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用于单细胞电穿孔的电微流控通道的制作。

Fabrication of electro-microfluidic channel for single cell electroporation.

机构信息

Department of Systems Design Engineering, University of Waterloo, Waterloo, Canada.

出版信息

Biomed Microdevices. 2013 Oct;15(5):759-66. doi: 10.1007/s10544-013-9761-0.

DOI:10.1007/s10544-013-9761-0
PMID:23494596
Abstract

The point of this paper is to demonstrate the use of a quick and cheap fabrication method to realize a laser-ablated microfluidic channel for single cell electroporation. Traditional lithography of microchannel with electrode in MEMS applications has always been complicated. Here, we introduce a new methodology of fabricating microchannel with electrical functionalities achieved through a fast and cheap process. In the present methodology, the microchannel pattern is cut out of polyimide, bonded to two ITO-coated substrates using Teflon as an adhesion layer. ITO as conductive material enables electric field in the channel and its optical transparency allows microscopy techniques to be utilized in characterizing the behavior of the microfluidic chip. The performance of the chip was tested on irreversible single-cell scale electroporation which requires relatively high voltages. CHO cells, as mammalian cells, were passed through the microchannel to experience electric field. Cells were loaded with a fluorogenic dye, Calcein AM, and the electroporation of each was individually recorded in real-time via fluorescent microscopy. The results show promising performance of the electric microchannel in electroporation. By customizing of ITO electrodes and the design of microchannel pattern, utilization and integration of the proposed electrical microchannel in variety of other MEMS-based devices are achievable.

摘要

本文旨在展示一种快速廉价的制造方法,以实现用于单细胞电穿孔的激光烧蚀微流控通道。在 MEMS 应用中,传统的微通道光刻与电极一直很复杂。在这里,我们介绍了一种通过快速廉价的工艺来制造具有电功能的微通道的新方法。在目前的方法中,微通道图案从聚酰亚胺中切割出来,使用聚四氟乙烯作为粘附层将其粘合到两个 ITO 涂层基底上。ITO 作为导电材料可以在通道中产生电场,其光学透明度允许使用显微镜技术来表征微流控芯片的行为。该芯片的性能在不可逆的单细胞规模电穿孔上进行了测试,这需要相对较高的电压。CHO 细胞作为哺乳动物细胞,通过微通道经历电场。将荧光染料 Calcein AM 加载到细胞中,并通过荧光显微镜实时记录每个细胞的电穿孔情况。结果表明,该电微通道在电穿孔方面具有良好的性能。通过定制 ITO 电极和微通道图案的设计,可以实现所提出的电微通道在各种其他基于 MEMS 的设备中的应用和集成。

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