A Microfluidic Mixer for Single-Molecule Kinetics Experiments

doi:10.3866/PKU.WHXB20110804

CORC > 化学研究所 > 中国科学院化学研究所

	A Microfluidic Mixer for Single-Molecule Kinetics Experiments
	Zhi Ze-Yong 1,3; Liu Peng-Cheng 1,3; Huang Yan-Yi 2,3; Zhao Xin-Sheng 1,3
刊名	ACTA PHYSICO-CHIMICA SINICA
	2011-08-01
卷号	27 期号:8 页码:1990-1995
关键词	Microfluidic Mixing Single-molecule Detection Fluorescence Resonance Energy Transfer Protein Folding Staphylococcal Nuclease
ISSN号	1000-6818
DOI	10.3866/PKU.WHXB20110804
英文摘要	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 sample and allows for precise and stable flow velocity control. The combination of microfluidic mixer and single-molecule fluorescence resonance energy transfer (smFRET) allows us to measure the time course of the distribution of the smFRET efficiency in protein folding. We used the fact that denatured protein collapses much faster than the mixing process to characterize the mixing time using donor and acceptor dyes labeled staphylococcal nuclease (SNase) as an smFRET efficiency indicator. By monitoring the smFRET efficiency of denatured SNase during the course of mixing, we determined that the mixing time was 150 ms under conditions suitable for single-molecule detection.
语种	英语
出版者	PEKING UNIV PRESS
WOS记录号	WOS:000292877400032
内容类型	期刊论文
源URL	[http://ir.iccas.ac.cn/handle/121111/72613]
专题	中国科学院化学研究所
通讯作者	Zhao Xin-Sheng
作者单位	1.Peking Univ, Coll Chem & Mol Engn, State Key Lab Struct Chem Unstable & Stable Speci, Beijing Natl Lab Mol Sci,Dept Biol Chem, Beijing 100871, Peoples R China 2.Peking Univ, Coll Engn, Beijing 100871, Peoples R China 3.Peking Univ, Biodynam Opt Imaging Ctr, Beijing 100871, Peoples R China
推荐引用方式 GB/T 7714	Zhi Ze-Yong,Liu Peng-Cheng,Huang Yan-Yi,et al. A Microfluidic Mixer for Single-Molecule Kinetics Experiments[J]. ACTA PHYSICO-CHIMICA SINICA,2011,27(8):1990-1995.
APA	Zhi Ze-Yong,Liu Peng-Cheng,Huang Yan-Yi,&Zhao Xin-Sheng.(2011).A Microfluidic Mixer for Single-Molecule Kinetics Experiments.ACTA PHYSICO-CHIMICA SINICA,27(8),1990-1995.
MLA	Zhi Ze-Yong,et al."A Microfluidic Mixer for Single-Molecule Kinetics Experiments".ACTA PHYSICO-CHIMICA SINICA 27.8(2011):1990-1995.