Gierke Sarah, Kumar Praveen, Wittmann Torsten
Department of Cell & Tissue Biology, University of California, San Francisco, California 94143-0512, USA.
Methods Cell Biol. 2010;97:15-33. doi: 10.1016/S0091-679X(10)97002-7.
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.
细胞内微管聚合动力学的时空调节，通过众多微管相关蛋白和其他机制，是许多细胞过程的核心。在这里，我们概述并提供实用指南，介绍如何通过荧光标记整个微管或利用仅与生长中的微管末端特异性结合的蛋白质来获取和分析活细胞中动态微管的延时序列，并总结不同方法的优缺点。我们给出成像条件的实用建议，并讨论这种分析的重要局限性，这些局限性由可实现的最大空间和时间采样频率决定。
