种方法的成像深度仅限于200 µm。这种限制可以通过使用OCT来解决，它允许对组织进行毫米级的三维（3D）成像，尽管这是以较低的空间分辨率（2-10 µm）为代价的。我们在这里描述如何OCT可以应用于发育中的小鼠胚胎的结构分析和胚胎深部血管的血流动力学分析。这些互补的方法可用于分析突变动物的心血管缺陷，以了解调节人类发育的遗传信号通路。
Early development of the mammalian cardiovascular system is a highly dynamic process. Live imaging is an essential tool for analyzing normal and abnormal cardiovascular development and dynamics. This article describes two optical approaches for live dynamic imaging of mouse embryonic cardiovascular development: confocal microscopy and optical coherence tomography (OCT). Confocal microscopy, used in combination with fluorescent protein reporter lines, enables visualization of the developing and remodeling cardiovascular system with submicron resolution and even allows visualization of subcellular details of labeled structures. We describe mouse transgenic lines that can be used to image the developing vasculature and characterize hemodynamics by tracking individual blood cells. Confocal microscopy of vital fluorescent markers reveals unique details about cell morphogenesis and movement; however, the imaging depth of this method is limited to ∼200 µm. This limitation can be addressed by using OCT, which allows three-dimensional (3D) imaging millimeters into tissue, although this is achieved at the expense of lower spatial resolution (2-10 µm). We describe here how OCT can be applied to the structural analysis of developing mouse embryos and hemodynamic analysis in deep embryonic vessels. These complementary approaches can be used to analyze cardiovascular defects in mutant animals to understand genetic signaling pathways regulating human development.