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Studying Kinetochores In Vivo Using FLIM-FRET

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The Mitotic Spindle

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1413))

Abstract

Kinetochores play essential roles in coordinating mitosis, as a mechanical connector between chromosome and microtubule and as a source of numerous biochemical signals. These mechanical and biochemical behaviors of kinetochores change dynamically in cells during mitosis. Therefore, understanding kinetochore function requires an imaging tool that quantifies the protein–protein interactions or biochemical changes with high spatiotemporal resolution. FRET has previously been used in combination with biosensors to probe protein–protein interactions and biochemical activity. In this chapter, we introduce FLIM-FRET, a lifetime-based method that quantifies FRET, and describe the use of FLIM-FRET as a method for studying dynamic kinetochore behavior in cells with high spatiotemporal resolution.

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Author information

Authors and Affiliations

  1. School of Engineering and Applied Sciences, Harvard University, Cambridge, MA, USA

    Tae Yeon Yoo & Daniel J. Needleman

  2. Departments of Applied Physics, and Molecular and Cellular Biology, Harvard University, 365.1 Northwest Building, 52 Oxford St., Cambridge, MA, 02138, USA

    Daniel J. Needleman

Authors
  1. Tae Yeon Yoo
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  2. Daniel J. Needleman
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Corresponding author

Correspondence to Daniel J. Needleman .

Editor information

Editors and Affiliations

  1. Massachusetts Institute of Technolo, Cambridge, Massachusetts, USA

    Paul Chang

  2. Dept of Cell & Developmental Biolog, Vanderbilt University, Nashville, Tennessee, USA

    Ryoma Ohi

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© 2016 Springer Science+Business Media New York

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Yoo, T.Y., Needleman, D.J. (2016). Studying Kinetochores In Vivo Using FLIM-FRET. In: Chang, P., Ohi, R. (eds) The Mitotic Spindle. Methods in Molecular Biology, vol 1413. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-3542-0_11

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  • DOI: https://doi.org/10.1007/978-1-4939-3542-0_11

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-3540-6

  • Online ISBN: 978-1-4939-3542-0

  • eBook Packages: Springer Protocols

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