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Localizing Protein–Protein Interactions in Living Cells Using Fluorescence Lifetime Imaging Microscopy

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Advanced Fluorescence Microscopy

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1251))

Abstract

In the past decade, advances in fluorescence lifetime imaging have extensively applied in the life sciences, from fundamental biological investigations to advanced clinical diagnosis. Fluorescence lifetime imaging microscopy (FLIM) is now routinely used in the biological sciences to monitor dynamic signaling events inside living cells, e.g., Protein–Protein interactions. In this chapter, we describe the calibration of both time-correlated single-photon counting (TCSPC) and frequency domain (FD) FLIM systems and the acquisition and analysis of FLIM-FRET data for investigating Protein–Protein interactions in living cells.

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Acknowledgments

The authors acknowledge funding from the University of Virginia, National Heart, Lung, and Blood Institute (NHLBI) PO1HL101871 and National Center for Research Resources NCRR-NIH RR027409. The authors thank Ms. Kay Christopher (Biology, University of Virginia) for preparing the samples, Dr. Steven Vogel (NIH/NIAAA) for providing the FRET-standard constructs, and Dr. Richard Day (Indiana University School of Medicine) for providing the C/EBPα-bZip constructs.

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Authors and Affiliations

  1. W.M. Keck Center for Cellular Imaging, Biology, University of Virginia, B005 Physical and Life Sciences Building, White Head Road, Charlottesville, VA, 22904, USA

    Yuansheng Sun & Ammasi Periasamy

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  1. Yuansheng Sun
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  2. Ammasi Periasamy
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Correspondence to Ammasi Periasamy .

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  1. Department of Systemic Cell Biology, Max Planck Institute of Molecular Physiology, Dortmund, Germany

    Peter J. Verveer

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Sun, Y., Periasamy, A. (2015). Localizing Protein–Protein Interactions in Living Cells Using Fluorescence Lifetime Imaging Microscopy. In: Verveer, P. (eds) Advanced Fluorescence Microscopy. Methods in Molecular Biology, vol 1251. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-2080-8_6

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  • DOI: https://doi.org/10.1007/978-1-4939-2080-8_6

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