IRI Life Sciences, Humboldt-Universität zu Berlin, Berlin 10115, Germany.
Research School of Biology, The Australian National University, Canberra, ACT 2600, Australia.
STAR Protoc. 2020 Nov 24;1(3):100177. doi: 10.1016/j.xpro.2020.100177. eCollection 2020 Dec 18.
Dynamic microtubules are essential for many processes in the lives of eukaryotic cells. To study and understand the mechanisms of microtubule dynamics and regulation, reconstitution with purified components has proven a vital approach. Imaging microtubule dynamics can be instructive for a given species, isoform composition, or biochemical modification. Here, we describe two methods that visualize microtubule dynamics at high speed and high contrast: (1) total internal reflection fluorescence microscopy and (2) label-free interference reflection microscopy. For complete details on the use and execution of this protocol, please refer to Hirst et al. (2020).
动态微管对于真核细胞的许多生命过程都是必不可少的。为了研究和理解微管动力学和调节的机制，用纯化的成分进行重建已被证明是一种至关重要的方法。对特定物种、同工型组成或生化修饰的微管动力学进行成像可能具有指导意义。在这里，我们描述了两种可以高速高对比度可视化微管动力学的方法：（1）全内反射荧光显微镜和（2）无标记干涉反射显微镜。有关此方案使用和执行的完整详细信息，请参阅 Hirst 等人（2020 年）。
