Same-cell correlative video light microscopy/Electron Microscopy tomography: An approach to understanding kinetochore behavior during mitosis
The goal of structural biology is to determine structure, structural relationships, and changes in these parameters in an effort to comprehend mechanism of function. In this context same-cell correlative LM/EM provides a powerful approach for elucidating the mechanisms responsible for the behavior of cell components. In this method the event of interest is followed in vivo by video-LM, and the cell then fixed at a critical time during the observational period for a subsequent 3D EM analysis. In this manner the history of a particular event or response can be correlated with the 3D ultrastructure underlying the event.Biologists have long sought to elucidate the mechanism(s) that generate, control and coordinate the poleward and away-from-pole motion of sister kinetochores on each chromosome during mitosis. These “congression” movements require the association of microtubules with each kinetochore (kMTs), and ultimately align the chromosome on the spindle equator. Motion in each direction is distinct because the slow growing (−) ends of kMTs end near the spindle pole while the rapidly growing (+) ends terminate in the kinetochore plate.