Dosage-controlled intracellular delivery mediated by acoustofluidics for lab on a chip applications

Abstract

Biological research and many cell-based therapies rely on the successful delivery of cargo materials into cells. Intracellular delivery in an in vitro setting refers to a variety of physical and biochemical techniques developed for conducting rapid and efficient transport of materials across the plasma membrane. Generally, the techniques that are time-efficient (e.g., electroporation) suffer from heterogeneity and low cellular viability, and those that are precise (e.g., microinjection) suffer from low-throughput and are labor-intensive. Here, we present a novel in vitro microfluidic strategy for intracellular delivery, which is based on the acoustic excitation of adherent cells. Strong mechanical oscillations, mediated by Lamb waves, inside a microfluidic channel facilitate the cellular uptake of different size (e.g., 3–500 kDa, plasmid encoding EGFP) cargo materials through endocytic pathways. We demonstrate successful delivery of 500 kDa dextran to various adherent cell lines with unprecedented efficiency in the range of 65–85% above control. We also show that actuation voltage and treatment duration can be tuned to control the dosage of delivered substances. High viability (≥91%), versatility across different cargo materials and various adherent cell lines, scalability to hundreds of thousands of cells per treatment, portability, and ease-of-operation are among the unique features of this acoustofluidic strategy. Potential applications include targeting through endocytosis-dependant pathways in cellular disorders, such as lysosomal storage diseases, which other physical methods are unable to address. This novel acoustofluidic method achieves rapid, uniform, and scalable delivery of material into cells, and may find utility in lab-on-a-chip applications.