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Fluorescence Lifetime Imaging of Biosensor Peptide Phosphorylation in Single Live Cells

Authors

  • Nur P. Damayanti,

    1. Department of Agricultural and Biological Engineering, Bindley Bioscience Center, Purdue Center for Cancer Research, Purdue University, 225 S. University Street, West Lafayette, IN 47907 (USA)
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  • Prof. Laurie L. Parker,

    Corresponding author
    1. Department of Medicinal Chemistry and Molecular Pharmacology, College of Pharmacy, Purdue Center for Cancer Research, Purdue University, 201 S. University Street, West Lafayette, IN 47907 (USA)
    • Department of Medicinal Chemistry and Molecular Pharmacology, College of Pharmacy, Purdue Center for Cancer Research, Purdue University, 201 S. University Street, West Lafayette, IN 47907 (USA)
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  • Prof. Joseph M. K. Irudayaraj

    Corresponding author
    1. Department of Agricultural and Biological Engineering, Bindley Bioscience Center, Purdue Center for Cancer Research, Purdue University, 225 S. University Street, West Lafayette, IN 47907 (USA)
    • Department of Agricultural and Biological Engineering, Bindley Bioscience Center, Purdue Center for Cancer Research, Purdue University, 225 S. University Street, West Lafayette, IN 47907 (USA)
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  • We acknowledge financial support from the National Institutes of Health (R00CA127161 to L.L.P.) and grants from the Purdue Center for Cancer Research and Indiana Clinical and Translational Sciences Institute (NIH/NCRR TR000006) to J.M.K.I.

Abstract

original image

Better living through biochemistry: Phosphorylation of a Cy5-labeled Abl kinase peptide biosensor resulted in an extension of fluorescence lifetime in live cells. Time-correlated single photon counting fluorescence lifetime imaging (FLIM) enabled excellent signal-to-noise to visualize the subcellular patterns of this sensor (see picture). This strategy should be generalizable to other peptide-based kinase substrates for imaging of kinase activity in single cells.

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