STRUCTURAL SMART MICROGELS- ENHANCING THE SENSITIVITY FOR SINGLE-CELL SECRETOMIC ANALYSIS

Abstract

High-throughput single-cell protein secretion analysis platform with a novel signal enhancement capability was established. Currently, flow cytometry is widely employed for single-cell protein analysis. However, it lacks the ability to directly measure cellular secretions. Herein, a droplet-based platform where each cell was co-encapsulated with a temperature responsive microgel immunosensor was developed. Temperature-induced volume phase transition of the microgels allowed analyte concentration within the gel matrix, enabling rapid signal amplification of ~5-fold. Using this platform, single-cell secretions of the multiple cytokines were analysed for both suspended cells and adherent cells, revealing distinct single-cell secretion heterogeneity. The platform was further improved to enable multi-modal analysis such as simultaneous detection of surface markers and secreted proteins. Overall, this technology builds upon prior successes in single-cell encapsulation in droplets as well as antibody-based immunoassays while utilizing simplified schemes of sample handling and analysis, making it highly versatile for a range of biological sample profiling.