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亚微米级球形支撑脂质双层的高密度阵列。

High-density arrays of submicron spherical supported lipid bilayers.

机构信息

Laboratory of Nanostructures and Biosensing, Department of Electrical and Computer Engineering, University of Minnesota, Minneapolis, Minnesota 55455, USA.

出版信息

Anal Chem. 2012 Oct 2;84(19):8207-13. doi: 10.1021/ac3014274. Epub 2012 Sep 21.

Abstract

Lipid bilayer membranes found in nature are heterogeneous mixtures of lipids and proteins. Model systems, such as supported lipid bilayers (SLBs), are often employed to simplify experimental systems while mimicking the properties of natural lipid bilayers. Here, we demonstrate a new method to form SLB arrays by first forming spherical supported lipid bilayers (SSLBs) on submicrometer-diameter SiO(2) beads. The SSLBs are then arrayed into microwells using a simple physical assembly method that requires no chemical modification of the substrate nor modification of the lipid membrane with recognition moieties. The resulting arrays have submicrometer SSLBs with 3 μm periodicity where >75% of the microwells are occupied by an individual SSLB. Because the arrays have high density, fluorescence from >1000 discrete SSLBs can be acquired with a single image capture. We show that 2-component random arrays can be formed, and we also use the arrays to determine the equilibrium dissociation constant for cholera toxin binding to ganglioside GM1. SSLB arrays are robust and are stable for at least one week in buffer.

摘要

自然界中的脂质双层膜是脂质和蛋白质的不均匀混合物。模型系统,如支撑脂质双层(SLB),通常被用来简化实验系统,同时模拟天然脂质双层的性质。在这里,我们展示了一种新的方法,通过首先在亚微米直径的 SiO2 球上形成球形支撑脂质双层(SSLB)来形成 SLB 阵列。然后,使用一种简单的物理组装方法将 SSLB 排列到微井中,该方法不需要对基底进行化学修饰,也不需要用识别基团修饰脂质膜。所得的阵列具有亚微米级的 SSLB 和 3 μm 的周期性,其中>75%的微井被单个 SSLB 占据。由于阵列具有高密度,因此可以用单个图像捕获获取来自>1000 个离散 SSLB 的荧光。我们表明可以形成 2 组分随机阵列,并且我们还使用该阵列来确定霍乱毒素与神经节苷脂 GM1 结合的平衡解离常数。SSLB 阵列是稳定的,在缓冲液中至少稳定一周。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d917/3476481/b7b5904e41e9/nihms409976f1.jpg

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