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Photoswitching Proteins pp 1–17Cite as

Photoinduced Damage Resulting from Fluorescence Imaging of Live Cells

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Part of the book series: Methods in Molecular Biology ((MIMB,volume 1148))

Abstract

The widespread application of fluorescence microscopy to study live cells has led to a greater understanding of numerous biological processes. Many techniques have been developed to uniquely label structures and track metabolic pathways using fluorophores in live cells. However, the photochemistry of nonnative compounds and the deposition of energy into the cell during imaging can result in unexpected and unwanted side effects. Herein, we examine potential live cell damage by first discussing common imaging considerations and modalities in fluorescence microscopy. We then consider several mechanisms by which various photochemical and photophysical phenomena cause cellular damage and introduce techniques that have leveraged these phenomena to intentionally create damage inside cells. Reviewing conditions under which intentional damage occurs can allow one to better predict when unintentional damage may be important. Finally, we delineate ways of checking for and reducing photochemical and photophysical damage.

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Acknowledgment

Funding for this work was provided by the National Science Foundation under Grant No. PHY-1150017.

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Authors and Affiliations

  1. Department of Chemistry, University of North Carolina at Chapel Hill, Chapel Hill, NC, 27599-3290, USA

    Matthew K. Daddysman, Michael A. Tycon & Christopher J. Fecko

Authors
  1. Matthew K. Daddysman
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  2. Michael A. Tycon
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  3. Christopher J. Fecko
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Corresponding author

Correspondence to Christopher J. Fecko .

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Editors and Affiliations

  1. Institute of Anatomy and Cell Biology, University of Heidelberg, Heidelberg, Germany

    Sidney Cambridge

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Daddysman, M.K., Tycon, M.A., Fecko, C.J. (2014). Photoinduced Damage Resulting from Fluorescence Imaging of Live Cells. In: Cambridge, S. (eds) Photoswitching Proteins. Methods in Molecular Biology, vol 1148. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-0470-9_1

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  • DOI: https://doi.org/10.1007/978-1-4939-0470-9_1

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-0469-3

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