| 用于单分子动力学实验的微流控混合器 |
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| 引用本文： | 支泽勇,刘鹏程,黄岩谊,赵新生.用于单分子动力学实验的微流控混合器[J].物理化学学报,2011,27(8):1990-1995. |
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| 作者姓名： | 支泽勇 刘鹏程 黄岩谊 赵新生 |
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| 作者单位： | 1. Beijing National Laboratory for Molecular Sciences, State Key Laboratory for Structural Chemistry of Unstable and Stable Species, Department of Chemical Biology, College of Chemistry and Molecular Engineering, Peking University, Beijing 100871, P. R. China;
2. College of Engineering, Peking University, Beijing 100871, P. R. China;
3. Biodynamic Optical Imaging Center, Peking University, Beijing 100871, P. R. China |
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| 基金项目： | supported by the National Natural Science Foundation of China(20733001,20973015); National Key Basic Research Program of China(973)(2006CB910300,2010CB912302)~~ |
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| 摘 要： | 设计制作了用于单分子动力学实验的微流控混合器, 该混合器用聚二甲基硅氧烷(PDMS)芯片和石英载玻片密封而成, 具有低的荧光背景, 广泛的生物相容性, 结合激光共聚焦显微镜能够在非平衡态下进行单分子荧光探测. 我们设计的压力控制系统和进样流路方便而稳定, 保证了微流路中流形的长时间稳定, 从而实现了样品流速和流量的精准控制. 这些技术特点保证了单分子探测得到准确和高信噪比的结果. 利用蛋白质的塌缩过程远快于混合过程的特点，采用荧光标记的金黄色葡萄球菌核酸酶作为指示物，分辨出蛋白质变性态的特征峰，并利用变性态的荧光共振能量传递效率随时间的变化表征出混合器在适合于单分子探测条件下的混合时间为150 ms.
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| 关 键 词： | 微流控混合 单分子探测 荧光共振能量传递 蛋白质折叠 金黄色葡萄球菌核酸酶 |
| 收稿时间： | 2011-04-27 |
| 修稿时间： | 2011-06-09 |
A Microfluidic Mixer for Single-Molecule Kinetics Experiments |
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| ZHI Ze-Yong,LIU Peng-Cheng,HUANG Yan-Yi,ZHAO Xin-Sheng.A Microfluidic Mixer for Single-Molecule Kinetics Experiments[J].Acta Physico-Chimica Sinica,2011,27(8):1990-1995. |
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| Authors: | ZHI Ze-Yong LIU Peng-Cheng HUANG Yan-Yi ZHAO Xin-Sheng |
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| Institution: | 1. Beijing National Laboratory for Molecular Sciences, State Key Laboratory for Structural Chemistry of Unstable and Stable Species, Department of Chemical Biology, College of Chemistry and Molecular Engineering, Peking University, Beijing 100871, P. R. China;
2. College of Engineering, Peking University, Beijing 100871, P. R. China;
3. Biodynamic Optical Imaging Center, Peking University, Beijing 100871, P. R. China |
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| Abstract: | We designed and built a microfluidic mixer based on the principle of hydrodynamic focusing governed by Navier-Stokes equation for single-molecule kinetics experiments.The mixer is a cast of poly(dimethylsiloxane)(PDMS) sealed with transparent fused-silica coverglass,which results in low fluorescence background and broad biological compatibility and this enables single-molecule fluorescence detection under nonequilibrium conditions.The pressure regulated sample delivery system is convenient for loading a sam... |
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| Keywords: | Microfluidic mixing Single-molecule detection Fluorescence resonance energy transfer Protein folding Staphylococcal nuclease |
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