Laboratory of Immunology, Faculty of Pharmacy, Osaka Ohtani University, Tondabayashi, Osaka 584-8540, Japan.
Laboratory of Immunology, Faculty of Pharmacy, Osaka Ohtani University, Tondabayashi, Osaka 584-8540, Japan; Research Fellow of Japan Society for the Promotion of Science, Japan; Department of Immunology, Genetics and Pathology, Uppsala University, Uppsala, Sweden.
J Neurosci Methods. 2021 May 1;355:109127. doi: 10.1016/j.jneumeth.2021.109127. Epub 2021 Mar 17.
Cell migration and cell proliferation are the basic principles that make up a living organism, and both biologically and medically. In order to understand living organism and biological phenomena, it is essential to track the migration, proliferation, and fate of cells in living cells and animals and to clarify the properties and molecular expression of cells. Recent developments in novel fluorescent proteins have made it possible to observe cell migration and proliferation as the cell cycle at the single-cell level in living individuals and tissues. Here, we introduce cell cycle visualization of living cells and animals by Fucci (Fluorescent Ubiquitination-based Cell Cycle Indicator) system and in situ cell labeling of cells and tracking cell migration by photoactivatable and photoconvertible proteins. In addition, we will present our established methods as an example of combines above tools with single-cell molecular expression analysis to reveal the fate of migrating cells at single cell level.
细胞迁移和细胞增殖是构成生物体的基本原理，无论是在生物学还是医学领域。为了了解生物体和生物现象，有必要追踪活细胞和动物中细胞的迁移、增殖和命运，并阐明细胞的特性和分子表达。新型荧光蛋白的最新发展使得在活体个体和组织中以单细胞水平观察细胞周期中的细胞迁移和增殖成为可能。在这里，我们通过 Fucci（荧光泛素化细胞周期指示剂）系统介绍了活细胞和动物的细胞周期可视化，以及细胞的原位标记和通过光活化和光转化蛋白追踪细胞迁移。此外，我们将以我们建立的方法为例，将上述工具与单细胞分子表达分析相结合，以揭示单细胞水平迁移细胞的命运。
