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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

Besser leben durch Biochemie: Die Phosphorylierung eines Cy5-markierten Abl-Kinase-Biosensors führte zu einer verlängerten Fluoreszenzlebensdauer in lebenden Zellen. Zeitkorrelierte Fluoreszenzlebensdauer-Bildgebung (FLIM) mit Zählung einzelner Photonen ermöglichte die Visualisierung der subzellulären Muster des Sensors (siehe Bild). Die Strategie sollte auf andere Peptid-basierte Kinasen zur Bildgebung der Kinase-Aktivität in einzelnen Zellen übertragbar sein.

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