Electroporation of mammalian cells in a microfluidic channel with geometric variation
Electroporation of mammalian cells in a microfluidic channel with geometric variation
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DOI：
10.1021/ac060733n
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发表时间：
2006-07-15
影响因子：
7.4
通讯作者：
Lu, Chang
中科院分区：
文献类型：
--
作者：
Wang, Hsiang-Yu;Lu, Chang
电穿孔已被广泛用于将不渗透的外源分子加载到细胞中。基于电穿孔的快速电裂解也已用于分析单细胞水平的细胞内物质。人们越来越需要以微流体形式实施电穿孔作为从药物和基因筛选到细胞内动力学研究等应用的基本工具。在本报告中，我们开发了一种简单的技术，可以在微流体平台上以高通量电穿孔哺乳动物细胞。在我们的设计中，由于几何变化引起的局部场放大，电穿孔仅发生在微流体通道的限定部分。细胞暴露于高场的时间由细胞的速度和切片的长度决定。实时观察电穿孔过程中细胞形态的变化。我们确定，当局部场强增加到大约 400 V/cm 时，就会发生中国仓鼠卵巢细胞的电穿孔。还进行了保留细胞活力的膜非渗透性分子（SYTOX green）的内化，以证明瞬时电透化。确定了装置的操作参数对细胞活力的影响。当调整参数时，电穿孔后很大一部分细胞仍然存活。我们还研究了场强在 600-1200 V/cm 范围内时的快速细胞裂解。当场强为1200V/cm时，细胞膜在30ms内发生破裂。由于简单、高通量以及与其他设备的高兼容性，这种微流控电穿孔技术可能会增加微流控系统在药物和生物分子筛选以及化学细胞计数中的应用。
Electroporation has been widely used to load impermeant exogenous molecules into cells. Rapid electrical lysis based on electroporation has also been applied to analyze intracellular materials at single-cell level. There has been increasing demand to implement electroporation in a microfluidic format as a basic tool for applications ranging from screening of drugs and genes to studies of intracellular dynamics. In this report, we have developed a simple technique to electroporate mammalian cells with high throughput on a microfluidic platform. In our design, electroporation only happened in a defined section of a microfluidic channel due to the local field amplification by geometric variation. The time of exposure of the cells to this high field was determined by the velocity of the cells and the length of the section. The change in the cell morphology during electroporation was observed in real time. We determined that electroporation of Chinese hamster ovary cells occurred when the local field strength was increased to similar to 400 V/cm. The internalization of membrane-impermeant molecules ( SYTOX green) with cell viability preserved was also carried out to demonstrate transient electropermeabilization. The influence of the operational parameters of the device on cell viability was determined. A large percentage of cells remained viable after electroporation when the parameters were tuned. We also studied rapid cell lysis when the field intensity was in the range of 600-1200 V/cm. The rupture of cell membrane happened within 30 ms when the field strength was 1200 V/cm. Given the simplicity, high throughput, and high compatibility with other devices, this microfluidic electroporation technique may increase the application of microfluidic systems in screening of drugs and biomolecules and chemical cytometry.