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Type: Journal article
Title: Development of bright and biocompatible nanoruby and its application to background-free time-gated imaging of g-protein-coupled receptors
Author: Sreenivasan, V.
Wan Razali, W.
Zhang, K.
Pillai, R.
Saini, A.
Denkova, D.
Santiago, M.
Brown, H.
Thompson, J.
Connor, M.
Goldys, E.
Zvyagin, A.
Citation: ACS Applied Materials and Interfaces, 2017; 9(45):39197-39208
Publisher: ACS Publications
Issue Date: 2017
ISSN: 1944-8244
Statement of
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 and Andrei V. Zvyagin
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.
Keywords: GPCR
time-gated microscopy
Rights: © 2017 American Chemical Society
DOI: 10.1021/acsami.7b12665
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