Abstract
Cell communication is primarily regulated by secreted proteins, whose inhomogeneous secretion often indicates physiological disorder. Parallel monitoring of innate protein-secretion kinetics from individual cells is thus crucial to unravel systemic malfunctions. Here, we report a label-free, high-throughput method for parallel, in vitro, and real-time analysis of specific single-cell signaling using hyperspectral photonic crystal resonant technology. Heterogeneity in physiological thrombopoietin expression from individual HepG2 liver cells in response to platelet desialylation was quantified demonstrating how mapping real-time protein secretion can provide a simple, yet powerful approach for studying complex physiological systems regulating protein production at single-cell resolution.
Highlights
Cell communication is primarily regulated by secreted proteins, whose inhomogeneous secretion often indicates physiological disorder
The functionalized Photonic crystal resonant surfaces (PCRS) were subsequently exposed to BHKTPO cells suspended in the supporting culture media [concentration of 105 units per milliliter (U/mL), significantly below the 100% confluence level to avoid contact inhibition of inherent protein/gene expression] and the adhesion of a single baby hamster kidney cells (BHK)-TPO cell in our defined region of interest was recorded using hyperspectral imaging
This increase in resonant frequency around the cell is associated with the local increase in refractive index due to TPO secreted from the BHK cell binding to the anti-TPO antibodies immobilized on the PCRS
Summary
Cell communication is primarily regulated by secreted proteins, whose inhomogeneous secretion often indicates physiological disorder. We report a high-throughput method for parallel, in vitro, and real-time analysis of specific single-cell signaling using hyperspectral photonic crystal resonant technology without the need of adding any fluorescent label.
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