Droplet squeezing for highly effective gene delivery into human t lymphocytes

We present a novel droplet-based intracellular delivery method for highly effective internalization of external macromolecules into human T lymphocytes. First, we generated droplets that encapsulate single cells mixed with biomolecules, using a flow-focusing droplet generator. The droplets containing cells were then passed through a series of narrow constrictions, creating transient nanopores in the cell membrane. Consequently, external cargos in droplets can be transported directly into the cell cytosol throughout the nanopores' lifespan, achieving effective intracellular delivery. This process was able to achieve highly efficient and high-throughput delivery of different nanomaterials into human T lymphocytes, while maintaining high cell viability.