Previously we have demonstrated surface plasmon enhanced energy transfer between fluorophores and gold nanorods under two-photon excitation using fluorescence lifetime imaging microscopy (FLIM) in both solution and intracellular phases. These studies demonstrated that gold nanoparticle-dye energy transfer combinations are appealing, not only in Förster resonance energy transfer (FRET) imaging, but also energy transfer-based fluorescence lifetime sensing of bio-analytes. Here, we apply this approach to study the internalization of gold nanorods (GNRs) in HeLa cells using the early endosome labeling marker GFP. The observed energy transfer between GFP and the GNRs indicates the involvement of endocytosis in GNR uptake. Moreover, a novel nanoprobe based on oligonucleotide functionalized gold nanorods for nucleic acid sensing via dye-GNRs energy transfer is demonstrated, potentially opening up new possibilities in cancer diagnosis and prognosis. The influence of oligonucleotide design on such nanoprobe performance was studied for the first time using time-resolved fluorescence spectroscopy, bringing new insights to the optimization of the nanoprobe.
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