微纳流控芯片制作方法及其富集应用
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摘要
近年来,随着微流控芯片技术的不断深入和发展,纳流控芯片技术或微纳流控芯片技术也得到了快速的发展。微纳流控芯片在离子或分子的分离与富集的分析研究具有广阔的研究空间和应用前景。随着纳流控芯片通道尺寸的降低,纳米通道具有一些特殊的性质如表面电荷,双电层,粘度增加和电渗流降低等。这些特殊的性质使得微纳流控芯片在生化分析方面具有重要作用。本文重点研究了微纳流控芯片的制作方法,采用玻璃和聚二甲基硅氧烷(PDMS)制作微纳流控芯片,最后采用微纳流控芯片进行离子和蛋白的富集实验。
由于玻璃材料具有很好的光学性能和电学性能,刻蚀的各向同向性和制造精度高等特点,因此本文优先采用玻璃材料来制作微纳流控芯片。结合微纳流控芯片结构特点,本文通过两块基片上采用光刻和湿法刻蚀的方法分别制作微米通道和纳米通道。通道形成以后,采用专门的对准装配装置对准,然后进行预联结,最后通过高温键合的方法永久性键合芯片。
PDMS具有价格便宜,制作工艺比较简单,制作周期比较短的特点,因此PDMS在微流控芯片的制作材料上具有很大的优势。本文首先制作了PDMS的微米通道芯片的SU-8胶模具,通过软光刻的方法制作微米通道芯片。由合作单位进行键合,制作了上下两层是PDMS的微米通道芯片,中间一层是纳米孔膜的三明治结构的PDMS微纳流控芯片。
最后,本文结合国内外富集原理初步分析了微纳流控芯片的特性:表面电荷,双电层和尺寸效应。本文在电驱动下的微纳流控芯片进行离子的富集实验,得到了高于富集溶液多倍的离子浓度。采用牛血清白蛋白(BSA/FITC)溶液作为富集溶液进行大分子的富集实验,也取得了很好的效果。
In recent years, with the development of fabricating technologies of microfluidic chip, fabricating technologies of nanofluidic chip or micro-nanofluidic Chip has also been rapidly developed. Micro-nanofluidic chip is used in the research such as ion or molecular analysis, separation and enrichment, so it has broad research space and application prospect. With an increasing smaller size of Nanochannel flow, nanofluidic chip has some special properties such as surface electrical charge, the electrical double layer, increased viscosity and decreased electroosmotic flow. These special properties make micro-nanofluidic chip play an important role in biochemical analysis. This thesis focuses on the fabrication methods of micro-nanofluidic chip and has fabricated glass and PDMS micro-nanofluidic chip. This thesis uses micro-nanofluidic chip to do the efficient enrichment experiments of ions and protein at last.
In this thesis, glass materials are used to fabricate micro-nanofluidic chip, because the glass material has excellent optical properties and electrical properties, and other good characteristics like all to the same anisotropic etching and high precision manufacture. Adding to the structural features of micro-nanofluidic chip, this thesis uses photolithography and wet etching process to fabricate microchannel and nanochannel chips respectively. Afer the channels are formed, the upper plate and the bottom plate can be assembled together by alignment, preconnection and thermal bonding sequently. At last, the bottom plate and the upper one are irreversibly linked together with thermal bonding.
As PDMS material is a cheap and relatively simple production process with the feature of short production cycle. PDMS material has great advantages in microfluidic chip fabrication. The author fabricates a SU-8 mold of the PDMS microfluidic chip, and then employs the casting process to make a microfluidic chip. With the help of cooperating party, the PDMS micro-nanofluidic chip is finally fabricated where the bottom layer and the upper layer are both PDMS microfluidic chips while the middle layer is a mesoporous membrane. This is a micro-nanofluidic chip with sandwich structure.
Finally, this thesis makes preliminary study of the characteristics of the micro-nanofluidic chip, surface charge, electrical double layer and the size effect. The ionic concentration of the solution is several times higher than before. The author also does a lot of experiments to explore the method of using BSA/FITC solution as a protein enrichment solution and achieved good results.
由于玻璃材料具有很好的光学性能和电学性能,刻蚀的各向同向性和制造精度高等特点,因此本文优先采用玻璃材料来制作微纳流控芯片。结合微纳流控芯片结构特点,本文通过两块基片上采用光刻和湿法刻蚀的方法分别制作微米通道和纳米通道。通道形成以后,采用专门的对准装配装置对准,然后进行预联结,最后通过高温键合的方法永久性键合芯片。
PDMS具有价格便宜,制作工艺比较简单,制作周期比较短的特点,因此PDMS在微流控芯片的制作材料上具有很大的优势。本文首先制作了PDMS的微米通道芯片的SU-8胶模具,通过软光刻的方法制作微米通道芯片。由合作单位进行键合,制作了上下两层是PDMS的微米通道芯片,中间一层是纳米孔膜的三明治结构的PDMS微纳流控芯片。
最后,本文结合国内外富集原理初步分析了微纳流控芯片的特性:表面电荷,双电层和尺寸效应。本文在电驱动下的微纳流控芯片进行离子的富集实验,得到了高于富集溶液多倍的离子浓度。采用牛血清白蛋白(BSA/FITC)溶液作为富集溶液进行大分子的富集实验,也取得了很好的效果。
In recent years, with the development of fabricating technologies of microfluidic chip, fabricating technologies of nanofluidic chip or micro-nanofluidic Chip has also been rapidly developed. Micro-nanofluidic chip is used in the research such as ion or molecular analysis, separation and enrichment, so it has broad research space and application prospect. With an increasing smaller size of Nanochannel flow, nanofluidic chip has some special properties such as surface electrical charge, the electrical double layer, increased viscosity and decreased electroosmotic flow. These special properties make micro-nanofluidic chip play an important role in biochemical analysis. This thesis focuses on the fabrication methods of micro-nanofluidic chip and has fabricated glass and PDMS micro-nanofluidic chip. This thesis uses micro-nanofluidic chip to do the efficient enrichment experiments of ions and protein at last.
In this thesis, glass materials are used to fabricate micro-nanofluidic chip, because the glass material has excellent optical properties and electrical properties, and other good characteristics like all to the same anisotropic etching and high precision manufacture. Adding to the structural features of micro-nanofluidic chip, this thesis uses photolithography and wet etching process to fabricate microchannel and nanochannel chips respectively. Afer the channels are formed, the upper plate and the bottom plate can be assembled together by alignment, preconnection and thermal bonding sequently. At last, the bottom plate and the upper one are irreversibly linked together with thermal bonding.
As PDMS material is a cheap and relatively simple production process with the feature of short production cycle. PDMS material has great advantages in microfluidic chip fabrication. The author fabricates a SU-8 mold of the PDMS microfluidic chip, and then employs the casting process to make a microfluidic chip. With the help of cooperating party, the PDMS micro-nanofluidic chip is finally fabricated where the bottom layer and the upper layer are both PDMS microfluidic chips while the middle layer is a mesoporous membrane. This is a micro-nanofluidic chip with sandwich structure.
Finally, this thesis makes preliminary study of the characteristics of the micro-nanofluidic chip, surface charge, electrical double layer and the size effect. The ionic concentration of the solution is several times higher than before. The author also does a lot of experiments to explore the method of using BSA/FITC solution as a protein enrichment solution and achieved good results.
引文
[1]林柄承,秦建华.微流控芯片实验室[M].北京:中国林业出版社,1993.
[2]方肇伦.微流控芯片[M].北京:科学出版社,2003.
[3]杨秀娟,李想,童艳丽等.微流控芯片在细胞分析中的应用[J].细胞生物学杂志,2008,30(6):721-726.
[4]Brandt B H,Schmidt H,Angelis G D,et al.Predictive laboratory diagnostics in oncology utilizing blood-borne cancer cells-current best practice and unmet eeds[J].Cancer Lett,2001,162:11-16.
[5]刘海军,杜立群,刘冲等.一种玻璃微流控芯片的实用制作工艺研究[J].分析科学学报,2006,22(6):671-674.
[6]杨波,唐飞,王晓浩等.一种微流控检测芯片的设计与工艺研究[J].仪器仪表学报,2009,30(7):1464-1468.
[7]陈实,张卫平,陈文元等.用于微流控芯片的PDMS微混合器工艺和数值分析[J].微细加工技术,2006,12(6):48-52.
[8]Andrew G H,Stephen C J,and J.Michael Ramsey.Microfluidic Assays of Acetylcholinesterase Inhibitor[J].Anal Chem,1999,71(22):5206-5212.
[9]Dongeun H,Mills K L,Xiaoyue Zhu et al.Tuneable elastomeric nanochannels for nanofluidic manipulation[J].nature materials,2007,6(6):424-428.
[10]曹炳阳,陈民,过增元.纳米通道内液体流动的滑移现象[J].物理学报,2006,55(10):5305-5310.
[11]Wang Y C,Stevens A L,Jongyoon H.Million-fold preconcentration of proteins and peptides by nanofluidic filter[J].Anal Chem,2005,77(14):4293-4299.
[12]Kuo T C,Donald M C,Chen Y N et al.Gateable Nanofluidic Interconnects for Multilayered Microfluidic Separation Systems[J].Anal Chem,2003,5(8):1861-1867.
[13]Datta A,Gangopadhyay S,Temkin H et al.Nanofluidic channels by anodic bonding of amorphous silicon to glass to study ion-accumulation and ion-depletion effect [J].Talanta,2006,68(3):659-665.
[14]Thomas S H,Nicolas F R,Staufer U.Fabrication and electroosmotic flow measurements in micro- and nanofluidic channels[J].Microfluid Nanofluidid,2006,2(2):117-124.
[15]Liu J,zhuang G S,Yang M Set al.Efficiency controllable sample preconcentration chips based on nano ion depletion bridge[C],10th μ TAS Conference,2006,Japan.
[16]Zheng Xu,Jin-kai Wen,Chong Liu et al.Research on forming and application of U-form glass micro-nanofluidic chip with long nanochannels[J].Microfluidic Nanofluidics,2009,7(3):423-429.
[17]廖裕评,陆瑞强.Tanner Pro集成电路设计与布局实战指导[M].北京:科学出版社,2004.
[18]程莉莉,余冬冬,邓晓清等.玻璃-PDM8微流控芯片制备工艺[J].武汉工程大学学报,2009,31(1):58-61.
[19]David S S,Roger T H.SACRIFICIAL LAYER Si02 WET ETCHING FOR MICROMACHINING APPLICATIONS[J].Transducers,IEEE,1991,647-650.
[20]Tenney A S,Ghezzo M.Etch Rates of Doped Oxides in Solutions of Buffered HF[J].Slid-State Science and Technology,1973,120(8):1091-1095.
[21]杜立群,李璞,刘军山.基于元胞自动机算法的三维玻璃微流控芯片湿法刻蚀模拟研究[J].计算机学报,2008,31(5):868-874.
[22]陈双.玻璃纳流控芯片的制作及应用研究[D]:(硕士学位论文).杭州:浙江大学,2007.
[23]乔红超.基于等离子体技术的聚合物微纳流控芯片制作[D]:(硕士学位论文).大连:大连理工大学,2008.
[24]罗国安,陈令新,刘科辉等.一种在普通实验室条件下制作玻璃芯片的方法[P].中国,发明专利,200410049750.4.2004,06,25.
[25]殷学锋,沈宏,方肇伦.制造玻璃微流控芯片的简易加工技术[J].分析化学(FENXI HUAXUE)仪器装置与实验技术,2003,31(1):116-119.
[26]罗怡,娄志峰,褚德南等.玻璃微流控芯片的制作[J].纳米技术与精密工程,2004,2(1):22-23.
[27]刘娜.微结构观测镜光学结构研究与设计[D]:(硕士学位论文).大津:天津大学,2003.
[28]顾书龙,张宏彬.扫描近场光学显微镜的技术与应用[J].阜阳师范学院学报自然科学版,2003,20(1):20-24.
[29]徐征,刘冲,温金开等.一种玻璃微纳流控芯片、制作组装方法及其辅助装配装置[P].中国,发明专利,200910302903.4.2009,6,4.
[30]徐征,王德佳,刘冲等.一种玻璃微纳流控芯片对准装配方法及装置[P].中国,发明专利,200910307123.9.2009,9,17.
[31]朱学林,郭育华,刘刚等.微流控芯片透光性基底SU8曝光工艺[J].功能材料与器件学报,2006,12(13):426-430.
[32]王彬,陈翔,许宝建等.基于SU-8负胶的微流体器件的制作及研究[J].功能材料与器件学报,2008,114(12):215-219.
[33]Bilenberg B,Nielsen T,Clausen B.PMMA to SU-8 bonding for polymer based lab-on-a-chip systemswith integrated optics[J].JMicromech Microeng,2004,14(6):814-818.
[34]秦江.SU-8胶紫外光刻理论与实验研究[D]:(硕士学位论文),大连:大连理工大学,2006.
[35]Luo Y,Wang X D,Liu C et al.Swelling of SU-8 structure in Ni mold fabrication by UV-LIGA technique[J].Microsyst Technol,2005,11(12):1272-1275.
[36]李丹.PDMS微流控芯片加工技术及微萃取系统的实验研究[D]:(硕士学位论文),天津:天津大学,2006.
[37]黄道君.PDMS芯片粘接性能及微萃取系统的实验研究[D]:(硕士学位论文),天津:天津大学,2007.
[38]余群,郑旭,王旭伟等.微纳组合管道连接处富集速率观测[C].第二届国际微流控学会议(第五届全国微全分析系统学术会议).中国.南京,2008.11:78-79.
[39]Kim,S M,Hasselbrink,E F,Jr.Micro Total Analysis Systems 2003;Northrup,M.A.,Jensen,K.F.,Harrison,D.J,Eds.Transducers Research Foundation[J],Inc.:Cleveland,2003,1171-1174.
[40]Reto Bruno SCHOCH.Transport phenomena in nanofluidics[D].form ionic studies to proteomic applications,Massachusetts:Univ.of Cambridge,2006.
[41]Qiaosheng P,Jongsin Y,Henryk T et al.Ion-Enrichment and Ion-Depletion Effect of Nanochannel Structures[J].NANO LETTERS,2004,4(6):1099-1103.
[2]方肇伦.微流控芯片[M].北京:科学出版社,2003.
[3]杨秀娟,李想,童艳丽等.微流控芯片在细胞分析中的应用[J].细胞生物学杂志,2008,30(6):721-726.
[4]Brandt B H,Schmidt H,Angelis G D,et al.Predictive laboratory diagnostics in oncology utilizing blood-borne cancer cells-current best practice and unmet eeds[J].Cancer Lett,2001,162:11-16.
[5]刘海军,杜立群,刘冲等.一种玻璃微流控芯片的实用制作工艺研究[J].分析科学学报,2006,22(6):671-674.
[6]杨波,唐飞,王晓浩等.一种微流控检测芯片的设计与工艺研究[J].仪器仪表学报,2009,30(7):1464-1468.
[7]陈实,张卫平,陈文元等.用于微流控芯片的PDMS微混合器工艺和数值分析[J].微细加工技术,2006,12(6):48-52.
[8]Andrew G H,Stephen C J,and J.Michael Ramsey.Microfluidic Assays of Acetylcholinesterase Inhibitor[J].Anal Chem,1999,71(22):5206-5212.
[9]Dongeun H,Mills K L,Xiaoyue Zhu et al.Tuneable elastomeric nanochannels for nanofluidic manipulation[J].nature materials,2007,6(6):424-428.
[10]曹炳阳,陈民,过增元.纳米通道内液体流动的滑移现象[J].物理学报,2006,55(10):5305-5310.
[11]Wang Y C,Stevens A L,Jongyoon H.Million-fold preconcentration of proteins and peptides by nanofluidic filter[J].Anal Chem,2005,77(14):4293-4299.
[12]Kuo T C,Donald M C,Chen Y N et al.Gateable Nanofluidic Interconnects for Multilayered Microfluidic Separation Systems[J].Anal Chem,2003,5(8):1861-1867.
[13]Datta A,Gangopadhyay S,Temkin H et al.Nanofluidic channels by anodic bonding of amorphous silicon to glass to study ion-accumulation and ion-depletion effect [J].Talanta,2006,68(3):659-665.
[14]Thomas S H,Nicolas F R,Staufer U.Fabrication and electroosmotic flow measurements in micro- and nanofluidic channels[J].Microfluid Nanofluidid,2006,2(2):117-124.
[15]Liu J,zhuang G S,Yang M Set al.Efficiency controllable sample preconcentration chips based on nano ion depletion bridge[C],10th μ TAS Conference,2006,Japan.
[16]Zheng Xu,Jin-kai Wen,Chong Liu et al.Research on forming and application of U-form glass micro-nanofluidic chip with long nanochannels[J].Microfluidic Nanofluidics,2009,7(3):423-429.
[17]廖裕评,陆瑞强.Tanner Pro集成电路设计与布局实战指导[M].北京:科学出版社,2004.
[18]程莉莉,余冬冬,邓晓清等.玻璃-PDM8微流控芯片制备工艺[J].武汉工程大学学报,2009,31(1):58-61.
[19]David S S,Roger T H.SACRIFICIAL LAYER Si02 WET ETCHING FOR MICROMACHINING APPLICATIONS[J].Transducers,IEEE,1991,647-650.
[20]Tenney A S,Ghezzo M.Etch Rates of Doped Oxides in Solutions of Buffered HF[J].Slid-State Science and Technology,1973,120(8):1091-1095.
[21]杜立群,李璞,刘军山.基于元胞自动机算法的三维玻璃微流控芯片湿法刻蚀模拟研究[J].计算机学报,2008,31(5):868-874.
[22]陈双.玻璃纳流控芯片的制作及应用研究[D]:(硕士学位论文).杭州:浙江大学,2007.
[23]乔红超.基于等离子体技术的聚合物微纳流控芯片制作[D]:(硕士学位论文).大连:大连理工大学,2008.
[24]罗国安,陈令新,刘科辉等.一种在普通实验室条件下制作玻璃芯片的方法[P].中国,发明专利,200410049750.4.2004,06,25.
[25]殷学锋,沈宏,方肇伦.制造玻璃微流控芯片的简易加工技术[J].分析化学(FENXI HUAXUE)仪器装置与实验技术,2003,31(1):116-119.
[26]罗怡,娄志峰,褚德南等.玻璃微流控芯片的制作[J].纳米技术与精密工程,2004,2(1):22-23.
[27]刘娜.微结构观测镜光学结构研究与设计[D]:(硕士学位论文).大津:天津大学,2003.
[28]顾书龙,张宏彬.扫描近场光学显微镜的技术与应用[J].阜阳师范学院学报自然科学版,2003,20(1):20-24.
[29]徐征,刘冲,温金开等.一种玻璃微纳流控芯片、制作组装方法及其辅助装配装置[P].中国,发明专利,200910302903.4.2009,6,4.
[30]徐征,王德佳,刘冲等.一种玻璃微纳流控芯片对准装配方法及装置[P].中国,发明专利,200910307123.9.2009,9,17.
[31]朱学林,郭育华,刘刚等.微流控芯片透光性基底SU8曝光工艺[J].功能材料与器件学报,2006,12(13):426-430.
[32]王彬,陈翔,许宝建等.基于SU-8负胶的微流体器件的制作及研究[J].功能材料与器件学报,2008,114(12):215-219.
[33]Bilenberg B,Nielsen T,Clausen B.PMMA to SU-8 bonding for polymer based lab-on-a-chip systemswith integrated optics[J].JMicromech Microeng,2004,14(6):814-818.
[34]秦江.SU-8胶紫外光刻理论与实验研究[D]:(硕士学位论文),大连:大连理工大学,2006.
[35]Luo Y,Wang X D,Liu C et al.Swelling of SU-8 structure in Ni mold fabrication by UV-LIGA technique[J].Microsyst Technol,2005,11(12):1272-1275.
[36]李丹.PDMS微流控芯片加工技术及微萃取系统的实验研究[D]:(硕士学位论文),天津:天津大学,2006.
[37]黄道君.PDMS芯片粘接性能及微萃取系统的实验研究[D]:(硕士学位论文),天津:天津大学,2007.
[38]余群,郑旭,王旭伟等.微纳组合管道连接处富集速率观测[C].第二届国际微流控学会议(第五届全国微全分析系统学术会议).中国.南京,2008.11:78-79.
[39]Kim,S M,Hasselbrink,E F,Jr.Micro Total Analysis Systems 2003;Northrup,M.A.,Jensen,K.F.,Harrison,D.J,Eds.Transducers Research Foundation[J],Inc.:Cleveland,2003,1171-1174.
[40]Reto Bruno SCHOCH.Transport phenomena in nanofluidics[D].form ionic studies to proteomic applications,Massachusetts:Univ.of Cambridge,2006.
[41]Qiaosheng P,Jongsin Y,Henryk T et al.Ion-Enrichment and Ion-Depletion Effect of Nanochannel Structures[J].NANO LETTERS,2004,4(6):1099-1103.
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