Development of Bright and Biocompatible Nanoruby and Its Application to Background-Free Time-Gated Imaging of G-Protein-Coupled Receptors

Varun K.A. Sreenivasan, Wan Aizuddin Wan Razali, Kai Zhang, Rashmi R. Pillai, Avishkar Saini, Denitza Denkova, Marina Santiago, Hannah Brown, Jeremy Thompson, Mark Connor, Ewa M. Goldys, Andrei V. Zvyagin

    Research output: Contribution to journalArticlepeer-review

    7 Citations (Scopus)

    Abstract

    At the forefront of developing fluorescent probes for biological imaging applications are enhancements aimed at increasing their brightness, contrast, and photostability, especially toward demanding applications of single-molecule detection. In comparison with existing probes, nanorubies exhibit unlimited photostability and a long emission lifetime (∼4 ms), which enable continuous imaging at single-particle sensitivity in highly scattering and fluorescent biological specimens. However, their wide application as fluorescence probes has so far been hindered by the absence of facile methods for scaled-up high-volume production and molecularly specific targeting. The present work encompasses the large-scale production of colloidally stable nanoruby particles, the demonstration of their biofunctionality and negligible cytotoxicity, as well as the validation of its use for targeted biomolecular imaging. In addition, optical characteristics of nanorubies are found to be comparable or superior to those of state-of-the-art quantum dots. Protocols of reproducible and robust coupling of functional proteins to the nanoruby surface are also presented. As an example, NeutrAvidin-coupled nanoruby show excellent affinity and specificity to μ-opioid receptors in fixed and live cells, allowing wide-field imaging of G-protein coupled receptors with single-particle sensitivity.

    Original languageEnglish
    Pages (from-to)39197-39208
    Number of pages12
    JournalACS Applied Materials and Interfaces
    Volume9
    Issue number45
    DOIs
    Publication statusPublished or Issued - 15 Nov 2017

    Keywords

    • GPCR
    • nanoruby
    • opioid
    • single-particle
    • time-gated microscopy

    ASJC Scopus subject areas

    • Materials Science(all)

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