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Biochemistry

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JoVE Journal Biochemistry
Simultaneous Visualization of the Dynamics of Crosslinked and Single Microtubules In Vitro by TIRF Microscopy
 

Simultaneous Visualization of the Dynamics of Crosslinked and Single Microtubules In Vitro by TIRF Microscopy

Article DOI: 10.3791/63377-v • 07:20 min • February 18th, 2022
February 18th, 2022 •
*1,2, *1, 1,2
1Molecular Biology, Massachusetts General Hospital, 2Department of Genetics, Harvard Medical School
* These authors contributed equally

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Summary
Automatic Translation

Here, a TIRF microscopy-based in vitro reconstitution assay is presented to simultaneously quantify and compare the dynamics of two microtubule populations. A method is described to simultaneously view the collective activity of multiple microtubule-associated proteins on crosslinked microtubule bundles and single microtubules.

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TIRF Microscopy Simultaneous Visualization Dynamics Crosslinked Microtubules Single Microtubules In Vitro Microtubule Arrays Dynamic Properties Regulators Proximal Microtubule Subpopulations Bottom-up Restitution Assay Organization Self-organization Multi-component Reconstitution Experimental Parameters PH Ionic Strength Protein Concentration Systematic Analysis BRB ADDTT Microscope Slide Coverslip Clear Nail Polish VALAP Sealant TIRF Objective Fluorescence Labeling Dilution Of Microtubules Laser Intensity
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