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Studying cell behavior in whole zebrafish embryos by confocal live imaging: Application to hematopoietic stem cells

机译:共聚焦实时成像研究整个斑马鱼胚胎中的细胞行为:在造血干细胞中的应用

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摘要

Confocal live imaging is a key tool for studying cell behavior in the whole zebrafish embryo. Here we provide a detailed protocol that is adaptable for imaging any progenitor cell behavior in live zebrafish embryos. As an example, we imaged the emergence of the first hematopoietic stem cells from the aorta. We discuss the importance of selecting the appropriate zebrafish transgenic line as well as methods for immobilization of embryos to be imaged. In addition, we highlight the confocal microscopy acquisition parameters required for stem cell imaging and the software tools we used to analyze 4D movies. The whole protocol takes 2 h 15 min and allows confocal live imaging from a few hours to several days.
机译:共聚焦实时成像是研究整个斑马鱼胚胎中细胞行为的关键工具。在这里,我们提供了详细的协议,适用于对活斑马鱼胚胎中的任何祖细胞行为进行成像。例如,我们对来自主动脉的第一个造血干细胞的出现进行了成像。我们讨论选择合适的斑马鱼转基因系的重要性以及固定要成像的胚胎的方法。此外,我们重点介绍了干细胞成像所需的共聚焦显微镜采集参数以及用于分析4D电影的软件工具。整个过程需要2小时15分钟,并且可以进行从几个小时到几天的共聚焦实时成像。

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