Flow-through Microfluidic Device for High-efficiency Transfection of Mammalian Cells through Combined Microelectroporation and Sonoporation

Flow-through Microfluidic Device for High-efficiency Transfection of Mammalian Cells through Combined Microelectroporation and Sonoporation. (May 2011) Whitney Leigh Longsine, B.S., Texas A&M University Chair of Advisory Committee: Dr. Arum Han In this study we are presenting a proof-of-concept microfluidic device that simultaneously applies the conditions required for microelectroporation and microsonoporation in a flow-through fashion that allows for high throughput, high efficiency transfection of mammalian cells. During the design stage, we developed a low-cost, high-resolution polymer microfabrication technique termed laser stenciling. While few other electro-sonoporation protocols have been reported, to the best of our knowledge, we are the first to incorporate microelectroporation, which has been well established in literature to be advantageous to conventional electroporation, with flow-through microsonoporation. When comparing transfection efficiency for our electro-sonoporation method to that of sonoporation or microelectroporation alone, we observed single batch improvements up to 20% and 17%, respectively. The average improvement in efficiency was approximately 15% greater than achieved with sonoporation and 10% greater than that of electroporation. Importantly, there was little difference in short term cell viability between the three methods (maintained at > 90%). The average transfection efficiency for electro-sonoporation was 81.25% and cell viability was 91.56%. Overall, we have presented a device and electro-sonoporation method that meets or outperforms the transfection efficiency and cell viability standards for HeLa cells set by other reported electroporation and sonoporation methods.