基于SPR技术的检测装置设计与实验研究
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摘要
基于表面等离子体共振(Surface Plasmon Resonance,SPR)的监测仪器具有灵敏度高、检测速度快、样品消耗量少、无需标记生物样品等特点,能够广泛应用于化学、生物、环境、食品、医疗、制药等领域。但是,目前国内外大部分SPR仪器都存在结构复杂、体积庞大、成本高等缺点,限制了其在现场监测、实时分析等方面的运用。因此,研制小体积低成本的SPR检测装置,必然能够推广SPR技术在各领域的应用,产生巨大的社会效益和经济效益。
本文在仔细分析SPR检测仪器应用领域、现状和发展趋势以及传感原理的基础上,自主开发了一套小型SPR检测装置。该装置主要包括SPR传感器、微流恒速进样系统、信号检测与控制电路以及计算机数据分析处理软件。传感器采用宽光束入射的非扫描式角度调制方式,简化了传统具有可动部件的角度扫描方式,简化了SPR传感器结构,减小了体积。微流恒速进样系统由高精度注射泵和智能多位阀组成,可满足极微量样品的选择以及控制样品进样速度的恒定,并可进行样品、缓冲液和再生液的自动选择进样,保证测量过程的自动完成。信号检测与控制系统和计算机数据分析处理软件为SPR仪器提供了强大的数据采集与分析处理功能,使其具有实用性强、操作方便等特点。
利用自行设计的小型SPR检测装置,搭建一套基于SPR技术的在线折射率计。对不同浓度的葡萄糖溶液进行测量,结果表明该装置具有较好的性能,其折射率检测灵敏度达到5×10~(-5)RIU(折射率单位),能够测量的葡萄糖溶液的最低浓度可达0.01%。
利用自行设计的小型SPR检测装置,搭建一套基于SPR技术的生物传感器。简要地分析SPR生物检测原理与检测过程,在此基础上,通过磺胺甲恶唑(sulfamethoxazole,SMX)抗原浓度的实验,表明该装置在生物分子相互作用方面的直接、实时在线观测的能力。
Surface plasmon resonance ( SPR ) monitor instrument has the advantage of high-sensitivity, fast detection, less sample needed and label-free analysis. It can be widely used in chemistry, biometrics, environmental monitor, food detection, clinic diagnosis, and medicine etc. However, most SPR instruments have the disadvantage of complex structure, huge physical volume and high cost. The disadvantage limits its application in field monitor and real-time analysis. As a result, developing a small and low-cost SPR monitor instrument will spread SPR technique using in each area. It will bring huge social benefits and economic benefits.
This research carefully analyses the application area, developing trend and sensor principle of the SPR monitor instrument. We developed a set of small SPR monitor instrument independently. The instrument consists of a SPR sensor, a sampling system with tiny and constant flow, a signal detection and control circuit and analysis processing software. The SPR sensor adopts non-scanning modulation structure with broad beam. This modulation is based on the traditional scanning modulation with moving parts. But it reduces the physical volume of SPR sensor. The sampling system with tiny and constant flow consists of high precision pump and multi-position micro-electric valve. This system can control the speed of sample and choose different samples. All the measuring process is auto finished. The signal detection and control circuit and analysis processing software provide the function of data acquisition and analysis processing for SPR instrument.
Dextrose solutions of different concentration are measured by our small SPR instrument. The result presents that the instrument has good performance. The sensitivity reaches approximately 5×10~(-5)RIU, the minimum concentration can be detected is 0.01%.
In the paper we also analyze the SPR principle of biological detection and detection process briefly. SMX is measured by our small SPR instrument. The result presents that the system's powerful ability on directly monitoring interactions between biomolecules.
本文在仔细分析SPR检测仪器应用领域、现状和发展趋势以及传感原理的基础上,自主开发了一套小型SPR检测装置。该装置主要包括SPR传感器、微流恒速进样系统、信号检测与控制电路以及计算机数据分析处理软件。传感器采用宽光束入射的非扫描式角度调制方式,简化了传统具有可动部件的角度扫描方式,简化了SPR传感器结构,减小了体积。微流恒速进样系统由高精度注射泵和智能多位阀组成,可满足极微量样品的选择以及控制样品进样速度的恒定,并可进行样品、缓冲液和再生液的自动选择进样,保证测量过程的自动完成。信号检测与控制系统和计算机数据分析处理软件为SPR仪器提供了强大的数据采集与分析处理功能,使其具有实用性强、操作方便等特点。
利用自行设计的小型SPR检测装置,搭建一套基于SPR技术的在线折射率计。对不同浓度的葡萄糖溶液进行测量,结果表明该装置具有较好的性能,其折射率检测灵敏度达到5×10~(-5)RIU(折射率单位),能够测量的葡萄糖溶液的最低浓度可达0.01%。
利用自行设计的小型SPR检测装置,搭建一套基于SPR技术的生物传感器。简要地分析SPR生物检测原理与检测过程,在此基础上,通过磺胺甲恶唑(sulfamethoxazole,SMX)抗原浓度的实验,表明该装置在生物分子相互作用方面的直接、实时在线观测的能力。
Surface plasmon resonance ( SPR ) monitor instrument has the advantage of high-sensitivity, fast detection, less sample needed and label-free analysis. It can be widely used in chemistry, biometrics, environmental monitor, food detection, clinic diagnosis, and medicine etc. However, most SPR instruments have the disadvantage of complex structure, huge physical volume and high cost. The disadvantage limits its application in field monitor and real-time analysis. As a result, developing a small and low-cost SPR monitor instrument will spread SPR technique using in each area. It will bring huge social benefits and economic benefits.
This research carefully analyses the application area, developing trend and sensor principle of the SPR monitor instrument. We developed a set of small SPR monitor instrument independently. The instrument consists of a SPR sensor, a sampling system with tiny and constant flow, a signal detection and control circuit and analysis processing software. The SPR sensor adopts non-scanning modulation structure with broad beam. This modulation is based on the traditional scanning modulation with moving parts. But it reduces the physical volume of SPR sensor. The sampling system with tiny and constant flow consists of high precision pump and multi-position micro-electric valve. This system can control the speed of sample and choose different samples. All the measuring process is auto finished. The signal detection and control circuit and analysis processing software provide the function of data acquisition and analysis processing for SPR instrument.
Dextrose solutions of different concentration are measured by our small SPR instrument. The result presents that the instrument has good performance. The sensitivity reaches approximately 5×10~(-5)RIU, the minimum concentration can be detected is 0.01%.
In the paper we also analyze the SPR principle of biological detection and detection process briefly. SMX is measured by our small SPR instrument. The result presents that the system's powerful ability on directly monitoring interactions between biomolecules.
引文
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[15]Sunil K.Arya,Pratima R.Solanki,S.P.Singh.Poly-(3-hexylthiophene)self-assembled monolayer based cholesterol biosensor using surface plasmon resonance technique,Biosensors and Bioelectronics.2007,22(11):2516-2524.
[16]Limin Cao,Hong Lin and Vladimir M.Mirsky.Surface plasmon resonance biosensor for enrofloxacin based on deoxyribonucleic acid.Analytica Chimica Acta.2007,589(1):1-5.
[17]John Waswa,Joseph Irudayaraj and Chitrita DebRoy.Direct detection of E.Coli O157:H7 in selected food systems by a surface plasmon resonance biosensor.LWT - Food Science and Technology.2007,40(2):187-192.
[18]Louis A.Obando,Darcy J.Gentleman,John R.Holloway and Karl S.Booksh Manufacture of robust surface plasmon resonance fiber optic based dip-probes.Sensors and Actuators B:Chemical.2004,100(3):439-449.
[19]Futao Kaneko,Takayuki Nakano,Mitsuru Terakado,Kazunari Shinbo.Emission light from prism/silver/rhodamine-B LB film and multiple surface plasmon excitations in the ATR Kretschmann configuration.Materials Science and Engineering:C.2002,22(2):409-412.
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[23]F.C.Chien,S.J.Chen,W.P.Hu,G.Y.Lin,et al.Nanoparticle-enhanced ultrahigh-resolution surface plasmon resonance biosensors.Plasmonics in Biology and Medicine,Proceedings of SPIE,Bellingham,WA,004.5327:140-147.
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[28]E.Kretschmann,H.Raether.Radiative decay of non-radiative surface plasmons excited by light.Z.Naturforsch,1968,23A:2135-2136.
[29]A.Otto.Excitation of surface plasma waves in silver by the method of frustrated totalreflection.Z.Physik,1968,216:398-410.
[30]聂奕.光波导水质分析仪的研究.华中科技大学图书馆,2001:12-19.
[1]Texas Instruments.SprcetaTM Evaluation Kit.Texas Instruments Incorporated,1999.
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[4]张迎新等编著,C8051系列SOC单片机原理及应用,国防工业出版社2005年8月第一版.
[5]康华光,电子技术基础模拟部分(第四版),高等教育出版社,1996年6月.
[6]http://www.analog.com
[7]http://www.maxim-ic.com
[8]徐爱钧,彭秀华 编著,单片机高级语言C51 Windows环境编程与应用,电子工业出版社,2001年7月第1版.
[9]谭浩强,C程序设计,清华大学出版社.1999.
[1]柳春郁,余有龙,高应俊.基于菲涅尔原理的光纤型折射率测量装置光电子.激光.2004,15(12):1464-1466.
[2]秦书乐,施健,李卫,邬丁宝.光纤传感式材料折射仪,上海计量测试.2004,31(5):22-23.
[3]胡晓伟,倪旭翔,朱育君,徐进基于CMOS图像传感器的液体折射率计研究,光学仪器.2005,27(4):69-72.
[4]宋大千,赵晓君,陈焕文等.表面等离子体子共振传感器 Ⅱ:实验装置与仪器,分析仪器.2001,1:1-6
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[7]Kazuyoshi Kurihara and Koji Suzuki.Theoretical Understanding of an Absorption-Based Surface Plasmon Resonance Sensor Based on Kretchmann's Theory,Anal.Chem.2002,74:696-701.
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[9]李大勇,曹振新,吴乐南.光纤表面等离子体共振传感检测系统的数据处理,测控技术 2005,12.
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[4]G Bischof,R.Bischoff,E.Birch-Hirschfeld,U.Gromann,S.Lindau,W.V.Meister,S.Bambirra,C.Bohley,S.Hoffmann,J.Biomol.Struct.Dyn1998,16:187.
[5]W.P.Chen,J.M.Chen,J.Opt.Soc.Am.1981,71(2):189.
[6]C.R.Lawrence,A.S.Martin,J.R.Sambles,Thin Solid Films,1992,208:269.
[7]A.H.Severs,R.B.M.Schasfoort Biosens.Bioelectron.,1993(8):365.
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[9]W.Knoll,M.Zizlsperger etc.Streptavidin Arrays as Supra-molecular Architectures in Surface Plasmon,Optical Sensor Formats.1999:57-59.
[10]Waveguide sensor detects trace levels of herbicidein water,OLE.1995:39
[11]缪璐,张水华,刘仲明.表面等离子体共振技术在食品工业中的应用,食品安全与检测,2006,3:136-138.
[12]缪璐,张水华,刘仲明.SPR传感器在食品检测中的应用,食品科技.2006(8):266-268.
[13]陈焕文,牟颖,赵晓君等.表面等离子体子共振传感器Ⅲ:应用与发展,分析仪器.2001,2:3-8.
[14]程慧,黄朝峰,段子渊.SPR生物传感器及其应用进展.中国生物工程杂志.2003,5.
[15]Sunil K.Arya,Pratima R.Solanki,S.P.Singh.Poly-(3-hexylthiophene)self-assembled monolayer based cholesterol biosensor using surface plasmon resonance technique,Biosensors and Bioelectronics.2007,22(11):2516-2524.
[16]Limin Cao,Hong Lin and Vladimir M.Mirsky.Surface plasmon resonance biosensor for enrofloxacin based on deoxyribonucleic acid.Analytica Chimica Acta.2007,589(1):1-5.
[17]John Waswa,Joseph Irudayaraj and Chitrita DebRoy.Direct detection of E.Coli O157:H7 in selected food systems by a surface plasmon resonance biosensor.LWT - Food Science and Technology.2007,40(2):187-192.
[18]Louis A.Obando,Darcy J.Gentleman,John R.Holloway and Karl S.Booksh Manufacture of robust surface plasmon resonance fiber optic based dip-probes.Sensors and Actuators B:Chemical.2004,100(3):439-449.
[19]Futao Kaneko,Takayuki Nakano,Mitsuru Terakado,Kazunari Shinbo.Emission light from prism/silver/rhodamine-B LB film and multiple surface plasmon excitations in the ATR Kretschmann configuration.Materials Science and Engineering:C.2002,22(2):409-412.
[20]R.Levy,A.Peled and S.Ruschin.Waveguided SPR sensor using a Mach-Zehnde interferometer with variable power splitting ratio.Sensors and Actuators B:Chemical.2006,119(1):20-26.
[21]A.Suarez-Garcia,R.del Coso,R.Serna,et al.Controlling the transmission at the surface plasmon resonance of nanocomposite films using photonic structures.Applied Physics Letters.2003,83:1842-1844.
[22]Kyung Min Byun,Donghyun Kim,and Sung June Kim.Investigation of the sensitivity enhancement of nanoparticle based surface plasmon resonance biosensors using rigorous coupled wave analysis Plasmonics in Biology and Medicine Ⅱ,Proceedings of SPIE Vol.5703,SPIE,Bellingham,WA.2005,61-70.
[23]F.C.Chien,S.J.Chen,W.P.Hu,G.Y.Lin,et al.Nanoparticle-enhanced ultrahigh-resolution surface plasmon resonance biosensors.Plasmonics in Biology and Medicine,Proceedings of SPIE,Bellingham,WA,004.5327:140-147.
[24]Borja Sepulveda,Ana Calle,Laura M.Lechuga,and Gaspar Armelles.Highly sensitive detection of biomolecules with the Magneto-Optic Surface Plasmon Resonance sensor.Optics Letters.2006.
[25]顾秉林,王喜坤.固体物理学.第1版,清华大学出版社,1989:1-7.
[26]H.E.Hall.固体物理学.第1版,刘志远,张增顺译.高等教育出版社,1983:243-248.
[27]赵建林.高等光学.第1版.国防工业出版社,2002:41-44.
[28]E.Kretschmann,H.Raether.Radiative decay of non-radiative surface plasmons excited by light.Z.Naturforsch,1968,23A:2135-2136.
[29]A.Otto.Excitation of surface plasma waves in silver by the method of frustrated totalreflection.Z.Physik,1968,216:398-410.
[30]聂奕.光波导水质分析仪的研究.华中科技大学图书馆,2001:12-19.
[1]Texas Instruments.SprcetaTM Evaluation Kit.Texas Instruments Incorporated,1999.
[2]Multipositon Microelectric Valve Actuators,VICI Valco Instruments Co.Inc.
[3]ConfluentTM PVM Instructions for Use,Sapphire Engineering:A unit of IDEX corporation,2006.
[4]张迎新等编著,C8051系列SOC单片机原理及应用,国防工业出版社2005年8月第一版.
[5]康华光,电子技术基础模拟部分(第四版),高等教育出版社,1996年6月.
[6]http://www.analog.com
[7]http://www.maxim-ic.com
[8]徐爱钧,彭秀华 编著,单片机高级语言C51 Windows环境编程与应用,电子工业出版社,2001年7月第1版.
[9]谭浩强,C程序设计,清华大学出版社.1999.
[1]柳春郁,余有龙,高应俊.基于菲涅尔原理的光纤型折射率测量装置光电子.激光.2004,15(12):1464-1466.
[2]秦书乐,施健,李卫,邬丁宝.光纤传感式材料折射仪,上海计量测试.2004,31(5):22-23.
[3]胡晓伟,倪旭翔,朱育君,徐进基于CMOS图像传感器的液体折射率计研究,光学仪器.2005,27(4):69-72.
[4]宋大千,赵晓君,陈焕文等.表面等离子体子共振传感器 Ⅱ:实验装置与仪器,分析仪器.2001,1:1-6
[5]Texas Instruments.SpreetaTM Evaluation Kit.Texas Instruments Incorporated,1999.
[6]江秀明,陈志春,杨绍明.光纤表面等离子体共振传感器研究进展,传感技术学报.2003,1:74-75.
[7]Kazuyoshi Kurihara and Koji Suzuki.Theoretical Understanding of an Absorption-Based Surface Plasmon Resonance Sensor Based on Kretchmann's Theory,Anal.Chem.2002,74:696-701.
[8]Multipositon Microelectric Valve Actuators,VICI Valco Instruments Co.Inc.
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