Cytokine diffusion and dynamics are central to cellular signaling, activation and migration, yet few, if any, technologies are currently able to quantify cytokine protein behavior in real-time, in a label-free fashion and, most importantly, with a spatial resolution that allows pinpointing individual cells. Photonic crystal resonant imaging offers high spatial and temporal resolution so it is an ideal platform for studying this mammalian extracellular process. Unlike other photonic biosensors, the resonant imaging technique offers very high spatial resolution (of order of a few μm), so it can be understood as an imaging array of highly sensitive biosensors. Here we propose and demonstrate a method of quantifying the secreted chemokine gradients from individual cells. Two Baby Hamster Kidney fibroblast cells lines (i.e. BHK cells) were employed in our studies, engineered to overexpress the chemokine interleukin 3 (IL-3) and the human thrombopoietin growth factor hormone (HTPO), respectively. Localized free cytokine/hormone levels were quantified permitting formation of real-time maps of localized secreted protein levels. The technology has been proven to be protein-specific, greatly minimizing non-specific binding from other carbohydrates secreted from the cells and therefore improving the signal-to-noise ratio of the maps. Ultimately, this system allows the identification of specific cells from mixed systems where diverse types of cells are present. This technique represents a powerful tool to noninvasively and nondestructively study cell behavior in-vitro, label-free, and in real-time; overcoming the inherent challenges of label-based methods.
José Juan-Colás, Steven Johnson, and Thomas F. Krauss, "Real-time mapping of cytokine gradients photonic resonant imaging technology (Conference Presentation)," Proc. SPIE 10497, Imaging, Manipulation, and Analysis of Biomolecules, Cells, and Tissues XVI, 104971C (Presented at SPIE BiOS: January 31, 2018; Published: 14 March 2018); https://doi.org/10.1117/12.2286853.5751467560001.
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