集成生化低电压芯片电泳系统的基础理论及关键技术研究
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摘要
微型全分析芯片系统是微型分析仪器发展的重要方向,以芯片电泳技术为主流的微型全分析系统(μ-TAS),以其高效、快速、微量、易自动化等优点,正以强劲的势头向生命科学等各相关领域渗透。特别是生化应用领域的集成微流控芯片电泳分析系统,已成为芯片技术的主要研究领域,高集成度化、微型化和便携化的芯片分析系统是人们目前研发的重点和热点。
本博士学位论文针对目前微流控芯片电泳存在的系统分离电压高、检测系统分离、芯片与外界的接口匹配、芯片系统一体化集成加工技术等问题,提出低电压运动梯度场电泳理论、集成芯片电泳的低电压分离模型与控制方法。通过对电场和流场的模拟计算和分析,确定了低电压电泳芯片的整体结构,并对系统的进样、分离和检测进行了结构和参数优化的设计;提出了在芯片微通道上双检测器的集成思想;提出了实现集成生化低电压芯片电泳分离的控制方法,并研制了实现低电压运动梯度场的控制电路;提出了以SOI-MEMS加工技术实现含阵列微电极的硅基低电压电泳芯片的加工工艺方法,实现了芯片的一体化集成;采用氨基酸样品体系对研制出的集成生化低电压芯片电泳系统进行了实验,实验结果有效验证了本论文研制的集成生化低电压芯片电泳系统的可行性。
本文主要工作如下:
①本文在查阅大量文献、资料和总结前期研究工作的基础上,在研究集成芯片电泳的低电压分离理论及其控制方法的基础上,提出集成低电压电泳芯片总体方案;
②通过集成低电压电泳芯片模型的建立,采用ANAS和ConvertorWare软件分析系统进行了集成低电压电泳芯片管道中电场和流场的模拟分析,确定低电压电泳芯片整体结构和主要参数;
③提出了在芯片分离微通道尾端进行非接触式高频电导检测和光学检测的双检测器集成的思想;实现了硅基电泳芯片非接触式高频电导检测方法;
④提出了实现集成生化低电压芯片电泳分离的控制方法,成功研制了实现低电压运动梯度场的控制电路;
⑤采用SOI-MEMS加工技术,探索了Si-PDMS电泳芯片加工兼容性,实现了芯片的一体化集成加工;
⑥采用氨基酸样品体系对集成生化低电压芯片电泳系统进行分离检测实验,验证了集成低电压电泳芯片系统和相应电路控制方法的有效性和实用性。
本论文的工作得到了国家自然科学基金半导体集成芯片系统重大基础研究计划和重庆市自然科学基金重点项目的资助。
The Microfluidic microchip is the most importmant development of miniaturized total analytical system in the field of micro analytical instrument. As a branch of the development, the integrated electrophoresis microchip, with the characteristics of high performance, fast speed, low reagent comsumption and easy automation, is more and more frequently being employed in life science and other relevant fields and is becoming the focus in the research of microfluidic analytical system. The microfluidic analytical system with high integration, easy automation and convenient portability is being pursued by researchers all over the world.
However, some difficulties still remain in microfluidic chip electrophoresis, such as high working voltage, separation of the large detecting system, inaccessible exterior connection, sophisticated fabricaton and so forth. This doctoral thesis develops on the basis of the proposed theory of, low voltage separate model and its controlling methodology. A theory of Moving Gradient Electric Field Electrophoresis on Microchip is porposed in this paper, and a low voltage separate model and its controlling methodology are discussed in detail as well. On this basis, an integrated bio-chemical low voltage electrophoresis microchip system is developed as a result. The overall structural design of the chip was optiumed by the simulation analysis of electric and fluidic in the microchannel on the chip. Operational parameters of sampling, separating and detecting process were also obtained and selected as a result. The concept of double detectors, high frequence connectless conductivity detector and photo-electric detector integrated on the designed chip were proposed. The circuit controlling methodology was put forword and discussed in detail. As a result, the controlling circuit was designed and produced, which was easy for using and controlling. Based on the SOI material, the MEMS fabricating process was proposed to make the low voltage micochip with electrode array. The integrating fabrication on the chip was successfully carried out, and the microchip with microchannels, controlling circuits and detectors was obtained. With the selecting amino acids sample system, electrophoresis on microchip was carried on the integrated microfluidic analytical system, and experimental results have shown that the system was effective and practical in use.
The main investigations in this thesis are as the fellowing.
①On the basis of a large amount reference and pervious work, the theory of Moving Gradient Electric Field Electrophoresis on Microchip and relative controlling methodology were presented. The concept of total design of the integrated low voltage CE chip was proposed.
②The solid mode of the integrated low voltage CE chip was set up at first. Then, ANAS and ConvertorWare finite element software was applied to simulate the electric and fluidic fields in the icrochannel on the integrated low voltage CE chip. According to the simulation results, structural and operational parameters were optimized and selected.
③The double detectors, high frequence connectless conductivity detector and photo-electric detector, were proposed to integrate at the end of the separating channel on the designed CE chip. In this thesis, the high frequence connectless conductivity detecting electrodes were fabricated on the silicon CE chip.
④The circuit controlling methodology of low voltage CE separation and analysis was put forword and discribed in detail. On this basis, the controlling circuit was successfully designed and produced.
⑤According to the charactors of SOI material, the MEMS fabrication process was proposed to produce the integrated low voltage CE chip. Compatibility between Silicon and PDME in processing and bonding was investigated. The integrated microchip sample with microchannels, controlling circuit,detecting system and information obtaining system was figured out.
⑥Amino acids were selected as the sample to be applied in the experiments of the low voltage electrophoresis on the integrated microchip. The experimental results had shown that the integrated low voltage CE chip analytical systemand relative controlling methodology was effective and practical.
The research work in this thesis was supported by National Nature and Science Fund and by Chongqing Nature and Science Fund.
本博士学位论文针对目前微流控芯片电泳存在的系统分离电压高、检测系统分离、芯片与外界的接口匹配、芯片系统一体化集成加工技术等问题,提出低电压运动梯度场电泳理论、集成芯片电泳的低电压分离模型与控制方法。通过对电场和流场的模拟计算和分析,确定了低电压电泳芯片的整体结构,并对系统的进样、分离和检测进行了结构和参数优化的设计;提出了在芯片微通道上双检测器的集成思想;提出了实现集成生化低电压芯片电泳分离的控制方法,并研制了实现低电压运动梯度场的控制电路;提出了以SOI-MEMS加工技术实现含阵列微电极的硅基低电压电泳芯片的加工工艺方法,实现了芯片的一体化集成;采用氨基酸样品体系对研制出的集成生化低电压芯片电泳系统进行了实验,实验结果有效验证了本论文研制的集成生化低电压芯片电泳系统的可行性。
本文主要工作如下:
①本文在查阅大量文献、资料和总结前期研究工作的基础上,在研究集成芯片电泳的低电压分离理论及其控制方法的基础上,提出集成低电压电泳芯片总体方案;
②通过集成低电压电泳芯片模型的建立,采用ANAS和ConvertorWare软件分析系统进行了集成低电压电泳芯片管道中电场和流场的模拟分析,确定低电压电泳芯片整体结构和主要参数;
③提出了在芯片分离微通道尾端进行非接触式高频电导检测和光学检测的双检测器集成的思想;实现了硅基电泳芯片非接触式高频电导检测方法;
④提出了实现集成生化低电压芯片电泳分离的控制方法,成功研制了实现低电压运动梯度场的控制电路;
⑤采用SOI-MEMS加工技术,探索了Si-PDMS电泳芯片加工兼容性,实现了芯片的一体化集成加工;
⑥采用氨基酸样品体系对集成生化低电压芯片电泳系统进行分离检测实验,验证了集成低电压电泳芯片系统和相应电路控制方法的有效性和实用性。
本论文的工作得到了国家自然科学基金半导体集成芯片系统重大基础研究计划和重庆市自然科学基金重点项目的资助。
The Microfluidic microchip is the most importmant development of miniaturized total analytical system in the field of micro analytical instrument. As a branch of the development, the integrated electrophoresis microchip, with the characteristics of high performance, fast speed, low reagent comsumption and easy automation, is more and more frequently being employed in life science and other relevant fields and is becoming the focus in the research of microfluidic analytical system. The microfluidic analytical system with high integration, easy automation and convenient portability is being pursued by researchers all over the world.
However, some difficulties still remain in microfluidic chip electrophoresis, such as high working voltage, separation of the large detecting system, inaccessible exterior connection, sophisticated fabricaton and so forth. This doctoral thesis develops on the basis of the proposed theory of, low voltage separate model and its controlling methodology. A theory of Moving Gradient Electric Field Electrophoresis on Microchip is porposed in this paper, and a low voltage separate model and its controlling methodology are discussed in detail as well. On this basis, an integrated bio-chemical low voltage electrophoresis microchip system is developed as a result. The overall structural design of the chip was optiumed by the simulation analysis of electric and fluidic in the microchannel on the chip. Operational parameters of sampling, separating and detecting process were also obtained and selected as a result. The concept of double detectors, high frequence connectless conductivity detector and photo-electric detector integrated on the designed chip were proposed. The circuit controlling methodology was put forword and discussed in detail. As a result, the controlling circuit was designed and produced, which was easy for using and controlling. Based on the SOI material, the MEMS fabricating process was proposed to make the low voltage micochip with electrode array. The integrating fabrication on the chip was successfully carried out, and the microchip with microchannels, controlling circuits and detectors was obtained. With the selecting amino acids sample system, electrophoresis on microchip was carried on the integrated microfluidic analytical system, and experimental results have shown that the system was effective and practical in use.
The main investigations in this thesis are as the fellowing.
①On the basis of a large amount reference and pervious work, the theory of Moving Gradient Electric Field Electrophoresis on Microchip and relative controlling methodology were presented. The concept of total design of the integrated low voltage CE chip was proposed.
②The solid mode of the integrated low voltage CE chip was set up at first. Then, ANAS and ConvertorWare finite element software was applied to simulate the electric and fluidic fields in the icrochannel on the integrated low voltage CE chip. According to the simulation results, structural and operational parameters were optimized and selected.
③The double detectors, high frequence connectless conductivity detector and photo-electric detector, were proposed to integrate at the end of the separating channel on the designed CE chip. In this thesis, the high frequence connectless conductivity detecting electrodes were fabricated on the silicon CE chip.
④The circuit controlling methodology of low voltage CE separation and analysis was put forword and discribed in detail. On this basis, the controlling circuit was successfully designed and produced.
⑤According to the charactors of SOI material, the MEMS fabrication process was proposed to produce the integrated low voltage CE chip. Compatibility between Silicon and PDME in processing and bonding was investigated. The integrated microchip sample with microchannels, controlling circuit,detecting system and information obtaining system was figured out.
⑥Amino acids were selected as the sample to be applied in the experiments of the low voltage electrophoresis on the integrated microchip. The experimental results had shown that the integrated low voltage CE chip analytical systemand relative controlling methodology was effective and practical.
The research work in this thesis was supported by National Nature and Science Fund and by Chongqing Nature and Science Fund.
引文
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