Fabrication and assemblage of plasmonic nanoparticles: approaches to design highly uniform and sensitive SERS substrates and applications in biomedical science

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研究生：	Agaje Bedemo Beyene Agaje Bedemo Beyene
論文名稱：	Fabrication and assemblage of plasmonic nanoparticles: approaches to design highly uniform and sensitive SERS substrates and applications in biomedical science Fabrication and assemblage of plasmonic nanoparticles: approaches to design highly uniform and sensitive SERS substrates and applications in biomedical science
指導教授：	蘇威年 Wei-Nien Su 蔡協致 Hsieh-Chih Tsai
口試委員:	Hsisheng Teng Hsisheng Teng Di-Yan Wang Di-Yan Wang Ching-Hsiang Chen Ching-Hsiang Chen Bing Joe Hwang Bing Joe Hwang Wei-Nien Su Wei-Nien Su Hsieh-Chih Tsai Hsieh-Chih Tsai
學位類別：	博士 Doctor
系所名稱：	應用科技學院 - 應用科技研究所 Graduate Institute of Applied Science and Technology
論文出版年：	2021
畢業學年度：	109
語文別：	英文
論文頁數：	156
中文關鍵詞：	SERS mapping 、gold nanoflower 、silver nanoparticle array 、electrophoretic deposition 、in-situ nanoparticle synthesis 、pancreatic cancer marker MUC4 、SERS-based bacterial detection
外文關鍵詞：	SERS mapping, gold nanoflower, silver nanoparticle array, electrophoretic deposition, in-situ nanoparticle synthesis, pancreatic cancer marker MUC4, SERS-based bacterial detection
相關次數：	點閱：245 下載：0
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Surface-enhanced Raman scattering (SERS) is an extremely sensitive analytical tool with the detection limit ranging down to a single molecule. Moreover, SERS is a minimally invasive analytical technique that involves little or no sample preparation and has high molecular specificity. Interestingly, in recent years, the miniaturization of Raman instrumentation down to smartphone size has proved its suitability for point-of-care. Owing to these attractive features, SERS has become a promising analytical technique for rapid and accurate disease detection in recent years. Excellent uniformity and sensitivity of SERS substrates are the two key requirements to get reproducible and reliable signals from a probe. The assemblage of nanoparticles into ordered 2D and 3D arrays and thereby engineering SERS hotspots is the main strategy in designing highly uniform and ultrasensitive SERS substrates. Ultra-uniform SERS substrates can be produced using top-down nanofabrication technologies such as high-resolution electron-beam lithography. However, such top-down methods involve highly expensive and sophisticated techniques that need a tightly controlled environment. Moreover, the yield from such methods is extremely low that these methods are not suitable for mass production of low-cost SERS substrates for routine analysis. As a result, cost-effective and facile fabrication of highly sensitive and uniform SERS substrates is at the center of the practical aspect of SERS-related researches. In this thesis, three different approaches are presented to fabricate highly uniform and sensitive SES substrates and applied for cancer and bacterial detections.
In the first approach, methods to improve the reliability and sensitivity of SERS-based sandwich immunoassay have been described. AuNF was used to design both extrinsic Raman label and capture surface and thereby to improve detection sensitivity. First, a highly uniform capture surface was designed by self-assembly of gold nanoflower (AuNF) onto a thiol-functionalized silicon wafer. Then, AuNF-based SERS immunoassay was carried out for the detection of pancreatic cancer marker MUC4 by tagging it with extrinsic Raman label constructed with AuNF, Raman reporter molecule and anti-MUC4 antibody. In addition to designing uniform SERS substrate, we employed Raman mapping over a large area to minimize the effect of spot-to-spot SERS signal variation. Furthermore, the use of a microwell plate for incubation of capture substrate minimized false positive error significantly. The designed method is sensitive enough to detect MUC4 down to 0.1 ng mL-1.
In the second approach, a SERS substrate with excellent uniformity and sensitivity has been designed by simple electrophoretic deposition of gold nanoflowers onto acid-etched Al-foil and into nanopores of anodized aluminum oxide (AAO). The SEM image and SERS mapping measurement over quite a large area confirmed the excellent uniformity of the substrates. Interestingly, the capping agent used to control the surface charge of nanoparticle can be wisely chosen so that it will also function as a linker molecule to capture a substance of interest onto the substrate. As aluminum has surface Plasmon property, LSPR coupling of gold nanoflowers with anodized aluminum nanoporous structure contributed about one order of magnitude to the overall enhancement compared to that of the silicon wafer.
In the last approach, a novel simplest ever method for fabrication of ultrasensitive and highly uniform SERS substrate and its application for label-free bacterial detection has been proposed. The idea of galvanic replacement reaction in combination with a seed-mediated particle-growth approach in the presence of hydroquinone has been employed, and Ag nanoparticle array was fabricated on Cu-foil in less than 3 min. Label-free detection of bacterial species has shown that the substrate is superior to highly acclaimed SERS substrates such as silver nanocubes. In contrast to most reported SERS detection of pathogens, the detection of bacteria was carried out by directly probing liquid samples. Interestingly, direct liquid bacteria sample analysis showed six-fold higher detection sensitivity than that of completely dried samples.

Abstract…………………………………………………………………………………………….i
Acknowledgement………………………………………………………………………………...v
List of acronyms………………………………………………………………………………….xi
List of figures………………………………………………………………………………….....xv
List of tables……………………………………………………………………………………..xxi
Chapter 1 Introduction .................................................................................................................................. 1
1. Background and rationale of the study .................................................................................................. 1
2. Objective of the study ............................................................................................................................ 4
Chapter 2 Literature Review on surface-enhanced Raman spectroscopy ..................................................... 5
1. Raman spectroscopy .............................................................................................................................. 5
1.1. Basics of Raman spectroscopy and related techniques .............................................................. 5
1.2. Raman instrumentation .............................................................................................................. 8
1.3. Raman active molecular vibrations ............................................................................................. 9
1.4. Laser wavelength selection ....................................................................................................... 13
2. Basics of surface-enhanced Raman spectroscopy (SERS) ................................................................... 14
2.1. Historical background and prominent advances in SERS .......................................................... 14
2.2. Enhancement mechanisms of SERS .......................................................................................... 16
2.3. SERS enhancement factor (EF) .................................................................................................. 20
2.4. SERS probes and SERS tags ....................................................................................................... 21
3. SERS substrate fabrication .................................................................................................................. 22
3.1. Materials for SERS substrate fabrication .................................................................................. 23
3.2. Top-down methods of SERS substrate fabrication ................................................................... 24
3.3. Bottom-up methods of SERS substrate fabrication .................................................................. 26
3.4 Shape and size-controlled synthesis of nanoparticles ............................................................... 27
3.5. Core-shell nanoparticles ........................................................................................................... 29
3.6. Assembled nanoparticles .......................................................................................................... 30
4. SERS for cancer detection ................................................................................................................... 36
viii
4.1. Cancer: general information, risk factors and symptoms ......................................................... 36
4.2. Biomarkers for SERS-based cancer detection ........................................................................... 38
5. SERS for bacterial detection ................................................................................................................ 39
Chapter 3 Experimental section .................................................................................................................. 41
1. Chemicals and materials ...................................................................................................................... 41
2. Fabrication of plasmonic nanoparticles ............................................................................................... 42
2.1. Synthesis and optimization of gold nanoflower (AuNF) ........................................................... 42
2.2. Synthesis of gold nanobipyramids (AuNB) ................................................................................ 42
2.3. Synthesis of silver nanocube (AgNC) ........................................................................................ 43
2.4. In situ synthesis of gold nanoparticles (AuNPs) on PEI modified filter paper .......................... 44
2.5. Fabrication of Ag nanoparticle array on Cu-foil (Cu/AgNP) ...................................................... 44
3. Thiol functionalization of silicon wafer and immobilization of AuNF ............................................... 45
4. Anodization of Al-foil and electrophoretic deposition of AuNF ......................................................... 46
5. Preparation of extrinsic Raman label (ERL) for sandwich immunoassay ........................................... 48
6. Preparation of capture surface and sandwich immunoassay procedures ............................................. 48
7. Bacteria sample preparation ................................................................................................................. 49
8. Characterizations .................................................................................................................................. 49
9. SERS measurements ............................................................................................................................ 50
Chapter 4 Detection of pancreatic cancer by AuNF-based SERS mapping immunoassay......................... 51
1. Scope of the study ................................................................................................................................ 51
2. Synthesis, characterization and SERS performance test of AuNFs ..................................................... 52
3. Quantitative estimation Raman reporter molecule (RRM) .................................................................. 57
4. Capture surface and experimental setup for sandwich immunoassay .................................................. 58
5. Incubation of capture substrates using micro-well plate ...................................................................... 60
6. SERS mapping for detection of MUC4 ............................................................................................... 62
7. Summary .............................................................................................................................................. 68
Chapter 5 Designing highly uniform SERS substrate by simple electrophoretic deposition of AuNF onto
Al-foil .......................................................................................................................................................... 69
1. Scope of the study ................................................................................................................................ 69
ix
2. Anodization of Al-foil in sulfuric acid and oxalic acid ........................................................................ 70
3. Internalizing gold nanoflowers into nanopores of AAO ...................................................................... 73
4. Electrophoretic deposition of AuNF onto etched smooth Al-foil ........................................................ 77
5. Summary .............................................................................................................................................. 80
Chapter 6 Facile fabrication of ultrasensitive and uniform SERS substrate (Cu/AgNP) and rapid bacterial
detection ...................................................................................................................................................... 83
1. Scope of the study ................................................................................................................................ 83
2. In situ fabrication of silver nanoparticle array on copper-foil (Cu/AgNP) .......................................... 85
3. Optimization of reaction conditions for fabrication of Cu/AgNP ........................................................ 88
4. Label-free fast detection of bacteria using Cu/AgNP .......................................................................... 95
4.1. SERS detection of liquid bacteria sample directly .................................................................... 95
4.2. Differentiation of S. aureus and V. parahaemolyticus bacteria species ................................... 98
4.3. Concentration dependence of bacterial SERS spectra ............................................................ 101
4.4. Comparison Cu/AgNP with other substrates for bacterial detection ..................................... 102
5. Comparison of Cu/AgNP with reported works on S. aureus detection .............................................. 105
6. Summary ............................................................................................................................................ 106
Chapter 7 Conclusion and perspectives .................................................................................................... 109
1. Conclusion ......................................................................................................................................... 109
2. Perspectives........................................................................................................................................ 111
References………………………………………………………………………………………115
Appendix: ....................................................................................................................................156
                                

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全文公開日期 2024/08/13 (校內網路)
全文公開日期 2028/08/13 (校外網路)
全文公開日期 2028/08/13 (國家圖書館：臺灣博碩士論文系統)

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