用于电穿孔快速优化的对称双曲格式微芯片

Mengxi Wu, Deyao Zhao, Hao Yan, Zicai Liang, Zhihong Li
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引用次数: 1

摘要

我们开发了一种新型微芯片,旨在快速优化电穿孔的电参数。电极被设计成对称的双曲格式,以产生线性递减的电场。细胞在微芯片上培养,贴壁电穿孔和原位观察。在不同的电场强度下,细胞活力和转染效率表现出不同的变化,其中三个区域的变化较为明显。通过在电极上设计标记,确定最佳电穿孔性能区域的位置和相应的电场强度范围。利用所设计的电穿孔芯片,获得了HEK-293A细胞系电穿孔的最佳电场谱,约为0.75 ~ 1.25 × 105 V/m。通过细胞活力和转染效率对电场强度的依赖性来评价电穿孔性能的多样性。在参考电场值的指导下,我们设计了一个特定的环形交叉微芯片来验证电穿孔特性。最后,我们利用自制的电穿孔芯片在最佳条件下获得了优异的电穿孔性能。
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A symmetrical hyperbolic formatted microchip for rapid optimization of electroporation
We have developed a novel microchip aiming to rapidly optimize of electric parameters for electroporation. The electrodes were designed as symmetrical hyperbolic formatted to generate linearly decrescent electric field. Cells were cultured on the microchip for adherent electroporation and in-situ observation. Exhibited diversely in cell viability and transfection efficiency, three areas were observed obviously since the intensity of electric field was varied. Assisted by markers designed on electrodes, the location of optimal electroporation performance area and the corresponding range of electric field intensity were determined. Using the proposed electroporation chip, we obtained the optimal electric field spectrum for electroporation of HEK-293A cell line, that was about 0.75 ~ 1.25 × 105 V/m. The diversity of electroporation performance was evaluated by the dependence of cell viability and transfection efficiency on electric field intensity. With the guidance of reference electric field value, we designed a specific annular-interdigitated microchip to testify the electroporation characteristics. At last, we achieved excellent electroporation performance under the optimal conditions using self-made electroporation microchip.
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