An integrated platform for high-throughput analysis and sorting of single mammalian cells

Single mammalian cell heterogeneity plays a critical role in biological research. However, current microfluidic platforms for single-cell analysis still face the challenge of simultaneously achieving high-efficiency capture, cost-effective target cell sorting, and functional integration. To overcome these challenges, we developed a novel microfluidic platform for high-throughput single-cell analysis and sorting. This platform integrates multiple functions, including high-efficiency capture, online incubation, in situ detection, and target sorting. The chip employs a cost-effective polydimethylsiloxane-indium tin oxide (PDMS-ITO) glass configuration with arrayed capture microstructures, achieving average capture efficiency exceeding 93% for mammalian cells (RAW264.7, HCT116, and MCF7) across a size range of approximately 10–20 μm. The platform allows online incubation and artificial intelligence (AI)-assisted detection using both bright-field and fluorescent imaging. Cell viability rates consistently exceeded 80% after 24 h incubation and live/dead staining. A laser-induced microbubble strategy enabled precise releasing target single cells which were subsequently collected in a 96-well plate. In addition, the platform was applied to evaluate heterogeneity in reactive oxygen species (ROS) responses among macrophages, revealing intercellular differences under stimulation. This high-throughput, cost-effective platform combines high capture efficiency, advanced detection, and cell sorting functionality, demonstrating its utility for single mammalian cell heterogeneity studies and downstream applications.