Imaging the environment of green fluorescent protein

Klaus Suhling, Jan Siegel, David Phillips, Paul M W French, Sandrine Lévêque-Fort, Stephen E D Webb, Daniel M. Davis

    Research output: Contribution to journalArticlepeer-review

    Abstract

    An emerging theme in cell biology is that cell surface receptors need to be considered as part of supramolecular complexes of proteins and lipids facilitating specific receptor conformations and distinct distributions, e.g., at the immunological synapse. Thus, a new goal is to develop bioimaging that not only locates proteins in live cells but can also probe their environment. Such a technique is demonstrated here using fluorescence lifetime imaging of green fluorescent protein (GFP). We first show, by time-correlated single-photon counting, that the fluorescence decay of GFP depends on the local refractive index. This is in agreement with the Strickler Berg formula, relating the Einstein A and B coefficients for absorption and spontaneous emission in molecules. We then quantitatively image, by wide-field time-gated fluorescence lifetime imaging, the refractive index of the environment of GFP. This novel approach paves the way for imaging the biophysical environment of specific GFP-tagged proteins in live cells.
    Original languageEnglish
    Pages (from-to)3589-3595
    Number of pages6
    JournalBIOPHYSICAL JOURNAL
    Volume83
    Issue number6
    DOIs
    Publication statusPublished - 1 Dec 2002

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