SPR芯片的生化传感及光纤SPR传感技术的研究
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摘要
表面等离子体共振(SPR)传感技术具有高灵敏度、高精度、免标记、无损伤、实时快速检测等优点,在生物传感、药物开发、临床诊断、环境检测以及食品安全等领域具有广阔的应用前景,是国内外近年来研究的热点之一。本论文从理论和实验上研究了基于棱镜结构的金银双金属SPR传感器、MgF2介质层均匀包覆、单面镀膜月牙型包覆及D型结构的光纤SPR传感器的传感特性。
本论文的主要研究工作和成果如下：
1.利用传输矩阵理论对棱镜结构的SPR传感器进行理论模拟分析,比较了不同金属的SPR传感性质,根据银共振峰窄、易氧化和金的稳定性好、生物修饰技术成熟的特点,提出并设计了金银双金属结构的SPR传感器。分析表明金膜/银膜厚度为10nm/40nm时,传感器的性质最好。提出了基于金银双金属结构的双角度强度检测法,可以简化测量装置和算法并消除光源波动带来的测量误差,同时提高信号强度随折射率的变化。
2.实验研究了金银双金属结构的SPR传感器,并在商业化的Biacore3000SPR仪器上进行了生物检测。结果表明,金银双金属结构的共振曲线更窄,其半高宽约为单层金膜的2/3,可以提高传感器的测量精度；利用竞争抑制法对磺胺甲噁唑抗生素和雌二醇激素进行了测量,实验结果表明,双金属芯片具有很好的稳定性和再生能力,磺胺甲噁唑和雌二醇的检测限分别下降到1.78ng/ml和0.31ng/ml,并对牛奶样品中的磺胺甲噁唑浓度进行了实际检测。利用双角度强度检测法,对雌二醇激素进行了测量,检测限达到0.30ng/ml,与角度检测法基本一致。
3.对均匀包覆的银膜光纤SPR传感器进行了理论和实验研究,分析了光纤直径、传感区域长度、数值孔径及环境折射率等参数对传感器传感特性的影响,结果表明,折射率越大,共振波长越长,灵敏度越高,环境折射率为1.3945时,传感器灵敏度为5.3*103nm/RIU。制备了低成本、高灵敏度、高稳定性的MgF2介质层均匀包覆银膜光纤SPR传感器,分析了介质层材料和厚度对传感器的影响。实验结果表明,MgF2介质层厚度为10nnm时,对传感器影响最小,折射率为1.3945时,传感器灵敏度为4.8*103nm/RIU。
4.制备并研究了加工工艺简单的单面镀膜的月牙型包覆银膜光纤SPR传感器,分析了不同银膜厚度对传感器性质的影响,结果表明,银膜厚度为40nm时,传感器具有最佳的传感性质,环境折射率为1.3901时,灵敏度为4.8*10.nm/RIU.利用不同银膜厚度对共振波长的调节作用,通过对入射光偏振和传感区域长度的优化,设计并实现了20nm和50nm银膜厚度的双通道光纤SPR传感器。同时研究了单面镀膜的D型单模光纤SPR传感器,当折射率为1.3998时,传感器灵敏度为4.4*103nm/RIU
本文创新点主要包括：
1.设计并优化了金银双金属芯片,首次将其与商业化SPR仪器结合并展开对生物分子的检测。有效的提高了传感器的测量精度并降低其生物分子检测限。提出了基于金银双金属芯片的双角度强度检测法并对生物分子进行了实际检测。该结果对提高商业化SPR生物传感器的探测精度、灵敏度具有非常重要的意义。
2.研制了高稳定性、高灵敏度、低成本的MgF2介质层均匀包覆银膜光纤SPR传感器,折射率为1.3945时,灵敏度达到4.8*10.nm/RIU,高于单层金膜和金银双金属结构的光纤SPR传感器。提出并制备了了加工工艺简单的单面镀膜月牙型包覆银膜光纤SPR传感器,同均匀包覆结构的灵敏度基本一致,折射率为1.3901时,灵敏度达到4.8*10.nm/RIU。
3.提出并实现了基于不同银膜厚度的双通道光纤SPR传感器,通过入射光偏振和传感区域长度优化,实现了银膜厚度为20nm和50nm的双通道光纤SPR传感器。
For great advantages of high sensitivity, high accuracy, label-free, non-destructive and real-time detection, the surface plasmon resonance (SPR) sensing technology has been widely used in the areas of biosensing, drug discovery, clinical diagnosis, environmental monitoring, and food analysis and so on, which becomes a very active topics in sensing area. In this dissertation, prism-coupled SPR sensor with Au/Ag bimetallic film, Fiber-optic SPR sensor with dielectric film of MgF2, crescent-coated Ag film and D-type Fiber-optic SPR sensor are studied both in theory and experiment.
The main research works and conclusions in this dissertation are as following:
1. The prism-coupled SPR sensor is simulated by transfer matrix method. The SPR characteristics of different metal film are compared and discussed. The Ag film has a narrow SPR curve and poor chemical stability while the Au film has a good characteristic in chemical stability and biological modification. A bimetallic film with Au/Ag film is proposed and optimized which can combine the advantages of silver and gold. The SPR sensor with bimetallic film of10nm Au film and40nm Ag film shows the best sensing performance. After that, a double-measure-angle intensity interrogation method with simple detection device and algorism is proposed, which both can eliminate the light intensity fluctuation and improve the sensitivity of the sensor.
2. The Au/Ag bimetallic film is prepared and employed in the commercial surface plasmon resonance (SPR) instrument, Biacore3000for biological test. The bimetallic film shows a sharper resonant curve than common Au film which can improve the sensing accuracy. After that, the bimetallic sensor chip is tested by detection of sulfamethoxazole (SMX) and estradiol (E2C) using competitive inhibition method. The experiment result shows that the bimetallic sensor chip can work stably and regenerate repeatedly. The limit of detection (LOD) of SMX and E2C are decreased to1.78ng/ml and0.31ng/ml. Then, SMX concentrations in ten commercial milk samples are determined. Double-measure-angle intensity interrogation method is tested by detection of E2C, which have a LOD of0.30ng/ml, the same as traditional method.
3. Fiber-optic SPR sensor with uniform-coated silver film and MgF2dielectric film is studied theoretically and experimentally. The sensor with different structure parameters is investigated, like the numerical aperture (NA), diameter of fiber core, length of the sensing region and refractive index (RI) of the environment, which shows that the sensitivity of the sensor is increased monotonously with the environment RI. The sensitivity can reach5.3*103nm/RIU at the RI of1.3945. An MgF2coated fiber-optic sensor with uniform-coated silver film is designed and fabricated, which has the advantages of stable, high sensitivity and low cost. Different dielectric film and thickness are discussed and optimized. The result shows that the sensor with10nm MgF2has the best sensing performance which can reach the sensitivity of4.8*103nm/RIU at environment RI of1.3945.
4. To simplify the preparation process of uniform-coated fiber-optic SPR sensor, a fiber-optic SPR sensor with crescent-coated Ag film is fabricated and studied by one-side coating. The sensor with different Ag film is tested, which shows that the optimized thickness of the sensor is40nm. The sensitivity of the sensor can reach4.8*103nm/RIU at environment RI of1.3901, nearly the same as the uniform-coated sensor. By control the polariton of incident light and length of sensing region, a double-channel fiber optical sensor is proposed and realized based on sensing region with different silver thicknesses of20nm and50nm. At last, D-type fiber-optic SPR sensor is tested which have a sensitivity of4.4*103nm/RIU at RI of13998.
Highlights of the dissertation are as follows:
1. An Au/Ag bimetallic film is designed and employed in Biacore3000system for the first time, which shows a higher sensing accuracy and lower LOD than common Au film. A double-measure-angle intensity interrogation method is proposed and tested for biological detection. The result is significant to improve the detection accuracy and sensitivity of commercial SPR instrument.
2. An MgF2coated fiber-optic sensor with uniform-coated silver film with advantages of stable, low cost and high sensitivity is studied. It has a sensitivity of4.8*103nm/RIU at RI of1.3945, which higher than Au and Au/Ag coated fiber-optic sensor. A fiber-optic sensor with crescent-coated Ag film is proposed and fabricated, at RI of1.3945, the sensitivity can reach4.8*103nm/RIU at RI of1.3901, nearly the same as the uniform-coated sensor.
3. A double-channel fiber-optic sensor base on different Ag thickness is studied. By control the polariton of incident light and length of sensing region, the double-channel sensor with Ag thickness of20nm and50nm is realized which shows a good sensing performance.
本论文的主要研究工作和成果如下：
1.利用传输矩阵理论对棱镜结构的SPR传感器进行理论模拟分析,比较了不同金属的SPR传感性质,根据银共振峰窄、易氧化和金的稳定性好、生物修饰技术成熟的特点,提出并设计了金银双金属结构的SPR传感器。分析表明金膜/银膜厚度为10nm/40nm时,传感器的性质最好。提出了基于金银双金属结构的双角度强度检测法,可以简化测量装置和算法并消除光源波动带来的测量误差,同时提高信号强度随折射率的变化。
2.实验研究了金银双金属结构的SPR传感器,并在商业化的Biacore3000SPR仪器上进行了生物检测。结果表明,金银双金属结构的共振曲线更窄,其半高宽约为单层金膜的2/3,可以提高传感器的测量精度；利用竞争抑制法对磺胺甲噁唑抗生素和雌二醇激素进行了测量,实验结果表明,双金属芯片具有很好的稳定性和再生能力,磺胺甲噁唑和雌二醇的检测限分别下降到1.78ng/ml和0.31ng/ml,并对牛奶样品中的磺胺甲噁唑浓度进行了实际检测。利用双角度强度检测法,对雌二醇激素进行了测量,检测限达到0.30ng/ml,与角度检测法基本一致。
3.对均匀包覆的银膜光纤SPR传感器进行了理论和实验研究,分析了光纤直径、传感区域长度、数值孔径及环境折射率等参数对传感器传感特性的影响,结果表明,折射率越大,共振波长越长,灵敏度越高,环境折射率为1.3945时,传感器灵敏度为5.3*103nm/RIU。制备了低成本、高灵敏度、高稳定性的MgF2介质层均匀包覆银膜光纤SPR传感器,分析了介质层材料和厚度对传感器的影响。实验结果表明,MgF2介质层厚度为10nnm时,对传感器影响最小,折射率为1.3945时,传感器灵敏度为4.8*103nm/RIU。
4.制备并研究了加工工艺简单的单面镀膜的月牙型包覆银膜光纤SPR传感器,分析了不同银膜厚度对传感器性质的影响,结果表明,银膜厚度为40nm时,传感器具有最佳的传感性质,环境折射率为1.3901时,灵敏度为4.8*10.nm/RIU.利用不同银膜厚度对共振波长的调节作用,通过对入射光偏振和传感区域长度的优化,设计并实现了20nm和50nm银膜厚度的双通道光纤SPR传感器。同时研究了单面镀膜的D型单模光纤SPR传感器,当折射率为1.3998时,传感器灵敏度为4.4*103nm/RIU
本文创新点主要包括：
1.设计并优化了金银双金属芯片,首次将其与商业化SPR仪器结合并展开对生物分子的检测。有效的提高了传感器的测量精度并降低其生物分子检测限。提出了基于金银双金属芯片的双角度强度检测法并对生物分子进行了实际检测。该结果对提高商业化SPR生物传感器的探测精度、灵敏度具有非常重要的意义。
2.研制了高稳定性、高灵敏度、低成本的MgF2介质层均匀包覆银膜光纤SPR传感器,折射率为1.3945时,灵敏度达到4.8*10.nm/RIU,高于单层金膜和金银双金属结构的光纤SPR传感器。提出并制备了了加工工艺简单的单面镀膜月牙型包覆银膜光纤SPR传感器,同均匀包覆结构的灵敏度基本一致,折射率为1.3901时,灵敏度达到4.8*10.nm/RIU。
3.提出并实现了基于不同银膜厚度的双通道光纤SPR传感器,通过入射光偏振和传感区域长度优化,实现了银膜厚度为20nm和50nm的双通道光纤SPR传感器。
For great advantages of high sensitivity, high accuracy, label-free, non-destructive and real-time detection, the surface plasmon resonance (SPR) sensing technology has been widely used in the areas of biosensing, drug discovery, clinical diagnosis, environmental monitoring, and food analysis and so on, which becomes a very active topics in sensing area. In this dissertation, prism-coupled SPR sensor with Au/Ag bimetallic film, Fiber-optic SPR sensor with dielectric film of MgF2, crescent-coated Ag film and D-type Fiber-optic SPR sensor are studied both in theory and experiment.
The main research works and conclusions in this dissertation are as following:
1. The prism-coupled SPR sensor is simulated by transfer matrix method. The SPR characteristics of different metal film are compared and discussed. The Ag film has a narrow SPR curve and poor chemical stability while the Au film has a good characteristic in chemical stability and biological modification. A bimetallic film with Au/Ag film is proposed and optimized which can combine the advantages of silver and gold. The SPR sensor with bimetallic film of10nm Au film and40nm Ag film shows the best sensing performance. After that, a double-measure-angle intensity interrogation method with simple detection device and algorism is proposed, which both can eliminate the light intensity fluctuation and improve the sensitivity of the sensor.
2. The Au/Ag bimetallic film is prepared and employed in the commercial surface plasmon resonance (SPR) instrument, Biacore3000for biological test. The bimetallic film shows a sharper resonant curve than common Au film which can improve the sensing accuracy. After that, the bimetallic sensor chip is tested by detection of sulfamethoxazole (SMX) and estradiol (E2C) using competitive inhibition method. The experiment result shows that the bimetallic sensor chip can work stably and regenerate repeatedly. The limit of detection (LOD) of SMX and E2C are decreased to1.78ng/ml and0.31ng/ml. Then, SMX concentrations in ten commercial milk samples are determined. Double-measure-angle intensity interrogation method is tested by detection of E2C, which have a LOD of0.30ng/ml, the same as traditional method.
3. Fiber-optic SPR sensor with uniform-coated silver film and MgF2dielectric film is studied theoretically and experimentally. The sensor with different structure parameters is investigated, like the numerical aperture (NA), diameter of fiber core, length of the sensing region and refractive index (RI) of the environment, which shows that the sensitivity of the sensor is increased monotonously with the environment RI. The sensitivity can reach5.3*103nm/RIU at the RI of1.3945. An MgF2coated fiber-optic sensor with uniform-coated silver film is designed and fabricated, which has the advantages of stable, high sensitivity and low cost. Different dielectric film and thickness are discussed and optimized. The result shows that the sensor with10nm MgF2has the best sensing performance which can reach the sensitivity of4.8*103nm/RIU at environment RI of1.3945.
4. To simplify the preparation process of uniform-coated fiber-optic SPR sensor, a fiber-optic SPR sensor with crescent-coated Ag film is fabricated and studied by one-side coating. The sensor with different Ag film is tested, which shows that the optimized thickness of the sensor is40nm. The sensitivity of the sensor can reach4.8*103nm/RIU at environment RI of1.3901, nearly the same as the uniform-coated sensor. By control the polariton of incident light and length of sensing region, a double-channel fiber optical sensor is proposed and realized based on sensing region with different silver thicknesses of20nm and50nm. At last, D-type fiber-optic SPR sensor is tested which have a sensitivity of4.4*103nm/RIU at RI of13998.
Highlights of the dissertation are as follows:
1. An Au/Ag bimetallic film is designed and employed in Biacore3000system for the first time, which shows a higher sensing accuracy and lower LOD than common Au film. A double-measure-angle intensity interrogation method is proposed and tested for biological detection. The result is significant to improve the detection accuracy and sensitivity of commercial SPR instrument.
2. An MgF2coated fiber-optic sensor with uniform-coated silver film with advantages of stable, low cost and high sensitivity is studied. It has a sensitivity of4.8*103nm/RIU at RI of1.3945, which higher than Au and Au/Ag coated fiber-optic sensor. A fiber-optic sensor with crescent-coated Ag film is proposed and fabricated, at RI of1.3945, the sensitivity can reach4.8*103nm/RIU at RI of1.3901, nearly the same as the uniform-coated sensor.
3. A double-channel fiber-optic sensor base on different Ag thickness is studied. By control the polariton of incident light and length of sensing region, the double-channel sensor with Ag thickness of20nm and50nm is realized which shows a good sensing performance.
引文
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