In Vitro Reconstitution and Imaging of Microtubule Dynamics by Fluorescence and Label-free Microscopy.
In Vitro Reconstitution and Imaging of Microtubule Dynamics by Fluorescence and Label-free Microscopy.
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DOI：
10.1016/j.xpro.2020.100177
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发表时间：
2020-12-18
期刊：
影响因子：
--
通讯作者：
Reber S
中科院分区：
文献类型：
--
作者：
Hirst WG;Kiefer C;Abdosamadi MK;Schäffer E;Reber S
动态微管在真核细胞生命的许多过程中都是必不可少的。为了研究和理解微管动力学和调控的机制，纯化组分的体外重构已被证明是一种重要的方法。成像微管动力学可以指导一个给定的物种，异构体组成，或生化修饰。在这里，我们描述了两种在高速和高对比度下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。有关使用和执行本协议的完整详情，请参阅。微管动力学的体外重建微管动力学在微管蛋白数量有限时用IRM无标记成像标记微管蛋白方案用TIRFM荧光微管蛋白成像动态微管在真核细胞生命的许多过程中是必不可少的。为了研究和理解微管动力学和调控的机制，纯化组分的体外重构已被证明是一种重要的方法。成像微管动力学可以指导一个给定的物种，异构体组成，或生化修饰。在这里，我们描述了两种在高速和高对比度下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。
Dynamic microtubules are essential for many processes in the lives of eukaryotic cells. To study and understand the mechanisms of microtubule dynamics and regulation, in vitro 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. In vitro reconstitution of microtubule dynamics Label-free imaging by IRM when tubulin amounts are limiting Labeling tubulin protocol Imaging of fluorescent tubulin by TIRFM Dynamic microtubules are essential for many processes in the lives of eukaryotic cells. To study and understand the mechanisms of microtubule dynamics and regulation, in vitro 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.
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影响因子：
3.3
作者：
HYMAN, AA;SALSER, S;MITCHISON, TJ
通讯作者：
MITCHISON, TJ
影响因子：
16.2
作者：
Ori-McKenney KM;McKenney RJ;Huang HH;Li T;Meltzer S;Jan LY;Vale RD;Wiita AP;Jan YN
通讯作者：
Jan YN
DOI：
10.1007/978-1-61779-252-6_2
发表时间：
2011-01-01
期刊：
MICROTUBE DYNAMICS: METHODS AND PROTOCOLS
影响因子：
--
作者：
Gell, Christopher;Friel, Claire T.;Howard, Jonathon
通讯作者：
Howard, Jonathon
影响因子：
3.4
作者：
Aitken, Colin Echeverria;Marshall, R. Andrew;Puglisi, Joseph D.
通讯作者：
Puglisi, Joseph D.
影响因子：
--
作者：
Asbury CL
通讯作者：
Asbury CL