Advances in gene editing are leading to new medical interventions where patients’ own cells are used for stem cell therapies and immunotherapies. One of the key limitations to translating these treatments to the clinic is the need for scalable technologies for engineering cells efficiently and safely. Toward this goal, microfluidic strategies to induce membrane pores and permeability have emerged as promising techniques to deliver biomolecular cargo into cells. As these technologies continue to mature, there is a need to achieve efficient, safe, nontoxic, fast, and economical processing of clinically relevant cell types. We demonstrate an acoustofluidic sonoporation method to deliver plasmids to immortalized and primary human cell types, based on pore formation and permeabilization of cell membranes with acoustic waves. This acoustofluidic-mediated approach achieves fast and efficient intracellular delivery of an enhanced green fluorescent protein-expressing plasmid to cells at a scalable throughput of 200,000 cells/min in a single channel. Analyses of intracellular delivery and nuclear membrane rupture revealed mechanisms underlying acoustofluidic delivery and successful gene expression. Our studies show that acoustofluidic technologies are promising platforms for gene delivery and a useful tool for investigating membrane repair.
Gold nanostars (AuNSTs) are biocompatible, have large surface areas, and are characterized by high near-infrared extinction, making them ideal for integration with technologies targeting biological applications. We have developed a robust and simple microfluidic method for the direct growth of anisotropic AuNSTs on oxide substrates including indium tin oxide and glass. The synthesis was optimized to yield AuNSTs with high anisotropy, branching, uniformity, and density in batch and microfluidic systems for optimal light-to-heat conversion upon laser irradiation. Surface-enhanced Raman scattering spectra and mesoscale temperature measurements were combined with spatially correlated scanning electron microscopy to monitor nanostar and ligand stability and microbubble formation at different laser fluences. The capability of the platform for generating controlled localized heating was used to explore hyperthermia-assisted detachment of adherent glioblastoma cells (U87-GFP) grafted to the capillary walls. Both flow and laser fluence can be tuned to induce different biological responses, such as ablation, cell deformation, release of intracellular components, and the removal of intact cells. Ultimately, this platform has potential applications in biological and chemical sensing, hyperthermia-mediated drug delivery, and microfluidic soft-release of grafted cells with single-cell specificity.
Self-assembled monolayers (SAMs) of carborane isomers with different dipole moments passivate germanium to modulate surface work function while maintaining chemical environment and surface energy. To identify head groups capable of monolayer formation on germanium surfaces, we studied thiol-, hydroxyl-, and carboxyl-terminated carboranes. These films were successfully formed with carboxylic acid head groups instead of the archetypal thiol, suggesting that the carborane cluster significantly affects headgroup reactivity. Film characterization included X-ray and ultraviolet photoelectron spectroscopies as well as contact angle goniometry. Using these carboranes, the germanium surface work function was tailored over 0.4 eV without significant changes to wetting properties.
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