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类型 基础研究 预答辩日期 2018-03-08
开始(开题)日期 2016-05-31 论文结束日期 2017-12-29
地点 逸夫科技馆三楼会议室 论文选题来源 973、863项目     论文字数 6.2 (万字)
题目 有序纳米碗阵列的制备及其潜在SERS应用研究
主题词 胶体晶,有序金纳米碗阵列,表面增强拉曼散射,癌胚抗原,甲胎蛋白
摘要 表面增强拉曼散射(surface-enhanced Raman scattering, SERS)是纳米尺度的粗糙表面所产生的非线性光学增强现象,它可以提供分子独特的振动光谱信号,已广泛地应用于生物、化学、医学、物理以及环境监测等领域。增强能力强、均一性好、制备简易的活性基底在未来的SERS快速灵敏检测中具有广泛的应用前景。贵金属周期性孔纳米材料如纳米碗阵列等能极大地增强结构表面的电磁场强度,具有更多的“热点”,还可克服传统金属纳米颗粒易团聚的缺点,有利于增强SERS信号,是实现SERS生物传感检测的重要基底选择。金属纳米碗阵列还具有较好的稳定性和生物相容性,非常适合开展生物医学研究,具有广阔的应用前景。 本论文构筑了单金属和双金属的有序纳米碗阵列,用于SERS活性检测并筛选出用于SERS分析的拉曼标记信号分子,以有序纳米碗阵列作为SERS基底,实现癌症标志物的联合检测。 本论文具体开展了如下工作: 1. 高质量SiO2胶体晶膜的制备 运用垂直沉降自组装法,经毛细管力驱使将单分散于无水乙醇中的SiO2胶体微球组装成高质量、大面积的SiO2胶体晶薄膜,利用扫描电子显微镜和紫外-可见分光光度计对胶体晶的显微形貌和光学性质进行了表征,直径190nm左右的SiO2胶体微球经垂直沉降自组装得到的胶体晶膜上SiO2胶体微球呈周期性排列,且紧密有序,并呈现出明亮的结构色彩。利用薄膜测厚仪对SiO2胶体晶的厚度进行测量,其结果与扫描电镜所测厚度一致。高质量SiO2胶体晶膜的制备为后续有序金纳米碗阵列和有序金/银双金属纳米碗阵列制备奠定基础。 2. 有序金纳米碗阵列制备 利用γ-氨基丙基三乙氧基硅烷(APTES)对SiO2胶体晶膜进行氨基化修饰,然后在其上静电吸附沉积金纳米粒子,通过H2O2将生长液(K2CO3-HAuCl4)中的AuCl4-还原成的单质金沉积到SiO2胶体晶膜表面,制备出金纳米壳阵列,用透明胶带揭下表面一层结构,氢氟酸去除SiO2胶体制备出有序金纳米碗阵列。通过扫描电子显微镜、共聚焦拉曼光谱仪对有序金纳米碗阵列的形貌、SERS效应进行表征。所制备的有序金纳米碗阵列具有形貌均一,可重复性和稳定性好的特性,具有周期性的孔结构,并呈六角形紧密堆积,能够产生了较强的热点效应,进而表现出较好的SERS效果,可以应用于SERS分析领域。此外有序金纳米碗阵列由于多孔结构使其具有较大的比表面积,为生物分子的固定提供更多的位点。 3. 有序金/银双金属纳米碗阵列制备 在SiO2胶体晶膜上静电吸附金纳米粒子,制备具有裂纹的有序金壳阵列,而后采用溅射方法在其上面沉积银,腐蚀掉SiO2胶体晶膜后获得了形貌可控的有序金/银双金属纳米碗阵列。由于金纳米材料的稳定性比较好,而银纳米材料的SERS增强效果佳,因此所制备的有序金/银双金属纳米碗阵列较好的稳定性和较强的SERS增强效果。采用有序金/银双金属纳米碗阵列作为SERS基底,对抗癌药物6-MP进行检测,检测限低至10-10 M,该方法简单,灵敏度高,可以将其应用于其它生物分子的检测,为生物传感、检测、成像等应用提供可能。 4. 基于有序金纳米碗阵列的癌症标志物检测新方法 以修饰有捕获抗体的有序金纳米碗阵列作为捕获基底,修饰标记抗体的金纳米壳和金纳米星作为标记探针,在抗原存在的情况下形成“三明治”结构,通过检测其SERS信号的强弱来定量癌症标志物的浓度。这种SERS方法不仅能够检测单个癌症标志物(CEA),而且能同时检测两种癌症标志物(CEA和AFP)。由于周期性孔的存在,增大了阵列的表面积,可以有效提高癌症标志物抗体的固载量。结果表明,该方法简单,固化抗体分子保持良好、灵敏度高、精确性高、抗干扰性强等,可用于分析检测癌症标志物。
英文题目 Fabrication of ordered gold nanobowl arrays and potential application in SERS
英文主题词 colloidal crystal, ordered gold nanobowl arrays, surface-enhanced Raman scattering, CEA, AFP
英文摘要 Surface enhancement Raman scattering (SERS) is a nonlinear optical enhancement produced on the rough surface in nanometer scale. It can provide the unique vibration spectrum signal for molecules and has been widely used in biology, chemistry, medicine, physics and environmental monitoring. A better SERS active substrate has the characteristics of strong enhancement, good uniformity, easy preparation and storage, and convenient use. Through the design and construction of nanostructures with SERS activity, it can be used for rapid and sensitive detection research. Nano material with periodic pore structure, such as nanobowl, with special morphology, overcomes shortcomings of agglomeration of nanoparticles. Nanobowl not only has more "hot spots" which can enhance the electric field around greatly, resulted in enhanced SERS effect, but also has good stability and biocompatibility. Nanobowl arrays can strengthen SERS signals, it is an important substrate for realizing biological sensing detection via SERS. In addition, the metal nanobowl arrays with good stability and biocompatibility is very suitable for biomedical research and has wide application prospect. In this work, we constructed ordered nanobowl arrays which used as SERS active substrates. By screening reporters with distinguished SERS peaks, SERS technique was design for detection of one or two cancer markers based on ordered nanobowl array. The work has been carried out as followings. 1. Preparation of SiO2 colloidal crystal film with high quality Using vertical self-assembly method, driven by the capillary force, monodisperse SiO2 microspheres assemble into high quality, large areas of SiO2 colloidal crystal film. The microscopic morphology and optical properties of SiO2 colloidal crystal film have been characterized by the scanning electron microscope (SEM), UV-vis spectrophotometer. The colloidal crystal film made of SiO2 microspheres with diameter of 190 nm has close-packed array with bright color. The thickness of SiO2 colloidal crystals was measured by the thin film analyzer. The results were in accordance with the thickness measured by the scanning electron microscope. The preparation of high quality SiO2 colloidal crystals film lay the foundation for fabrication of ordered nanobowl arrays. 2. Fabrication of ordered gold nanobowl arrays. Firstly, SiO2 arrays were modified by APTES, which then immersed into gold nanoparticles solution to form SiO2/GNPs. The gold nanoparticles that adsorbed on the surface of SiO2/GNPs arrays were used as nucleation sites to template the growth of a gold nanoshells layer by H2O2, which reduce AuCl4- to Au0. With excessive growth solution, uniform gold nanoshell arrays formed. Finally, a monolayer of hexagonal close-packed gold nanoshell arrays was lifted up by tape from the gold nanoshell arrays, and the silica spheres were removed by hydrofluoric acid, producing a monolayer ordered gold nanobowl arrays adsorbed on the tape. Ordered gold nanobowl arrays were characterized by scanning electron microscopy (SEM) and confocal Raman spectrometer, and they showed strong SERS effect. The ordered gold nanobowl arrays also have uniform morphology, good repeatability with rich "hot spots" and exhibits excellent SERS enhancement performance. In addition, the periodic hole in the ordered gold nanobowl arrays can enlarge specific surface area, which could facilitate the immobilization of biomolecule. 3. Construction of ordered gold/silver bimetallic nanobowl arrays. Based on ordered gold nanobowl arrays, the ordered gold/silver bimetallic nanobowl arrays were prepared. At first, gold nanoparticles were absorb on the SiO2 colloidal crystal film via electrostatic adsorption to fabricated ordered gold shell arrays with crack, which then coated by silver through sputter deposition. Finally, the ordered gold/silver bimetallic nanobowl arrays were prepared by etching SiO2 colloidal crystal film. Because the gold nanomaterial has high stability, silver nanomaterials possess stong SERS enhancement effect, the prepared ordered gold/silver bimetallic nanobowl arrays with both advantages of good stability and strong SERS enhancement effect. It can be seen that ordered gold/silver bimetallic nanobowl arrays is an SERS active substrate, and it was explored to detect 6-MP with high sensitivity, the detection limit is 10-10 M. Ordered gold/silver bimetallic nanobowl arrays can also be applied in detection of other biomolecules. What’s more, it show potential in biological sensing, detection and imaging. 4. Cancer biomarker detection based on ordered gold nanobowl arrays Based on the ordered gold nanobowl arrays, a new method for detecting cancer biomarker (CEA and AFP) has been developed. The ordered gold nanobowl arrays and the gold nanoshells (gold nanostars) were used as the capture substrate and labeling tags, respectively. In the presence of antigen, capture substrate and labeling tags formed sandwich structures. The concentration of cancer biomarker is quantified by detecting the strength of the SERS signal. This technique can not only detect single cancer biomarker, but also detect two cancer biomarkers simultaneously. Due to the large specific surface area of ordered gold nanobowl arrays, the antibody loading capacity can be improved effectively. The results showed that this SERS method is simple with high sensitivity and strong interference resistance, which can be used for the analysis and detection of cancer biomarker.
学术讨论
主办单位时间地点报告人报告主题
东南大学生物电子学国家重点实验室(逸夫科技馆408) 2015.07 东南大学逸夫科技馆南408 李里 有序金纳米碗阵列的制备及SERS应用
东南大学生物电子学国家重点实验室(逸夫科技馆408) 2015.12 东南大学逸夫科技馆南408 李里 基于有序金纳米碗阵列的癌胚抗原的检测研究
东南大学生物电子学国家重点实验室(逸夫科技馆408) 2016.06 东南大学逸夫科技馆南408 李里 基于有序金纳米碗阵列的癌胚抗原和甲胎蛋白双检测研究
东南大学生物电子学国家重点实验室(逸夫科技馆408) 2017.6 东南大学逸夫科技馆南408 李里 有序纳米碗阵列的制备及SERS应用
     
学术会议
会议名称时间地点本人报告本人报告题目
第四届全国“生物与医学纳米技术”博士生学术论坛 2015.07 中国, 苏州 Fabrication of Functional Gold Nanobowl Arrays and Their Application for Sensitive CEA Detection by SERS
第五届全国“生物与医学纳米技术”博士生学术论坛 2016.06 中国, 苏州 Determination of CEA and AFP by Surface Enhanced Raman Spectroscopy Using Gold Nanobowl Aarrays
International Conference on Chemical and Biochemical Engineering 2015.07 Paris, France Determination of Carcinoembryonic Antigen by Surface Enhanced Raman Spectroscopy Using Gold Nanobowl Arrays
     
代表作
论文名称
Determination of Carcinoembryonic Antigen by SERS Using Gold Nanobowl Arrays
Two Types of Immunoassay Based on Nile Blue Labeling Polydopamine Nanospheres
Multiplexing determination of cancer-associated biomarkers by surface-enhanced Raman scattering
 
答辩委员会组成信息
姓名职称导师类别工作单位是否主席备注
徐静娟 正高 教授 博导 南京大学
肖守军 正高 教授 博导 南京大学
袁春伟 正高 教授 博导 东南大学
徐春祥 正高 教授 博导 东南大学
王雪梅 正高 教授 博导 东南大学
      
答辩秘书信息
姓名职称工作单位备注
董健 副高 副教授 东南大学