Smart Hydrogel Microfluidics for Single‐Cell Multiplexed Secretomic Analysis with High Sensitivity

: Secreted proteins determine a range of cellular functionalities correlated with human health and disease progression. Because of cell heterogeneity, it is essential to measure low abundant protein secretions from individual cells to determine single-cell activities. In this study, an integrated platform consisting of smart hydrogel immunosensors for the sensitive detection of single-cell secretions is developed. A single cell and smart hydrogel microparticles are encapsulated within a droplet. After incubation, target secreted proteins from the cell are captured in the smart hydrogel particle for immunoassay. The temperature-induced volume phase transition of the hydrogel biosensor allows the concentration of analytes within the gel matrix to increase, enabling high-sensitivity measurements. Distinct heterogeneity for live cell secretions is determined from 6000 cells within 1 h. This method is tested for low abundant essential secretions, such as interleukin-6, interleukin-8, and monocyte chemoattractant protein-1 secretions of both suspended cells (HL60) and adherent cells (MCF7 and MDA-MB-231). This platform is highly flexible and can be used to simultaneously measure a wide range of clinically relevant cellular secretions; it thus represents a novel tool for precise biological assays.