陳炳煇臺灣大學：機械工程學研究所張天立Chang, Tien-LiTien-LiChang2007-11-282018-06-282007-11-282018-06-282006http://ntur.lib.ntu.edu.tw//handle/246246/61382nThe main purpose of this research is to develop a biochip that employs a novel approach for electrical detection of DNA or protein using nanogap electrodes and nanoparticles. Besides, the fluid flow characteristics of microfluidic chip are discussed from the experimental results. This study presents fabrication of nanogap electrodes, preparation of nanoparticles, electrical detection principle of DNA and protein using self-assembled multilayer gold nanoparticles, and bio-bar-code amplification (BCA) technique for improving detection sensitivity of sensing biochip. The nanogap electrodes of biochip are fabricated by using electron-beam lithography technique. The nanogap distance between the electrodes is around 300 nm and the height of electrodes is 65 nm. Gold nanoparticles (AuNPs) and magnetic nanoparticles (MNPs) are synthesized by using chemical reduction method and co-precipitation method, respectively. The average diameter of AuNPs and MNPs is 12 ± 2.1 nm and 27 ± 3.6 nm, respectively. In addition to these, the microfluidic devices are fabricated utilizing the micromolding of SU-8 photoresist and based on the replica molding technique of polydimethylsiloxane (PDMS). In this work, the study of electrical detection of DNA and protein is based on nanogap electrodes and self-assembled multilayer AuNPs to amplify the electrical signal. Unfortunately, the current-voltage (I-V) behavior of target DNA and target antigen at some low concentration still can not obey Ohm’s law. Furthermore, a novel ultrasensitive detection based on BCA technique is adopted to integrate above mentioned. In this way, MNPs and bio-bar-code DNA are used to amplify obtainable current through nanogap electrodes from extremely low concentration of target DNA and antigen. The detected concentration of target DNA and antigen with electrical DNA and protein biosensor is lower than 1 fM and 1 pg/µL, respectively. From these results we could find effectively improved electrical signals. Albeit, the results of flow behavior in microfluidic chips including microjet, micromixer and microseparator are discussed. Finally we say that the study of an ultrasensitive electrical detection for nanogap sensing on-chip system and microfluidic chip system will be helpful to improve commercially potential novel biochip platform and development of an integrated Lab-on-a-Chip application.Acknowledgements I Abstract III Contents VI List of Tables XI List of Figures XII Chapter 1. Introduction 1 1.1 General Remarks 1 1.2 Literatures Review 4 1.2.1 Chip-Based Electrical Detection of DNA 4 1.2.2 Chip-Based Electrical Detection of Protein 7 1.2.3 Gold Nanoparticles Material for Biochip 10 1.2.4 Magnetic Nanoparticles Material Biochip 13 1.2.5 Chip-based Microfluidic Devices in Biotechnology Applications 14 1.3 Motivation and Goals 15 1.4 Thesis Organization 18 Chapter 2. Experimental Principle and Phenomena 20 2.1 Principle of Single-Electron Charging Transport for Nanostructure Device 20 2.2 Electrical Detection of Biomolecular On-Chip Sensing System Principle 27 2.3 Properties of Gold Nanoparticles 29 2.4 Properties of Magnetic Nanoparticles 31 2.5 Principle for Different Gold Nanoparticle Sizes to Build an Electrical Detection DNA between Nanogap Electrodes 33 2.6 Principle for Electrical Detection of DNA Using Self-Assembled Multilayer AuNPs between Nanogap Electrodes 34 2.7 Principle for Electrical Detection of Protein Using Self-Assembled Multilayer AuNPs between Nanogap Electrodes 37 2.8 Principle for Ultrasensitive Electrical Detection of DNA Using Nanogap Electrodes and Nanoparticle-Based DNA Amplification 38 2.9 Principle for Ultrasensitive Electrical Detection of Protein Using Nanogap Electrodes and Nanoparticle-Based DNA Amplification 41 2.10 Analyses of Flow Behaviors in Microfluidic Devices Principle 42 Chapter 3. Experimental Design and Fabrication 45 3.1 Fabrication of Nanogap Gold Electrodes 45 3.2 Fabrication of Microfluidic Devices 49 3.3 Preparation of Gold Nanoparticles 51 3.4 Preparation of Magnetic Nanoparticles 53 Chapter 4. Experimental Apparatus and Procedures 55 4.1 Experimental Apparatus 55 4.1.1 Electron-Beam Lithography System 55 4.1.2 Electrical Characterization System 57 4.1.3 Other Major Common Facilities 58 4.2 Experimental Materials 59 4.3 Experimental Procedures 62 4.3.1 Experimental Procedures for Effect of Different Gold Nanoparticle Sizes to Build an Electrical Detection DNA between Nanogap Electrodes 62 4.3.2 Experimental Procedures for DNA Using Self-Assembled Multilayer AuNPs between Nanogap Electrodes 64 4.3.3 Experimental Procedures for Protein Using Self-Assembled Multilayer AuNPs between Nanogap Electrodes 66 4.3.4 Experimental Procedures for DNA Using Nanogap Electrodes and Nanoparticle-Based DNA Amplification 69 4.3.5 Experimental Procedures for Protein Using Nanogap Electrodes and Nanoparticle-Based DNA Amplification 72 4.3.6 Experimental Procedures for Microfluidic Devices Using an Image Analysis 75 Chapter 5. Results and Discussions 77 5.1 Properties of Gold Nanoparticles Analysis 77 5.2 Properties of Magnetic Nanoparticles Analysis 79 5.3 Effect of Different Gold Nanoparticle Sizes to Build an Electrical Detection DNA between Nanogap Electrodes 81 5.4 Electrical Detection of DNA Using Self-Assembled Multilayer AuNPs between Nanogap Electrodes 85 5.5 Electrical Detection of Protein Using Self-Assembled Multilayer AuNPs between Nanogap Electrodes 88 5.6 Ultrasensitive Electrical Detection of DNA Using Nanogap Electrodes and Nanoparticle-Based DNA Amplification 90 5.7 Ultrasensitive Electrical Detection of Protein Using Nanogap Electrodes and Nanoparticle-Based DNA Amplification 95 5.8 Analysis of Flow Behaviors in Microfluidic Devices 99 Chapter 6. Conclusions and Future Prospects 103 6.1 Conclusions 103 6.2 Recommendations for Future Study 106 References 1756450072 bytesapplication/pdfen-USnNanogap ElectrodesMicrofluidic ChipMagnetic NanoparticlesGold NanoparticlesSelf-AssemblyElectrical Detectionthesishttp://ntur.lib.ntu.edu.tw/bitstream/246246/61382/1/ntu-95-D91522011-1.pdf