The Graduate School, Syracuse University, Syracuse, NY, USA.
The Laboratory of Cell and Molecular Biology, National Institutes of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Bethesda, MD, USA.
Methods Mol Biol. 2020;2150:11-23. doi: 10.1007/7651_2019_225.
The maintenance of many adult tissues depends on stem cell systems, which must balance proliferation and differentiation. To understand the properties of adult stem cell systems, one powerful tool is visualization of the cell dynamics in vivo. Here we describe a protocol to track cells in the germline progenitor zone (which includes germline stem cells) in live C. elegans adult worms. Tracking is achieved by using a genetically encoded photoconvertible fluorescent protein, where photoconversion is used to mark cells of interest and their descendants. Individual worms are immobilized, the cells of interest are selected for photoconversion, and the worms are then recovered to plates and imaged at later timepoints. This allows longitudinal studies of individual worms, providing valuable information regarding germline stem cell dynamics.
许多成人组织的维持依赖于干细胞系统，干细胞系统必须平衡增殖和分化。为了了解成人干细胞系统的特性，一种强大的工具是在体内可视化细胞动力学。在这里，我们描述了一种在活体 C. elegans 成虫中追踪生殖系祖细胞区（包括生殖干细胞）细胞的方案。通过使用遗传编码的光可转化荧光蛋白进行追踪，其中光转化用于标记感兴趣的细胞及其后代。将单个蠕虫固定，选择感兴趣的细胞进行光转化，然后将蠕虫回收至平板上，并在稍后的时间点进行成像。这允许对单个蠕虫进行纵向研究，提供有关生殖干细胞动力学的有价值信息。
