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微流控装置中巨囊泡的电形成和电融合。

Electroformation and electrofusion of giant vesicles in a microfluidic device.

机构信息

Key Laboratory of Biorheological Science and Technology (Chongqing University), Ministry of Education, and Key Lab of Visual Damage and Regeneration & Restoration of Chongqing, Chongqing, Bioengineering College, Chongqing University, Chongqing 400030, China.

出版信息

Colloids Surf B Biointerfaces. 2013 Oct 1;110:81-7. doi: 10.1016/j.colsurfb.2013.04.042. Epub 2013 Apr 30.

Abstract

Electroformation and electrofusion of giant vesicles with diameters of 10-20μm have been performed in a microfluidic device with high-density microelectrodes forming the sidewalls of the microchannel. Electroformation of giant vesicles by a solution mixture of phosphatidylcholine (PC) and cholesterol (Chol) with different concentrations under AC electric field was investigated. Under the conditions of 0.5-12mg/mL PC and 0.1-2.4mg/mL Chol, vesicles were electroformed by the AC electric field imposed. About 60% electroformed vesicles were giant (unilamellar) vesicles with diameters 10-20μm. The eletroformed vesicles were collected from the chip, re-suspended in fresh buffer, and then separated by centrifugation to segregate the ones with desired diameters (10-20μm). Electrofusion of the giant vesicles was conducted in the same chip. Vesicles were aligned to form pairs under AC electric field due to positive dielectrophoresis, and the paired vesicles were subsequently fused upon the application of high strength electrical pulses. The alignment and fusion efficiencies were, respectively, about 50% and 20%.

摘要

在具有高密度微电极的微流控设备中进行了直径为 10-20μm 的巨大囊泡的电成形和电融合,这些微电极形成微通道的侧壁。研究了在交流电场下用不同浓度的磷脂酰胆碱(PC)和胆固醇(Chol)的溶液混合物进行巨大囊泡的电成形。在 0.5-12mg/mL PC 和 0.1-2.4mg/mL Chol 的条件下,通过施加交流电场形成囊泡。大约 60%的电成形囊泡是直径为 10-20μm 的巨大(单层)囊泡。从芯片中收集电成形的囊泡,重新悬浮在新鲜缓冲液中,然后通过离心分离以分离所需直径(10-20μm)的囊泡。在同一芯片中进行巨大囊泡的电融合。由于正介电泳,囊泡在交流电场下排列成对,并且在施加高强度电脉冲后,成对的囊泡随后融合。对齐和融合效率分别约为 50%和 20%。

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