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Biology subjects

Kashani, M. N.

Publications and source records attributed to Kashani, M. N..

2 recordsLinked to original sources

Microbial stir bars: light activated rotation of tethered bacterial cells to enhance mixing in stagnant fluids.

Microfluidics devices are gaining significant interest in biomedical applications. However, in a micron-scale device, reaction speed is often limited by the slow rate of diffusion of the reagents. Several active and passive micro-mixers have been fabricated to enhance mixing in microfluidic devices. Here, we demonstrate external control of mixing by rotating a rodshaped bacterial cell. This rotation is driven by ion transit across the bacterial flagellar stator complex. We first measured the flow fields generated by rotating a single bacterial cell rotationally locked to rotate either clockwise (CW) or counterclockwise (CCW). Micro-Particle Image Velocimetry (PIV) and Particle Tracking Velocimetry results showed that a bacterial cell of ~ 2.75 m long, rotating at 5.75 {+/-} 0.39 Hz in a counterclockwise direction could generate distinct micro-vortices with circular flow fields with a mean velocity of 4.72 {+/-} 1.67 m/s and maximum velocity of 7.90 m/s in aqueous solution. We verified our experimental data with a numerical simulation at matched flow conditions which revealed vortices of similar dimensions and speed. We observed that the flow-field diminished with increasing z-height above the plane of the rotating cell. Lastly, we showed we could activate and tune rotational mixing remotely using strains engineered with Proteorhodopsin (PR), where rotation could be activated by controlled external illumination using green laser light (561 nm).

microbiology↗

Intracellular delivery of mRNA to human primary T cells with microfluidic vortex shedding

Intracellular delivery of functional macromolecules, such as DNA and RNA, across the cell membrane and into the cytosol, is a critical process in both biology and medicine. Herein, we develop and use microfluidic chips containing post arrays to induce microfluidic vortex shedding, or VS, for cell membrane poration that permits delivery of mRNA into primary human T lymphocytes. We demonstrate transfection with VS by delivery of a 996-nucleotide mRNA construct encoding enhanced green fluorescent protein (EGFP) and assessed transfection efficiencies by quantifying levels of EGFP protein expression. We achieved high transfection efficiency (63.6 {+/-} 3.44% EGFP+ viable cells) with high cell viability (77.3 {+/-} 0.58%) and recovery (88.7 {+/-} 3.21%) in CD3+ T cells 19 hrs after VS processing. Importantly, we show that processing cells via VS does not negatively affect cell growth rates or alter cell states. We also demonstrate processing speeds of greater than 2.0 x 106 cells s-1 at volumes ranging from 0.1 to 1.5 milliliters. Altogether, these results highlight the use of VS as a rapid and gentle delivery method with promising potential to engineer primary human cells for research and clinical applications.

bioengineering↗