Department of Biological Chemistry, Medical Scientist Training Program, David Geffen School of Medicine, University of California Los Angeles, Los Angeles, CA, USA.
Department of Neurobiology, Stanford University, Stanford, CA, USA.
Methods Mol Biol. 2021;2274:25-35. doi: 10.1007/978-1-0716-1258-3_3.
Visualizing progression through the cell cycle provides valuable information for the study of development, tissue maintenance, and dysregulated growth in proliferative diseases, such as cancer. Developments in fluorescent biosensors have facilitated dynamic tracking of molecular processes, including the cell cycle. The genetically encoded set of fluorescent indicators, Fucci4, enables the visualization of transitions between each cell cycle phase. Here, we describe a method to track progression through each cell cycle phase using Fucci4 in live epifluorescence imaging. In principle, this approach can be adapted to in vitro time-lapse imaging of any four spectrally resolvable fluorescent indicators.
通过细胞周期的可视化提供了有价值的信息，可用于研究发育、组织维持和增殖性疾病（如癌症）中的失调生长。荧光生物传感器的发展促进了分子过程的动态跟踪，包括细胞周期。荧光指示剂的遗传编码集 Fucci4 能够可视化细胞周期各阶段之间的转变。在这里，我们描述了一种使用 Fucci4 在活荧光显微镜成像中跟踪每个细胞周期阶段进展的方法。原则上，这种方法可以适用于任何四种光谱可分辨荧光指示剂的体外延时成像。
