A microfluidic flow-through device for high throughput electrical lysis of bacterial cells based on continuous dc voltage
A microfluidic flow-through device for high throughput electrical lysis of bacterial cells based on continuous dc voltage
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DOI：
10.1016/j.bios.2006.01.032
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发表时间：
2006-12-15
影响因子：
12.6
通讯作者：
Lu, Chang
中科院分区：
文献类型：
--
作者：
Wang, Hsiang-Yu;Bhunia, Arun K.;Lu, Chang
人们对微流控平台上生物细胞的电裂解越来越感兴趣，因为它允许在不引入裂解剂的情况下快速恢复细胞内的内容物。在这项研究中，我们展示了一种简单的微流控流动装置，它可以在连续的直流电压下裂解大肠杆菌细胞。此前，大肠杆菌细胞已被修饰为表达绿色荧光蛋白(GFP)。在我们的设计中，由于几何修饰的局域场放大，细胞裂解只发生在微流控通道的指定截面上。几何修改还有效地将裂解所需的电压降低了几倍。我们发现，几乎100%的细胞死亡需要1000-1500V/cm的局部磁场强度。这一阈值场强远低于文献中报道的值，可能是由于场的持续时间较长[Lee，S.W.，Tai，Y.C.，1999]。感官。执行器A：物理。73、74-79]。平板计数和荧光光谱分析检测细胞溶解情况。当电场强度大于2000V/cm时，微流控装置裂解段细胞膜完全解体。该器件采用低成本软光刻技术制造，其沟道宽度远大于单元尺寸，以避免堵塞并确保稳定的性能。我们的工具将是高通量处理细菌细胞的理想工具，用于细胞内内容物的化学分析，如DNA和蛋白质。与使用电脉冲用于类似目的的设备相比，连续直流电压的应用极大地简化了仪器。原则上，同样的方法也可以应用于哺乳动物细胞的裂解和电穿孔转染法。(C)2006爱思唯尔B.V.保留所有权利。
Interest in electrical lysis of biological cells on a microfludic platform has increased because it allows for the rapid recovery of intracellular contents without introducing lytic agents. In this study we demonstrated a simple microfluidic flow-through device which lysed Escherichia coli cells under a continuous dc voltage. The E. coli cells had previously been modified to express green fluorescent protein (GFP). In our design, the cell lysis only happened in a defined section of a microfluidic channel due to the local field amplification by geometric modification. The geometric modification also effectively decreased the required voltage for lysis by several folds. We found that local field strength of 1000-1500 V/cm was required for nearly 100% cell death. This threshold field strength was considerably lower than the value reported in the literature, possibly due to the longer duration of the field [Lee, S.W., Tai, Y.C., 1999. Sens. Actuators A: Phys. 73, 74-79]. Cell lysis was detected by both plate count and fluorescence spectroscopy. The cell membrane was completely disintegrated in the lysis section of the microfluidic device, when the field strength was higher than 2000 V/cm. The devices were fabricated using low-cost soft lithography with channel widths considerably larger than the cell size to avoid clogging and ensure stable performance. Our tool will be ideal for high throughput processing of bacterial cells for chemical analysis of intracellular contents such as DNA and proteins. The application of continuous dc voltage greatly simplified the instrumentation compared to devices using electrical pulses for similar purposes. In principle, the same approach can also be applied for lysis of mammalian cells and electroporative transfection. (c) 2006 Elsevier B.V. All rights reserved.