Thesis (Master of Applied Sciences in Chemistry)--Cape Peninsula University of Technology, 2018. / Biosensors have been on the forefront to provide clinical diagnosis tools for various diseases. Proper selection of biomarkers as well as chemical electrode modification is key in the fabrication of electrochemical biosensors. Hence, electrode modified with nanomaterials devices to improve electroanalytical applications. These nanomaterials were functionalized to improve conductivity, accelerate signal transduction and amplify biorecognition events. Thus, resulting in novel sensing platforms that are highly sensitive and selective towards the target analyte. In this study, gold nanorods (Au NRs) capped with CTAB, zeolitic imidazole framework were synthesised using the seed mediated and hydrothermal method respectively. Composites of gold nanorods with cysteine, ZIF-8 or both were also synthesised. All synthesised materials were characterized using ultraviolet–visible (UV-Vis) spectroscopy, Fourier transform infrared spectroscopy (FTIR), scanning electron microscopy (SEM), X-Ray diffraction (XRD) and cyclic voltammetry (CV) techniques. The obtained results confirmed the synthesis of the nanomaterials and composites. Identification of the ideal platform for fabrication of a transducer with the best electrochemical response was determined by studying the combinations of the synthesised nanomaterials and composites. The studied parameters were surface coverage, conductivity, rate of electron transfer constant. Cysteine-Au NRs composites platforms, had exceptional properties hence its synthesis optimisation of was undertaken. The effect of CTAB, reaction time, volume and concentration ratio of Au:Cysteine, temperature and pH on the composite properties were assessed. However, this composite’s electrochemical properties decreased when bioconjugated with the antibodies. Hence, the choice of Au NRs CTAB functionalised ZIF-8 (CTABAu/ZIF-8) as the transducer for biosensor applications due to a more favourable biocompatibility. Biosensor fabrication was done by drop coating glassy carbon electrode with the CTABAu/ZIF-8 forming a transducer followed by immobilisation of the antibody (Ab) using a covalent attachment method with glutaraldehyde (GA) as a cross linker. The target analyte, epidermal growth factor (EGF) was interacted with the Ab binding sites via electrostatic forces. All the fabrication steps were optimized for biosensor components, immobilization technique (drop coating and immersion), concentration and incubation time of linker and bioreceptor, as well as the synthesis of the CTABAu-ZIF-8 composites where in situ and ex situ techniques were compared together with the effect of the concentration ratio of Au: ZIF-8. There was also an analysis of optimum pH. Optimum conditions were found to be immersion in 3 % GA and 2 μg/ml Ab, with incubation times of 8, 10 and 5 minutes for GA, Ab and EGF respectively at a pH of 6. The following electroanalytical techniques: cyclic voltammetry (CV), differential pulse voltammetry (DPV) and square wave voltammetry (SWV) were utilised for EGF detection. The DPV showed better reproducibility, higher currents and better resolution hence; it was the method of choice. The technique’s optimisation involved assessments of the effect of step potential, starting potential and pulse amplitude. The optimum response for pulse amplitude, step potential and starting potential were 60 mV, 20 mV and 0.5 V respectively. The biosensor analytical parameters were linear towards EGF in the concentration range from 2 to 100 nM with a detection limit of 0.58 nM. Reproducibility and repeatability tests were acceptable, and the biosensor had a stability over 80 % within 15 days. There was no interference observed in the presence of glucose and creatine. The EGF biosensor was successfully applied in urine and saliva analysis, obtaining 67.5 and 3.12 nM respectively. This biosensor’s positive outcome strongly suggests its potential as a diagnosis tool for early detection of kidney disease as it was able to detect EGF concentration within physiological levels of EGF in normal kidney function.

Heavy metals from natural and man-made sources can be a great threat to human and animal life. As small inorganic ions they are challenging to detect, usually requiring expensive and complicated machinery. Several heavy metals can accumulate in the human body, leading to long term toxic effects on the nervous system. Many bacteria have developed strategies to survive in heavy metal rich environments. One of these strategies is a bacterial operon containing genes for detoxification mechanisms controlled by a promoter and a regulatory protein. In this work some of these promoter-protein pairs, Pars-ArsR, PcopA-CueR, PmerTPAD-MerR and PzntA-ZntR from Escherichia coli have been employed in the design and construction of a set of biosensors aimed at the detection of heavy metals in drinking water. Biosensors usually employ biological recognition elements, transducing the signal from these to produce an output that can be integrated into electronic circuitry. The sensors presented in this work focus on reducing complexity and on providing a controlled sensor reaction. The arsenic biosensor ‘AsGard’ is based on the Pars-ArsR pair and functions by making the dissociation of an ArsR-mCherry fusion protein from its binding site in the Pars promoter visible. In the cell, ArsR dissociates from Pars upon binding of trivalent arsenic ions. Immobilising the relevant part of the Pars sequence on a solid plastic support allows for the mobilisation of previously bound ArsR-mCherry proteins in the presence of arsenic to become the sensor output. The AsGard sensor detects arsenic within minutes in a concentration range overlapping with the arsenic thresholds for drinking water as set by the World Health Organisation. Additional prototype sensors are presented bringing a reporter gene under the control of the aforementioned promoters. These sensors have been tested in vivo and in vitro in a cell free transcription translation system and partially detect metal concentrations close to relevant ranges. The Pars based sensor is tuneable in vitro by modifying the ratio of the supplied regulatory protein ArsR and is able to detect arsenic well within the relevant range. Spinach2, a fluorescent RNA aptamer, may make future designs independent from translation, drastically reducing complexity of cell free biosensors based on cis-trans transcriptional regulation.

Em trabalhos anteriores nós apresentamos um sensor sólido de óxido nítrico (NO) preparado pelo método sol-gel utilizando como aprisionadores complexos de Fe 2+ -DETC. Neste trabalho, reportamos o efeito de alguns surfactantes (CTAB-catiônico, SDS-aniônico, Triton X100-neutro e o plurônico F127) sobre o sensor. A ressonância paramagnética eletrônica (RPE) foi usada para quantificar o número de moléculas de NO-Fe 2+ -DETC. A presença dos surfactantes aumenta a resistência mecânica dos sensores sólidos, sendo mais pronunciada nos sensores contendo CTAB e Triton. Sem os surfactantes não há sinal do complexo NO-Fe 2+ - DETC. O tempo de secagem foi otimizado para 30 min. O sinal mais intenso foi obtido com os sensores contendo os surfactantes a 12 mM. Os surfactantes de um modo geral aumentam a quantidade de NO aprisionado. A difusão do NO foi estimada através do tempo de aprisionamento do NO sendo maior na presença do F127 e SDS. A saturação do sinal de NO nos sensores acontece em 10 min. Uma maior mobilidade dos complexos NO-Fe 2+ -DETC foi encontrada nos sensores contendo SDS e F127 e foi estimada pela forma de linha. Para os sensores no estado sólido o limite de detecção foi de 2 µM utilizando o F127 como aditivo, para o SDS, CTAB e Triton o limite foi de 6 µM, 8 µM e 10 µM respectivamente. Na solução coloidal a menor quantidade detectada foi de 0,1µM também com o F127. A sensibilidade dos sensores aumenta em pelo menos 5 vezes nos sensores sólidos preparados com o F127 em detrimento aos demais e pode ser melhorada utilizando surfactantes mistos. / In previous work we present a solid sensor for nitric oxide (NO) prepared by sol-gel method using the trapped complex of Fe 2+ -DETC. In this work, we report the effect of some surfactants (CTAB, cationic, anionic, SDS, Triton-X100 and neutral pluronic F127) on the sensor. The electron paramagnetic resonance (EPR) was used to quantify the number of molecules of NO-Fe 2+ -DETC. The use of surfactant increases the mechanical strength of solid sensors, being more pronounced in sensors containing CTAB and Triton. Without the surfactant no EPR signal of the complex NO-Fe 2+ -DETC was observed. The drying time was optimized to 30 min. The strongest EPR signal was obtained with the sensors containing the surfactant to 12 mM. The surfactants in general increase the amount of NO trapped. The diffusion of NO was estimated by the time of trapping of the NO that it was higher in the presence of F127 and SDS. The signal saturation of the NO sensors occurs in 10 min. A higher mobility of the complex NO-Fe 2+ -DETC was found in sensors containing SDS and F127 and was estimated by lineshapes. For the solid state sensors the detection limit was 2 µM using the F127 as additive and for the SDS, CTAB and Triton the limit was 6 µM, 8 µM and 10 µM respectively. In the colloidal solution the least amount detected was 0.1 µM also with the F127. The sensitivity of the sensors increases at least 5 times in the solid sensors prepared with the F127 in detriment to the other and can be improved by using mixed surfactants.

Esta tese de doutoramento relata estudos sobre a interação de nanotubos de carbono com nanomateriais, biomoléculas e células, com o propósito de obter informações relevantes para o desenvolvimento de biossensores e para o campo da nanotoxicologia. No primeiro estudo, foram produzidos e caracterizados três tipos de eletrodos modificados com filmes multicamadas obtidos através da técnica de automontagem. Estes filmes continham nanotubos de carbono de parede simples (SWNT), ftalocianina tetrasulfonada de níquel (NiTsPc) e o dendrímero poli(amidoamina) de geração 2 (PAMAM G2), e estes polieletrólitos foram organizados nos seguintes sistemas: (PAMAM G2/NiTsPc), (PAMAM G2/SWNT) e (nanocompósito SWNT/PAMAM G2 / NiTsPc). Medidas de voltametria cíclica com a sonda ferrocianeto de potássio revelaram que os três sistemas podem ser aplicados como eletrodos descartáveis por serem instáveis e que os dois sistemas com NiTsPc apresentam um amplo intervalo de potencial para detecção de analitos sem a interferência dos picos redox da Pc. Também foram conduzidos ensaios de citometria de fluxo para avaliação da toxicidade de nanocompósitos contendo nanotubos de carbono e poli(amidoamina) de gerações 2, 4 e 6 em células F C3H, correspondentes a fibroblastos saudáveis de fígado humano. Os resultados mostraram que o contato com os nanomateriais provoca uma queda significativa na viabilidade deste tipo de célula, e apontam para a necessidade de aprofundar a investigação sobre os efeitos biológicos deste nanocompósito para que ele seja aplicado com segurança como um vetor de drogas e material genético. A última parte da tese é dedicada a explorar ferramentas computacionais para elucidar os mecanismos de formação do nanocompósito SWNT/PAMAM G2 e sua interação com modelos de membrana celular. Simulações por dinâmica molecular revelaram que a estabilidade do nanocompósito é mantida por interações entre as paredes apolares dos nanotubos e as cadeias internas não polares do dendrímero. O estudo envolvendo bicamadas lipídicas sugeriu que a presença de espécies aniônicas, como as fosfatidilserinas, é crucial para iniciar a ligação desta nanopartícula à membrana celular. O contato do nanocompósito com a bicamada resultou na extração destrutiva de lipídeos da membrana, um efeito deletério que pode causar danos às células. / This thesis describes the interaction between carbon nanotubes and nanomaterials, biomolecules and cells, to obtain relevant information for the development of biosensors and the progress of the nanotoxicology field. In a first study, we produced and characterized three types of modified electrodes made from layer-by-layer films. These nanostructures had single-walled carbon nanotubes (SWNT), nickel tetrasulfonated phthalocyanine (NiTsPc) and poly(amidoamine) dendrimer, generation 2 (PAMAM G2). These polyelectrolytes were organized in the following multilayers: (PAMAM G2/NiTsPc), (PAMAM G2/SWNT) and (SWNT/PAMAM G2 nanocomposite / NiTsPc). Cyclic voltammetry measurements with a potassium ferrocyanide probe revealed that the three systems can be used as disposable electrodes for being unstable and that the two systems with NiTsPc exhibit a wide useful potential interval for detection without the interference of the Pc redox peaks. We also performed flow cytometry experiments to evaluate the toxicity of nanocomposites containing carbon nanotubes and poly(amidoamine) generations 2, 4 and 6 in F C3H cells, derived from healthy human liver fibroblasts. The results showed that the contact with these nanomaterials decreases the viability of this type of cell and point to the need to further determine the biological effects of this nanocomposite before it can be safely applied as a vector for drugs and genetic material. The last part of the thesis explores computational tools to unravel the mechanisms behind the formation of the nanocomposite SWNT/PAMAM G2 and its interaction with cell membrane models. Molecular dynamics simulations revealed that the stability of the nanocomposite is kept mainly by interactions between the apolar nanotube walls and the inner non polar chains of the dendrimer. The study involving lipid bilayers suggested that the presence of anionic species, such as phosphatidylserines, is crucial to trigger the binding of this nanoparticle to the cell membrane. The contact of the nanocomposite with the bilayer resulted in the destructive extraction of lipids from the membrane, an effect that ultimately causes cell damage.

Neste projeto estudou-se a interação da enzima peroxidase de raiz forte (HRP) em interfaces nanoestruturadas e sua possível aplicação em biossensores de peróxido de hidrogênio. Foram utilizadas as técnicas de Langmuir, Langmuir-Blodgett (LB) e automontagem por adsorção física para formar filmes nanoestruturados. A interação da enzima com espécies em interfaces foi investigada com materiais que serviram de matrizes de adsorção, ou seja, a quitosana (Ch) e o fosfolipídio 1,2-dipalmitoil-sn-glicero-3-[fosfatidil-rac-(1-glicerol)] (sal de sódio) (DPPG). Os filmes de Langmuir foram caracterizados com medidas de pressão e potencial de superfície, espectroscopia no infravermelho, e tensão superficial dinâmica. Para os filmes LB e automontados, empregaram-se espectroscopias de fluorescência, ultravioleta-visível e infravermelho e microgravimetria por cristal de quartzo. A peroxidase de raiz forte apresentou forte interação com DPPG, confirmada em filmes de Langmuir por medidas de pressão de superfície, elasticidade dinâmica e de espectroscopia de reflexão e absorção no infravermelho, com modulação por polarização (PM-IRRAS). A massa de peroxidase transferida em filmes Langmuir-Blodgett (LB) mistos com DPPG foi de aproximadamente 200 ng, de acordo com medidas com uma microbalança de cristal de quartzo. A atividade da HRP foi mantida no filme LB, inclusive com atividade catalítica maior do que em meio homogêneo e nos filmes automontados com quitosana. As medidas de atividade não afetaram a morfologia dos filmes LB, estudada com microscopia de força atômica (AFM), ao contrário dos filmes automontados. Conclui-se que a imobilização de HRP é mais eficiente num filme LB, com matriz fosfolipídica, apresentando boas perspectivas de emprego em biossensores de peróxido de hidrogênio. / A study has been performed on the interaction of the enzyme horseradish peroxidase (HRP) in nanostructured interfaces and their possible application in biosensors for hydrogen peroxide. The nanostructured films were obtained with the Langmuir, Langmuir-Blodgett (LB) and layer-by-layer (LbL) methods. The interaction between HRP and species at interfaces was investigated using materials that served as matrix for immobilization, viz. chitosan (Ch) and the phospholipid 1,2-dipalmytoil-sn-glycero-3-[phosphatidyl-rac-(1-glycerol)] (sodium salt) (DPPG). The Langmuir films were characterized with surface pressure, surface potential, elasticity measurements and polarization-modulation reflection and absorption infrared spectroscopy (PM-IRRAS). For LB and LbL films, use was made of fluorescence, absorption in the UV-vis. and infrared spectroscopy. HRP displayed strong interaction with DPPG, which was confirmed in Langmuir films with measurements of surface pressure, dynamic elasticity and PM-IRRAS. The mass of HRP transferred onto a solid support in a mixed LB film with DPPG was 200 ng, according to data from a quartz crystal microbalance. The HRP activity was preserved in the mixed LB film, with a catalytic activity that was even higher than in solution or in LbL films of HRP/Ch. The catalytic activity measurements did not affect the morphology of the LB films, studied with atomic force microscopy (AFM), in contrast to the LbL films. The main conclusion is that HRP immobilization is more efficient in an LB film with a phospholipid matrix, with good prospects for developing biosensors for hydrogen peroxide.

Este trabalho apresenta o desenvolvimento de biossensores de pH, ureia e glicose, utilizando óxidos como plataforma para a parte seletiva. Os filmes finos de óxidos condutores foram produzidos por diferentes técnicas de deposição, como spin-coat, dip-coat, spray-pyrolysis e casting. Os materiais fabricados foram AZO e TiO2, ambos depositados sobre substratos de FTO, ITO ou vidro hidroflilizado. O número de camadas foi variado para cada técnica e as caracterizações morfológicas e estruturais foram feitas por MEV, DRX e FTIR. As caracterizações elétricas foram feitas por EGFET e voltametria cíclica. Os filmes foram testados como sensores de pHs na faixa de 2 a 8. O filme depositado com AZO em substrato de FTO pela técnica de spray-pyrolysis apresentou melhor resposta, com sensibilidade de 31,7 mV/pH entre toda a faixa de pHs do 2 ao 8. Já para os filmes de TiO2, o filme produzido por dip-coat com 5 camadas em substrato de FTO apresentou sensibilidade de 37,8 mV/pH entre a faixa de pHs de 2 a 8. Paralelamente, os filmes tiveram suas superfícies funcionalizadas com proteínas como urease ou glicose oxidase. Neste caso, os dispositivos foram testados entre as concentrações de 5 a 200 mg/dL de ureia e glicose. Como biossensor de ureia, o filme de TiO2 depositado por spin-coat com 5 camadas em substrato de FTO apresentou a maior sensibilidade, com valor 3,32 mV/(mg/dL) entre as concentrações de 5 a 120 mg/dL. Para os filmes estudados como biossensores de glicose, o melhor resultado também foi obtido pelo filme de TiO2 depositado por spin-coat com 5 camadas em substrato de FTO, apresentando sensibilidade em torno de 6,18 mV/(mg/dL) entre as concentrações de 5 a 200 mg/dL. Alguns resultados encontrados foram iguais ou melhores aos encontrados na literatura vigente, mesmo que os dispositivos ainda são passíveis de otimização. / This study presents the development of pH, urea and glucose biosensors, by the use of oxides as the base of the sensing membrane. Conducting oxide thin films were prepared by different deposition techniques, such as spin-coat, dip-coat, spray-pyrolysis, and casting. AZO and TiO2 were produced and coated onto FTO, ITO and glass substrates. The number of layers were changed for each deposition technique, and the surface characterizations were made by SEM, XRD and FTIR, while the electrical characterizations were performed with an EGFET device and cyclic voltammetry. The samples were used as pH sensors in the pH range from 2 to 8. The AZO thin film onto FTO substrate deposited by spray-pyrolysis deposition technique presented sensitivity as 31.7 mV/pH for the total pH range from 2 to 8. TiO2 thin films, produced by dip-coat deposition technique with 5 layers onto FTO substrate presented 37.8 mV/pH as sensitivity, for the same pH range. All the films have their surfaces immobilized with proteins as urease or glucose oxidase. In this case, the samples were performed in different concentrations of urea or glucose, respectively, from 5 to 200 mg/dL, for both. Urea biosensors presented good results for measurements performed with the same sample as well as performed with individuals samples, for all the concentrations range. Spin-coated TiO2 thin films with 5 layers onto FTO substrates presented 3.32 mV/(mg/dL) as sensitivity from 5 to 120 mg/dL urea concentrations, while for glucose biosensors presented 6.18 mV/(mg/dL) for the same film between the glucose concentration range from 5 to 20 mg/dL. Some results were equal or better than the results in current literature, even the films are still capable of optimization.

Fung, Kar Man. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2002. / Includes bibliographical references (leaves 110-114). / Abstracts in English and Chinese. / 摘要 --- p.i / ABSTRACT --- p.ii / ACKNOWLEDGMENTS --- p.iii / TABLE OF CONTENTS --- p.iv / LIST OF FIGURES --- p.vi / Chapter 1. --- Introduction --- p.1 / Chapter 1.1 --- Background and Motivation --- p.1 / Chapter 1.2 --- Objective of the project --- p.4 / Chapter 1.3 --- Organization of the thesis --- p.5 / Chapter 2. --- Literature Review --- p.7 / Chapter 2.1 --- Control on Micro-Manipulation --- p.7 / Chapter 2.1.1 --- Visual Feedback Control --- p.8 / Chapter 2.1.2 --- Sensor-Based Feedback Control --- p.9 / Chapter 2.1.3 --- Bilateral Control --- p.9 / Chapter 2.2 --- Force Sensing System on Micro-Manipulation --- p.10 / Chapter 2.3 --- PVDF Sensor --- p.11 / Chapter 2.4 --- Summary of the Literature Review --- p.12 / Chapter 3. --- Micro-Manipulation --- p.14 / Chapter 3.1 --- Introduction of Micro-Manipulation --- p.14 / Chapter 3.2 --- Probe Station --- p.14 / Chapter 3.2.1 --- Micromanipulators --- p.15 / Chapter 3.2.2 --- Microscopes --- p.15 / Chapter 4. --- Piezoelectric Polyvinylidence Fluoride (PVDF) Sensor --- p.16 / Chapter 4.1 --- Charteristic of PVDF Sensor --- p.16 / Chapter 4.1.1 --- Piezoelectric Properties --- p.16 / Chapter 4.1.2 --- Dimensions of the PVDF Sensor --- p.18 / Chapter 4.2 --- Comparison of Piezoelectric Materials --- p.19 / Chapter 5. --- Theoretical Analysis of PVDF Sensor --- p.21 / Chapter 5.1 --- Sensitivity of PVDF Sensor --- p.21 / Chapter 5.2 --- Relationship between the Deflection and the Force of the PVDF --- p.22 / Chapter 5.3 --- Calculation of the Spring Constant K of the PVDF --- p.23 / Chapter 5.4 --- Simulation on the output from the PVDF Sensor --- p.23 / Chapter 6. --- Experimental Analysis of PVDF Sensor --- p.26 / Chapter 6.1 --- Force-Deflection Diagram --- p.26 / Chapter 6.2 --- Frequency Response of the PVDF Sensor --- p.27 / Chapter 7. --- 1-D PVDF-Based Sensing System --- p.30 / Chapter 7.1 --- Original Design of the Sensing System --- p.30 / Chapter 7.1.1 --- Plastic pipe and adapter --- p.31 / Chapter 7.1.2 --- PVDF Sensor --- p.32 / Chapter 7.1.3 --- Probe-tip holder --- p.32 / Chapter 7.2 --- Current Design of the Sensing System --- p.32 / Chapter 7.3 --- Analysis of the Sensing System --- p.34 / Chapter 7.3.1 --- Frequency Response of the Sensing System --- p.34 / Chapter 7.3.2 --- Sensitivity of the Sensing System --- p.41 / Chapter 8. --- Experiments on 1-D PVDF Sensing System --- p.49 / Chapter 8.1 --- Experimental Setup of the 1-D Sensing System --- p.49 / Chapter 8.1.1 --- Programmable Micromanipulator --- p.50 / Chapter 8.1.2 --- Charge Amplifier --- p.51 / Chapter 8.2 --- Calibration of the 1-D Sensing System --- p.53 / Chapter 8.2.1 --- Noise Signal from the system --- p.53 / Chapter 8.2.2 --- Signal from vibration --- p.55 / Chapter 8.3 --- Experimental Results on touching a substrate --- p.60 / Chapter 8.3.1 --- Description --- p.60 / Chapter 8.3.2 --- Results from touching a substrate --- p.62 / Chapter 8.3.3 --- Analysis of the deflection after touched a substrate --- p.67 / Chapter 8.4 --- Experimental Results on touching a micro mirror --- p.68 / Chapter 8.4.1 --- Description --- p.68 / Chapter 8.4.2 --- Results --- p.70 / Chapter 8.5 --- Experimental Results on lifting a mass platform --- p.74 / Chapter 8.5.1 --- Description --- p.74 / Chapter 8.5.2 --- Results --- p.76 / Chapter 9. --- Modification of 1-D Sensing System --- p.79 / Chapter 9.1 --- Design of the system --- p.79 / Chapter 9.2 --- Experimental Setup of the system --- p.80 / Chapter 9.3 --- Experimental Results on lifting a mass platform --- p.81 / Chapter 10. --- 2-D PVDF-Based Sensing System --- p.90 / Chapter 10.1 --- Design of the Sensing System --- p.90 / Chapter 10.2 --- Experimental Setup --- p.91 / Chapter 10.3 --- Calibration of the 2-D Sensing System --- p.92 / Chapter 10.3.1 --- Noise Signal from the system --- p.92 / Chapter 10.4 --- Experiments Results on touching a substrate --- p.94 / Chapter 11. --- Experimental Analysis --- p.97 / Chapter 11.1 --- Data Acquisition --- p.97 / Chapter 11.2 --- Spectrum Analysis of the Experimental Data --- p.101 / Chapter 12. --- Conclusion --- p.103 / Chapter 13. --- Future Work --- p.105 / Chapter 13.1 --- Control of the Sensing System --- p.105 / Chapter 13.2 --- Tele-operation System on force feedback sensing system --- p.107 / Chapter A. --- Appendix --- p.109 / Chapter A. 1 --- Procedures in using probe station --- p.109 / Bibliography --- p.110

近年來感測器、集成電路及無線通信的科技迅速發展，促使IEEE802.15工作小組6（TG6）致力硏究一個新的無線通信標準─人體區域網路（BAN）。這個新標準特別考量在人體上、人體內或人體周邊的應用。雖然BAN至今還未達成最後定案，不同類型的應用方案已被廣泛提出。這些方案可分為醫療應用（例如：生命徵象感測和植入式治療）及非醫療應用（例如：消費性電子、個人娛樂和遙遠控制）。無線感測節點〈WSN）的基本要求包括輕巧、廉價及低耗電量。因此，本論文提出了一個符合以上要求的注入式鎖態發射機。此外，我們設計了三個發射機的內部模組。由於BAN的物理層例如調變方式和頻譜配置還未完全製訂，本文的電路設計將基於IEEE802.15 TG6的初步建議。 / 第一個模組是一個利用同相位雙路輸入及電流再使用技術的次毫瓦、第一次諧波LC注入式鎖態振盪器〈ILO）。該振盪器操作範圍在醫療植入式通訊服務〈MICS）頻段，並已採用了0.13-μm CMOS工藝實現而僅佔有200 m x 380 m芯片面積。實驗結果表明，在輸入動力0 dBm時，其鎖定範圍可達800 MHz (150 950 MHz) 。最重要的是，該ILO擁有-30 dBm的高輸入靈敏度，同時在1-V供電下只消耗660 A靜態電流。超低的靜態電流使WSN能從人體收集能量而變得完全自主。 / 第二個模組是一個低功耗MICS非整數型頻率合成器，其目的在於選擇信道。雖然整數鎖相環由於其低複雜性而被廣泛使用，對MICS頻段而言並不是一項良好方案。主要原因在於其信道寬只有300 kHz，速度、頻率解析度和相位雜訊變得很難平衡。為此，我們採用0.13-μm CMOS製程設計了一個4階第二型和差積分〈Σ-）調變器分數鎖相環。為了抑制混附單頻信號，二階單迴路數字Σ-調變器加入了抖動。仿真結果顯示該頻率合成器能在15 s內鎖定，同時在1.5-V供電下只消耗4 mW功耗。 / 第三個模組是一個高效能、完全集成的E類功率放大器〈PA）。該PA採用了自給偏壓反相器作為前置放大器，操作範圍在MICS頻段及工業、科學和醫學〈ISM）頻段。在0.18-m CMOS工藝下實現的該PA佔有0.9 mm x 0.7 mm芯片面積。實驗結果表明，在1.2-V供電下及操作頻率是433 MHz時，該PA的漏極效率及輸出功率分別可達40.2 %和14.7 dBm。當操作頻率從380 MHz 到460 MHz，該PA仍能保侍最少34.7 %的漏極效率。此設計適用於低數據傳輸率、固定振幅調變，例如：QPSK、OQPSK等。 / Recent technological advances in sensors, integrated circuits and wireless communication enable miniature devices located on, in or around the human body to form a new wireless communication standard called wireless Body Area Network (BAN). Although BAN is still being investigated by the IEEE 802.15 Task Group 6 (TG6), a vast variety of applications has been proposed which can be categorized into medical applications (e.g. vital signs monitoring and implantable therapeutic treatment) and non-medical applications (e.g. consumer electronics and remote control). The basic requirements of each Wireless Sensor Node (WSN) include light weight, small form-factor, low cost and low power consumption. This thesis proposes an injection-locked transmitter which is a potential candidate to minimize the power consumption of the RF transmitter in WSNs. Three circuit blocks in the proposed injection-locked transmitter are designed and implemented. Since the physical layer of BAN, such as modulation scheme and frequency allocation, has still not been finalized yet, the prototypes in this thesis are designed based on the preliminary suggestions made by the IEEE 802.15 TG6. / The first circuit block is a sub-mW, current-reused first-harmonic LC injection-locked oscillator (ILO) using in-phase dual-input injection technique, operating in the Medical Implantable Communications Service (MICS) band from 402MHz to 405 MHz for medical implants. It has been fabricated in a standard 0.13-m CMOS technology; occupying 200 m x 380 m. Measurement results show that the proposed ILO features a wide locking range of 800 MHz (150-950 MHz) at input power of 0 dBm. More importantly, it has a high input sensitivity of -30 dBm to lock the 3-MHz bandwidth of the MICS band, while consuming only 660 W at 1-V supply. This ultra-low power consumption enables autonomous WSNs by energy harvested from the human body. / The second circuit block is a low power MICS fractional-N frequency synthesizer for channel selection. Although integer-N phase-locked loop (PLL) is widely used due to its low circuit complexity, it is not considered as a good solution for MICS band where the channel spacing is just 300 kHz, due to the severe trade-off between speed, frequency resolution and phase noise performance. To solve this issue, a 4th-order type-II Σ- fractional-N PLL is designed using a standard 0.18-m CMOS technology. A 2nd-order single-loop digital Σ- modulator with dither is designed to eliminate the spurious tones. Simulation results verify that the synthesizer achieves 15 s locking time and consumes 4 mW at a power supply of 1.5 V. / Finally, a power-efficient fully-integrated class-E power amplifier with a self-biased inverter used as a preamplifier stage has been implemented in a standard 0.18-m CMOS process, with 0.9 mm x 0.7 mm active area. It operates in both MICS band for implantable devices and Industrial, Scientific and Medical (ISM) band for wearable devices. Experimental results shows that it achieves 40.2 % drain efficiency while output power is 14.7 dBm at 433 MHz under 1.2-V supply. Moreover, the drain efficiency maintains at least 34.7 % over the frequency range from 380 MHz to 460 MHz. This design is suitable for low data-rate, constant envelope modulation, such as QPSK, OQPSK, etc. / Detailed summary in vernacular field only. / Detailed summary in vernacular field only. / Detailed summary in vernacular field only. / Detailed summary in vernacular field only. / Li, Kwan Wai. / Thesis (Ph.D.)--Chinese University of Hong Kong, 2012. / Includes bibliographical references. / Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Abstract also in Chinese. / Abstract of thesis entitled: --- p.I / 摘要 --- p.IV / Contents --- p.VI / List of Figures --- p.XI / List of Tables --- p.XVII / Acknowledgement --- p.XVIII / Chapter CHAPTER 1. --- Introduction --- p.1 / Chapter 1.1 --- Motivation for body area network (BAN) --- p.1 / Chapter 1.2 --- Standardization of BAN and its positioning between different communication technologies --- p.3 / Chapter 1.3 --- Classification of BAN and its potential applications --- p.5 / Chapter 1.4 --- Requirements and challenges of BAN --- p.7 / Chapter 1.5 --- Research objectives and organization of this dissertation --- p.9 / References --- p.11 / Chapter CHAPTER 2. --- Background information of biomedical transceivers --- p.12 / Chapter 2.1 --- MICS band --- p.12 / Chapter 2.1.1 --- Frequency allocation --- p.12 / Chapter 2.1.2 --- Output power --- p.13 / Chapter 2.1.3 --- Transmit spectral mask --- p.14 / Chapter 2.1.4 --- Transmit center frequency tolerance --- p.14 / Chapter 2.1.5 --- Channel model --- p.15 / Chapter 2.1.6 --- Link budget --- p.17 / Chapter 2.2 --- Fundamental figure of merits for transceivers --- p.18 / Chapter 2.2.1 --- Noise figure, noise floor and receiver sensitivity --- p.18 / Chapter 2.2.2 --- Transmitter energy efficiency --- p.19 / References --- p.20 / Chapter CHAPTER 3. --- Review of transmitter architectures --- p.21 / Chapter 3.1 --- Overview --- p.21 / Chapter 3.2 --- Architectures --- p.22 / Chapter 3.2.1 --- Quadrature --- p.22 / Chapter 3.2.2 --- Polar --- p.23 / Chapter 3.2.3 --- PLL-based --- p.24 / Chapter 3.2.4 --- Injection-locked --- p.26 / Chapter 3.3 --- Radio architecture selection for biomedical systems in BAN --- p.27 / Chapter 3.3.1 --- Data-rate --- p.27 / Chapter 3.3.2 --- Modulation scheme --- p.28 / Chapter 3.3.3 --- Proposed transmitter architecture --- p.28 / References --- p.31 / Chapter CHAPTER 4. --- Design of sub-mW injection-locked oscillator --- p.33 / Chapter 4.1 --- Introduction --- p.34 / Chapter 4.2 --- Circuit design and analysis --- p.34 / Chapter 4.3 --- Experimental results --- p.47 / Chapter 4.4 --- Summary --- p.55 / References --- p.56 / Chapter CHAPTER 5. --- Design of low-power fractional-N frequency synthesizer --- p.58 / Chapter 5.1 --- Synthesizer architectures --- p.59 / Chapter 5.2 --- PLL design fundamentals --- p.63 / Chapter 5.2.1 --- Stability --- p.63 / Chapter 5.2.2 --- Phase noise --- p.65 / Chapter 5.3 --- Proposed architecture --- p.67 / Chapter 5.4 --- System design --- p.68 / Chapter 5.4.1 --- Stability --- p.68 / Chapter 5.4.2 --- Phase noise --- p.73 / Chapter 5.5 --- Σ modulation in fractional-N synthesis --- p.75 / Chapter 5.5.1 --- Basic operating principles --- p.76 / Chapter 5.5.2 --- An accumulator as a first-order Σ- modulator --- p.78 / Chapter 5.5.3 --- Noise analysis --- p.80 / Chapter 5.5.4 --- Architectures --- p.84 / Chapter 5.5.5 --- Design and modeling --- p.87 / Chapter 5.5.6 --- Digital circuit implementation --- p.99 / Chapter 5.5.7 --- Measurement results --- p.104 / Chapter 5.6 --- Time domain behavioral modeling --- p.104 / Chapter 5.7 --- Design of building blocks --- p.106 / Chapter 5.7.1 --- VCO --- p.107 / Chapter 5.7.1.1 --- Principles --- p.107 / Chapter 5.7.1.2 --- Circuit design --- p.111 / Chapter 5.7.2 --- PFD --- p.131 / Chapter 5.7.2.1 --- Principles --- p.131 / Chapter 5.7.2.2 --- Circuit design --- p.133 / Chapter 5.7.3 --- CP --- p.136 / Chapter 5.7.3.1 --- Principles --- p.136 / Chapter 5.7.3.2 --- Circuit design --- p.137 / Chapter 5.7.4 --- Frequency divider --- p.138 / Chapter 5.7.4.1 --- Principles --- p.138 / Chapter 5.7.4.2 --- Circuit design --- p.145 / Chapter 5.7.5 --- Loop filter --- p.148 / Chapter 5.8 --- Layout issues --- p.149 / Chapter 5.9 --- Overall simulation results --- p.150 / Chapter 5.1 --- Summary --- p.152 / References --- p.153 / Chapter CHAPTER 6. --- Design of high-efficient power amplifier --- p.154 / Chapter 6.1 --- Classification of PAs --- p.154 / Chapter 6.2 --- Circuit design considerations --- p.158 / Chapter 6.3 --- Experimental results --- p.160 / Chapter 6.4 --- Summary --- p.164 / References --- p.166 / Chapter CHAPTER 7. --- Conclusions and future work --- p.167 / Chapter 7.1 --- Conclusions --- p.167 / Chapter 7.2 --- Future work --- p.168 / References --- p.171

The introduction of prostate-specific antigen (PSA) testing about 3 decades ago led to the possibility of early detection of prostate cancer (PCa). Although PSA testing reduced the mortality rate, it is also associated with high risk of over diagnosis in patients with and without PCa. Despite the current drawbacks, it would be a challenge to replace PSA testing entirely. Instead, there is a need to develop parallel testing of other potential biomarkers that can complement the results from PSA tests. To address alternative biomarker sensing, this thesis highlights on the development of oligonucleotide-based biosensors for the detection of different biomarkers of PCa. Using PSA as a gold standard, the first study of this dissertation investigates the use of DNA aptamers to detect PSA using electrochemical impedance spectroscopy (EIS). The study compares 6-mercapto 1-hexanol chemistry with sulfo-betaine chemistry for the development of PSA aptasensor in terms of performance and selectivity. The second study focuses on glycoprofiling in order to complement PSA quantification as an additional information for reliable PCa diagnosis. This strategy was developed in a microfluidic channel with an optical read out using chemiluminescence. This study addresses one of the major problems of cross-reactivity with lectins in glycoprofiling, which can be solved using DNA aptamers. A third study concentrates on the development of an aptasensor for Alpha-Methylacyl-CoA Racemase (AMACR). AMACR has been reported for its high specificity and sensitivity to PCa. For the fabrication of the biosensor, a new strategy using polyethylene glycol was developed by electrochemical grafting it to a polypyrrole film. Since PCa diagnosis can be improved by looking at different biomarkers, an electrochemical platform for miRNA/DNA detection using a gold nanoparticle amplification strategy was also investigated. The sensor was fabricated using peptide nucleic acids (PNA) probes on gold electrodes. The study presents non-Faradaic EIS and amperometric techniques in order to exploit the inherent charges of nucleic acids. In conclusion, this thesis wants to serve as a potential orientation for overcoming the shortcomings of the current PCa testing and contribute towards the development of oligonucleotide-based biosensors for PCa biomarker detection and hopefully enhance the diagnosis and prognosis of PCa.

"Recently, nanomaterials have been vigorously studied for the development of biosensors. Among them, carbon nanotubes (CNTs) have stimulated enormous interest for constructing biosensors due to their unique physical and chemical properties such as high surface-to-volume ratio, high conductivity, high strength and chemical inertness. Our study is to develop a general design of biosensors based on vertically aligned CNT arrays. Glucose biosensor is selected as the model system to verify the design of biosensors. In the preliminary design, glucose oxidase (GOx) is attached to the walls of the porous alumina membrane by adsorption. Porous highly ordered anodized aluminum oxide (AAO) prepared by two-step anodization are used as templates. Deposited gold on both sides of template surfaces serve as a contact and prevent non-specific adhesion of GOx on the surface. In order to find out optimized thickness of gold coating, the oxidation and reduction (redox) reaction in [Fe(CN)6]3¨C /[Fe(CN)6]4¨C system is monitored by Cyclic Voltammetry (CV). Subsequently, enzymatic redox reaction in glucose solutions is also attempted by CV. We expect protein layers with GOx form a conductive network. However, no obvious enzymatic redox reaction is detected in the voltammogram. To take advantage of the attractive properties of CNTs, the design of enzyme electrodes is modified by attaching CNT onto the sidewalls of AAO template nanopores and then immobilizing GOx to the sidewalls and tips of CNTs. AAO templates provided vertical, parallel, well separated and evenly spacing nanochannels for CNT growth. Cobalt is used as a catalyst to fabricate CNTs. As a result, multi-walled carbon nanotubes (MWCNTs) are fabricated inside the AAO templates by catalytic chemical vapor deposition (CCVD). Characterization of AAO templates and cobalt electrochemical deposition are employed by scanning electron microscope (SEM), and energy dispersive X-ray spectrometry (EDS). Detailed structure and texture of CNTs are examined by transmission electron microscope (TEM). "