Single‐Target Pairing System (StarPair) for Large‐Scale Interrogation of Cell–Cell Interactions

Coordination of cell populations through intercellular interactions is essential to the formation and function of living organisms. However, technologies enabling systematic dissection of cell-cell communications or manipulation of cell-cell interactions at the single-cell resolution remain largely out of reach. To solve this, a single-target pairing system, termed StarPair, is proposed, for the high-throughput and high-precision combination of desired single targets in droplets. How target buoyancy, target concentration, sorting parameters, droplet flow rates, and electric stimulation collectively determine the single-target pairing performance is comprehensively explored. Upon optimization of these parameters, pairing efficiencies over 95% can be achieved with an operation frequency reaching one million single-target pairs per 9.5 h for two-target assembly. Leveraging StarPair, the immune cell-cancer cell interactions are assessed between 4 × 10⁵ cell pairs and the NK-92MI cells with high secretion capability are further enriched. It is envisioned that StarPair will open new perspectives for characterizing and manipulating cell-cell interactions at scale.