乳糖传感检测与癌细胞现场红外分析研究
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摘要
20世纪80年代以高技术群为代表的新技术革命,把生物技术、新材料和信息技术并列为新技术革命的重要标志。人口、地球环境、食物、能源与健康等重大问题的解决,莫不寄希望于生命科学的发展和材料的创新。本论文针对生命科学领域中的相关问题,开发了无线磁弹性传感技术,测定了与人类健康息息相关的食品中的乳糖含量;研究了现场红外-衰减全反射傅立叶变换红外光谱技术在人乳腺癌细胞生长过程分析中的应用;此外,文章还针对材料开发相关问题,探索了碳纤维的催化石墨化及其机理。
(1)牛奶中乳糖的无线传感分析:本文基于无线无源传感器的原理,在磁弹性传感器表面固定一层pH聚合物,制备了pH传感器,然后再在该pH传感器的表面固定β-半乳糖苷酶、葡萄糖氧化酶和过氧化氢酶的复合酶,获得了乳糖传感器。利用底物在多酶作用下水解最终生成H+,使溶液pH值变小,导致传感器表面pH敏感膜发生收缩,从而使表面质量负载减少导致传感器共振频率增加的原理,分析了乳糖标准溶液的浓度,线性范围为2～16 mmol/L,检测下限达0.72 mmol/L。将该传感器应用于实际牛奶样品中的检测,效果与光谱分析数据相当。
(2)乳腺癌细胞生长过程的现场红外分析:基于红外光谱能够反映物质结构的特点,采用衰减全反射傅立叶变换红外(ATR-FTIR)光谱,在线分析人乳腺癌细胞生长全过程的物质变化,实现了对人乳腺癌细胞生长的在线实时无损监测。通过比较正常人乳腺细胞与癌细胞的区别,为红外光谱诊断癌症的工作提供了数据材料;通过探索癌细胞的生长行为,为癌细胞的生长过程机理研究做出了贡献,同时该方法的建立为进一步研究抗癌药物对癌细胞作用机制提供了可行性依据。
(3)磁场诱导化学镀Ni-P对碳纤维催化石墨化的影响研究:针对聚丙烯腈(PAN)碳纤维难石墨化的问题,采用磁场诱导的方法,在化学镀镍的过程中获得了颗粒均一的镀层表面。考察了镀层均匀程度对碳纤维石墨化度的影响。通过扫描电子显微镜(SEM)图比较了两种不同的化学镀层,通过X射线衍射(XRD)表征研究了石墨化度跟镀层的均匀程度的关系。
Since 1980s, biotechnology, new materials and information technology have been regarded as the symbols of technical innovation, which are important to the areas of population, environment, food, energy, health and so on. This dissertation focuses on the areas of life science and materials. We developed a magnetoealstic biosensor to determine the lactose in milk sample, studied the application of in situ attenuated total reflectance Fourier transform infrared (ATR-FTIR) spectroscopy in analysis of cancer cells. Finally, we investigated the graphitization of carbon fibers.
(1) Analysis of lactose in milk samples by wireless sensing: We developed a wireless biosensor for analysis of lactose in milk by co-immobilizingβ-galactosidase (GLA), glucose oxidase (GOD) and catalase onto a magnetoelastic ribbon-like sensor which was pre-coated with a layer of pH-sensitive polymer. The lactose can be hydrolyzed to produce acid finally by the multi-enzyme, resulting in the pH-responsive polymer shrinking, and consequently the resonance frequency (?r) of the sensor increasing. The shifts in frequency were linearly proportional to the concentration of lactose from 2 mmol/L to 16 mmol/L with a detection limit of 0.72 mmol/L. The sensor was applied to the milk samples, and the results were the same as that of spectroscopic analysis.
(2) In situ evaluation of breast cancer cell growth with ATR-FTIR Spectroscopy: Base on the powerful ability of Fourier-transform infrared (FTIR) spectroscopy to elucidate the structure of biological samples. We introduced ATR-FTIR to in situ evaluate the growth of human breast cancer cell, and elucidated the distinguishing phase of the cell cycle with 3D ATR-FTIR spectroscopy. The different spectroscopy between cancer cell and normal cell offered information to the cancer screening. This study not only provides a time sketch of cancer events but also shows the potential to the mechanism of the anti-cancer drugs acting on the cells.
(3) The investigation of effect on the graphitization of carbon fibers with electroless deposition of Ni-P catalyst under a magnetic filed: In the process of electroless deposition of Ni, a magnetic filed was added to obtain a homogenous, smooth and orientated coating. SEM images determined the surface of two different coatings. A conclusion that the graphitization is independent on the degree of smooth of coating surfaces was achieved by XRD data.
(1)牛奶中乳糖的无线传感分析:本文基于无线无源传感器的原理,在磁弹性传感器表面固定一层pH聚合物,制备了pH传感器,然后再在该pH传感器的表面固定β-半乳糖苷酶、葡萄糖氧化酶和过氧化氢酶的复合酶,获得了乳糖传感器。利用底物在多酶作用下水解最终生成H+,使溶液pH值变小,导致传感器表面pH敏感膜发生收缩,从而使表面质量负载减少导致传感器共振频率增加的原理,分析了乳糖标准溶液的浓度,线性范围为2～16 mmol/L,检测下限达0.72 mmol/L。将该传感器应用于实际牛奶样品中的检测,效果与光谱分析数据相当。
(2)乳腺癌细胞生长过程的现场红外分析:基于红外光谱能够反映物质结构的特点,采用衰减全反射傅立叶变换红外(ATR-FTIR)光谱,在线分析人乳腺癌细胞生长全过程的物质变化,实现了对人乳腺癌细胞生长的在线实时无损监测。通过比较正常人乳腺细胞与癌细胞的区别,为红外光谱诊断癌症的工作提供了数据材料;通过探索癌细胞的生长行为,为癌细胞的生长过程机理研究做出了贡献,同时该方法的建立为进一步研究抗癌药物对癌细胞作用机制提供了可行性依据。
(3)磁场诱导化学镀Ni-P对碳纤维催化石墨化的影响研究:针对聚丙烯腈(PAN)碳纤维难石墨化的问题,采用磁场诱导的方法,在化学镀镍的过程中获得了颗粒均一的镀层表面。考察了镀层均匀程度对碳纤维石墨化度的影响。通过扫描电子显微镜(SEM)图比较了两种不同的化学镀层,通过X射线衍射(XRD)表征研究了石墨化度跟镀层的均匀程度的关系。
Since 1980s, biotechnology, new materials and information technology have been regarded as the symbols of technical innovation, which are important to the areas of population, environment, food, energy, health and so on. This dissertation focuses on the areas of life science and materials. We developed a magnetoealstic biosensor to determine the lactose in milk sample, studied the application of in situ attenuated total reflectance Fourier transform infrared (ATR-FTIR) spectroscopy in analysis of cancer cells. Finally, we investigated the graphitization of carbon fibers.
(1) Analysis of lactose in milk samples by wireless sensing: We developed a wireless biosensor for analysis of lactose in milk by co-immobilizingβ-galactosidase (GLA), glucose oxidase (GOD) and catalase onto a magnetoelastic ribbon-like sensor which was pre-coated with a layer of pH-sensitive polymer. The lactose can be hydrolyzed to produce acid finally by the multi-enzyme, resulting in the pH-responsive polymer shrinking, and consequently the resonance frequency (?r) of the sensor increasing. The shifts in frequency were linearly proportional to the concentration of lactose from 2 mmol/L to 16 mmol/L with a detection limit of 0.72 mmol/L. The sensor was applied to the milk samples, and the results were the same as that of spectroscopic analysis.
(2) In situ evaluation of breast cancer cell growth with ATR-FTIR Spectroscopy: Base on the powerful ability of Fourier-transform infrared (FTIR) spectroscopy to elucidate the structure of biological samples. We introduced ATR-FTIR to in situ evaluate the growth of human breast cancer cell, and elucidated the distinguishing phase of the cell cycle with 3D ATR-FTIR spectroscopy. The different spectroscopy between cancer cell and normal cell offered information to the cancer screening. This study not only provides a time sketch of cancer events but also shows the potential to the mechanism of the anti-cancer drugs acting on the cells.
(3) The investigation of effect on the graphitization of carbon fibers with electroless deposition of Ni-P catalyst under a magnetic filed: In the process of electroless deposition of Ni, a magnetic filed was added to obtain a homogenous, smooth and orientated coating. SEM images determined the surface of two different coatings. A conclusion that the graphitization is independent on the degree of smooth of coating surfaces was achieved by XRD data.
引文
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