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Continuous Lysis of Cells in a Locally Concentrated DC Field on a Microfluidic Chip

机译:在微流体芯片上局部浓缩DC场中的细胞连续裂解

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Recently, on-chip electrical lysis of biological cells has been raising a lot of interests due to its widespread applications in recovering the contents of cells without introducing lysing agents which may interfere with subsequent biological assays. Applying a DC field for cell lysis typically requires the overall voltage as high as thousands of volts to generate an field strength of 1-10 kV/cm [1, 2]. Bubble generation and Joule heating can often be induced under such high field strengths and continuous operation becomes difficult. Decreasing the gap between the electrodes can decrease the overall voltage [3, 4]. However the fabrication of high density microscale electrodes increases the cost. Furthermore, AC or pulsed electrical fields are typically applied and they are not compatible with bioanalysis methods such as electrophoresis. In this study, we demonstrated that GFP-expressing E. coli cells were lysed continuously while flowing through a microfluidic device. The DC field was intensified in a defined section of the microfluidic channel by altering the channel geometry. The overall applied voltage was lowered to several hundred volts to general a local field of 1.5-7 kV/cm. This permitted 3-5V/cm of lysis field strength per volt applied. We found that local lysis field strength of 1500V/cm was required for the lysis of 95-100% of GFP-expressing E. coli. This field strength was substantially lower than the lysis field needed in the case of pulsed electrical fields (around 7kV/cm). The lysis was witnessed by the plate count and the fluorescence spectroscopy. Our design enabled spatially confined electrolysis with a moderate overall voltage. Operating continuously under DC field makes our approach a high throughput method, which can be easily coupled with on-chip electrophoresis. The same method can also be applied for the lysis of mammalian cells and for the electroporation and transfection.
机译:最近,由于其在不引入可能干扰随后的生物学测定的裂解剂的情况下,在恢复细胞含量的广泛应用,生物细胞的片上电裂解已经提高了许多兴趣。施加用于细胞裂解的DC场通常需要高达数千伏的总电压,以产生1-10kV / cm [1,2]的场强。在这种高场强度下,通常可以诱导泡​​沫生成和焦耳加热,并且难以进行连续操作。减小电极之间的间隙可以降低总电压[3,4]。然而,高密度微观电极的制造增加了成本。此外,通常施加AC或脉冲电场,它们与诸如电泳的生物分析方法不相容。在这项研究中,我们证明,在流过微流体装置的同时连续裂解GFP表达的大肠杆菌细胞。通过改变通道几何形状,在微流体通道的定义部分中加强DC场。将整体施加的电压降低到几百伏特,以一般为1.5-7 kV / cm的局部场。这允许其施加3-5V / cm的裂解场强度。我们发现,裂解局部裂解场强度为1500V / cm,裂解95-100%的GFP表达的大肠杆菌。该场强基本上低于脉冲电场(约7kV / cm)所需的裂解场。通过板数和荧光光谱目睹裂解。我们的设计使空间限制电解具有适中的整体电压。在直流场下连续运行,使我们的方法成为高通量法,其可以容易地与片上电泳。还可以应用于哺乳动物细胞的裂解和电穿孔和转染的方法。

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