生物亲和性核壳纳米颗粒SERS标记物的合成及其在生物分析中的应用
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摘要
纳米尺度上的生物分析是当今生物和化学领域研究的前沿及发展方向,是分析科学关注的研究热点。近年来,将纳米技术应用于生物分析化学,在生物检测、纳米生物传感器、纳米生物芯片等方面显示出光明的发展前景。本文主要从事纳米颗粒SERS标记物的研究,针对纳米颗粒SERS标记物的前沿问题,从纳米颗粒SERS标记物的研究现状出发,提出了多种制备生物相容性核壳纳米颗粒SERS标记物的新型合成方法,并把这些合成出来的核壳型纳米颗粒SERS标记物应用于免疫分析和DNA检测。具体内容如下:
1.核壳型纳米颗粒SERS标记物的合成
我们采用两种新型的合成方法分别合成包埋了染料的Au@SiO_2和Ag@SiO_2两种核壳型纳米颗粒SERS标记物。从而既丰富了核壳型纳米颗粒SERS标记物的种类,又简化了实验过程。文献中报道的制备核壳型纳米颗粒SERS标记物的方法要求先用偶联剂即氨基或巯基硅烷化试剂处理,以使金属纳米颗粒表面具有亲玻璃性。
(1)在第二章中,我们提出了改进的方法即不采用偶联剂处理的合成方法,这种制备方法避免了偶联剂与拉曼活性物之间的竞争,因而,常用的有机染料分子像结晶紫和罗丹明6G等都可成功地制成相应的SERS标记物,从而大大地丰富了SERS标记物的种类。我们发现,这种合成方法所需时间比文献中报道的常规方法显著减少。对于Au@SiO_2纳米颗粒SERS标记物形成过程中所涉及的硅化学,我们提出了可能的机理即正乙氧基硅烷的水解形成末端羟基,通过与吸附在纳米金颗粒上的柠檬酸根作用而导致硅壳层的形成。并且,我们通过实验验证了这个机理。
(2)在第三章中,利用微乳液可以用作纳米反应器的特点,我们采用油包水微乳液技术合成了包埋异硫氰罗丹明染料的Ag@SiO_2核壳纳米颗粒SERS标记物。采用这种方法合成时,纳米银的形成、染料在纳米银表面上的键合和二氧化硅壳层在纳米银与染料上面的形成都是在一个微反应器中进行的。因此,采用这种方法合成时,也不需要把纳米金属进行亲玻璃性预处理。我们采用这两种新型的合成方法制备出来的Au@SiO_2和Ag@SiO_2两种核壳型纳米颗粒SERS标记物具有很强的表面增强拉曼散射信号并且具有很好的稳定性。
2. Fe_3O_4@Ag@TiO_2纳米颗粒复合物SERS基质的合成
在第四章中,我们制备了一种既能与常见的拉曼活性物产生SERS信号,本身
Bioanalysis on nanometer scale has been a topic of significant interest in the developments of current biological and chemical research. Recently, the integration of nanotechnology with biology and analytical chemistry has broad implications in a variety of areas including nano-biosensor and nano-biochips. This dissertation describes the research on biocompatible core-shell nanoparticle-based surface-enhanced Raman scattering (SERS) tags including the novel synthesis methods and their applications in immunoassays and DNA detection. The details are summarized as follows:
1. The synthesis of core-shell nanoparticle-based SERS tags
Two novel synthesis methods have been developed in the preparation of core-shell nanoparticle-based SERS tags including Au@SiO_2 and Ag@SiO_2 based SERS tags. To enrich the kinds of the nanoparticle-based SERS tags and reduce the reaction time during the synthesis of the core-shell nanoparticle-based SERS tags have been the goal in designing these two novel synthesis methods. In the previous literature, it was necessary to pretreat the metal nanoparticles with siliane agent with the group of amino or mercapto to make their surface vitrophilic.
(1) In chapter 2, we employed improved synthesis method without the need of coupling agent to prepare Au@SiO_2 nanoparticle-based SERS tags. This synthesis method obviates the competition between the coupling agents and the Raman reporters. Thus, common dyes such as crystal violet and rhodamine 6G can be successfully encapsulated in the Au@SiO_2 core-shell nanoparticle-based SERS tags which efficiently enriched the kinds of SERS tags. Compared to the previous methods reported in literature, the whole synthesis procedure was greatly simplified and the reaction time was significantly reduced. Furthermore, Our experiments have demonstrated the hypothesis that the glass encapsulation chemistry is based on the reaction between silanol groups hydrolyzed by TEOS and citrate anion groups adsorbed onto the gold nanoparticles leading to the formation of a silica layer through further reaction with silanol groups of other hydrolyzed TEOS.
(2) In chapter 3, we synthesized Ag@SiO_2 core-shell nanoparticle-based SERS labels using reverse micelle technique or microemulsion being used as a nanoreactor. Using the proposed approach, different processes including the preparation of silver
1.核壳型纳米颗粒SERS标记物的合成
我们采用两种新型的合成方法分别合成包埋了染料的Au@SiO_2和Ag@SiO_2两种核壳型纳米颗粒SERS标记物。从而既丰富了核壳型纳米颗粒SERS标记物的种类,又简化了实验过程。文献中报道的制备核壳型纳米颗粒SERS标记物的方法要求先用偶联剂即氨基或巯基硅烷化试剂处理,以使金属纳米颗粒表面具有亲玻璃性。
(1)在第二章中,我们提出了改进的方法即不采用偶联剂处理的合成方法,这种制备方法避免了偶联剂与拉曼活性物之间的竞争,因而,常用的有机染料分子像结晶紫和罗丹明6G等都可成功地制成相应的SERS标记物,从而大大地丰富了SERS标记物的种类。我们发现,这种合成方法所需时间比文献中报道的常规方法显著减少。对于Au@SiO_2纳米颗粒SERS标记物形成过程中所涉及的硅化学,我们提出了可能的机理即正乙氧基硅烷的水解形成末端羟基,通过与吸附在纳米金颗粒上的柠檬酸根作用而导致硅壳层的形成。并且,我们通过实验验证了这个机理。
(2)在第三章中,利用微乳液可以用作纳米反应器的特点,我们采用油包水微乳液技术合成了包埋异硫氰罗丹明染料的Ag@SiO_2核壳纳米颗粒SERS标记物。采用这种方法合成时,纳米银的形成、染料在纳米银表面上的键合和二氧化硅壳层在纳米银与染料上面的形成都是在一个微反应器中进行的。因此,采用这种方法合成时,也不需要把纳米金属进行亲玻璃性预处理。我们采用这两种新型的合成方法制备出来的Au@SiO_2和Ag@SiO_2两种核壳型纳米颗粒SERS标记物具有很强的表面增强拉曼散射信号并且具有很好的稳定性。
2. Fe_3O_4@Ag@TiO_2纳米颗粒复合物SERS基质的合成
在第四章中,我们制备了一种既能与常见的拉曼活性物产生SERS信号,本身
Bioanalysis on nanometer scale has been a topic of significant interest in the developments of current biological and chemical research. Recently, the integration of nanotechnology with biology and analytical chemistry has broad implications in a variety of areas including nano-biosensor and nano-biochips. This dissertation describes the research on biocompatible core-shell nanoparticle-based surface-enhanced Raman scattering (SERS) tags including the novel synthesis methods and their applications in immunoassays and DNA detection. The details are summarized as follows:
1. The synthesis of core-shell nanoparticle-based SERS tags
Two novel synthesis methods have been developed in the preparation of core-shell nanoparticle-based SERS tags including Au@SiO_2 and Ag@SiO_2 based SERS tags. To enrich the kinds of the nanoparticle-based SERS tags and reduce the reaction time during the synthesis of the core-shell nanoparticle-based SERS tags have been the goal in designing these two novel synthesis methods. In the previous literature, it was necessary to pretreat the metal nanoparticles with siliane agent with the group of amino or mercapto to make their surface vitrophilic.
(1) In chapter 2, we employed improved synthesis method without the need of coupling agent to prepare Au@SiO_2 nanoparticle-based SERS tags. This synthesis method obviates the competition between the coupling agents and the Raman reporters. Thus, common dyes such as crystal violet and rhodamine 6G can be successfully encapsulated in the Au@SiO_2 core-shell nanoparticle-based SERS tags which efficiently enriched the kinds of SERS tags. Compared to the previous methods reported in literature, the whole synthesis procedure was greatly simplified and the reaction time was significantly reduced. Furthermore, Our experiments have demonstrated the hypothesis that the glass encapsulation chemistry is based on the reaction between silanol groups hydrolyzed by TEOS and citrate anion groups adsorbed onto the gold nanoparticles leading to the formation of a silica layer through further reaction with silanol groups of other hydrolyzed TEOS.
(2) In chapter 3, we synthesized Ag@SiO_2 core-shell nanoparticle-based SERS labels using reverse micelle technique or microemulsion being used as a nanoreactor. Using the proposed approach, different processes including the preparation of silver
引文
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