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STAR Protoc
2020 Dec 18;13:100177. doi: 10.1016/j.xpro.2020.100177.
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In Vitro Reconstitution and Imaging of Microtubule Dynamics by Fluorescence and Label-free Microscopy.
Hirst WG
,
Kiefer C
,
Abdosamadi MK
,
Schäffer E
,
Reber S
.
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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).
Aitken,
An oxygen scavenging system for improvement of dye stability in single-molecule fluorescence experiments.
2008, Pubmed
Aitken,
An oxygen scavenging system for improvement of dye stability in single-molecule fluorescence experiments.
2008,
Pubmed
Asbury,
Data Analysis for Total Internal Reflection Fluorescence Microscopy.
2016,
Pubmed
Axelrod,
Total internal reflection fluorescence.
1984,
Pubmed
Bormuth,
LED illumination for video-enhanced DIC imaging of single microtubules.
2007,
Pubmed
Bugiel,
The Kinesin-8 Kip3 Depolymerizes Microtubules with a Collective Force-Dependent Mechanism.
2020,
Pubmed
CURTIS,
THE MECHANISM OF ADHESION OF CELLS TO GLASS. A STUDY BY INTERFERENCE REFLECTION MICROSCOPY.
1964,
Pubmed
Caplow,
Evidence that a single monolayer tubulin-GTP cap is both necessary and sufficient to stabilize microtubules.
1996,
Pubmed
Chaaban,
The Structure and Dynamics of C. elegans Tubulin Reveals the Mechanistic Basis of Microtubule Growth.
2018,
Pubmed
Dixit,
Studying plus-end tracking at single molecule resolution using TIRF microscopy.
2010,
Pubmed
Gardner,
Rapid microtubule self-assembly kinetics.
2011,
Pubmed
Gell,
Microtubule dynamics reconstituted in vitro and imaged by single-molecule fluorescence microscopy.
2010,
Pubmed
Gell,
Purification of tubulin from porcine brain.
2011,
Pubmed
Guo,
Mechanism and dynamics of breakage of fluorescent microtubules.
2006,
Pubmed
Helenius,
The depolymerizing kinesin MCAK uses lattice diffusion to rapidly target microtubule ends.
2006,
Pubmed
Hirst,
Differences in Intrinsic Tubulin Dynamic Properties Contribute to Spindle Length Control in Xenopus Species.
2020,
Pubmed
,
Xenbase
Hyman,
Role of GTP hydrolysis in microtubule dynamics: information from a slowly hydrolyzable analogue, GMPCPP.
1992,
Pubmed
Kapoor,
MTrack: Automated Detection, Tracking, and Analysis of Dynamic Microtubules.
2019,
Pubmed
Mahamdeh,
Label-free high-speed wide-field imaging of single microtubules using interference reflection microscopy.
2018,
Pubmed
Mitchison,
Dynamic instability of microtubule growth.
,
Pubmed
Nguyen,
Using supported bilayers to study the spatiotemporal organization of membrane-bound proteins.
2015,
Pubmed
,
Xenbase
Ori-McKenney,
Phosphorylation of β-Tubulin by the Down Syndrome Kinase, Minibrain/DYRK1a, Regulates Microtubule Dynamics and Dendrite Morphogenesis.
2016,
Pubmed
Rasnik,
Nonblinking and long-lasting single-molecule fluorescence imaging.
2006,
Pubmed
Reusch,
Affinity Purification of Label-free Tubulins from Xenopus Egg Extracts.
2020,
Pubmed
,
Xenbase
Schindelin,
Fiji: an open-source platform for biological-image analysis.
2012,
Pubmed
Simmert,
LED-based interference-reflection microscopy combined with optical tweezers for quantitative three-dimensional microtubule imaging.
2018,
Pubmed
Szkop,
A simple and reproducible protocol of glass surface silanization for TIRF microscopy imaging.
2018,
Pubmed
Ti,
Human β-Tubulin Isotypes Can Regulate Microtubule Protofilament Number and Stability.
2018,
Pubmed
Vemu,
Tubulin isoform composition tunes microtubule dynamics.
2017,
Pubmed
Zanic,
Measuring the Effects of Microtubule-Associated Proteins on Microtubule Dynamics In Vitro.
2016,
Pubmed
