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脂质囊泡的可控电融合:仿生容器内反应的引发与分析

Controllable electrofusion of lipid vesicles: initiation and analysis of reactions within biomimetic containers.

作者信息

Robinson T, Verboket P E, Eyer K, Dittrich P S

机构信息

Department of Chemistry and Applied Biosciences, ETH Zurich, Zurich, 8093, Switzerland.

出版信息

Lab Chip. 2014 Aug 7;14(15):2852-9. doi: 10.1039/c4lc00460d. Epub 2014 Jun 9.

Abstract

We present a microfluidic device that is able to trap multiple giant unilamellar vesicles (GUVs) and initiate electrofusion via integrated microelectrodes. PDMS posts were designed to trap and isolate two or more vesicles. Electrodes patterned onto the glass surface of the microchannels are able to apply a short, high voltage pulse across the traps for controllable electrofusion of the GUVs. The entire array of traps and electrodes are designed such that an average of 60 individual fusion experiments can be performed on-chip. An assay based on Förster resonance energy transfer (FRET) is performed to show successful lipid mixing. Not only can the device be used to record the dynamics of lipid membrane fusion, but it can be used for reaction monitoring by fusing GUVs containing reactants. We demonstrate this by fusing vesicles encapsulating femtolitre volumes of cobalt chloride or EDTA and monitoring the amount of the complexation product over time.

摘要

我们展示了一种微流控装置,它能够捕获多个巨型单层囊泡(GUVs)并通过集成微电极启动电融合。聚二甲基硅氧烷(PDMS)柱被设计用于捕获和隔离两个或更多囊泡。图案化在微通道玻璃表面上的电极能够在捕获区域施加短暂的高压脉冲,以实现GUVs的可控电融合。整个捕获区域和电极阵列的设计使得平均可以在芯片上进行60次单独的融合实验。基于Förster共振能量转移(FRET)的检测方法用于显示脂质混合成功。该装置不仅可用于记录脂质膜融合的动力学,还可通过融合含有反应物的GUVs用于反应监测。我们通过融合封装飞升至体积的氯化钴或乙二胺四乙酸(EDTA)的囊泡并随时间监测络合产物的量来证明这一点。

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