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Single-molecule imaging of cytoplasmic dynein in vivo

Ananthanarayanan, Vaishnavi; Tolić, Iva Marija (2015) Single-molecule imaging of cytoplasmic dynein in vivo. In: Biophysical Methods in Cell Biology. Elsevier: Monograph Series, pp. 1-12. ISBN 978-0-12-801103-4

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While early fluorescence microscopy experiments employing fluorescent probes afforded snapshots of the cell, the power of live-cell microscopy is required to understand complex dynamics in biological processes. The first successful cloning of green fluorescent protein in the 1990s paved the way for development of approaches that we now utilize for visualization in a living cell. In this chapter, we discuss a technique to observe fluorescently tagged single molecules in fission yeast. With a few simple modifications to the established total internal reflection fluorescence microscopy, cytoplasmic dynein molecules in the cytoplasm and on the microtubules can be visualized and their intracellular dynamics can be studied. We illustrate a technique to study motor behavior, which is not apparent in conventional ensemble studies of motors. In general, this technique can be employed to study single-molecule dynamics of fluorescently tagged proteins in the cell interior.

Item Type: Book Section
Uncontrolled Keywords: Diffusion; Dynein; Fission yeast; HILO; Live-cell imaging; Microtubules; Motor proteins; Single-molecule observation; TIRF; Total internal reflection fluorescence microscopy
Subjects: NATURAL SCIENCES > Physics > Biophysics and Medical Physics
NATURAL SCIENCES > Biology > Biochemistry and Molecular Biology
Divisions: Division of Molecular Biology
Depositing User: Patrik Risteski
Date Deposited: 26 Oct 2015 07:49
DOI: 10.1016/bs.mcb.2014.10.001

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