Analysis of microtubule polymerization dynamics in live cells.
The spatiotemporal regulation of intracellular microtubule polymerization dynamics, by numerous microtubule-associated proteins and other mechanisms, is central to many cell processes. Here, we give an overview and practical guide on how to acquire and analyze time-lapse sequences of dynamic microtubules in live cells by either fluorescently labeling entire microtubules or by utilizing proteins that specifically associate only with growing microtubule ends and summarize the strengths and weaknesses of different approaches. We give practical recommendations for imaging conditions, and discuss important limitations of such analysis that are dictated by the maximum achievable spatial and temporal sampling frequencies.
Gierke
Sarah
S
Department of Cell & Tissue Biology, University of California, San Francisco, California 94143-0512, USA.
Kumar
Praveen
P
Wittmann
Torsten
T
D000256:Adenoviridae / Q000502:physiology; D000818:Animals; D002468:Cell Physiological Phenomena; D049109:Cell Proliferation; D019411:Clinical Laboratory Techniques; D006801:Humans; D008856:Microscopy, Fluorescence / Q000379:methods; D008870:Microtubules / Q000737:chemistry / Q000378:metabolism*; D008954:Models, Biological; D055503:Protein Multimerization / Q000502:physiology*