Altan-Bonnet Nihal
Rutgers University, Department of Biological Sciences, Newark, New Jersey, USA.
Methods Mol Biol. 2007;390:309-28. doi: 10.1007/978-1-59745-466-7_21.
Live-cell imaging is a powerful tool which allows the observation of dynamic cellular processes while maintaining the native organization of the cell. Its advantages over other methods that disrupt cell integrity are abundantly evident in the study of cell division, where multiple subcellular organelles and molecules are involved in dynamic, spatio-temporally regulated processes such as Golgi and nuclear envelope disassembly/reassembly, spindle apparatus formation, chromosome condensation and segregation, and cytoplasmic division. This chapter will describe practical methods for cell synchronization, selection of fluorescent markers for transfection, and setting up imaging conditions and microscope parameters for acquiring time-lapse images of the Golgi apparatus in mitotic cells. These are general methods that can be applied to the study of many different types of organelles and molecules in dividing cells.
活细胞成像技术是一种强大的工具，它能够在维持细胞天然组织结构的同时观察动态细胞过程。与其他破坏细胞完整性的方法相比，其优势在细胞分裂研究中极为明显，在细胞分裂过程中，多个亚细胞器和分子参与了动态的、时空调节的过程，如高尔基体和核膜的解体/重组、纺锤体形成、染色体浓缩和分离以及胞质分裂。本章将介绍细胞同步化的实用方法、用于转染的荧光标记物的选择，以及为有丝分裂细胞中的高尔基体获取延时图像而设置成像条件和显微镜参数的方法。这些都是通用方法，可应用于研究分裂细胞中许多不同类型的细胞器和分子。
