Mohammad Aghaamoo

Acoustic microstreaming vortices enable high-throughput cell trapping and sequential delivery of plasmid DNA and Cas9 RNP, boosting transfection up to sevenfold by eliminating cargo competition and/or interaction.

Dielectrophoresis (DEP) is a powerful tool for label-free sorting of cells, even those with subtle differences in morphological and dielectric properties. Nevertheless, a major limitation is that most existing DEP techniques can efficiently sort cells only at low throughputs (150,000 cells min⁻¹) with 90% separation purity, 85% cell recovery, and no negative impact on cell viability.

A high‐throughput non‐viral intracellular delivery platform is introduced for the transfection of large cargos with dosage‐control. This platform, termed Acoustic‐Electric Shear Orbiting Poration (AESOP), optimizes the delivery of intended cargo sizes with poration of the cell membranes via mechanical shear followed by the modulated expansion of these nanopores via electric field. Furthermore, AESOP utilizes acoustic microstreaming vortices wherein up to millions of cells are trapped and mixed uniformly with exogenous cargos, enabling the delivery of cargos into cells with targeted dosages. Intracellular delivery of a wide range of molecule sizes (90%), cell viability (>80%), and uniform dosages (−1 per single chip is demonstrated. AESOP is successfully applied to two gene editing applications that require the delivery of large plasmids: i) enhanced green fluorescent protein (eGFP) plasmid (6.1 kbp) transfection, and ii) clustered regularly interspaced short palindromic repeats (CRISPR)‐Cas9‐mediated gene knockout using a 9.3 kbp plasmid DNA encoding Cas9 protein and single guide RNA (sgRNA). Compared to alternative platforms, this platform offers dosage‐controlled intracellular delivery of large plasmids simultaneously to large populations of cells while maintaining cell viability at comparable delivery efficiencies.

In this work, we describe the mechanism of particle trapping and release at the flow-focusing microfluidic droplet generation junction, utilizing the hydrodynamic microvortices generated in the dispersed phase. This technique is based solely on our unique flow-focusing geometry and the flow control of the two immiscible phases and, thus, does not require any on-chip active components. The effectiveness of this technique to be used for particle trapping and the subsequent size selective release into the droplets depends on the fundamental understanding of the nature of the vortex streamlines. Here, we utilized theoretical, computational, and experimental fluid dynamics to study in detail these microvortices and parameters affecting their formation, trajectory, and magnitude.

In this work, we describe the mechanism of particle trapping and release at the flow-focusing microfluidic droplet generation junction, utilizing the hydrodynamic microvortices generated in the dispersed phase. This technique is based solely on our unique flow-focusing geometry and the flow control of the two immiscible phases and, thus, does not require any on-chip active components. The effectiveness of this technique to be used for particle trapping and the subsequent size selective release into the droplets depends on the fundamental understanding of the nature of the vortex streamlines. Here, we utilized theoretical, computational, and experimental fluid dynamics to study in detail these microvortices and parameters affecting their formation, trajectory, and magnitude.

In this work, we describe the mechanism of particle trapping and release at the flow-focusing microfluidic droplet generation junction, utilizing the hydrodynamic microvortices generated in the dispersed phase. This technique is based solely on our unique flow-focusing geometry and the flow control of the two immiscible phases and, thus, does not require any on-chip active components. The effectiveness of this technique to be used for particle trapping and the subsequent size selective release into the droplets depends on the fundamental understanding of the nature of the vortex streamlines. Here, we utilized theoretical, computational, and experimental fluid dynamics to study in detail these microvortices and parameters affecting their formation, trajectory, and magnitude.

In this work, we describe the mechanism of particle trapping and release at the flow-focusing microfluidic droplet generation junction, utilizing the hydrodynamic microvortices generated in the dispersed phase. This technique is based solely on our unique flow-focusing geometry and the flow control of the two immiscible phases and, thus, does not require any on-chip active components. The effectiveness of this technique to be used for particle trapping and the subsequent size selective release into the droplets depends on the fundamental understanding of the nature of the vortex streamlines. Here, we utilized theoretical, computational, and experimental fluid dynamics to study in detail these microvortices and parameters affecting their formation, trajectory, and magnitude.

In this work, we describe the mechanism of particle trapping and release at the flow-focusing microfluidic droplet generation junction, utilizing the hydrodynamic microvortices generated in the dispersed phase. This technique is based solely on our unique flow-focusing geometry and the flow control of the two immiscible phases and, thus, does not require any on-chip active components. The effectiveness of this technique to be used for particle trapping and the subsequent size selective release into the droplets depends on the fundamental understanding of the nature of the vortex streamlines. Here, we utilized theoretical, computational, and experimental fluid dynamics to study in detail these microvortices and parameters affecting their formation, trajectory, and magnitude.

In this work, we describe the mechanism of particle trapping and release at the flow-focusing microfluidic droplet generation junction, utilizing the hydrodynamic microvortices generated in the dispersed phase. This technique is based solely on our unique flow-focusing geometry and the flow control of the two immiscible phases and, thus, does not require any on-chip active components. The effectiveness of this technique to be used for particle trapping and the subsequent size selective release into the droplets depends on the fundamental understanding of the nature of the vortex streamlines. Here, we utilized theoretical, computational, and experimental fluid dynamics to study in detail these microvortices and parameters affecting their formation, trajectory, and magnitude.

Intracellular delivery of cargos for cell engineering plays a pivotal role in transforming medicine and biomedical discoveries. Recent advances in microfluidics and nanotechnology have opened up new avenues for efficient, safe, and controllable intracellular delivery, as they improve precision down to the single-cell level. Based on this capability, several promising micro- and nanotechnology approaches outperform viral and conventional non-viral techniques in offering dosage-controlled delivery and/or intracellular delivery of large cargos. However, to achieve this level of precision and effectiveness, they are either low in throughput, limited to specific cell types (e.g., adherent vs. suspension cells), or complicated to operate with. To address these challenges, here we introduce a versatile and simple-to-use intracellular delivery microfluidic platform, termed Acoustic-Electric Shear Orbiting Poration (AESOP). Hundreds of acoustic microstreaming vortices form the production line of the AESOP platform, wherein hundreds of thousands of cells are trapped, permeabilized, and mixed with exogenous cargos. Using AESOP, we show intracellular delivery of a wide range of molecules (from 6kbp) with viabilities over 80% and comparable delivery efficiencies, but also is an order of magnitude higher in throughput, compatible with both adherent and suspension cell lines, and simple to operate.