Understanding how individual cells behave is essential for uncovering the complexity of biology and disease, yet most single-cell technologies provide only static molecular snapshots. We present the High-throughput Single-Cell Omni-functional Profiling Engine (HiSCOPE), a microfluidic platform that directly investigates functional dynamics of ≥10,000 single cells. HiSCOPE integrates a hydrodynamic single-cell trapping array with customized modular assay chambers to reproducibly trap single cells, pairs, and trios of cells originating from distinct populations and to support diverse functional assays—including viability, proliferation, cytotoxicity, cytokine secretion, and contact-dependent and -independent cell–cell interactions. Importantly, live-cell retrieval enables downstream analysis of selected cells. We demonstrate HiSCOPE’s utility by evaluating leukemia cells’ drug susceptibility, profiling natural killer (NK) cell cytotoxicity and cytokine secretion, and dissecting T cell–macrophage crosstalk. Our results reveal striking functional diversity that is masked in bulk assays. HiSCOPE offers a scalable solution for functional profiling of single cells, with broad implications for biology, immunotherapy, and personalized medicine.
Shao et al. (Mon,) studied this question.