Bioscience Department, Veritas Corporation, Minato-ku, Tokyo, Japan.
Methods Mol Biol. 2023;2519:27-40. doi: 10.1007/978-1-0716-2433-3_4.
Cell cycle-dependent regulation of chromosome is a dynamic event. After replication in S phase, sister chromatids show dynamic behavior including condensation, alignment, and segregation in M phase. These beautiful behaviors of chromosomes observed through the microscope have fascinated people since more than 100 years ago, and now we can sketch the dynamics of regulatory proteins and their posttranscriptional modifications through the fluorescent microscope. The purpose of this chapter is describing the basic methods of immunofluorescence analysis of mitotic cells and chromosomes. Besides, the key ideas for improving the preparation of the specimen are also described. Because the characteristic of the proteins of your interest differs one by one, modifying the method might cause the crucial improvement in the observation.
细胞周期中染色体的调节是一个动态事件。在 S 期复制后,姐妹染色单体在 M 期表现出动态行为,包括浓缩、对齐和分离。自 100 多年前以来,通过显微镜观察到的这些染色体的美丽行为一直令人们着迷,现在我们可以通过荧光显微镜描绘出调节蛋白及其转录后修饰的动态。本章旨在描述有丝分裂细胞和染色体免疫荧光分析的基本方法。此外,还描述了改善标本制备的关键思路。由于您感兴趣的蛋白质的特性各不相同,因此修改方法可能会对观察结果产生至关重要的影响。