MIPS Molecular Imaging Program at Stanford

W. E. Moerner

Publication Details

  • Azido push-pull fluorogens photoactivate to produce bright fluorescent labels.

    Lord SJ, Lee HL, Samuel R, Weber R, Liu N, Conley NR, Thompson MA, Twieg RJ, Moerner WE. J Phys Chem B. 2010; 114 (45): 14157-67

    Dark azido push-pull chromophores have the ability to be photoactivated to produce bright fluorescent labels suitable for single-molecule imaging. Upon illumination, the aryl azide functionality in the fluorogens participates in a photochemical conversion to an aryl amine, thus restoring charge-transfer absorption and fluorescence. Previously, we reported that one compound, DCDHF-V-P-azide, was photoactivatable. Here, we demonstrate that the azide-to-amine photoactivation process is generally applicable to a variety of push-pull chromophores, and we characterize the photophysical parameters including photoconversion quantum yield, photostability, and turn-on ratio. Azido push-pull fluorogens provide a new class of photoactivatable single-molecule probes for fluorescent labeling and super-resolution microscopy. Lastly, we demonstrate that photoactivated push-pull dyes can insert into bonds of nearby biomolecules, simultaneously forming a covalent bond and becoming fluorescent (fluorogenic photoaffinity labeling).

    PubMedID: 19860443

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