TY - JOUR
T1 - Development of Bright and Biocompatible Nanoruby and Its Application to Background-Free Time-Gated Imaging of G-Protein-Coupled Receptors
AU - Sreenivasan, Varun K.A.
AU - Wan Razali, Wan Aizuddin
AU - Zhang, Kai
AU - Pillai, Rashmi R.
AU - Saini, Avishkar
AU - Denkova, Denitza
AU - Santiago, Marina
AU - Brown, Hannah
AU - Thompson, Jeremy
AU - Connor, Mark
AU - Goldys, Ewa M.
AU - Zvyagin, Andrei V.
N1 - Publisher Copyright:
© 2017 American Chemical Society.
PY - 2017/11/15
Y1 - 2017/11/15
N2 - 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.
AB - 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.
KW - GPCR
KW - nanoruby
KW - opioid
KW - single-particle
KW - time-gated microscopy
UR - http://www.scopus.com/inward/record.url?scp=85034759170&partnerID=8YFLogxK
U2 - 10.1021/acsami.7b12665
DO - 10.1021/acsami.7b12665
M3 - Article
C2 - 29022702
AN - SCOPUS:85034759170
SN - 1944-8244
VL - 9
SP - 39197
EP - 39208
JO - ACS Applied Materials and Interfaces
JF - ACS Applied Materials and Interfaces
IS - 45
ER -