基于琼脂糖凝胶集成样品预富集功能的微流控芯片
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摘要
提出了一种基于微注模技术的琼脂糖凝胶微流控富集检测芯片,该芯片利用与传统电泳技术兼容的琼脂糖凝胶作为芯片主体材料,并利用微注模的方法在特定区域形成琼脂糖凝胶纳孔膜结构,以此纳孔膜结合电泳作用实现样品预富集。初步富集实验实现了DNA样品在10 s内8.2倍的浓缩。
A novel method for integrating a nanoporous membrane in microfluidic devices was introduced.The nanoporous agarose gel membrane can be fabricated in-situ on PDMS microchips via micromolding techniques.The membrane is just a thin gel diaphragm perpendicularly embedded in the separation channel and close to the junction of the sample injection and separation channels,which enables electrokinetic concentration of DNA samples by a size exclusion mechanism prior to separation.This design allows a concentrated and well-defined sample to inject into the separation channel for sensitive high-resolution electrophoresis analysis.Experiments show that the DNA fragments can be concentrated up to nearly 1 order of magnitude greater than initial sample concentration within 10 s.
A novel method for integrating a nanoporous membrane in microfluidic devices was introduced.The nanoporous agarose gel membrane can be fabricated in-situ on PDMS microchips via micromolding techniques.The membrane is just a thin gel diaphragm perpendicularly embedded in the separation channel and close to the junction of the sample injection and separation channels,which enables electrokinetic concentration of DNA samples by a size exclusion mechanism prior to separation.This design allows a concentrated and well-defined sample to inject into the separation channel for sensitive high-resolution electrophoresis analysis.Experiments show that the DNA fragments can be concentrated up to nearly 1 order of magnitude greater than initial sample concentration within 10 s.
引文
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[2]Meldrum D.,Holl,Tech.Sight.Microfluidics.Microscale bioanalytical systeims[J].M.Science,2002,297:1197-1198.
[3]Levy M H,Goswami S,Plawsky J,Cramer S M.Parameters Governing the Formation of Photopolymerized Silica Sol-Gel Monoliths in PDMSMicrofluidic Chips[J].Chromatographia,2013,76:993-1002.
[4]Shih-Hao Yeh,Kuang-Hua Chou,Ruey-Jen Yang.Sample pre-concentration with high enrichment factors at a fixed location in paper-based microfluidic devices[J].Lab on a chip,2016,16:925-931.
[5]L.Malic,X.Zhang,D.Brassard,L.Clime,et al.Polymer-based microfluidic chip for rapid and efficient immunomagnetic capture and release of Listeria monocytogenes[J].Lab on a chip,2015,15:3994-4007.
[6]Levy M H,Plawsky J,Cramer S M.Photopolymerized sol-gel monoliths for separations of glycosylated proteins and peptides in microfluidic chips[J].J.Sep.Sci,2013,36:2358-2365.
[7]李刚,陈强,赵建龙.一种集成微孔膜的微流控芯片的制备方法:CN,101158694A[P].
[8]Song S.,Singh A.On-chip sample preconcentration for integrated microfluidic analysis[J].Anal Bioanal Chem.,2006,384:41-43.
[9]Song S.,Singh A.,Kirby B.Electrophoretic concentration of proteins at laser-patterned nanoporous membranes in microchips[J].Anal Chem,2004,76:4589-4592.
[10]Foote R.,Khandurina J.,Jacobson S.,et al.Preconcentration lf Proteins on Microfluidic Devices Using Porous Silica Membranes[J].Anal Chem.,2005,77:57-63.
[11]Badal M.,Wong M.,Chiem N.,et al.Protein separation and surfactant control of electroosmotic flow in poly(dimethylsiloxane)-coated capillaries and microchips[J].J Chromatogr A.,2002,947:277-286.
[12]Lu J.,Liu S.A robust cross-linked polyacrylamide coating for microchip electrophoresis of dsDNA fragments[J].Electrophoresis.,2006,27:3764-3771.
[13]C Wang,J Ouyang,DK Ye.Rapid protein concentration,efficient fluorescence labeling and purification on a micro/nanofluidics chip[J].Lab on a chip,2012,12:2664-2671.
[14]马玖彤,江丹丹,祁瑞芳,等.整体柱富集技术在微流控芯片系统中的应用[J].分析测试学报,2015,34(03):289-295.
[15]杜敏,叶雄英,冯金扬,等.一种基于微流控芯片的生物样品循环给样富集方法[J].分析化学,2012,40(11):1668-1673.
[16]张鹏,林阳,赵昕,等.场放大进样法在毛细管电泳样品富集方面的应用[J].当代化工,2013,42(12):1768-1772.
[17]H YU,Y Lu,YG Zhou,etc,et al.A simple,disposable microfluidic device for rapid protein concentration and purification via direct-printing[J].Lab on a chip,2008,8:1496-1501.
[18]Alzahrani E,Welham K.Fabrication of an octadecylated silica monolith inside a glass microchip for protein enrichment[J].Anal.Methods,2014,6:558-568.