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Live-Cell Migration and Adhesion Turnover Assays

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Cell Imaging Techniques

Part of the book series: Methods in Molecular Biology ((MIMB,volume 931))

Abstract

Fluorescence microscopy has revolutionized the way live-cell imaging is achieved. At the same time, it is also potentially harmful to a living specimen. Therefore, the specimen must be monitored for viability and health before, during, and after imaging sessions. Methods for monitoring cell viability and health will be discussed in this chapter. Another key to successful live-cell imaging is to minimize light exposure as much as possible. A summary of strategies for minimizing light exposure including maximizing the light throughput of the microscope and the sensitivity of light detection is presented. Various fluorescence microscopy techniques are presented with a focus on how the light is delivered to the sample (i.e., light density) and pros and cons for use with living specimens. The reader is also directed to other publications that go into these topics in more detail. Methods are described on how to prepare samples for single cell migration assays, how to measure cell migration rates (e.g., bright-field, semi-automated, and automated), and how to measure focal adhesion turnover rates. Details of how to correct images for background intensity and field-illumination uniformity artifacts for quantitative imaging are also described. Overall, this chapter will be helpful to scientists who are interested in imaging live specimens using fluorescence microscopy techniques. It will be of particular interest to anyone wanting to perform quantitative fluorescence imaging, and wanting to measure cell migration rates, and focal adhesion dynamics.

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Acknowledgments

Images for this chapter were collected at the McGill University Life Sciences Complex Imaging Facility funded by the Canadian Foundation for Innovation (CFI) and the Ministère du Développement économique, Innovation et Exportation—Québec (MDEIE) and the Cell Imaging and Analysis Network (CIAN), also funded by the CFI. Thank you to users and colleagues of CIAN, the McGill Imaging facility, MIA Cellavie, the Association of Biomolecular Resource Facilities, The Canadian Cytometry and Microscopy Association (CCMA) and corporate application scientists with whom we constantly interact. Thank you to everyone who read the chapter and gave us valuable constructive feedback. To Ji-Sook Lee for critical reading of the chapter.

Author information

Authors and Affiliations

  1. Department of Biology, Cell Imaging and Analysis Network, McGill University, Montreal, Canada

    J. Lacoste

  2. Microscopy Imaging Analysis Cellavie Inc., Montreal, Canada

    J. Lacoste

  3. Department of Physiology, McGill University, Montreal, Canada

    K. Young

  4. Department of Physiology, Life Sciences Complex Imaging Facility, McGill University, Montreal, Canada

    Claire M. Brown

Authors
  1. J. Lacoste
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  2. K. Young
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  3. Claire M. Brown
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Corresponding author

Correspondence to Claire M. Brown .

Editor information

Editors and Affiliations

  1. College of Medicine, Professor, Department of Pathology, University of Vermont, Beaumont Ave 89, Burlington, 05405-1742, Vermont, USA

    Douglas J. Taatjes

  2. , Abt. Zell- und Molekularpathologie, Universität Zürich, Schmelzbergstr. 12, Zürich, 8091, Switzerland

    Jürgen Roth

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Lacoste, J., Young, K., Brown, C.M. (2012). Live-Cell Migration and Adhesion Turnover Assays. In: Taatjes, D., Roth, J. (eds) Cell Imaging Techniques. Methods in Molecular Biology, vol 931. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-056-4_3

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  • DOI: https://doi.org/10.1007/978-1-62703-056-4_3

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  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-62703-055-7

  • Online ISBN: 978-1-62703-056-4

  • eBook Packages: Springer Protocols

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