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.
Highlights
Binding dynamics for the advantages of simple procedure, low cost and non-invasive[6,7,8,9,10,11,12,13,14,15,16,17]
It is used in two configurations. i) The gold film is placed in a petri dish immersed in cell culture medium, with its top surface fully open to the culture medium environment (Fig. 1b)
This paper reports on a new method employing a micro-nano-device for label-free and real-time monitoring of single cell attachment on template-stripped plasmonic nano-holes
Summary
Binding dynamics for the advantages of simple procedure, low cost and non-invasive[6,7,8,9,10,11,12,13,14,15,16,17]. The optically thick gold film perforated with large-area nano-holes fabricated by template-stripping constitutes the basic sensing unit and cell attachment substrate in experiment. I) The gold film is placed in a petri dish immersed in cell culture medium, with its top surface fully open to the culture medium environment (Fig. 1b) This configuration was adopted initially to develop and demonstrate the constant measurement capability of single cell attachment. Ii) The gold film is enclosed by top PDMS microfluidic channels with single cell trap units (Fig. 1c) This configuration was adopted later to trap and culture an array of single cells on-site for collection of data, which corresponded to the start and end points of measurement. The spectrum data were exported to MATLAB to obtain the fitting curve and fitting function
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