Micropatterned fluid lipid bilayer arrays created using a continuous flow microspotter.
Micropatterned fluid lipid bilayer arrays created using a continuous flow microspotter.
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使用连续流微点样器创建的微图案流体脂质双层阵列。
DOI：
10.1021/ac800860u
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发表时间：
2008
影响因子：
7.4
通讯作者：
Conboy,JohnC
中科院分区：
文献类型：
--
作者：
Smith,KathrynA;Gale,BruceK;Conboy,JohnC
我们已经开发了一种新的方法，用于创建微图案化的脂质双层阵列（MLBAs）使用3D微流体系统。使用连续流动微量点样仪将流体脂质膜阵列图案化到玻璃基底上。荧光显微镜实验用于验证在玻璃基板上形成双层结构。光漂白实验后的荧光恢复表明，双层膜的流动性保持，而被单独地围在基板上。通过膜荧光与融合到表面之前掺入囊泡中的罗丹明官能化脂质的摩尔%的线性响应来证明阵列内双层形成的再现性。使用三种不同的荧光标记的脂质来产生多组分脂质阵列，证明了MLBA的高度可定制性质。最后，使用霍乱毒素B/神经节苷脂GM 1、抗二硝基苯基（DNP）抗体/DNP和NeutrAvidin/生物素蛋白-配体系统来模拟MLBA上的多蛋白-配体结合。用GM 1、DNP和生物素脂质将多组分图案化的双层功能化，并通过记录表面荧光与增加的膜结合配体浓度的关系来产生结合曲线。
We have developed a new method for creating micropatterned lipid bilayer arrays (MLBAs) using a 3D microfluidic system. An array of fluid lipid membranes was patterned onto a glass substrate using a Continuous Flow Microspotter. Fluorescence microscopy experiments were used to verify the formation of a bilayer structure on the glass substrate. Fluorescence recovery after photobleaching experiments demonstrated the bilayersʼ fluidity was maintained while being individually corralled on the substrate. The reproducibility of bilayer formation within an array was demonstrated by the linear response of membrane fluorescence versus mol % rhodamine functionalized lipids incorporated into the vesicles prior to fusion to the surface. The highly customizable nature of the MLBAs was demonstrated utilizing three different fluorescently labeled lipids to generate a multiple component lipid array. Finally, the cholera toxin B/ganglioside GM1, antidinitrophenyl (DNP) antibody/DNP, and NeutrAvidin/biotin protein−ligand systems were used to model multiple protein−ligand binding on the MLBAs. The multicomponent patterned bilayers were functionalized with GM1, DNP, and biotin lipids, and binding curves was generated by recording surface fluorescence versus increasing concentration of membrane bound ligands.