Spelling suggestions: "subject:"microfluidic""
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[en] PORESCALE ANALYSIS OF OIL DISPLACEMENT BY POLYMER SOLUTION / [pt] ANÁLISE DO DESLOCAMENTO DE ÓLEO POR SOLUÇÕES POLIMÉRICAS EM MICROESCALANICOLLE MIRANDA DE LIMA 12 May 2016 (has links)
[pt] A injeção de água é o método de recuperação secundária mais utilizado na indústria do petróleo. No entanto, a alta razão de mobilidade entre a água e o óleo limita a quantidade de óleo deslocada. Uma alternativa para minimizar este problema é a aplicação de tecnologias que agem como agentes de controle da mobilidade. Soluções poliméricas podem ser utilizadas para aumentar a viscosidade da água e consequentemente reduzir a razão de mobilidade. Evidências experimentais têm mostrado que o comportamento elástico de soluções poliméricas pode além de diminuir a razão de mobilidade, contribuir para um melhor deslocamento de óleo em escala de poro, reduzindo a saturação de óleo residual. Esse comportamento em escala de poro ainda não está completamente entendido. Nesse trabalho, um micromodelo de vidro formado por uma rede bi-dimensional de canais foi utilizado como meio poroso. Esse dispositivo tem algumas características importantes de meios porosos e permite a visualização do fluxo em escala de poro. A evolução do deslocamento de óleo pela fase aquosa é acompanhada no microscópio e são obtidas imagens dos perfis de saturação. Três diferentes fases aquosas foram usadas: água deionizada, uma solução de poli(óxido de etileno) de alto peso molecular e uma mistura de água com glicerina com a mesma viscosidade do poli(óxido de etileno). A visualização do fluxo no micromodelo permite obter informações específicas sobre a presença de óleo preso por forças capilares e o movimento da interface óleo/água no interior da rede. Resultados mostraram que as forças viscoelásticas modificam a distribuição de fluidos no meio poroso, melhorando a eficiência de deslocamento em escala de poro e consequentemente a saturação de óleo residual. / [en] Water flooding is the most commonly used oil recovery method in the oil industry. However, the high mobility ratio between the water and oil phases limits the amount of oil displaced by the water phase. An effective alternative to minimize this problem is the application of technologies that act as mobility control agents. Polymer solution is used in many cases as a way to increase the water phase viscosity and consequently reduce the mobility ratio. Experimental evidences have shown that the elastic behavior of some polymer solution may not only improve the mobility ratio but also contribute to a better pore level oil displacement, reducing the residual oil saturation. This pore level behavior is not clearly understood. In this work, a glass microfluidic chip made of a 2-D array of channels is used as a two-dimensional porous space. This device has the principal features of a porous media and provides means for pore level flow visualization. A microscopic is used to monitor the evolution of the water phase as it displaces oil and images of the saturation profiles can be made. Three different water phases were used: pure water, a high molecular weight poly(ethylene oxide) solution and a glycerol-water mixture with the same viscosity of the polymer solution. Flow visualization provides specific information about the presence of the trapped oil phase and the movement of the oil/water interface in the network. Results show that the viscoelastic forces modify the liquid distribution in the porous media, improving the displacement efficiency at pore scale and consequently the residual oil saturation.
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Stress-Strain Behavior of Single Vimentin Intermediate FilamentsBlock, Johanna Lena 23 April 2018 (has links)
No description available.
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[en] THREE-DIMENSIONAL VISUALIZATION OF OIL DISPLACEMENT BY FLEXIBLE MICROCAPSULES SUSPENSIONS IN POROUS MEDIA / [pt] VISUALIZAÇÃO TRIDIMENSIONAL DO DESLOCAMENTO DE ÓLEO POR SUSPENSÕES DE MICROCÁPSULAS FLEXÍVEIS EM MEIOS POROSOSJOSE RONALDO VIMIEIRO JUNIOR 24 October 2017 (has links)
[pt] Em um mundo globalizado, a demanda por energia está sempre crescendo. Uma vez que a indústria de óleo e gás é responsável pela entrega da maior parte desta demanda, isso faz dos hidrocarbonetos componentes cada vez mais importantes no mercado mundial. Entretanto tais recursos são finitos, portanto, uma exploração consciente, buscando sempre o máximo desempenho se faz necessária. Como os reservatórios de petróleo, logo após a aplicação das técnicas de recuperação primária e secundária, geralmente ainda possuem cerca de 65 por cento do volume de óleo originalmente contido em seus poros, métodos que visam a redução dessa porcentagem estão ganhando um papel cada vez mais importante na indústria energética. Nesse contexto, esse trabalho apresenta um micromodelo tridimensional representativo de um meio poroso que será utilizado para a análise do escoamento de fluidos na escala de poro. A microscopia confocal será adotada para visualizar os diferentes fenômenos que ocorrem em microescala, permitindo a obtenção de informações específicas sobre a dinâmica dos gânglios de óleo, em relação a sua formação, mobilização e aprisionamento, e assim, ao final do experimento quantificar a saturação residual de óleo em diferentes condições de escoamento. Os resultados obtidos mostram que o uso das suspensões de microcápsulas flexíveis como agente de controle de mobilidade, modifica a distribuição dos fluidos no meio poroso, o que melhora a eficiência de deslocamento do fluido deslocante na escala de poro, e consequentemente diminui a saturação de óleo residual. / [en] In a globalized world, the demand for energy is always growing. Since the oil and gas industry is responsible for delivering most of this demand, this makes hydrocarbon components increasingly important in the worldwide economy. However, such resources are finite, so a conscious exploration always seeking the maximum performance is required. As oil reservoirs after the application of primary and secondary recovery techniques usually still have about 65 percent of the original oil volume contained in their pores, methods that aim its reduction are gaining an increasingly important role in the energy industry. In this context, this work presents a three-dimensional micromodel representative of a porous medium that is used for pore-scale flow analysis. Confocal microscopy is used to visualize the microscale phenomena, leading to specific information about ganglia dynamics, related to its formation, mobilization and entrapment. The residual oil saturation, an important value to measure the amount of oil produced in a given reservoir is determined for different flow conditions. The results show that the suspensions composed by flexible microcapsules could be used as a mobility control agent, since it modifies the fluid distribution in the porous media, improving the pore-scale displacement efficiency, and consequently reducing the residual oil saturation.
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Synthèse de polymères poreux à base de PEG-DA par voie microfluidique et pour une application en tant qu'isolant acoustique / Synthesis of porous polymers made of PEG-DA by microfluidics and for an application of acoustic insulatorTurani-I-Belloto, Alexandre 19 October 2018 (has links)
La réalisation de revêtements fins, solides et isolants dans le domaine ultrasonore,contenant de petites particules poreuses, représente le challenge de ces travaux. Plusieurs points sont déterminants pour arriver à cet objectif final. Tout d’abord, il faut trouver le matériau adéquat pour les particules poreuses, notre choix s’est porté sur un polymère mou, le PEG-DA. Une fois le matériau choisi, il faut déterminer une voie de synthèse pour en faire un matériau poreux, une technique par photopolymérisation UV etpar dissolution d’un porogène sacrificiel (CaCO3) a été utilisée. Ensuite, la microfluidiquenous a permis de réaliser des microparticules de ce polymère poreux de tailles contrôlées et monodisperses. La dernière étape consiste à disperser ces particules dans une matrice pour en faire un revêtement. Les mesures acoustiques ont montré une bonne atténuation du PEG-DA dans les ultrasons.Par ailleurs, ces travaux ont permis d’étudier la photopolymérisation frontale de milieux transparents et très diffusants, et de présenter une méthode à partir de simulations Monte-Carlo pour décrire les profils de conversion du polymère en fonction de l’intensité initiale et de l’épaisseur traversée. D’autres études se sont focalisées sur le séchage d’hydrogels de PEG-DA et sur la dissolution de CaCO3 à travers ces mêmes hydrogels. Un modèle physique décrivant la compétition entre cinétique de dissolution et diffusion de l’acide a été proposé. / The fabrication of thin, solid and insulating coatings containing small porous particles is the major challenge of these works. Several key points need to be raised to succeed. First, a good material for the particles has to be found, we chose a soft polymer, PEG-DA. Then, the choice of the synthesis routine to make porous polymers is crucial. A technique using UV photopolymerization and dissolution of sacrificial porogen (CaCO$_3$) will be used. Next, we will decide to make porous particles by microfluidics to obtain well controlled sizes and good monodispersity. The last step will be about the dispersion of those particles in a matrix to create coatings. Acoustics measurements will show the acoustic response of PEG-DA in ultrasounds. Moreover, this work will also permit us to study the frontal photopolymerization of transparent and scattering media and to present a method with Monte-Carlo simulations to describe conversion profiles of polymer as a function of intensity and path length. Other studies will present drying of PEG-DA hydrogels and dissolution of CaCO$_3$ through those hydrogels. A physical model will be proposed to describe the competition between kinetics and acid diffusion.
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Medium sized molecules clearance through artificial kidneys / Clairance de molécules de taille moyenne à travers un rein artificielSnisarenko, Dmytro 07 November 2016 (has links)
Malgré une longue histoire de développement, l'hémodialyse (rein artificiel) possède encore quelques limitations, telles que la perte des propriétés initiales de la membrane en cours de traitement à cause du colmatage et la mauvaise élimination des toxines urémiques de taille moyenne. La présente étude fait partie d'un projet européen nommé BioArt dont le but est d'apporter des solutions à ces limites. Dans cet objectif, l'un des partenaires du projet a proposé le développement d'un nouveau concept de membrane double couche au sein de laquelle sont incorporées des particules adsorbantes. Une caractérisation complète de cette nouvelle membrane était alors nécessaire, plus précisément l'impact de la matrice mixte sur l'élimination des toxines urémiques de divers groupes devait être évalué, ainsi que la propension du matériau membranaire à se colmater. Les études des phénomènes de colmatage sont classiquement menées à l'échelle macroscopique (faisceau de fibres creuses) sans analyse à l'échelle d'une fibre isolée. Le but premier de la présente thèse a alors été de proposer un dispositif permettant une étude du colmatage membranaire induit par la protéine à l'échelle microscopique. Un dispositif microfluidique transparent dans lequel la membrane polymère est insérée a été élaboré et mis en œuvre pour la filtration des protéines modèles : l'albumine de sérum bovin (BSA) et l'a-lactalbumine. Grâce au couplage avec la microscopie de fluorescence, différents modes d'adsorption des protéines sur la surface de la membrane ont été observés et liés aux variations des conditions hydrodynamiques à l'intérieur de la puce. Il a été constaté, sous certaines conditions, une différence dans l'accumulation de protéines entre l'entrée, le centre et la sortie du canal tandis que dans d'autres conditions cet effet s'annule. En outre, un phénomène inattendu, l'agrégation de l'a-lactalbumine, a été observé au cours de la filtration. La localisation dans le canal et la forme des agrégats dépendent également des conditions hydrodynamiques et de la pression transmembranaire appliquée. Dans le but d'optimiser la conception de la membrane vis à vis de son aptitude à éliminer des molécules de taille moyenne de la circulation sanguine, un modèle mathématique a été proposé. L'objectif du modèle était, en prenant en compte la présence de particules adsorbantes à l'intérieur de la membrane double couche, de rendre compte de la combinaison des trois mécanismes d'élimination du soluté : la convection, la diffusion et l'adsorption. Le modèle permet de prédire l'influence de divers paramètres tels que la diffusivité de la molécule, l'épaisseur de la membrane, la présence de la convection, la charge en particules adsorbantes, sur l'intensification des flux à travers la membrane. Le modèle semble être un outil utile pouvant être appliqué à l'optimisation de membranes pour l'élimination des toxines. / Despite a long history of development, the hemodialysis procedure (artificial kidney) still possesses some limitations, such as loss of the initial properties of the membrane due to fouling and poor removal of the middle sized uremic toxins. The present study is part of an European project named BioArt the aim of which was to overcome these limitations. In that objective, one of the partners of BioArt project reported on the development of the novel promising concept of double layer membrane with embedded adsorptive particles. A thorough characterization of the new membrane was then necessary, more precisely the extent to which mixed matrix layer can improve the removal of the uremic toxins from various groups needed to be evaluated, as well as the propensity of the membrane material to become fouled. The studies of the fouling phenomena are conventionally performed at the macro scale (bundle of hollow fibers) without insights of what is happening at the scale of an isolated fiber. Therefore, the primary aim of the present Thesis was to transfer the research of the protein-induced membrane fouling from the macro to the micro scale. A novel transparent microfluidics device with the polymeric membrane inside has been developed and applied for the filtration of model proteins: bovine serum albumin (BSA) and a-lactalbumin. Thanks to the coupling of the microchip with the fluorescent microscopy, different patterns of protein deposition on the membrane surface were observed and related to the variations in the hydrodynamic conditions inside the microchip. It was found that at certain conditions one may observe the difference in protein accumulation in the inlet, the middle, and the outlet of the channel while at other conditions this effect vanishes. Additionally, the unexpected phenomena of a-lactalbumin aggregation was observed over the course of filtration. The location and shape of the aggregates were also dependent on the hydrodynamic conditions and the applied transmembrane pressure. Aiming to address the problem of membrane design optimization for the enhancement of the middle molecules elimination from the bloodstream, a mathematical model, which accounts for the presence of adsorptive particles inside the complex double-layer membrane, has been proposed. The objective of the model was to understand the interplay of three solute removal mechanisms: convection, diffusion, and adsorption. The model allows predicting the influence of various parameters such as molecule diffusivity, membrane thickness, the presence of convection, content of adsorptive particles on the flux intensification across the membrane. The developed model seems to be a useful tool, which may be applied to design optimized membranes for the removal of toxins.
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Interaction et diffusion hydrodynamiques dans une suspension de vésicules et globules rouges / Hydrodynamic interactions and diffusion in vesicle and red blood cell suspensionsSrivastav, Aparna 26 January 2012 (has links)
Blood is a complex suspension of deformable particles, red blood cells, which exhibits a sophisticated dynamics when flowing in the microvasculature. Most of these complex phenomena, non-linear rheology, structuration of the suspension, heterogeneities of the hematocrit distribution, are directly connected to the rich microscopic dynamics of individual red blood cells, and their hydrodynamics interactions. We investigate a few aspects of the dynamics of red blood cells and giant vesicles - a simple model for RBCs. A study on the dynamics of very deflated vesicles, with shapes similar to those of red blood cells, shows that these objects which haven't received a lot of attention so far can exhibit richer than expected dynamics. We then mainly focus on the still unexplored problem of hydrodynamic interactions between vesicles or red blood cells and their consequences at the scale of the suspension. An experimental study of the interaction of two identical vesicles in shear flow shows that there is a net repulsion between the cells that leads to an increase of the distance between vesicles in a suspension. Scaling arguments are proposed for this interaction and a comparison with numerical results is performed and a quantitative estimation of a shear induced diffusion coefficient obtained by averaging the results for pair interactions is found. Finally, we investigate the diffusion of a cloud of red blood cells in Poiseuille flow in order to directly determine diffusion coefficients. The experiment shows that the cloud widens when traveling along the channel with a power law behaviour indicating sub-diffusion. This effect is confirmed by a theoretical analysis of a few limit cases. / Blood is a complex suspension of deformable particles, red blood cells, which exhibits a sophisticated dynamics when flowing in the microvasculature. Most of these complex phenomena, non-linear rheology, structuration of the suspension, heterogeneities of the hematocrit distribution, are directly connected to the rich microscopic dynamics of individual red blood cells, and their hydrodynamics interactions. We investigate a few aspects of the dynamics of red blood cells and giant vesicles - a simple model for RBCs. A study on the dynamics of very deflated vesicles, with shapes similar to those of red blood cells, shows that these objects which haven't received a lot of attention so far can exhibit richer than expected dynamics. We then mainly focus on the still unexplored problem of hydrodynamic interactions between vesicles or red blood cells and their consequences at the scale of the suspension. An experimental study of the interaction of two identical vesicles in shear flow shows that there is a net repulsion between the cells that leads to an increase of the distance between vesicles in a suspension. Scaling arguments are proposed for this interaction and a comparison with numerical results is performed and a quantitative estimation of a shear induced diffusion coefficient obtained by averaging the results for pair interactions is found. Finally, we investigate the diffusion of a cloud of red blood cells in Poiseuille flow in order to directly determine diffusion coefficients. The experiment shows that the cloud widens when traveling along the channel with a power law behaviour indicating sub-diffusion. This effect is confirmed by a theoretical analysis of a few limit cases.
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Simulation de globules rouges modèles et analyse analytique de modèles de suspensions très concentrées / Simulation models of red blood cells in microcirculation, and analytical model analysis of highly concentrated suspensionsTahiri, Najim 11 October 2013 (has links)
L'objectif principal de cette thèse est consacré à l'étude de la dynamique et la rhéologie d'une suspension de particules denses qui se comportent comme des fluides complexes. La premier partie de cette thèse est consacrée à l'étude de la déformation, le comportement dynamique et la rhéologie d'une suspension de vésicule (un modèle simple pour les globules rouges) sous l'action d'un écoulement externe appliqué (cisaillement simple et Poiseuille confiné) dans la limite de faible nombre de Reynolds. L'étude basée sur des simulations numériques en utilisant la méthode des intégrales de frontière. Cette étude est inspirée par le comportement des globules rouges dans le système microvasculaire. Notre étude est ensuite consacrée aux effets du confinement et du nombre capillaire sur la forme, le comportement dynamique et la viscosité effective d'une suspension de vésicules. Nous avons montré que pour des membranes rigides (nombre capillaire petit), on peut observer en plus de la forme parachute et pantoufle, les formes suivantes : (i) forme d'oscillation centrée, (ii) forme d'oscillation décentrée et (iii) la forme cacahuète. Egalement, nous avons examiné l'influence du contraste de viscosité sur la dynamique et la rhéologie d'une vésicule. Nous avons montré qu'il existe une phase de "coexistence" entre la forme pantoufle et la forme parachute. Dans la deuxième partie de cette thèse, nous avons proposé un modèle analytique et une étude numérique pour étudier les propriétés dynamiques et rhéologiques d'une suspension de particules rigides sous écoulement de Poiseuille confiné. Le débit, la dissipation et la viscosité apparente sont étudiés en fonction de la structure des plaques dans le canal. Egalement, l'étude numérique d'une suspension de particules sphériques (formes des chaînes de particules) est en accord qualitatif avec le modèle analytique qui considère les longues plaques. Cette étude numérique est basée sur une méthode de la dynamique des particules du fluide, où les particules sont représentées par un champ scalaire ayant une viscosité élevée à l'intérieur. / The principal objective of this thesis is devoted to study the dynamics and rheology of a suspension of dense particles which behave as complex fluids. The first part of this thesis is dedicated to the study of the deformation, the dynamic behavior and rheology of a single neutrally buoyant suspended vesicle (a closed phospholipid membrane), as a response to external applied flows (simple shear and confined Poiseuille flows), is studied in the limit of small Reynolds. The study based on numerical simulations using the boundary integral method. This study is inspired by the behavior of red blood cells in the microvasculature. Our study is then dedicated on the effects of confinement and capillary number on the shape, the dynamic behavior and the effective viscosity of a vesicles suspension. We have shown that for rigid membranes (small capillary number) may be exist other shapes parachute and slipper, these shapes are : (i) Centered Snaking, (ii) Off-Centered Snaking and (iii) Peanut-shaped. Equally, we examined the influence of viscosity contrast on the dynamics and rheology of a vesicle. We have shown that there is a phase of "coexistence" between the slipper and parachute shape. The coexistence of shapes seems to be supported by experiments, though a systematic experimental study is lacking. In the second part of this thesis, we proposed an analytical model and a numerical study for study the dynamics and rheology properties of a rigid particles suspended in a confined Poiseuille flow. The flow rate, the dissipation and the apparent viscosity are studied as a function of the underlying structure. Equally, the numerical study is in good qualitative agreement with the analytical theory considering long plates. The numerical study is based on a fluid dynamic particle method where the particles are represented by a scalar field having high viscosity inside.
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Thermal Actuation and Fluidic Characterization of a Fluorescence-Based Multiplexed Detection SystemJanuary 2018 (has links)
abstract: This work describes efforts made toward the development of a compact, quantitative fluorescence-based multiplexed detection platform for point-of-care diagnostics. This includes the development of a microfluidic delivery and actuation system for multistep detection assays. Early detection of infectious diseases requires high sensitivity dependent on the precise actuation of fluids.
Methods of fluid actuation were explored to allow delayed delivery of fluidic reagents in multistep detection lateral flow assays (LFAs). Certain hydrophobic materials such as wax were successfully implemented in the LFA with the use of precision dispensed valves. Sublimating materials such as naphthalene were also characterized along with the implementation of a heating system for precision printing of the valves.
Various techniques of blood fractionation were also investigated and this work demonstrates successful blood fractionation in an LFA. The fluid flow of reagents was also characterized and validated with the use of mathematical models and multiphysics modeling software. Lastly intuitive, user-friendly mobile and desktop applications were developed to interface the underlying Arduino software. The work advances the development of a system which successfully integrates all components of fluid separation and delivery along with highly sensitive detection and a user-friendly interface; the system will ultimately provide clinically significant diagnostics in a of point-of-care device. / Dissertation/Thesis / Masters Thesis Biomedical Engineering 2018
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Controle das características geométricas de nanopartículas de prata através da conformação temporal de pulsos ultracurtos utilizando algorítimos genéticos / Control of the geometric characteristics of silver nanoparticles by ultrashort pulses temporal shaping using genetic algorithmsThiago da Silva Cordeiro 12 August 2013 (has links)
Este trabalho utilizou pulsos laser ultracurtos para modificar, de forma controlada, as características dimensionais de nanopartículas de prata em solução aquosa. Para atingir este objetivo foram empregados algoritmos genéticos e circuitos microfluídicos. Utilizou-se um conformador temporal de pulsos ultracurtos para criar diversos perfis temporais de pulsos que irradiaram soluções de nanopartículas de prata. Estes perfis temporais foram ajustados em tempo real, visando otimizar o resultado do experimento, quantificada pela diminuição do diâmetro médio das nanopartículas nas soluções irradiadas. Uma vez que cada experimento de minimização do diâmetro das nanopartículas exigiu centenas de medidas, sua realização foi possível em decorrência da utilização de um circuito microfluídico construído especialmente para este trabalho. Neste circuito é possível utilizar pequenas quantidades de amostra, levando a curtos tempos de irradiação e medição, além da evidente economia de amostras. Para a realização deste trabalho foi elaborado e testado um algoritmo genético interfaceado a diversos equipamentos, incluindo um filtro acustóptico dispersivo programável que modifica as características temporais dos pulsos ultracurtos, através da introdução de componentes de fases espectrais nestes pulsos. Utilizando o algoritmo genético e o filtro acustóptico dispersivo programável foram realizados experimentos de encurtamento da duração temporal dos pulsos ultracurtos provenientes do sistema laser, resultando na obtenção de pulsos com durações próximas às limitadas por transformada de Fourier. Além disso, foram realizados experimentos para a otimização do processo evolutivo do algoritmo genético escrito em Labview. Os experimentos de irradiação de soluções de nanopartículas de prata mostraram que, ao conformar a duração dos pulsos utilizados nas irradiações, pôde-se controlar as dimensões destas nanopartículas, diminuindo seu tamanho médio por um fator 2. Esses experimentos caracterizam a irradiação de nanopartículas por lasers de pulsos ultracurtos como uma importante técnica de controle de características de nanopartículas. / This work used ultrashort laser pulses to modify, in a controlled way, the dimensional characteristics of silver nanoparticles in aqueous solution. To reach this goal, genetic algorithm and microfluidic circuits were used. A pulse shaper was used to create different temporal profiles for the ultrashort pulses used to irradiate the silver nanoparticle solutions. These temporal profiles were conformed in real time, aiming to optimize the experiment result, quantified by the decrease of the average diameter of the nanoparticles in the irradiated solutions. Since each nanoparticle diameter minimization experiment demanded hundreds of measurements, its achievement was possible by the use of a microfluidic circuit specially built for this work. This circuit enables the use of small sample quantities, leading to short irradiation and measurement intervals, besides evident sample savings. To make this work possible, a genetic algorithm was created and tested. This genetic algorithm was interfaced to several equipments, including an acustooptic programmable dispersive filter that modifies the ultrashort pulses temporal characteristics by the introduction of spectral phases in the pulses. The genetic algorithm and the acustooptic programmable dispersive filter were used in conjunction in experiments to temporally shorten the ultrashort pulses from the laser system, generating pulses durations close to the Fourier transform limited ones. Besides, experiments were performed with the Labview coded genetic algorithm to optimize its evolutionary process. The silver nanoparticles irradiation experiments showed that the ultrashort pulses temporal conformation allowed the control of these particles dimensions, decreasing its mean size by a factor of 2. These experiments characterize the nanoparticles irradiation by ultrashort pulses as an important technique to control the nanoparticles characteristics.
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Desenvolvimento de uma técnica para seleção de espermatozoides em amostra seminal não processada para utilização na injeção intracitoplasmática de espermatozoides / Development of a technique for selection of spermatozoa for use in intracytoplasmic sperm injection without previous semen processingHamilton de Martim 27 June 2017 (has links)
Durante a injeção intracitoplasmática de espermatozoides (ICSI) seleciona-se um espermatozoide móvel e morfologicamente normal para injeção em ovócito maduro. Evidências recentes indicam que mesmo espermatozoides aparentemente normais podem ocultar defeitos em nível molecular. O principal objetivo deste trabalho foi a descrição de um novo método capaz de selecionar espermatozoides maduros a partir de amostras não processadas em procedimento de ICSI. Em um estudo comparativo e prospectivo testou-se uma gota estendida modificada. A \"gota estendida com mecanismo contracorrente\" - GEMC foi montada em uma placa de ICSI padrão a partir de seis gotas de meio de cultivo (10 ?L). O posicionamento e união precisa das gotas deram origem a dois reservatórios e um canal, resultando em um fluxo de líquido através do canal. A adição de uma solução de PVP (polivinilpirrolidona) gerou um gradiente de viscosidade no final do circuito. Amostras seminais foram obtidas de 40 pacientes inférteis. Cada amostra seminal foi dividida em 4 alíquotas: uma alíquota para o processamento por centrifugação em gradiente de densidade (CGD), uma alíquota para a GEMC utilizando amostras não processadas, uma alíquota para a GEMC utilizando amostras processadas e uma alíquota para controle. Nos grupos GEMC uma média de 200 espermatozoides foram consecutivamente coletados, sem seleção, no reservatório de captura utilizando-se uma micropipeta de injeção como no procedimento convencional de ICSI. A morfologia espermática foi avaliada e demonstrou melhora, comparando-se com os controles, em todos os tratamentos utilizados. A imaturidade da cromatina foi avaliada utilizando-se o teste do azul de anilina. Em relação à imaturidade, 100% dos homens obtiveram melhores resultados tanto após o preparo por CGD quanto utilizando o método GEMC. Isto se refletiu em uma redução das formas imaturas de 28.65 ± 8.97% no sêmen fresco para 17.29 ± 7.72% após processamento por CGD (P < 0.01). Uma redução ainda maior nas formas imaturas foi obtida após o método GEMC quando comparado com o processamento por CGD: 0.89 ± 1.31% (P < 0.01) utilizando-se sêmen fresco e 1.05 ± 1.63% (P < 0.01) utilizando-se amostras processadas. Um novo método unindo seleção e captura de espermatozoides em um mesmo procedimento de ICSI foi descrito e testado. O método GEMC seleciona espermatozoides de forma fácil e permite o uso direto de amostras não processadas em procedimentos de ICSI. Este método seleciona espermatozoides maduros com mais eficiência do que o processamento por CGD. Novos estudos são necessários para se analisar o impacto do método nas taxas de fertilização, desenvolvimento embrionário, gravidez, implantação e abortamento / During intracytoplasmic sperm injection (ICSI) a motile spermatozoon with normal morphology is visually selected for insemination of an oocyte. Recent evidence indicates that even though the sperm appears morphologically normal, a possibility of defects at the molecular level still exists. The main objective of this work was to describe a novel approach capable of selecting mature spermatozoa from unprocessed semen sample in a one-step ICSI procedure. A modified extended drop was tested in a prospective comparative study. The \"Gota Estendida com Mecanismo Contracorrente - GEMC\" (Positive Rheotaxis Extended Drop - PRED) was assembled on a standard ICSI dish and consisted of six culture medium droplets (10 ?L). The precise merging of the drops created two reservoirs and a channel therefore the fluid flew through the channel. The addition of a PVP solution created a viscosity gradient in the final sector of the circuit. Semen samples were taken from 40 subfertile men. Each semen sample was divided into four aliquots: one aliquot for density gradient centrifugation (DGC), one aliquot for GEMC using fresh semen, one aliquot for GEMC using processed semen and one aliquot for the control. In GEMC a mean of 200 spermatozoa were collected consecutively, without selection, from the outlet reservoir with an injecting pipette as for conventional ICSI procedure. Sperm morphology was assessed and resulted in improvement compared to controls in all treatments. Chromatin immaturity was assessed using aniline blue assay. Regarding to chromatin immaturity, 100% of men had better results after DGC preparation and GEMC approach. This was reflected in a mean reduction from 28.65 ± 8.97% uncondensed chromatin in the native ejaculates to 17.29 ± 7.72% in DGC processed semen (P < 0.01). An even greater reduction was achieved after GEMC approach showing a mean of 0.89 ± 1.31% uncondensed chromatin compared to DGC processed sample (P < 0.01). A novel one-step ICSI approach joining sperm selection and recovery was developed and tested. This GEMC approach can select sperm easily and permits the direct use of native semen in ICSI. This approach can select sperm with lower chromatin immaturity than DGC method. Further studies need to access its relation to fertilization, embryo development, pregnancy, implantation and miscarriage rates
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