Microfluidic fabrication of asymmetric giant lipid vesicles.
Microfluidic fabrication of asymmetric giant lipid vesicles.
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DOI：
10.1021/am101191d
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发表时间：
2011-05
影响因子：
9.5
通讯作者：
Malmstadt, Noah
中科院分区：
文献类型：
--
作者：
Hu, Peichi C.;Li, Su;Malmstadt, Noah
我们已经开发了一种微流控技术的组成不对称的巨型单层囊泡（GUV）的制造。囊泡在两个独立的步骤中组装。在每个步骤中，在水-油界面处形成脂质单层。第一单层在微流体装置内部形成，该微流体装置具有多相液滴流构造，该多相液滴流构造由其中形成水滴的连续油流组成。这些液滴被分配到一个容器中，该容器包含一层油，一层水。通过离心将液滴转移通过该第二油-水界面来形成第二脂质单层。通过将不同的脂质组合物溶解在不同的油相中，可以控制所得脂质双层的每个小叶的组成。我们已经证明了膜的不对称性，通过显示每个小叶中标记的脂质的差异荧光猝灭，并通过证明不对称GUV将结合亲和素包被的表面，只有当生物素化的脂质靶向外小叶。此外，我们已经证明了磷脂酰丝氨酸脂质成功的不对称靶向每个小叶，产生具有仿生和生理相关成分不对称性的膜。
We have developed a microfluidic technology for the fabrication of compositionally asymmetric giant unilamellar vesicles (GUVs). The vesicles are assembled in two independent steps. In each step, a lipid monolayer is formed at a water-oil interface. The first monolayer is formed inside of a microfluidic device with a multiphase droplet flow configuration consisting of a continuous oil stream in which water droplets are formed. These droplets are dispensed into a vessel containing a layer of oil over a layer of water. The second lipid monolayer is formed by transferring the droplets through this second oil-water interface by centrifugation. By dissolving different lipid compositions in the different oil phases, the composition of each leaflet of the resulting lipid bilayer can be controlled. We have demonstrated membrane asymmetry by showing differential fluorescence quenching of labeled lipids in each leaflet and by demonstrating that asymmetric GUVs will bind an avidin-coated surface only when biotinylated lipids are targeted to the outer leaflet. In addition, we have demonstrated the successful asymmetric targeting of phosphatidylserine lipids to each leaflet, producing membranes with a biomimetic and physiologically relevant compositional asymmetry.
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发表时间：
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影响因子：
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发表时间：
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影响因子：
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