Fluorescence microscopy applied to the analysis of mitotic spindle dynamics

The mitotic spindle is a cellular structure that guides proper chromosome segregation to daughter cells, thus ensuring genomic stability during cell proliferation. The mitotic spindle is made of microtubules, polymers of alpha and beta tubulin, and it is highly dynamic during the cell cycle, indeed it is build de novo and it is destroyed at each cell division. The spindle form, position and function are tightly coordinated with other cell cycle events and with cell cycle progression. Recent advances in fluorescence microscopy are very useful to study several aspects of mitotic spindle cycle. In this chapter, we will describe the use of fluorescence microscopy and live cell imaging applied to the study of mitotic spindle dynamics in budding yeast, a unicellular eukaryotic organism. Despite the small size of yeast cells, their morphology, subcellular structures and protein localization can be successfully imaged using proper strategies.