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Live-Cell Imaging of Mitochondria and the Actin Cytoskeleton in Budding Yeast

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Cytoskeleton Methods and Protocols

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1365))

Abstract

Maintenance and regulation of proper mitochondrial dynamics and functions are necessary for cellular homeostasis. Numerous diseases, including neurodegeneration and muscle myopathies, and overall cellular aging are marked by declining mitochondrial function and subsequent loss of multiple other cellular functions. For these reasons, optimized protocols are needed for visualization and quantification of mitochondria and their function and fitness. In budding yeast, mitochondria are intimately associated with the actin cytoskeleton and utilize actin for their movement and inheritance. This chapter describes optimal approaches for labeling mitochondria and the actin cytoskeleton in living budding yeast cells, for imaging the labeled cells, and for analyzing the resulting images.

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Acknowledgments

This work was supported by awards from the Ellison Medical Foundation (AG-SS-2465) and the NIH (GM45735, GM45735S1, and GM096445) to L.A.P. GM45735S1 was issued from the NIH under the American Recovery and Reinvestment Act of 2009. The microscopes used for these studies were supported in part through a NIH⁄NCI grant (5 P30 CA13696) and obtained using funds from the NIH-NCRR (1S10OD014584) to L.A.P.

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Authors and Affiliations

  1. Department of Pathology and Cell Biology, College of Physicians and Surgeons, Columbia University, 630 W. 168th Street, New York, NY, 10032, USA

    Ryo Higuchi-Sanabria, Istvan R. Boldogh & Liza A. Pon

  2. Confocal and Specialized Microscopy Shared Resource, Herbert Irving Comprehensive Cancer Center, Columbia University, New York, NY, USA

    Theresa C. Swayne & Liza A. Pon

Authors
  1. Ryo Higuchi-Sanabria
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  2. Theresa C. Swayne
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  3. Istvan R. Boldogh
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  4. Liza A. Pon
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Corresponding author

Correspondence to Liza A. Pon .

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Editors and Affiliations

  1. Brooklyn College, Brooklyn, New York, USA

    Ray H. Gavin

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Higuchi-Sanabria, R., Swayne, T.C., Boldogh, I.R., Pon, L.A. (2016). Live-Cell Imaging of Mitochondria and the Actin Cytoskeleton in Budding Yeast. In: Gavin, R. (eds) Cytoskeleton Methods and Protocols. Methods in Molecular Biology, vol 1365. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-3124-8_2

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  • DOI: https://doi.org/10.1007/978-1-4939-3124-8_2

  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-3123-1

  • Online ISBN: 978-1-4939-3124-8

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