光子晶體特性結構製作及其癌症生醫檢測｜國立清華大學博碩士論文庫

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研究生：	鄭涵云 Han Yun Cheng
論文名稱：	光子晶體特性結構製作及其癌症生醫檢測 Fabrication of Photonic Crystal-based Structure and its Biomedical Application for Tumor Marker
指導教授：	董瑞安 Ruey-An Doong
口試委員:
學位類別：	碩士 Master
系所名稱：	原子科學院 - 生醫工程與環境科學系 Department of Biomedical Engineering and Environmental Sciences
論文出版年：	2007
畢業學年度：	95
語文別：	中文
論文頁數：	91
中文關鍵詞：	光子晶體、腫瘤標記分子、生物感測器
相關次數：	點閱：3 下載：0
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中文摘要
近數十年來，孔洞結構因具有高比表面積、大孔洞密度和可調控孔洞尺寸之優勢而吸引了眾多的目光。這類高規則度的孔洞結構可被應用於許多領域中，諸如過濾材料、分離層析材料、光催化基材、感測器或是光子晶體材料，都展現出亮眼的提升效果，而受到廣泛的重視。其中光子晶體材料可被應用於許多光電元件上。本研究的目的在於製作光子晶體特性結構，搭配流動注入系統，對於腫瘤標記分子進行連續性的檢測。在實驗過程中，首先對於模板濃度，介面活性劑濃度，乾燥溫度，雜合式溶膠凝膠反應和表面修飾流程進行最佳化的調控。最後再針對流動注入系統，進行流速和流量的最佳化調控。
研究發現利用2 wt% (w/w) 濃度聚苯乙烯溶液添加1 CMC Tween 20，於60 □C溫度乾燥下，藉自然重力堆積於玻璃基材上，可形成六方最密堆積的蛋白石結構。Tetraethoxy silicate (TEOS)和Methyltrimethoxy silicate (MTMS)混合液於酸性環境催化下進行溶膠凝膠反應，並填入蛋白時結構中。樣品於550 □C鍛燒移除聚苯乙烯模板後，再浸泡於5 % (3-aminopropyl)trimethoxy silicate (APTS)一個小時，使得孔洞表面接上胺基，而可以和生物分子產生胜肽鍵。將修飾完成的材料置入流動注入系統，依序以流速50 □L/min流入抗體20分鐘，小牛血清蛋白10分鐘，最後流入抗原10分鐘，即可有效作為生物分子感測元件。對於甲型胎兒蛋白(AFP)的偵測範圍可從10到5000 ng/mL，座落於生醫應用範圍，而B型肝炎核心抗體(anti-HBcAg)的檢測則是落於1到100 ng/mL之間，偵測極限分別為100 ng/mL和10 ng/mL。總體而言，此方法具有簡單，方便，便宜和快速的優勢，且由研究結果顯示，此高規則度結構材料具有生醫檢測之應用潛力。

Abstract
Porous materials have attracted much attention over the past decade on account of the advantages of high specific surface area, high volume density and tunable pore sizes. These hierarchically ordered materials can be applied to various fields, such as filters, separation, chromatography, catalytic supports, sensors and photonic band gap structure. The porous structures enhance the efficiency splendidly, and it receives much interest in fabricating the highly ordered porous materials. Among these applications, the photonic crystal structure can be designed for the optical device.
The purpose of this study is to fabricate a well-ordered porous material incorporated with flow injection system for tumor marker detection. The template concentration, surfactant concentration, drying temperature, the hybrid sol-gel reaction and surface modification reaction were investigate and optimized. Results showed that 2 wt% (w/w) polystyrene can be effectively used as a template to fabricate the highly ordered porous structures in the presence of 1 CMC Tween 20. After drying the opal structure at 60 □C, a hexagonally arranged opal structure by natural gravity was formed. The hybrid sol solution prepared with tetraethoxy silicate (TEOS) and methyltrimethosy silicate (MTMS) at pH 2 was then infiltrated into the interstitial voids between polystyrenes by spin-coating. The samples were aged at room temperature and were calcined at 550 □C for the removal of polystyrene. The amino-modified structures derived from soaking with 5 % (3-aminopropyl)trimethoxy silicate (APTS) for 1 hour were incorporated with the flow injection system for biomedical detection. The flow rate of 50 □L/min to 20 min of flow time was used to detect the cancer biomarkers. The detection range of alpha-fetoprotein (AFP) was from 10 to 5000 ng/mL, which locating on the biomedical application range, while the detection of anti-Hepatitis B core antigen (anti-HBcAg) was 1 to 100 ng/mL. The detection limits of AFP and anti-HBcAg are 100 and 10 ng/mL, respectively. In conclusion, the approach developed in this study is simple, convenient, cheap and rapid. Results obtained in this study indicate the high potential application for biomedical detection of cancer biomarker.

Contents
謝  誌    I
中文摘要    II
Abstract    III
Contents    V
Table index    VIII
Figure index    IX
Chapter 1  Introduction    1
1    Motivation    1
2    Objective    2
Chapter 2  Background and theory    4
1    Concepts of photonic crystals    8
2    three-dimensionally ordered structure methods    9
2.1    Gravimetric sedimentation    9
2.2    Spin-coating    10
2.3    Vertical deposition    11
2.4    Advanced vertical deposition    12
2.5    Ice crystallization    16
2.6    Sonication-assisted method    17
2.7    Physical confinement    18
3    Porous structure    19
3.1    Vertical lifting method    19
3.2    Photopolymerization    20
3.3    Electropolymerization    21
4    Applications    22
4.1    Photocatalysis    22
4.2    Biosensors    23
5    Sol-gel method    25
5.1    Hybrid sol-gel method    27
6    Tumor markers    28
6.1    Alpha-fetoprotein    29
6.2    Anti-hepatitis B core virus antigen    31
Chapter 3  Materials and Methods    32
1  Reagents and materials    32
2  Experimental flow chart    33
3  Preparation polystyrene solution and fabrication of ordered opal structure    35
4  Hybrid sol solution preparation    35
5  Infiltration sol solution into the voids and hierarchical inverse opal structure    36
6  Surface modification    36
7  Biomolecule detection    37
8  Characterization    37
8.1  Scanning electron microscopy (SEM)    38
8.2  Thermogravimetry and differential scanning calorimetry (TG/DSC)    38
8.3  X-ray photoelectron spectroscopy (XPS)    39
8.4  UV-visible spectrometry    39
Chapter 4  Results and discussion    40
1  Hierarchical opal structure    40
1.1  The relationship between latex concentrations and the number of layers    40
1.2    Opal structure dried at different temperatures    45
1.3    Opal structure with various critical micelle concentrations (CMC) of Tween 20…………………………………………………………………………………...48
2  Infiltration the voids between microspheres with hybrid sol-gel method    52
2.1  Hybrid sol solution adjusted with different pH value    52
3  Inverse opal structure fabrication    57
3.1  Thermo-gravimetric analysis (TGA) and differential scanning calorimeter analysis (DSC)    57
4  Surface modification by chemical binding    59
4.1  Bovine serum albumin-FITC (BSA-FITC) binding test    59
4.2  X-ray photoelectron spectroscopy (XPS) surface analysis    61
5    Biomolcule detection    67
5.1    Flow rate optimization    68
5.2    Flow time period optimization    71
5.3    Antibody-antigen binding period optimization    73
5.4    Antigen detection    75
Chapter 5  Conclusions    77
References    78
Appendix A    84
Appendix B    86

                                

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