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首页> 外文期刊>Electrophoresis: The Official Journal of the International Electrophoresis Society >Microfluidic cell disruption system employing a magnetically actuated diaphragm
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Microfluidic cell disruption system employing a magnetically actuated diaphragm

机译：采用磁驱动隔膜的微流细胞破碎系统

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摘要

A microfluidic cell lysis chip equipped with a micromixer and SPE unit was developed and used for quantitative analysis of intracellular proteins. This miniaturized sample preparation system can be employed for any purpose where cell disruption is needed to obtain intracellular constituents for the subsequent analysis. This system comprises a magnetically actuated micromixer to disrupt cells, a hydrophobic valve to manipulate the cell lysate, and a packed porous polymerized monolith chamber for SPE and filtering debris from the cell lysate. Using recombinant Escherichia coli expressing intracellular enhanced green fluorescent protein (EGFP) and lipase as model bacteria, we optimized the cell disruption condition with respect to the lysis buffer composition, mixing time, and the frequency of the diaphragm in the micromixer, which was magnetically actuated by an external magnetic stirrer in the micromixer chamber. The lysed sample prepared under the optimal condition was purified by the packed SPE in the microfluidic chip. At a frequency of 1.96 Hz, the final cell lysis efficiency and relative fluorescence intensity of EGFP after the cell disruption process were greater than 90 and 94%, respectively. Thus, this microfluidic cell disruption chip can be used for the efficient lysis of cells for further analysis of intracellular contents in many applications.
机译：开发了配有微型混合器和SPE单元的微流细胞裂解芯片，并将其用于细胞内蛋白质的定量分析。该小型化的样品制备系统可用于需要破坏细胞以获得细胞内成分用于后续分析的任何目的。该系统包括一个用于破坏细胞的磁力微混合器，一个用于控制细胞裂解物的疏水阀，一个用于SPE并过滤来自细胞裂解物的碎屑的填充多孔聚合整料腔室。使用表达细胞内增强型绿色荧光蛋白（EGFP）和脂肪酶的重组大肠杆菌作为模型细菌，我们针对裂解缓冲液组成，混合时间和微混合器中被电磁驱动的隔膜的频率优化了细胞破坏条件通过微型混合室中的外部磁力搅拌器。在最佳条件下制备的裂解样品通过装在微流控芯片中的SPE进行纯化。在1.96 Hz的频率下，细胞破裂过程后的最终细胞裂解效率和EGFP的相对荧光强度分别大于90％和94％。因此，在许多应用中，该微流细胞破坏芯片可用于细胞的有效裂解，以进一步分析细胞内内容物。

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