Orientador: Ivo Milton Raimundo Junior / Tese (doutorado) - Universidade Estadual de Campinas, Instituto de Quimica / Made available in DSpace on 2018-08-11T10:12:39Z (GMT). No. of bitstreams: 1 Fonseca_Alexandre_D.pdf: 7116553 bytes, checksum: dfa6bce9b276600fe0ac2ff916aa79e1 (MD5) Previous issue date: 2008 / Resumo: Este trabalho descreve a construção, caracterização e aplicação de microssistemas para análise em fluxo. A ablação por laser foi utilizada para o desenvolvimento de um sistema em poli(metilmetacrilato), cujos canais, com seções transversais retangulares medindo 200 mm de largura e 4,5 mm de profundidade, foram selados com um filme plástico adesivo. Este dispositivo suportou vazões de até 2,0 mL min sem apresentar vazamentos e a sua aplicação para a determinação fotométrica de Fe(II) em medicamentos proporcionou resultados que não diferiram significativamente daqueles obtidos por espectrometria de absorção atômica ao nível de 95 % de confiança. A fotolitografia profunda no ultravioleta foi empregada para a construção de microssistemas em resina a base de uretana-acrilato. Os canais com seções transversais triangulares e dimensões entre 200 e 600 mm foram selados com uma camada do próprio substrato, formando uma estrutura monolítica que suportou vazões de ate 3,0 mL min. Fibras ópticas plásticas e agulhas hipodérmicas foram acopladas aos dispositivos e utilizadas para a integração dos sistemas ópticos de detecção e para o acesso das soluções de trabalho, respectivamente. Um microssistema com célula integrada para detecção fluorimétrica e dois pontos de confluência foi aplicado a determinação de íons Ca e Mg em águas minerais. Outros dois dispositivos foram aplicados separadamente a determinação fotométrica de íons Cr(VI) em águas contaminadas e em ligas metálicas e a determinação de íons ions Cl em águas minerais. Para todas as aplicações, as concentrações determinadas através dos métodos com os microssistemas foram concordantes com aquelas determinadas através dos métodos em batelada (espectrofotometria e volumetria de complexação). A geração de volumes extremamente pequenos de resíduos (cerca de 25 mL de solução para uma jornada de 8 horas de trabalho) indicou que os dispositivos propostos cumprem adequadamente os requisitos estabelecidos pela Química Verde / Abstract: This work describes the construction, characterization and application of microsystems for flow analysis. Laser ablation was employed for the development of a system in poly(methylmethacrylate), whose channels, with rectangular cross sections measuring 200 mm width by 4,5 mm depth, were sealed with an adhesive plastic film. This device supported flow rates up to 2,0 mL min without leaking and its application for the photometric determination of Fe(II) in medicines provided results that did not differ significantly of those obtained by flame atomic absorption spectrometry at a confidence level of 95 %. The deep UV-photolithography was used for the construction of microsystems in urethane / acrylate based resin. The channels with triangular cross sections and dimensions between 200 e 600 mm were sealed with a layer of the same resin, forming a monolithic structure that supported flow rates up to 3.0 mL min. Plastic optical fibers and hypodermic needles were coupled to the devices and used for the integration of the optical detection systems and for the access of the working solutions, respectively. A microsystem with an integrated flow cell for fluorimetric detection and two confluence points was applied to the determination of Ca e Mg in mineral waters. Other two devices were separately applied to the photometric determination of Cr(VI) in contaminated waters and in metallic alloys, and to the determination of Cl in mineral waters. For all the applications, the concentrations obtained agreed with those determined by batch methods (spectrophotomety and complexometric titration). The generation of extremely small volumes of residues (ca. 25 mL of solution after an 8-h working day) indicates that the proposed devices appropriately accomplish the requirements established by the Green Chemistry / Doutorado / Quimica Analitica / Doutor em Ciências

Este trabalho de doutorado apresenta o desenvolvimento de novas tecnologias para fabricação de microsistemas analíticos e detecção eletroquímica. Primeiramente, a poliuretana elastomérica, derivada de uma fonte renovável, o óleo de mamona, foi utilizada como um novo e alternativo material para fabricação de microdispositivos. Foram avaliadas as características físicas dos microcanais formados por moldagem, a compatibilidade química com solventes e eletrólitos, as características de superfície através dos ângulos de contato, o EOF em diferentes pHs e a performance analítica em experimentos de eletroforese com detecção eletroquímica. A segunda parte do trabalho apresenta o desenvolvimento de um método para a determinação simultânea de azo-corantes comumente usados na indústria alimentícia. Amaranto, amarelo crepúsculo FCF, amarelo sólido AB, ponceu 4R e vermelho 2G, foram separados e quantificados através de eletroforese em microdispositivos com detecção eletroquímica. Foram estudados e otimizados vários parâmetros que influenciaram a separação eletroforética e detecção eletroquímica, em experimentos realizados usando microdispositivos de vidro e eletrodo de trabalho de carbono vítreo. Finalmente, a terceira parte desse trabalho apresenta o uso das propriedades magnéticas e eletrocatalíticas de nanofios de níquel no desenvolvimento de um detector adaptativo magneticamente modulável para eletroforese em microdispositivos. / The development of microfluidic analytical systems has witnessed an explosive growth during the last 15 years. Particular attention has been given to microchip electrophoresis because of their fast and efficient separation capabilities. Electrochemistry detection offers considerable promise for such microfluidic systems, with features that include remarkable sensitivity, inherent miniaturization and portability, low cost, and high compatibility with microfabrication technologies. This thesis shows the development of new fabrication technologies for miniaturized analytical systems with electrochemical detection and it is presented in four chapters, Chapter I shows an introductory view of the main aspects related to miniaturization of analytical systems and amperometric detection configurations commonly coupled to microchip electrophoresis. In Chapter II, the use of elastomeric polyurethane (PU), derived from castor oil (CO) biosource, as a new material for fabrication of microfluidic devices by rapid prototyping is presented. Including the irreversible sealing step, PU microchips were fabricated in less than 1 h by casting PU resin directly on the positive high-relief molds fabricated by standard photolithography and nickel electrodeposition. Physical characterization of microchannels was performed by scanning electron microscopy (SEM) and profilometry. Polymer surface was characterized using contact angle measurements and the results showed that the hydrophilicity of the PU surface increases after oxygen plasma treatment. The polymer surface demonstrated the capability of generating an electroosmotic flow (EOF) of 2.6 × 10-4 cm2 V-1 s-1 at pH 7 in the cathode direction, which was characterized by current monitoring method at different pH values. The compatibility of PU with a wide range of solvents and electrolytes was tested by determining its degree of swelling over a 24 h period of contact. The performance of microfluidic systems fabricated using this new material was evaluated by fabricating miniaturized capillary electrophoresis systems. We used catecholamines as model analytes that were separated in aqueous solutions and detected with end-channel amperometric detection. In Chapter III, a method based on microchip electrophoresis with electrochemical detection has been developed for the simultaneous determination of Yellow AB, Red 2G, Sunset Yellow, Ponceu 4R, and Amaranth which are azo-dyes frequently added to foodstuffs. Factors affecting both separation and detection processes were examined and optimized, with best performance achieved by using a 10 mM phosphate buffer (pH 11) as running buffer and applying a voltage of 2500 V both in the separation and in the electrokinetic injection (duration 4 s). Under these optimal conditions, the target dye analytes could be separated and detected within 300 s by applying a detection potential of -1,0 V (vs. Ag/AgCl) to the glassy carbon (GC) working electrode. The recorded peaks were characterized by a good repeatability (RSD = 1,8 - 3,2%), high sensitivity, and a wide linear range. Detection limits of 3.8, 3.4, 3.6, 9.1, 15.1 ?M were obtained for Yellow AB, Red 2G, Sunset Yellow, Ponceu 4R, and Amaranth, respectively. Fast, sensitive, and selective response makes the new microchip protocol very attractive for the quantitative analysis of commercial soft drinks and candies Finally, in Chapter IV, we demonstrate for the first time the use of adaptive functional nickel nanowires for switching on demand operation of microfluidic devices. Controlled reversible magnetic positioning and orientation of these nanowires at the microchannel outlet offers modulation of the detection and separation processes, respectively. The former facilitates switching between active and passive detection states to allow the microchip to be periodically activated to perform a measurement and reset it to the passive (\"off\") state between measurements. Fine magnetic tuning of the separation process (post channel broadening of the analyte zone) is achieved by reversibly modulating the nanowire orientation (i.e., detector alignment) at the channel outlet. The concept can be extended to other microchip functions and stimuli-responsive materials and holds great promise for regulating the operation of microfluidic devices in reaction to specific needs or unforeseen scenarios.

O presente trabalho teve como foco o desenvolvimento de um software para a simulação de processos de microfabricação em substrato e de microfabricação em superfície baseado em autômatos celulares, o simMEMS. Além disso, visando a futura incorporação de ferramentas para análise das estruturas geradas pelo programa, um módulo com funcionalidades básicas para a análise mecânica de estruturas também foi desenvolvido. No que tange à microfabricação em superfície, o software desenvolvido permite simular a corrosão anisotrópica úmida do Si em KOH e deep reactive ion etching (DRIE). O simulador de corrosão úmida utiliza um autômato celular conhecido como BCA. O simulador de DRIE usa um autômato próprio. Para a simulação dos processos de microfabricação em superfície o software fornece quatro processos: deposição de filmes, corrosão de filmes, fotolitografia e planarização. Para corrosão e deposição de filmes, diversos autômatos celulares da literatura foram analisados e os resultados dessas análises é aqui apresentado. Todos os simuladores, tanto de microfabricação em superfície como em substrato, podem ser utilizados em conjunto. Isso torna o software bastante útil e capaz de simular a fabricação de um grande número de dispositivos. / The main goal of this project is the development of a software capable of simulating both surface and bulk micromachining based on a cellular automata approach. This software has been called simMEMS. In order to enable future versions of the software to also be able to analyze the structures created by the software, a module capable of running a mechanical analysis through the finite element method is also developed. simMEMS allows the user to simulate two bulk micromachining processes: wet anisotropic KOH etching and deep reactive ion etching DRIE. The wet etching simulator uses a cellular automaton known as BCA. The DRIE simulator uses an automaton developed during this project. The surface micromachining simulator allows the user to simulate four types of processes: photolithography, film deposition, film etching and substrate planarization. Several automata for the deposition and etching of films are studied and the results of this study are presented here. All processes, be they for surface or bulk micromachining, can be used on the same substrate to simulate the entire fabrication process for a large array of devices. This makes simMEMS a very useful software.

Orientador: Ioshiaki Doi / Tese (doutorado) - Universidade Estadual de Campinas, Faculdade de Engenharia Elétrica e de Computação / Made available in DSpace on 2018-08-20T21:32:16Z (GMT). No. of bitstreams: 1 Neli_RobertoRibeiro_D.pdf: 20995056 bytes, checksum: 0072f9af377dc8c222632b4d9608b5b0 (MD5) Previous issue date: 2012 / Resumo: Este trabalho tem como objetivo a fabricação e caracterização de sensores térmicos descritos como bolométricos, que são dedicados a detecção da radiação infravermelha distante. Estes sensores são construídos a partir de técnicas de microfabricação, utilizando filmes finos seletivos a corrosão úmida. Estas microestruturas mecânicas são formadas sobre laminas de silício a partir de um ataque químico úmido sobre a superfície da mesma. Como estas estruturas são obtidas utilizando-se técnicas convencionais de fabricação de circuitos integrados, torna-se possível a integração monolítica de circuitos eletrônicos e dispositivos mecânicos, permitindo o desenvolvimento de microssistemas integrados. O ouro poroso ou "ouro negro" foi estudado e caracterizado, sendo utilizado como absorvedor de radiação e apresentou neste trabalho índices de absorção superiores a 80%. Foi desenvolvido também um processo para integrar este filme ao dispositivo. O silício policristalino, submetido a dopagem de boro, foi desenvolvido para se obter valores de TCR próximos a -2%K-1 e resistências abaixo de 1k'ômega'. Finalmente, foram desenvolvidos os layouts, fabricadas e testadas as microestruturas de diversas geometrias, como pontes, vigas, membranas, espiras, entre outras. Os dispositivos bolométricos testados apresentaram TCR de -2,54%K-1 , um tempo de resposta de aproximadamente 2 ms, uma responsividade de 0,35 V/W e uma detectividade específica de 6,04.109 mHz1/2W-1, quando submetido a uma radiação de 0,85 THz / Abstract: This work has as a main goal the fabrication and characterization of thermal sensors, described as bolometrics, which are dedicated to detection of far infrared radiation. These sensors are fabricated using microfabrication techniques and the thin films are selectives to wet etching. These mechanical microstructures are formed on silicon wafers using a surface wet etching. As these structures are obtained using conventional techniques for CI's manufacturing, it becomes possible perform a monolithic integration of electronics and mechanical devices, allowing the integrated microsystems development. The porous gold or "gold black" used as a radiation absorber, was studied and characterized, and this study showed absorption index greater than 80%. Was developed a process to integrate this film to device. The doped polycrystalline silicon was performed to obtain TCR values near to -2% K-1 and resistance less than 1k'omega'. Finally, the layouts are designed, performed and tested the microstructure of various geometries such as bridges, beams, membranes, coils, among others. The devices tested presented TCR about -2.54% K-1, a response time of approximately 2 ms, responsivity about 0.35 V / W and specific detectivity about 6.04x109 mHz1/2W-1 when subjected to a 0,85 THz radiation / Doutorado / Eletrônica, Microeletrônica e Optoeletrônica / Doutor em Engenharia Elétrica

Le travail de thèse a consisté à développer des voies originales de microfabrication pour laconception d’électrodes qui pourront être utilisées dans un biocapteur basé sur unetransduction électrochimique. Une des perspectives étant de pouvoir intégrer ce type decapteur dans un microsystème analytique à base microfluidique, nous avons fait le choix duverre comme matériau de base. Par ailleurs, nous avons privilégié les technologies de« lithographie douce » au détriment de voies classiques telles que la photolithographie afin derendre inutile l’accès à des salles à environnement contrôlé ou l’utilisation d’appareillagessophistiqués.Lors de ce travail, nous avons plus particulièrement travaillé sur le développement deméthodes combinant la technique de microtamponnage et la métallisation chimique de typeautocatalytique (electroless). Cette métallisation nécessitant des surfaces catalytiques pourfaire croître la couche métallique, nous avons développé des méthodes de traitements desurface afin de rendre le substrat de base catalytique sur toute sa surface. La technique demicrotamponnage a ensuite été utilisée afin de passiver les zones où la métallisation n’est pasdésirée et cela a mené à des microstructures métalliques en surface du verre présentant peu oupas de défauts. Notre approche nous a conduit à utiliser plusieurs types de catalyseurs sous laforme de nanoparticules métalliques à base d’argent, d’or ou de palladium et nous avonsdiscuté les différences entre les méthodes basées sur ces différents catalyseurs.Une autre voie a consisté à graver de façon localisée des couches minces métalliquesuniformes en protégeant les zones ne devant pas être gravées par la technique demicrotamponnage. Ceci a permis le développement de deux voies originales demicrostructuration sur couches minces métalliques uniformes (d’une part le pelage sélectif etd’autre part le procédé à double inversion).Dans l’ensemble de nos travaux, des caractérisations d’extrême surface par les techniquesSEM, AFM, ToF-SIMS, XPS et de mouillabilité ont été menées afin d’optimiser ledéveloppement des différents procédés. / This thesis work consisted in the development of original strategy for the microfabrication ofelectrodes which could be used in a biosensor as an electrochemical transducer. One of theprospects of this work is to insert this type of sensor into a microfluidic chip, We have madethe choice of using glass as a substrate. Moreover, we have favoured soft lithographictechnologies at the expense of conventional strategy like photolithography.In this work, we mainly worked on the development of methods which combines microcontact printing and autocatalytic metallisation (electroless). As this type of metallisationneeds catalytic surfaces to grow the metallic layer, we developed surface treatments methodsto make the surface of the substrate catalytic for the metallisation. To follow, the microcontact printing technique has been used to passivate areas where metallisation should notoccur and this leads to metallic microstructure with very few defects. Our approach leads uson the use of different catalyst like gold, silver or palladium nanoparticles and we havediscussed differences between the different methods.Another strategy consisted in the selective etching of thin metallic layer. Areas not to be etchare protected by the micro contact printing technique. This leads to the development of twooriginal strategies of microfabrication on thin metallic layer.In the whole work, extreme surface characterisation like SEM, AFM, ToF-SIMS, XPS andwettability have been carried out in order to optimize the development of the differentmethods.

Cell encapsulation is a promising concept in regenerative medicine and stem cell treatment of diseases. Cells encapsulated in hydrogels have shown to yield better therapeutic outcome over cells in suspension. Microfluidic platforms have facilitated the process of cell encapsulation through the controlled mixing of aqueous cell solution and hydrogel with an immiscible liquid to yield a monodispersed population of microcapsules at a high throughput. However, given that the microfluidic process of placing cells in microcapsules is completely random, yielded samples are often riddled with empty microcapsules, raising the need for a post-encapsulation purification step to sort empty microcapsules from cell-laden ones. Sorting of microcapsules can be achieved through several techniques, most desirable of which are electrokinetic such as dielectrophoresis (DEP) and electrophoresis (EP). The advantages of DEP and EP techniques are that they support label-free sorting and yield a high throughput. However to achieve true effective DEP or EP sorting, there is a need to understand how empty microcapsules react to these electrokinetic forces versus occupied microcapsules. This study developed microfluidic devices for characterising the electrokinetic effects on microcapsules using DEP and EP. Results of both characterization techniques showed notable differences in the response of empty microcapsules versus cell-laden ones, reinforcing their potentials for sorting. Furthermore, this study proposed designs for microcapsules sorting devices that leverage EP and DEP.

The inability of the myocardium to regenerate after injury plus the inadequate number of available hearts for transplantation have drawn attention to the creation of functional tissue constructs for implantation within the injured heart. In addition, there is an increasing interest in developing in vitro models to study heart physiology and pathology as well as to evaluate drug efficacy. Formation of these in vitro models and tissue constructs requires highly specific conditions to mimic the normal environment of cells in the body. Firstly, in this study, plasma lithography patterning of elastomeric substrates is exploited for creating microtissues composed of neonatal cardiomyocytes, and investigating their development in different mechanical microenvironments. Immunofluorescence microscopy and force spectroscopy show that the size and shape of the cardiomyocyte clusters, as well as the sarcomere length, fiber alignment, and beating amplitude and frequency of the cardiomyocytes, are regulated by microenvironmental cues. Computational analysis reveals that the mechanical stress at the cluster-substrate interface strongly correlates with the aforementioned characteristics of the cardiomyocytes. Taken together, our results underscore a collective mechanoadaptation scheme in cardiac development. Secondly, a silicone substrate with tunable elasticity is characterized for biological studies. Uniaxial tensile testing and microindentation show that these substrates could cover the biological range of stiffness for normal and pathological conditions. Spectrophotometry demonstrates that the transmittance of these substrates is comparable to those of glass and Sylgard 184. Atomic force microscopy shows that the surface roughness of samples is lower than that of widely-used Sylgard 184. Contact angle measurements before and after exposure to air plasma indicate that these samples are compatible with plasma lithography patterning. Thirdly, a new technique for cell patterning is developed which utilizes selective plasma lithography to modify protein adhesion on the substrate. This approach is based on controlling the conformation of Pluronic F-127 layer adsorbed on the surface by modifying surface wettability. Contact angle measurements show that both PDMS and plastic petri dish are compatible with this technique. X-ray photoelectron spectroscopy and atomic force microscopy confirm the adsorption of PF-127 layers with controlled conformation. Fluorescent and bright-field microscopy demonstrate selective adhesion of proteins and attachment of cells merely on plasma-treated areas. Finally, micropillar arrays are employed to determine the effects of two proteins associated with regulation of thin filament length, i.e. Lmod2 and Tmod1, on contractile force generation at the cellular level. Our results demonstrate that the contractile force of single isolated Lmod2-KO cardiomyocytes decreases compared to the wildtype control. Transduction of Lmod2 in the knockout cardiomyocytes restores their contractile force to the level of their WT counterparts, verifying that the observed contractile dysfunction is specific to the loss of Lmod2. Our data demonstrate that overexpression of Tmod1 in cardiomyocytes decreases their contractile force compared to the WT cells and confirm the effects of Lmod2 knockout on contractile force generation.

Complex-mask coronagraphs destructively interfere unwanted starlight with itself to enable direct imaging of exoplanets. This is accomplished using a focal plane mask (FPM); a FPM can be a simple occulter mask, or in the case of a complex-mask, is a multi-zoned device designed to phase-shift starlight over multiple wavelengths to create a deep achromatic null in the stellar point spread function. Creating these masks requires microfabrication techniques, yet many such methods remain largely unexplored in this context. We explore methods of fabrication of complex FPMs for a Phased-Induced Amplitude Apodization Complex-Mask Coronagraph (PIAACMC). Previous FPM fabrication efforts for PIAACMC have concentrated on mask manufacturability while modeling science yield, as well as assessing broadband wavelength operation. Moreover current fabrication efforts are concentrated on assessing coronagraph performance given a single approach. We present FPMs fabricated using several process paths, including deep reactive ion etching and focused ion beam etching using a silicon substrate. The characteristic size of the mask features is 5 mu m with depths ranging over 1 mu m. The masks are characterized for manufacturing quality using an optical interferometer and a scanning electron microscope. Initial testing is performed at the Subaru Extreme Adaptive Optics testbed, providing a baseline for future experiments to determine and improve coronagraph performance within fabrication tolerances.

The multidisciplinary field of microfluidics has shown great promise for research at the interface of biology, chemistry, engineering, and physics. Laminar flow, versatile fabrication, and small length scales have made microfluidics especially well-suited for single-cell characterization. In particular, the evaluation of single-cell growth rates is of fundamental interest for studying the cell cycle and the effects of environmental factors, such as drugs, on cellular growth. This work presents aspects in the development of a microfluidic cell impedance sensor for measuring the volumetric growth rate of single cells and covers its application in the investigation of a new discovery relating to multidrug resistance in S. cerevisiae. While there are many avenues for the utilization and interpretation of growth rates, this application focused on the quantitative assessment of biological fitness—an important parameter in population genetics and mathematical biology. Through a combination of growth measurements and optics, this work concludes a novel case of bet-hedging in yeast, as well as the first ever case of bet-hedging in eukaryotic multidrug resistance.

Endothelial cells, EC, are the cell type closest to the blood stream in vessel walls. These cells can affect the origin of atherosclerosis, plaques clogging the vessels. The behaviour of EC is affected by neighbouring smooth muscle cells and shear stress from the blood flow. The aim with this thesis was to fabricate a structure for an aorta-on-a-chip that can be used to study these two parameters and their influence on EC and vascular diseases. Previous research using a two-channel system resulted in leakage and low viability of the muscle cells. A three-channel system has therefore been made to include a middle channel with the muscle cells incorporated in a gel. Cell medium is flowed in the outer channels to provide the cells with nutrition. The flow in the channel with EC has been calculated to correspond to the shear stress in an aorta. Membranes of polyethylene terephthalate and polycarbonate were used to divide the channels and both were shown to be compatible with EC. Different bonding procedures were investigated to manufacture leakage-free chips. In the study, adhesive bonding clogged the channels and the parameters for thermal bonding of COC, cyclic olefin copolymer, were not fully optimised. This made chemical bonding with layers of PDMS, polydimethylsiloxane, the best alternative. APTES, (3-Aminopropyl)triethoxysilane, treatment in addition to plasma treatment on the surfaces improved the bonding strength. Polycarbonate membranes got better results in the bonding tests than polyethylene terephthalate. The resulting aorta-on-a-chip was therefore successfully fabricated in PDMS and polycarbonate membranes using plasma and APTES treatment for bonding.