近场表面增强拉曼散射实验技术与血清分析的研究
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摘要
本论文以近场表面增强拉曼散射技术及其应用为主要内容,分三部分论述。
首先介绍了一种改进的Meisel等人提出的银胶体粒子制备方法,采用微波加热方法制备银胶体粒子,该方法简单、快捷;制备出的银胶体粒子多数不规则的类球形粒子,通过对银胶体粒子表面形貌分析以及以染料分子罗丹明6G(R6G)为探针样品的表面增强拉曼散射光谱测试,证明微波加热方法制备的银胶体粒子具有高效的表面增强拉曼散射效应。
其次,利用微波加热方法制备的银胶体粒子,获得了小牛血清白蛋白和小鼠血清表面增强拉曼散射光谱,通过对拉曼光谱的分析,认为这种银胶体粒子对生物样品同样具有很强的表面增强效应,而且对生物活性没有明显的改变,文中还分析了小鼠血清表面增强拉曼散射光谱中,由于表面增强拉曼散射的作用,获得了血清中低浓度成分的表面增强拉曼散射光谱,为实现某些在血液中体现的疾病的早期诊断提供了一种分子水平上的检测手段。在乙肝病毒的检测方面,利用表面增强拉曼散射光谱的特点,得到感染乙肝病毒血清的拉曼光谱,通过拉曼光谱,获得乙肝病毒核心抗原DNA的拉曼光谱。由于常规血液检测中无法获得的乙肝病毒核心抗原DNA情况,利用表面增强拉曼散射光谱,为快速简单的获得血液中乙肝病毒核心抗原DNA的存在提供了一种新的可能实现的检测方法。
最后,我们设计一种新型的激励光为隐失光的高效的近场增强拉曼样品池,该样品池利用大数值孔径物镜形成隐失光,利用拉曼散射光的特点,不仅接收到可传输的拉曼散射光,还接收隐失光形式存在的拉曼散射光,同时根据拉曼散射光的特点,通过加盖反射半球,提高了拉曼散射信噪比。通过实验验证了隐失光激励和传输光激励的拉曼散射光谱的不同。利用该样品池获得了超低浓度样品的表面增强拉曼散射光谱。
In this doctoral article, application of near-field enhanced Raman spectrum (NERS) and surface enhance Raman spectrum (SERS) is the main contents, which is introduced by three parts.
Firstly, novel silver colloid was synthesized by microwave heated method which is fast and easily. The particles of this novel silver colloid is alike spherical. It is proved that the novel silver particles is high effect surfaced enhanced Raman substrate by detected R6G SERS , and its characteristic is analyzed by TEM and UV- vis extinction spectra.
Secondly, SERS of bovine serum albumin and rat serum was obtained by novel silver colloid substrate, which is same high effective surfaced enhanced Raman for biologic sampled. At same time, the active of biologic sample the silver particles was not changed by silver particles. Low concentration of component in serum was detected by SERS, so that SERS could be a method detected some disease in early stage, which some component changed in serum in molecule level. SERS of human serum affected hepatitis B virus. By the Raman spectrum of serum, Raman spectrum of DNA of hepatitis B Core Antigen was obtained, which could not be detected straightly in nomal clinical inspection. SERS could be a new fast and easy means to detect hepatitis B Core Antigen in serum.
Lastly, a novel near-field enhanced Raman sample cell which power light was evanescent wave entirely was brought out. The evanescent light was formed by large numerical aperture object lens and half-ball platinged sliver film reflected the Raman scatting light induced high signal to signal-to-noise Raman spectrum. It was show the different between Raman spetrum excited by evanescent wave and transmitted wave. At same time, SERS of super low concentration sample was acquired used by novle sample cell.
首先介绍了一种改进的Meisel等人提出的银胶体粒子制备方法,采用微波加热方法制备银胶体粒子,该方法简单、快捷;制备出的银胶体粒子多数不规则的类球形粒子,通过对银胶体粒子表面形貌分析以及以染料分子罗丹明6G(R6G)为探针样品的表面增强拉曼散射光谱测试,证明微波加热方法制备的银胶体粒子具有高效的表面增强拉曼散射效应。
其次,利用微波加热方法制备的银胶体粒子,获得了小牛血清白蛋白和小鼠血清表面增强拉曼散射光谱,通过对拉曼光谱的分析,认为这种银胶体粒子对生物样品同样具有很强的表面增强效应,而且对生物活性没有明显的改变,文中还分析了小鼠血清表面增强拉曼散射光谱中,由于表面增强拉曼散射的作用,获得了血清中低浓度成分的表面增强拉曼散射光谱,为实现某些在血液中体现的疾病的早期诊断提供了一种分子水平上的检测手段。在乙肝病毒的检测方面,利用表面增强拉曼散射光谱的特点,得到感染乙肝病毒血清的拉曼光谱,通过拉曼光谱,获得乙肝病毒核心抗原DNA的拉曼光谱。由于常规血液检测中无法获得的乙肝病毒核心抗原DNA情况,利用表面增强拉曼散射光谱,为快速简单的获得血液中乙肝病毒核心抗原DNA的存在提供了一种新的可能实现的检测方法。
最后,我们设计一种新型的激励光为隐失光的高效的近场增强拉曼样品池,该样品池利用大数值孔径物镜形成隐失光,利用拉曼散射光的特点,不仅接收到可传输的拉曼散射光,还接收隐失光形式存在的拉曼散射光,同时根据拉曼散射光的特点,通过加盖反射半球,提高了拉曼散射信噪比。通过实验验证了隐失光激励和传输光激励的拉曼散射光谱的不同。利用该样品池获得了超低浓度样品的表面增强拉曼散射光谱。
In this doctoral article, application of near-field enhanced Raman spectrum (NERS) and surface enhance Raman spectrum (SERS) is the main contents, which is introduced by three parts.
Firstly, novel silver colloid was synthesized by microwave heated method which is fast and easily. The particles of this novel silver colloid is alike spherical. It is proved that the novel silver particles is high effect surfaced enhanced Raman substrate by detected R6G SERS , and its characteristic is analyzed by TEM and UV- vis extinction spectra.
Secondly, SERS of bovine serum albumin and rat serum was obtained by novel silver colloid substrate, which is same high effective surfaced enhanced Raman for biologic sampled. At same time, the active of biologic sample the silver particles was not changed by silver particles. Low concentration of component in serum was detected by SERS, so that SERS could be a method detected some disease in early stage, which some component changed in serum in molecule level. SERS of human serum affected hepatitis B virus. By the Raman spectrum of serum, Raman spectrum of DNA of hepatitis B Core Antigen was obtained, which could not be detected straightly in nomal clinical inspection. SERS could be a new fast and easy means to detect hepatitis B Core Antigen in serum.
Lastly, a novel near-field enhanced Raman sample cell which power light was evanescent wave entirely was brought out. The evanescent light was formed by large numerical aperture object lens and half-ball platinged sliver film reflected the Raman scatting light induced high signal to signal-to-noise Raman spectrum. It was show the different between Raman spetrum excited by evanescent wave and transmitted wave. At same time, SERS of super low concentration sample was acquired used by novle sample cell.
引文
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