Imaging the Golgi apparatus in living mitotic cells

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.