AI-Guided Droplet Microreactors Enable Rapid and Reproducible Protein Crystallization

Protein crystallization is essential for macromolecular structure determination, which in turn enables advances in drug discovery, enzyme engineering, and functional biology. However, conventional crystallization methods require large sample volumes, extensive screening, and long incubation times, while often yielding irreproducible results. Microfluidic droplets reduce reagent use and enable high-throughput screening, but their sealed environments fix solute concentrations at formation, limiting control over supersaturation and crystal growth. Here, we present the Droplet Concentration Control and Vision (DCCV) platform, which integrates programmable osmotic modulation with automated computer vision. Using semi-permeable double emulsion droplets, DCCV allows post-formation, dynamic tuning of solute concentrations through engineered osmotic gradients. A deep learning-based imaging system provides high-throughput, label-free quantification of droplet size, permeability, and morphology over time. As a demonstration, DCCV produced X-ray-quality protein crystals within 20 h, supported by a predictive model of osmotic transport. Beyond crystallization, DCCV establishes a versatile framework for microscale reaction engineering, biomolecular self-assembly, and materials discovery.