Kim Samuel, Zare Richard N
Department of Chemistry, Stanford University, Stanford, California, USA.
Methods Enzymol. 2010;472:119-32. doi: 10.1016/S0076-6879(10)72013-9.
Microfluidics serves as a convenient platform for single-molecule experiments by providing manipulation of small amounts of liquids and micron-sized particles. An adapted version of capillary electrophoresis (CE) on a microchip can be utilized to separate chemical species with high resolution based on their ionic mobilities (i.e., charges and sizes), but identification of separated species is not trivial, especially for complex mixtures of sticky biomolecules. We describe here how to use a surfactant mixture system for CE on a poly(dimethylsiloxane) (PDMS) microchip, capture separated peaks within a 50-pl chamber using microvalves, analyze the fluorescence signals with correlation spectroscopy to extract molecular diffusion characteristics, and to identify the biomolecular clusters in a model immunocomplex system.
微流控技术通过对少量液体和微米级颗粒的操控，为单分子实验提供了一个便捷的平台。基于芯片的毛细管电泳（CE）的改进版本可用于根据化学物质的离子迁移率（即电荷和大小）对其进行高分辨率分离，但对分离出的物质进行鉴定并非易事，尤其是对于粘性生物分子的复杂混合物。我们在此描述如何在聚二甲基硅氧烷（PDMS）微芯片上使用表面活性剂混合体系进行CE，利用微阀在一个50皮升的腔室内捕获分离出的峰，用相关光谱分析荧光信号以提取分子扩散特性，并在一个模型免疫复合物系统中鉴定生物分子簇。
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