Triple-Color Live Imaging of Mouse Oocytes.

Meiotic division is a dynamic process that exhibits active interactive behaviors amongst different intracellular structures and components for spindle assembly and chromosome segregation. Understanding the mechanisms of meiotic spindle assembly and chromosome segregation therefore requires a quantitative analysis of spatiotemporal relationships among different structures and components. In this chapter, we describe a method for triple-color live imaging of meiotic division in mouse oocytes. This approach combines the microinjection of RNAs encoding proteins tagged with green and red fluorescent proteins and the visualization of microtubules with the fluorogenic far-red probe SiR-Tubulin. This method enables the simultaneous spatiotemporal mapping of three different components of the spindle and chromosomes, which opens the way to quantitative analysis of their interactive behaviors.