Investigating morphogenesis in Xenopus embryos: imaging strategies, processing, and analysis.
Investigating morphogenesis in Xenopus embryos: imaging strategies, processing, and analysis.
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DOI：
10.1101/pdb.top073890
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发表时间：
2013-04-01
影响因子：
--
通讯作者：
Davidson, Lance A
中科院分区：
文献类型：
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作者：
Kim, Hye Young;Davidson, Lance A
已经开发了用于可视化从非洲爪蟾胚胎分离的活多细胞组织内的形态发生期间的细胞运动和蛋白质动力学的方法。这些包括非洲爪蟾胚胎中报告基因构建体的制备和使用、分离胚胎组织的显微外科技术以及长期培养活组织的方法。在这篇文章中，我们提出了通过改善信号和最大限度地减少过度暴露对细胞和组织的损伤来成功成像大而厚的胚胎组织的策略。我们还描述了图像分析的策略，包括构建kymographs，使用时间和z投影共焦堆栈，以及使用感兴趣的区域分割图像的方法。有了这些成像工具，“剪切和粘贴”胚胎学的非洲爪蟾模型系统允许显着访问的力学细胞和组织，以及复杂的细胞生物学的粘附和细胞骨架在形态发生。
Methods have been developed for visualizing cell movement and protein dynamics during morphogenesis within live multicellular tissues isolated from Xenopus laevis embryos. These include the preparation and use of reporter constructs in Xenopus embryos, microsurgical techniques for isolating embryonic tissues, and methods for culturing live tissues for extended periods. In this article, we present strategies for successful imaging of large thick embryonic tissues by improving the signal and minimizing damage to cells and tissues from overexposure. We also describe strategies for image analysis, including construction of kymographs, use of time- and z-projected confocal stacks, and approaches to segment images using regions of interest. With these imaging tools, the "cut-and-paste" embryology of the Xenopus model system allows remarkable access to both the mechanics of cells and tissues as well as the complex cell biology of adhesion and the cytoskeleton during morphogenesis.