自组装核壳纳米复合材料和纳米结构的三金属作为高灵敏度的SERS基质的生物标志物的检测

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研究生：	Tran Thi Bich Quyen Tran - Thi Bich Quyen
論文名稱：	自组装核壳纳米复合材料和纳米结构的三金属作为高灵敏度的SERS基质的生物标志物的检测 Self-assembled Core-shell Nanocomposites and Trimetallic Nanostructure as Highly Sensitive SERS Substrates for the Detection of Biomarkers
指導教授：	黃炳照 Bing-Joe Hwang
口試委員:	陳生明 Shen-Ming Chen 楊明長 Ming-Chang Yang 杜景順 Jing-Shan Do
學位類別：	博士 Doctor
系所名稱：	工程學院 - 化學工程系 Department of Chemical Engineering
論文出版年：	2013
畢業學年度：	102
語文別：	英文
論文頁數：	144
中文關鍵詞：	Au@SiO2 核/壳纳米粒子、R6G 、纳米棒、三金属纳米立方体、Rd3B 、CEA抗原、SERS
外文關鍵詞：	SERS, Au@SiO2 core/shell nanoparticles, nanorods, R6G, trimetallic nanocubes, Rd3B, CEA antigen
相關次數：	點閱：189 下載：2
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最近科学家表明在疾病的诊断协议的发展极大的兴趣与检测极低限（LOD），高灵敏度和选择性。本研究的目标是开发一个表面增强拉曼散射通过使用金基核/壳纳米结构三金属（SERS ）的诊断协议与检出限低，灵敏度高，选择性。
首先，利用金@二氧化硅核/壳纳米颗粒组合与高度敏感的表面增强拉曼光谱研究了葡萄糖和尿酸在本研究的决心。罗丹明6G （ R6G ），染料分子被用来评估对合成区@二氧化硅的核/壳纳米粒子与各种二氧化硅壳的厚度的SERS增强因子。从R6G的SERS信号的增强，发现增加与壳厚度的减少。芯/壳组合为1-2纳米的二氧化硅层（用NaOH （0.1M）溶液的较厚的二氧化硅层2-3毫微米的烧蚀）在一个Au纳米颗粒的∼ 36 nm的显示出最高的SERS信号。我们的研究结果表明，表面增强拉曼光谱技术能够检测血糖和尿酸在较宽的浓度范围内，即20毫微克/升∼20毫克/分升（ 10-12 - 10-3 M）和16.8 ng / dL到2.9毫克/分升（ 10-11 - 1.72 × 10-4 M）分别为∼ 14.6纳克/升（ ∼ 0.73 × 10-12 M）和∼ 12.3纳克/升（ ∼ 0.73 × 10-11关联较低的检出限（LOD ） M）。
第二种方法是使用欧@二氧化矽核/壳纳米棒的癌胚抗原（ CEA）检测，以建立一个高度敏感的自聚焦表面增强拉曼散射（SERS ）方法。的SERS增强因子各向异性金@二氧化硅纳米棒与各种厚度，赖以罗丹明6G （ R6G ）染料应用的报道分子的定量测定CEA的二氧化硅壳进行了评价。最高R6G信号被获得了1-2纳米厚度的[使用HF （ 30 ％）溶液更厚的二氧化硅层（ 6-8纳米）的烧蚀]的二氧化硅层。自聚焦字符来源于抗体 - 抗原相互作用这有利于SERS探针组装和显著增加CEA的检测灵敏度。我们的结果表明，表面增强拉曼光谱技术能够很宽的浓度范围内检测CEA 。与检测的0.86蛋白原/毫升（ LOD）的非常低的限制，在Au @二氧化硅纳米探针使癌症的早期诊断。
最后的方法是开发基于电偶置换反应和共还原相应的离子的第一次的中空或多孔的银/金/铂三金属纳米粒子的简单和有效的合成。该纳米粒子的特点是UV-Vis光谱，透射电子显微镜（TEM），高分辨透射电子显微镜和X射线衍射。它也表明，银/金/铂三金属纳米粒子进行了非常SERS活性的灵敏度和稳定性的增强因子为单分子检测不够高。我们的结果可知，若丹明3B染料分子可以在很宽的浓度范围从10-15到10-8M被检测，具有检测被10-15 M的下限
在这项研究中，在Au纳米颗粒的和Au纳米棒的表面上，并且替换方法为银/金/铂三金属纳米粒子的合成分别极薄且均匀的二氧化硅外壳层的涂布技术提供了一种可行的方法，涂层和其它纳米材料architecturing可可以扩展到其他类似的技术应用

Recently scientists show great interest in the development of a disease diagnosis protocol with extremely low limit of detection (LOD), high sensitivity and selectivity. The goal of this research is to develop a surface-enhanced Raman scattering (SERS) diagnosis protocol with low LOD, high sensitivity, and selectivity by using Au-based core/shell and trimetallic nanostructures.
First of all, the use of Au@SiO2 core/shell nanoparticle assemblage with highly sensitive SERS was investigated for the determination of glucose and uric acid in this study. Rhodamine 6G (R6G) dye molecules were used to evaluate the SERS enhancement factor for the synthesized Au@SiO2 core/shell nanoparticles with various silica shell thicknesses. The enhancement of SERS signal from R6G was found to increase with a decrease in the shell thickness. The core/shell assemblage with silica layer of 1-2 nm (by the ablation of a thicker silica layer 2-3 nm using NaOH (0.1 M) solution) over a Au nanoparticle of ~36 nm showed the highest SERS signal. Our results show that the SERS technique is able to detect glucose and uric acid within wide concentration ranges, i.e. 20 ng/dL to 20 mg/dL (10-12 – 10-3 M) and 16.8 ng/dL to 2.9 mg/dL (10-11 – 1.72 × 10-4 M) respectively, with associated lower detection limits (LOD) of ~14.6 ng/dL (~0.73 × 10-12 M) and ~12.3 ng/dL (~0.73 × 10-11 M).
The second approach is to develop a highly sensitive self-focusing surface-enhanced Raman scattering (SERS) methodology by using Au@SiO2 core/shell nanorods for carcinoembryonic antigen (CEA) detection. The SERS enhancement factor was evaluated for anisotropic Au@SiO2 nanorods with silica shells of various thicknesses, upon which Rhodamine 6G (R6G) dye was applied as a reporter molecule for the quantitative determination of CEA. The highest R6G signal was attained with a silica layer of 1-2 nm thickness [by the ablation of a thicker silica layer (6-8 nm) using HF (30%) solution]. The self-focusing character originates from the antibody-antigen interaction which facilitates SERS probes assemble and significantly increases the detection sensitivity of CEA. Our results show that the SERS technique is able to detect CEA within a wide concentration range. With the extremely low limit of detection (LOD) of 0.86 fg/mL, the Au@SiO2 nanoprobes enable an early diagnosis of cancer.
The final approach is to develop a simple and effective synthesis of hollow or porous Ag/Au/Pt trimetallic nanocubes based on the galvanic replacement reaction and co-reduction of the corresponding ions for the first time. The nanocubes have been characterized by UV–vis spectroscopy, transmission electron microscopy (TEM), high-resolution TEM, and X-ray diffraction. It was also demonstrated that the Ag/Au/Pt trimetallic nanocubes were be extremely SERS-active sensitivity and stability with an enhancement factor high enough for single molecule detection. Our results that the Rhodamine 3B dye molecules could be detected over a wide concentration range from 10-15 to 10-8 M, with a lower limit of detection being 10-15 M.
In this study, the ultrathin and uniform a silica shell layer coating techniques on Au nanoparticle’s and Au nanorod’s surfaces, and replacement techniques for the synthesis of Ag/Au/Pt trimetallic nanocubes respectively provides a feasible method for coating and architecturing of other nanomaterials which can be extended to other similar technological applications.

Abstract	i
Acknowledgement	iii
Contents	iv
List of figures 	viii
List of schemes and equations	xiv
List of tables	xv
Acronyms	xvi
Chapter 1 Introduction	1
1 Information about Raman/SERS	1
2 SERS enhancement theories 	5
2.1 	Electromagnetic enhancement (EE)	8
2.2	Chemical enhancement (CE)	10
2.3 	Enhancement factor	12
3 Applications of SERS to bioanalysis	13
4 SERS based bioassay methods	18
5 Nanotechnology for cancer therapeutics	21
6 Biocompatibility of nanoparticles	22
7 Information about lung cancer	23
8 Overview of issues relating to material application in SERS	24
9 Advantages of silica shell and trimetallic compositions in SERS	25
10 Motivation and objectives	25
Chapter 2 Materials and Methods	28
1 Materials	28
2 Methods	28
2.1 	Analytical tools	28
2.1.1 	X-ray diffraction (XRD) measurements	28
2.1.2 	Scanning electron microscopy (SEM) measurements	29
2.1.3 	Transmission electron microscopy (TEM) measurements	29
2.1.4 	Determining of metal(s) loading	29
2.1.5 	Raman measurements	29
2.2 	Preparation of materials	30
2.2.1 	Antibody conjugation with Au@SiO2 core/shell nanorod	30
2.2.2	Preparation of CEA antigen	30
2.2.3	Preparation of SERS substrates	31
Chapter 3 Au@SiO2 core/shell nanoparticle assemblage used for highly sensitive SERS based determination of glucose and uric acid	32
1 Introduction	32
2 Experimental section	35
2.1 	Synthesis of Au nanoparticles	35
2.2 	Preparation of Au@SiO2 nanoparticles	35
3 Results and discussion	36
3.1. 	Morphology and optical absorption properties of the Au and Au@SiO2 core/shell nanoparticles	36
3.2. 	Effect of Au nanoparticles’ sizes on the SERS signal	38
3.3. 	Effect of the Au@SiO2 core/shell nanoparticles’ silica shell thicknesses on the SERS signal…	40
3.4. 	Quantitative detection of glucose using Au@SiO2 core/shell nanoparticles	42
3.5. 	Temporal stability of glucose on Au@SiO2 core/shell nanoparticles	43
3.6. 	Quantitative study of uric acid using Au@SiO2 core/shell nanoparticles	44
4 Summary	46
Chapter 4 Self-focusing Au@SiO2 nanorods with rhodamine 6G as highly sensitive SERS substrate for carcinoembryonic antigen detection	47
1 Introduction 	47
2 Experimental section	50
2.1 	Synthesis of Au nanorods	50
2.2 	Preparation of Au@SiO2 nanorods	50
3 Results and discussion	52
3.1 	Morphology and optical absorption properties of Au and Au@SiO2 core/shell nanorod	52
3.2 	Effect of the silica shell thicknesses on the SERS signal	55
3.3 	SERS spectra of CEA antigen on the Au@SiO2 core/shell nanorods	57
4 Summary	64
Chapter 5 Novel Ag/Au/Pt trimetallic nanocubes and their application in surface-enhanced Raman scattering	65
1 Introduction 	65
2 Experimental section	67
2.1 	Preparation of silver nanocubes	67
2.2 	Synthesis of Ag/Au/Pt trimetallic nanocubes	67
3 Results and discussion	68
3.1 	Characterization of the Ag nanocubes and Ag/Au/Pt trimetallic nanocubes	68
3.2 	The formation mechanism of the Ag/Au/Pt trimetallic porous nanocubes	72
3.3 	Application of the Ag/Au/Pt trimetallic porous nanocubes in SERS measurements	73
4 Summary	79
Chapter 6 Conclusions	80
Chapter 7 Future Perspectives	82
References	84
Appendix	119
Curriculum vitae	120
List of publications	121
List of conference papers	122
List of conferences/workshops	123

                                

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