Department of Biomedical Engineering, School of Medicine, Tsinghua University, Beijing 100084, China.
General hospital of Ningxia Medical University, Yinchuan 750004, China.
Bioelectrochemistry. 2015 Apr;102:35-41. doi: 10.1016/j.bioelechem.2014.10.002. Epub 2014 Oct 23.
Electroporation is a commonly used approach to rapidly introduce exogenous molecules into cells without permanent damage. Compared to classical electroporation protocols, microchip-based electroporation approaches have the advantages of high transfection efficiency and low consumption, but they also commonly rely on costly and tedious microfabrication technology. Hence, it is desirable to develop a novel, more affordable, and effective approach to facilitate cell electroporation. In this study, we utilized a standard printed circuit board (PCB) technology to fabricate a chip with an interdigitated array of electrodes for electroporation of suspended cells. The electrodes (thickness ~35 μm) fabricated by PCB technology are much thicker than the two-dimensional (2D) planar electrodes (thickness < 1 μm) fabricated by conventional microfabrication techniques and possess a smooth corner edge. Numerical simulations showed that the three-dimensional (3D) electrodes fabricated by PCB technology can provide a more uniformly distributed electric field compared to 2D planar electrodes, which is beneficial for reducing the electrolysis of water and improving cell transfection efficiency. The chip constructed here is composed of 18 individually addressable wells for high throughput cell electroporation. HeLa, MCF7, COS7, Jurkat, and 3T3-L1 cells were efficiently transfected with the pEGFP-N1 plasmid using individually optimal electroporation parameters. This work provides a novel method for convenient and rapid cell transfection and thus holds promise for use as a low-cost disposable device in biomedical research.
电穿孔是一种将外源分子快速导入细胞而不造成永久损伤的常用方法。与经典的电穿孔方案相比,基于微芯片的电穿孔方法具有高转染效率和低消耗的优点,但它们通常依赖于昂贵且繁琐的微制造技术。因此,开发一种新颖、更经济实惠且有效的方法来促进细胞电穿孔是很有必要的。在这项研究中,我们利用标准印刷电路板(PCB)技术制造了一种带有叉指式电极阵列的芯片,用于悬浮细胞的电穿孔。通过 PCB 技术制造的电极(厚度约 35μm)比通过传统微制造技术制造的二维(2D)平面电极(厚度<1μm)厚得多,并且具有光滑的拐角边缘。数值模拟表明,与 2D 平面电极相比,通过 PCB 技术制造的三维(3D)电极可以提供更均匀分布的电场,这有利于减少水的电解和提高细胞转染效率。这里构建的芯片由 18 个可单独寻址的孔组成,用于高通量细胞电穿孔。使用单独优化的电穿孔参数,成功地将 pEGFP-N1 质粒转染到 HeLa、MCF7、COS7、Jurkat 和 3T3-L1 细胞中。这项工作为方便快速的细胞转染提供了一种新方法,有望成为生物医学研究中低成本的一次性设备。