Fluorescence photoactivation provides a strategy for monitoring protein kinetics within living cells. In particular, fluorescence photoactivation of a subpopulation of microtubule subunits within the spindle using photoactivatable fluorescent tubulin constructs has proven useful for assessing a variety of features of spindle microtubule dynamics, including poleward microtubule movement, microtubule depolymerization, and microtubule turnover, in various cellular settings. The current chapter describes a method for monitoring microtubule dynamics within the mouse egg spindle by photoactivation of photoactivatable-GFP-tubulin, followed by time-lapse confocal imaging.
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