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TitleUltrasensitive optical imaging with lanthanide lumiphores
AuthorUkrae Cho, Daniel P Riordan, Paulina Ciepla, Kiranmai S Kocherlakota, James K Chen & Pehr B Harbury
Affiliation(s)Department of Biochemistry, Stanford University School of Medicine, Stanford, California, USA.
PublishedNature Chemical Biology doi:10.1038/nchembio.2513 06 November 2017
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AbstractIn principle, the millisecond emission lifetimes of lanthanide chelates should enable their ultrasensitive detection in biological systems by time-resolved optical microscopy. In practice, however, lanthanide imaging techniques have provided no better sensitivity than conventional fluorescence microscopy. Here, we identified three fundamental problems that have impeded lanthanide microscopy: low photon flux, inefficient excitation, and optics-derived background luminescence. We overcame these limitations with a new lanthanide imaging modality, transreflected illumination with luminescence resonance energy transfer (trLRET), which increases the time-integrated signal intensities of lanthanide lumiphores by 170-fold and the signal-to-background ratios by 75-fold. We demonstrate that trLRET provides at least an order-of-magnitude increase in detection sensitivity over that of conventional epifluorescence microscopy when used to visualize endogenous protein expression in zebrafish embryos. We also show that trLRET can be used to optically detect molecular interactions in vivo. trLRET promises to unlock the full potential of lanthanide lumiphores for ultrasensitive, autofluorescence-free biological imaging.

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