National Center for Nanoscience and Technology , Beijing 100190, China.
Anal Chem. 2014 Oct 21;86(20):10215-22. doi: 10.1021/ac502294e. Epub 2014 Oct 6.
Microfluidics based continuous cell electroporation is an appealing approach for high-throughput cell transfection, but cell viability of existing methods is usually compromised by adverse electrical or hydrodynamic effects. Here we present the validation of a flow-through cell electroporation microchip, in which dielectrophoretic force was employed to sort viable cells. By integrating parallel electroporation electrodes and dielectrophoresis sorting electrodes together in a simple straight microfluidic channel, sufficient electrical pulses were applied for efficient electroporation, and a proper sinusoidal electrical field was subsequently utilized to exclude damaged cells by dielectrophoresis. Thus, the difficulties for seeking the fine balance between electrotransfection efficiency and cell viability were steered clear. After careful investigation and optimization of the DEP behaviors of electroporated cells, efficient electrotransfection of plasmid DNA was demonstrated in vulnerable neuron cells and several hard-to-transfect primary cell types with excellent cell viability. This microchip constitutes a novel way of continuous cell transfection to significantly improve the cell viability of existing methodologies.
基于微流控的连续细胞电穿孔是一种高通量细胞转染的有吸引力的方法,但现有的方法的细胞活力通常因不良的电或流体力学效应而受到损害。在这里,我们验证了一种用于细胞电穿孔的微流控芯片,其中介电泳力用于分选活细胞。通过将平行的电穿孔电极和介电泳分选电极集成在一个简单的直微流道中,施加足够的电脉冲以实现高效的电穿孔,随后利用适当的正弦电场通过介电泳排除受损的细胞。因此,避免了在电转染效率和细胞活力之间寻求精细平衡的困难。在仔细研究和优化电穿孔细胞的介电泳行为后,我们在脆弱的神经元细胞和几种难以转染的原代细胞类型中实现了高效的质粒 DNA 电转染,同时保持了优异的细胞活力。这种微芯片构成了一种连续细胞转染的新方法,可以显著提高现有方法的细胞活力。