Label-free and real-time monitoring of single cell attachment on template-stripped plasmonic nano-holes

Abstract

Leveraging microfluidics and nano-plasmonics, we present in this paper a new method employing a micro-nano-device that is capable of monitoring the dynamic cell-substrate attachment process at single cell level in real time without labeling. The micro-nano-device essentially has a gold thin film as the substrate perforated with periodic, near-cm2-area, template-stripped nano-holes, which generate plasmonic extraordinary optical transmission (EOT) with a high sensitivity to refractive index changes at the metal-dielectric interface. Using this device, we successfully demonstrated label-free and real-time monitoring of the dynamic cell attachment process for single mouse embryonic stem cell (C3H10) and human tumor cell (HeLa) by collecting EOT spectrum data during 3-hour on-chip culture. We further collected the EOT spectral shift data at the start and end points of measurement during 3-hour on-chip culture for 50 C3H10 and 50 HeLa cells, respectively. The experiment results show that the single cell attachment process of both HeLa and C3H10 cells follow the logistic retarded growth model, but with different kinetic parameters. Variations in spectral shift during the same culture period across single cells present new evidence for cell heterogeneity. The micro-nano-device provides a new, label-free, real-time, and sensitive, platform to investigate the cell adhesion kinetics at single cell level.

Core–Shell-Structured Gold Nanocone Array for Label-Free DNA Sensing

“…Metal nanostructures have been applied for the detection of biomolecules using their interactions such as antigen−antibody reactions, 16 DNA hybridization, 17,18 and the analysis of cell dynamics. 19 Hence, label-free biosensor chips have the potential to be applied in medical diagnosis, especially in point of care testing (POCT). 20,21 For POCT applications, it is desirable that determination be completed in visible light due to the inexpensive equipment requirement and highly sensitive visual examination.…”

“…Label-free biosensing by metal nanostructures has been a focus area in the biosensor field due to its many advantages such as high sensitivity, inexpensive equipment, simple and easy operation, fast detection, and real-time monitoring capability. Metal nanostructures have been applied for the detection of biomolecules using their interactions such as antigen–antibody reactions, DNA hybridization, , and the analysis of cell dynamics . Hence, label-free biosensor chips have the potential to be applied in medical diagnosis, especially in point of care testing (POCT). , For POCT applications, it is desirable that determination be completed in visible light due to the inexpensive equipment requirement and highly sensitive visual examination.…”

“…Hence, label-free biosensor chips have the potential to be applied in medical diagnosis, especially in point of care testing (POCT). , For POCT applications, it is desirable that determination be completed in visible light due to the inexpensive equipment requirement and highly sensitive visual examination. Various metal nanostructures have been utilized for label-free sensing to date, including periodic metal nanostructure arrays, for example, nanodisk array, nanopillar array, double nanopillars with nanogap, nanohole array, , PCF-SPR structure, metal–insulator–metal structure, and metamaterial-based nanohole array . However, those nanostructures have several disadvantages in sensor applications such as low sensitivity, near-infrared wavelength of LSPR, complex fabrication process, and expensive equipment required.…”

Label‐Free Optofluidic Nanobiosensor Enables Real‐Time Analysis of Single‐Cell Cytokine Secretion

“…This detection scheme circumvents the tedious and costly molecular tagging and enables the elucidation of biomolecular interactions in a noninvasive and dynamic manner. Among various label‐free sensing platforms, optical biosensors are distinguished by their compatibility with physiological solutions, robust performance, and ability to perform quantitative detection . The recent advances in nanotechnology have inspired the development of extraordinarily improved optical biosensors based on photonic nanostructures.…”

Nanohole array plasmonic biosensors: Emerging point-of-care applications

“…Fig 11b shows the schematics of the cell and detection module and a resonance peak shift which was observed upon interactive binding of VEGF to the antibodies with a sensitivity of 145 pg/mL. In another study, (Tu et al 2017) demonstrated label-free cell monitoring of C3H10 and HeLa cells for understanding the cell kinetics of these two different cell types. A mathematical expression was used to fit the retarded growth function and deduce the difference in adhesion dynamics.…”