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In Vitro Reconstitution and Imaging of Microtubule Dynamics by Fluorescence and Label-free Microscopy

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 g...

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Detalles Bibliográficos
Autores principales: Hirst, William Graham, Kiefer, Christine, Abdosamadi, Mohammad Kazem, Schäffer, Erik, Reber, Simone
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7757405/
https://www.ncbi.nlm.nih.gov/pubmed/33377071
http://dx.doi.org/10.1016/j.xpro.2020.100177
Descripción
Sumario: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 Hirst et al. (2020).