修飾之氮化銦離子感測場效電晶體對DNA雜合反應偵測

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研究生：	林政毅 Lin, Cheng-Yi
論文名稱：	修飾之氮化銦離子感測場效電晶體對DNA雜合反應偵測 Functionalized InN Ion Sensitive Field Effect Transistor for DNA Hybridization Detection
指導教授：	葉哲良 Yeh, J. Andrew
口試委員:
學位類別：	碩士 Master
系所名稱：	電機資訊學院 - 電子工程研究所 Institute of Electronics Engineering
論文出版年：	2009
畢業學年度：	98
語文別：	英文
論文頁數：	93
中文關鍵詞：	氮化銦離子感測場效應電晶體、DNA 雜合反應、分子氣相沉積、3-硫醇基矽丙烷、生物分子感測器
外文關鍵詞：	InN ISFET, DNA Hybridization, Molecular Vapor Deposition (MVD), 3-mercaptopropyltrimethoxysilane (MPTMS), Biosensors
相關次數：	點閱：2 下載：0
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超薄膜 (~10 nm) 氮化銦 (InN) 離子感測場效應電晶體 (ISFET)，經分子氣相沉積(Molecular vapor deposition, MVD)技術，將 3-硫醇基矽丙烷 (3-Mercaptopropyltrimethoxysilane, MPTMS)以氣相法修飾在其閘極表面，以進行DNA雜和反應的偵測。超薄膜氮化銦離子感測場效應電晶體對於溶液中的陰離子，具有相當高的靈敏度與快速的反應時間，在化學與生物分子感測上展現出極大的應用可能。本研究以氣相沉積技術來矽烷化 (Silanization) 3-硫醇基矽丙烷，比起傳統的液相自組裝膜 (Self-assembled monolayer, SAM)，此技術能大大地縮短表面修飾的時間。藉水的接觸角驗證，首先在氧離子的電漿清潔後，氮化銦表面呈現0o角，而經1.5個小時的氣相 3-硫醇基矽丙烷修飾後，氮化銦表面可達68o。3-硫醇基矽丙烷的分子尾端具有硫醇 (-SH) 官能基，能用來固化在5端經丙烯酰胺 (Acrylic phosphoramidite) 修飾的DNA探針 (Probe DNA)。將表面覆有3-硫醇基矽丙烷的氮化銦離子感測場效應電晶體浸置到10 uM 的DNA探針溶液中持續12個小時後，修飾完成的氮化銦離子感測場效應電晶體便能與互補的單股DNA (5'-ATTGTTATTAGG-3') 進行雜合反應。我們觀察到當互補對DNA溶液滴入閘極表面時，氮化銦離子感測場效應電晶體的源汲極電流有明顯約6 uA的電流下降。此電流下降的原因可歸因為溶液中帶負電的互補DNA雜合黏附到閘極表面所引起。對於12個鹼基的寡聚核苷酸探針 (Oligonucleotide probe)，本研究已成功地偵測到 1 nM 的互補段目標 (Target) DNA，然而對帶有一個鹼基不匹配的非互補DNA，則偵測不到任何明顯的電流變化。

Ultrathin (~10 nm) InN ion sensitive field effect transistors (ISFETs) with gate region modified with 3-mercaptopropyltrimethoxysilane (MPTMS) by molecular vapor deposition (MVD) are used to detect hybridization of deoxyribonucleic acid (DNA). The ultrathin InN ISFETs have a high sensitivity and short response time for anion detection, showing a great potential for chemical and biological sensing applications. Vapor-phase silanization of MPTMS using MVD substantially shortens the response time for surface modification compared to the conventional self-assembled monolayer (SAM) techniques. The change of contact angle of water on InN surface was observed from 0o, indicating the O2 plasma cleaning, to 68o after 1.5 h vapor deposition. MPTMS with -SH terminal functional groups was used to immobilize probe DNA with acrylic phosphoramidite modification at 5'-end. After immersed in 10 uM DNA probes solution for 12 h, the functionalized InN ISFET was used to perform the hybridization with complementary single stranded (ss) DNA 5'-ATTGTTATTAGG-3'. A drain-source current decrease (~6 uA) was observed when a complementary DNA was introduced to the gate region of ISFETs. The current decrease is attributed to the attachment of negatively charged DNA. For a 12-mer oligonucleotide probe, the detection of 1 nM target DNA was accomplished, while the noncomplementary DNA with one base mismatch did not show any obvious current variation.

Abstract  I
摘要  II
誌謝  III
Table of Contents  IV
Figure Captions  VII
List of Tables  XII
List of Symbols  XIII
Chapter I: Introduction  1
1 Background  1
2 Ultrathin InN ISFET  2
2.1 Properties of InN Film  2
2.2 Detection Mechanism  3
2.3 Response to Anion  4
3 Motivation and Objective  5
4 Organization of this Thesis  6
Chapter II: Literature Survey  7
1 DNA Hybridization  7
1.1 Introduction  7
1.2 Biosensors for DNA Hybridization  9
2 Field Effect Transistor Sensors  11
2.1 Introduction  11
2.2 Functionalization for Biological Recognition  12
2.3 Detection Mechanism  15
3 FET-Based Biosensors  16
3.1 Si-Based Biosensors  18
3.2 Diamond-Based Biosensors  19
3.3 AlGaN/GaN-Based Biosensors  20
3.3 CNT-Based FET Biosensors  21
3.4 NW-Based FET Biosensors  22
3.5 Comparison  23
Chapter III: Fabrication and Functionalization  24
1 Fabrication  24
1.1 Design of Channel Width to Length Ratio  24
1.2 Electrode Design  25
1.3 Process Flow  26
1.4 PCB Design  29
1.5 Device Encapsulation  30
2 Functionalization  31
2.1 Molecular Vapor Deposition for Silaniztion  31
2.2 Covalent Immobilization of Probe DNA  34
2.3 Experimental Section  35
Chapter IV: Measurement  38
1 Confirmation of Functionalization  38
1.1 Water Contact Angle Measurement  38
1.2 XPS Measurement  42
1.2.1Summary  48
2 Current Measurement with DC Configuration  49
2.1 System setup  49
2.2 Response to Gate Voltage Modulation  50
3 Voltage Measurement with AC Configuration  53
3.1 System Setup  55
3.2 Response to pH Variation  57
3.3 Equivalent Current Response  58
4 Time-Resolved pH Measurement  60
4.1 System Setup  60
4.2 Response to Titration with NaOH  62
Chapter V: Results and Discussion  64
1 Electric Detection of Immobilization of Probe DNA  64
1.1 I-V Response  64
1.2 Resistance Measurement  66
2 Electric Detection of DNA Hybridization  67
2.1 Detection with DC Configuration  67
2.2 Detection with AC Configuration  69
2.2.1 Without Functionalization  71
2.2.2 With Functionalization  73
2.3 Detection of Different Concentration with AC Configuration  75
3 Discussion of DNA Assay  77
Chapter VI: Conclusion  79
Chapter VII: Future Work  80
References  81
Appendix  85
Curriculum Vitae  90

                                

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