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

*Corresponding author for this work

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 - 15 Nov 2017

Keywords

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

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