Parallelized droplet microfluidic mechanoporation enables robust and clogging-resistant intracellular gene delivery.

Microfluidic platforms have emerged as powerful tools for efficient intracellular delivery of exogenous cargo. While droplet microfluidics coupled with cell mechanoporation shows significant potential, its broader adoption is often hindered by channel clogging and limited scalability. To address these challenges, we developed a parallelized droplet-based cell mechanoporation platform with integrated bypass channels. This architecture stabilizes internal pressure and mitigates clogging-induced failure, ensuring robust and continuous operation. The platform achieves delivery efficiencies exceeding 98% and cell viabilities above 80% at a throughput of 2 × 10⁷ cells/h, enables highly efficient mRNA transfection (~98%), and supports CRISPR/Cas9-mediated CD3 knock-out. Collectively, these results establish parallelized droplet cell mechanoporation as a scalable and reliable strategy for intracellular delivery with applicability in cell engineering and therapeutic development.