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DOI: 10.3791/4350-v
Please note that some of the translations on this page are AI generated. Click here for the English version.
This article presents a method for tracking photo switched cells in living zebrafish embryos using photoconvertible fluorescent proteins. The technique allows for the monitoring of cell biological events during development through confocal microscopy.
在这里,我们提出了一种在活斑马鱼胚胎中光转化荧光蛋白 (PCFPs) 进行光激活开关的方法,并在发育过程中的特定时间点进一步跟踪光转化蛋白。该方法允许监测活体脊椎动物生物体中不同发育过程背后的细胞生物事件。
该程序的总体目标是追踪活斑马鱼胚胎中的光开关细胞。这是通过首先包埋胚胎,在低熔点温度下表达组织特异性光、可转换荧光蛋白来实现的。接下来,对感兴趣区域内的蛋白质进行完全光转化。
然后小心地将胚胎从 aros 中取出,并在 28.5 摄氏度的鸡蛋水中保持直到后期。最后,光转化的胚胎再次嵌入低熔融温度的 aros 中。最终,可以获得结果表明,由于通过共聚焦显微镜稳定存在光转化蛋白,可以在后期精确检测到光转化细胞。
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