Ultrasensitive and real-time optical detection of cellular oxidative stress using graphene-covered tunable plasmonic interfaces

Nano Convergence - Tập 9 - Trang 1-12 - 2022
Hakchun Kim1, Hyun Ji An1, Junhee Park1, Yohan Lee2, Min Seob Kim3, Seungki Lee1, Nam Dong Kim3, Jihwan Song2, Inhee Choi1
1Department of Life Science, University of Seoul, Seoul, Republic of Korea
2Department of Mechanical Engineering, Hanbat National University, Daejeon, Republic of Korea
3Institute of Advanced Composite Materials, Korea Institute of Science and Technology, Wanju-gun, Republic of Korea

Tóm tắt

Reactive oxygen species (ROS) regulate various physiological and pathological conditions in cells by interacting with signaling molecules and inducing oxidative stress. Therefore, sensitive monitoring of ROS levels in living cells is important to track cellular state and study the complex role of ROS in the development of various pathologies. Herein, we present an optically tunable plasmonic interface covered with graphene to monitor cellular ROS levels with superior sensitivity and cellular comfortability. As a sensing principle, we employed plasmon resonance energy transfer (PRET)-based spectral quenching dips modulated by redox-active cytochrome c for real-time monitoring. By transferring graphene layers to plasmonic nanoparticles immobilized on a glass substrate, the scattering profiles of the nanoprobes were adjusted in terms of the position, width, and intensity of the peaks to determine the optimal conditions for measuring the PRET signal. Using the optimized graphene-covered plasmonic nanoprobe, we obtained calibration curves over a wide concentration range from femtomoles to millimoles for hydrogen peroxide based on the change in the PRET signal. Before monitoring cellular ROS, we confirmed that a high density of cells adhered well to the graphene-covered plasmonic interface by observing immunofluorescence images of the cytoskeleton of the immobilized cells. Finally, we monitored the real-time ROS generated by the cells under oxidative stress conditions by directly measuring the spectral changes of the probes around the cells. We believe that the proposed graphene-covered tunable plasmonic interface has versatile applicability for investigating cellular stress and disease progression by monitoring ROS levels under various cellular conditions.

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