O objeto de estudo deste trabalho é caracterizar um transdutor magnético de posição absoluto de longo percurso e robusto quanto à sua resposta dinâmica. O transdutor foi concebido na dissertação de mestrado do autor e denominada de ¨Régua Magnética¨na qual foi estudada a sua resposta estática. Diferentemente de outros transdutores magnéticos este tem como princípio de funcionamento a divisão do fluxo magnético entre dois caminhos distintos, cujas dimensões variam em função da movimentação da bobinas de excitação. Neste estudo analisa-se então a resposta dinâmica do transdutor, pois, quando o seu carro se desloca com velocidade constante, há uma modificação na grandeza física que é responsável pelo sinal de saída correlacionado à posição daquele carro. O estudo foi desenvolvido em dois transdutores de aproximadamente 2 m de comprimento confeccionados com lâminas de Fe-Si cortadas a laser. Os sinais de tensão induzida nas culatras, corrente e fluxo magnético no eletro-ímã do carro bem como o sinal de posição deste são coletados por um dispositivo digital e processados em um computador de modo a se poder obter as curvas estáticas e dinâmicas e assim compará-las. Com este propósito, foi desenvolvida uma bancada original de ensaio de transdutores de posição lineares para a medição da diferença dos sinais entre o caso estático e o dinâmico, denominado de diferença dinâmica, na qual foi empregado um transdutor de posição resistivo para a referência de posição, um sistema de aquisição de sinais para a medição das grandezas envolvidas no processo e um conjunto movimentador e servomotor para controle da velocidade e posição da parte móvel do transdutor. No final deste trabalho são comparados os resultados analíticos e experimentais obtidos como antes descritos. / The aim of this work is to characterize a long-span absolute magnetic position transducer that presents a robust dynamic response. The transducer was conceived by the MSc studies of the author and is called named “Magnetic Ruler” by which its static response was studied. Unlike other magnetic transducers, the magnetic ruler is based on the principle of the division of magnetic flux between two distinct magnetic paths that depends on the linear position of its movable part along the span of the transducer. This present study examines the dynamic response of the transducer when its movable part or car is moving with constant speed. Under such conditions, there is a change in the signal output that is correlated to the position of that car. The study was conducted by means of two transducers of approximately 2-m long laminations of Si-Fe that were laser cut. The voltage signals induced through the search coils on the end yokes, current and magnetic flux in the excitation coil on car, and the position are acquired by a digital device and processed by a computer. That is done to obtain the static and the dynamic curves and, thus, to allow the comparison between the two behaviors. With this objective, a specially built and unique test bench was developed to test linear position transducers under static and dynamic cases. The difference between the two results is called dynamic difference. That test bench employs a resistive position transducer to provide position reference, a signal acquisition system for measuring the quantities involved in the process, and a controlled traction system based on a servo system to control the speed and the position of the moving part of the transducer under test. At the end this work the analytical and experimental results obtained as described above are compared.

O objeto de estudo deste trabalho é caracterizar um transdutor magnético de posição absoluto de longo percurso e robusto quanto à sua resposta dinâmica. O transdutor foi concebido na dissertação de mestrado do autor e denominada de ¨Régua Magnética¨na qual foi estudada a sua resposta estática. Diferentemente de outros transdutores magnéticos este tem como princípio de funcionamento a divisão do fluxo magnético entre dois caminhos distintos, cujas dimensões variam em função da movimentação da bobinas de excitação. Neste estudo analisa-se então a resposta dinâmica do transdutor, pois, quando o seu carro se desloca com velocidade constante, há uma modificação na grandeza física que é responsável pelo sinal de saída correlacionado à posição daquele carro. O estudo foi desenvolvido em dois transdutores de aproximadamente 2 m de comprimento confeccionados com lâminas de Fe-Si cortadas a laser. Os sinais de tensão induzida nas culatras, corrente e fluxo magnético no eletro-ímã do carro bem como o sinal de posição deste são coletados por um dispositivo digital e processados em um computador de modo a se poder obter as curvas estáticas e dinâmicas e assim compará-las. Com este propósito, foi desenvolvida uma bancada original de ensaio de transdutores de posição lineares para a medição da diferença dos sinais entre o caso estático e o dinâmico, denominado de diferença dinâmica, na qual foi empregado um transdutor de posição resistivo para a referência de posição, um sistema de aquisição de sinais para a medição das grandezas envolvidas no processo e um conjunto movimentador e servomotor para controle da velocidade e posição da parte móvel do transdutor. No final deste trabalho são comparados os resultados analíticos e experimentais obtidos como antes descritos. / The aim of this work is to characterize a long-span absolute magnetic position transducer that presents a robust dynamic response. The transducer was conceived by the MSc studies of the author and is called named “Magnetic Ruler” by which its static response was studied. Unlike other magnetic transducers, the magnetic ruler is based on the principle of the division of magnetic flux between two distinct magnetic paths that depends on the linear position of its movable part along the span of the transducer. This present study examines the dynamic response of the transducer when its movable part or car is moving with constant speed. Under such conditions, there is a change in the signal output that is correlated to the position of that car. The study was conducted by means of two transducers of approximately 2-m long laminations of Si-Fe that were laser cut. The voltage signals induced through the search coils on the end yokes, current and magnetic flux in the excitation coil on car, and the position are acquired by a digital device and processed by a computer. That is done to obtain the static and the dynamic curves and, thus, to allow the comparison between the two behaviors. With this objective, a specially built and unique test bench was developed to test linear position transducers under static and dynamic cases. The difference between the two results is called dynamic difference. That test bench employs a resistive position transducer to provide position reference, a signal acquisition system for measuring the quantities involved in the process, and a controlled traction system based on a servo system to control the speed and the position of the moving part of the transducer under test. At the end this work the analytical and experimental results obtained as described above are compared.

This master's thesis deals with the issue of temperature measuring with the use of contactless thermometer. The thesis examines the physical findings, methods and detectors for contactless temperature measurement. Is then selected a suitable detector, measuring methods and assembled a block diagram of the IR thermometer. The main section is a specific system design. This includes circuit solution, including printed circuit board design and simulation of the functional parts.

These master‘s thesis deals with the issues of contactless temperature measurement, designe and construction of the infrared contactless thermometer. The thesis is concerned with the IR physical magnitudes and laws, methods of measurment and types of detectors, which are used for contactless temperature measurment. In the practical part of the thesis is created system design and block diagram of the IR termometer. In the next part of the thesis is presented system design, drawing of the printed circuit board and software processing of the IR thermometer. In conclusion are summerized our achieved resultes.

Microfluidics has found numerous applications ranging from the life sciences industries for pharmaceuticals and biomedicine (drug design, delivery and detection, diagnostic devices) to industrial applications of combinational synthesis (such as rapid analysis and high throughput screening). Among all these, one of the intriguing exploitation of microfluidics or micro total analysis systems (µTAS) is the separation of circulating tumor cells (CTCs) from body fluids. Cancer cells spread from the initial site of a tumor by first invading the surrounding tissue, then by entering the blood or lymph vessels, and finally by crossing the vessel wall to exit the vasculature into distal organs. The September 2006 issue of the Journal of the National Cancer Institute (NCI) states: "The war on cancer was declared 40 years ago and cancer is still here," and "Technologies that capture enemy CTCs for further interrogation might prove useful in the war on cancer." CTCs cannot only become a new marker for cancer prognosis, but their detection can also be a valid new parameter for diagnosing cancer early, for monitoring disease progression and relapse, and for optimizing therapy. This research established a new method to manipulate rare cell types based on their electrical signatures using dielectrophoresis (DEP) without having direct contact between the electrodes and the sample, known as contactless dielectrophoresis (cDEP). DEP is the motion of a particle in a suspending medium due to its polarization in the presence of a non-uniform electric field. cDEP relies upon reservoirs filled with highly conductive fluid to act as electrodes and provide the necessary electric field. These reservoirs are placed adjacent to the main microfluidic channel and are separated from the sample by a thin barrier of a dielectric material as is shown in Figure 1h. The application of a high-frequency electric field to the electrode reservoirs causes their capacitive coupling to the main channel and an electric field is induced across the sample fluid. Similar to traditional DEP, cDEP exploits the varying geometry of the electrodes to create spatial non-uniformities in the electric field. However, by utilizing reservoirs filled with a highly conductive solution, rather than a separate thin film array, the electrode structures employed by cDEP can be fabricated in the same step as the rest of the device; hence the process is conducive to mass production. We demonstrated the ability to isolate human leukemia cancer cells (THP-1) cells from a heterogeneous mixture of live and dead cells using cDEP with more than 99% selectivity and 95% removal efficiency. Through numerical and experimental investigations, new generation of cDEP devices have been designed and tested to detect and isolate THP-1 cells from spiked blood samples with high selectivity and cell capture efficiency. Our experimental observations, using prototype devices, indicate that breast cancer cell lines at their different stages (MCF-7, MCF-10, and MDA-MB231) have unique electrical. Furthermore, through collaborations at the Wake Forest Comprehensive Center, we demonstrated that prostate tumor initiating cells (TICs) exhibit unique electrical signatures and DEP responses and cDEP technology can be exploited to isolate and enrich TICs for further genetic pathways investigations. / Ph. D.

Dielectrophoresis (DEP), an electrokinetic force, is the motion of a polarizable particle in a non-uniform electric field. Contactless DEP (cDEP) is a recently developed cell sorting and isolation technique that uses the DEP force by capacitavely coupling the electrodes across the channel. The cDEP platform sorts cells based on intrinsic biophysical properties, is inexpensive, maintains a sterile environment by using disposable chips, is a rapid process with minimal sample preparation, and allows for immediate downstream recovery. This platform is highly competitive compared to other cell sorting techniques and is one of the only platforms to sort cells based on phenotype, allowing for the isolation of unique cell populations not possible in other systems. The original purpose of this work was to determine differences in the bioelectrical fingerprint between several critical cancer types. Results demonstrate a difference between Tumor Initiating Cells, Multiple Drug Resistant Cells, and their bulk populations for experiments conducted on three prostate cancer cell lines and treated and untreated MOSE cells. However, three significant issues confounded these experiments and challenged the use of the cDEP platform. The purpose of this work then became the development of solutions to these barriers and presenting a more commercializable cDEP platform. An improved analysis script was first developed that performs whole cell detection and cell tracking with an accuracy of 93.5%. Second, a loading system for doing smart sample handling, specifically cell focusing, was developed using a new in-house system and validated. Experimental results validated the model and showed that cells were successfully focused into a tight band in the middle of the channel. Finally, a proof of concept for an on-chip micropump is presented and achieved 4.5% in-plane deformation. When bonded over a microchannel, fluid flow was induced and measured. These solutions present a stronger, more versatile cDEP platform and make for a more competitive commercial product. However, these solutions are not just limited to the cDEP platform and may be applicable to multitudes of other microfluidic devices and applications. / Master of Science

The effect of inorganic electrolytes on the determination of relative permittivity and concentration of water solutions of 1,4 dioxane was studied in this work. The method used was dielectrimetry, and NaCl, KCl, CsCl and LiCl at various concentrations were tested electrolytes. The interfering effect of these electrolytes was found for concentrations above 0.05 mmol l-1. It was also found that the interfering factor is purely the solution conductivity independent on the ion kind. The measurements were carried out with both the planar and the tubular detection cell having the electrodes insulated from the solution. The cell was connected to the differentiator (the voltage is the analytical signal) or multivibrator (the frequency is the analytical signal). A good agreement between experimental results and the modeled calculation for the detector with differentiator was also obtained.

This thesis deals with optimization of capillary electrophoresis conditions for the separation of unreacted substrate and products (cellobiose, glucose) generated by the enzymatic cleavage of cellotriose with enzyme cellobiase. The optimal separation electrolyte was shown to be a solution containing 30 mmol·dm-3 sodium hydroxide with addition of 1.0 mmol·dm-3 sodium phosphate and 1.0 mmol·dm-3 sodium phosphate dibasic (pH = 12.38). With this separation electrolyte calibration curves and repeatability were measured and the limits of detection and quantification were determined. The relative standard deviations of peak areas ranged up to 6.1 % and they did not exceed 0.2 % for the migration time. Limits of detection were determined in the range from 0.020 to 0.026 mmol·dm-3 and limits of quantification from 0.066 mmol·dm-3 to 0.085 mmol·dm-3 . Subsequently the enzyme reaction was monitored in offline setup. Furthermore, the offline procedure was automated and the relative standard deviation of cellodextrines after 30 minutes of hydrolysis was ranging up to 13.0 % of glucose and cellotriose and 3.0 % cellobiose using this automated method, which makes it suitable for semiquantitative and comparative measurements. The last part of the experiment was to carry out enzymatic reaction in online setup....

Um grande crescimento no interesse por microdispositivos eletroforéticos tem sido observado nos últimos anos. Vantagens atrativas como baixo consumo de reagentes e diminutos tempos de analise são verificados. Microdispositivos fabricados usando toner de impressora e folhas de transparência (poliéster) vem a somar valores como baixo custo e simplicidade de confecção à técnica de micro-separação eletroforética. Entretanto, a aplicação analítica utilizando esse tipo de microdispositivo tem sido pouco explorada. <br />Neste trabalho o uso de microchips eletroforéticos fabricados em poliéster-toner (PT) é utilizado para a determinação simultânea do herbicida glifosato e seu maior metabólito, AMPA (ácido aminometilfosfônico). Em um primeiro momento, o trabalho apresenta o desenvolvimento de uma metodologia no qual utiliza condutometria sem contato (C4D) como sistema de detecção, aliada à separação eletroforética em microchips de PT. Vários parâmetros que regem uma boa confiabilidade analítica, tanto quanto otimizam a sensibilidade do sistema foram avaliados. <br />Em um segundo momento, ainda aliada à microchips de PT, a técnica de cronoamperometria foi utilizada como sistema de detecção. Nessa etapa do projeto estudos eletroquímicos foram inicialmente realizados em eletrodos convencionais de ouro e cobre, buscando averiguar qual metal apresenta maior sensibilidade para o herbicida glifosato. Tendo o metal cobre como melhor escolha, eletrodos planares foram construídos a partir da combinação das tecnologias da produção de máscaras de toner, e placas de circuito impresso como fonte de cobre. <br />Em ambas as metodologias desenvolvidas, picos bem resolvidos foram encontrados para os analitos em estudo. Tempos menores que 80 s foram gastos entre o processo de separação e detecção; uma boa repetibilidade também foi encontrada. Os valores de limite de detecção (LD) utilizando C4D foram 60,8 e 74,8 &micro;mol L-1 para glifosato e AMPA respectivamente. Como esperado, menores valores de LD foram obtidos na detecção amperométrica, com 1,88 &micro;mol L-1 para glifosato e 16,45 &micro;mol L-1 para AMPA. A aplicabilidade dos métodos foi verificada através da analise do herbicida e seu metabólito em amostras de água. Etapas de derivatização e pré-concentração off-line não foram usadas nesse trabalho, dessa forma os dois métodos desenvolvidos apresentaram como principais vantagens o extremo baixo custo, e a simplicidade de uso. / A large increase in interest in electrophoretic microdevices has been observed in recent years. Advantages attractive as low reagent consumption and low analytical time are checked. Microdevices fabricated using printer toner and polyester transparency sheets are the sum values as low cost and simplicity of the technique of making micro-electrophoretic separation. However, the analytical application using this type of microdevice has been little explored. <br />In this work the use of microchip electrophoresis fabricated on polyester-toner (PT) is used for the simultaneous determination of the herbicide glyphosate and its major metabolite, AMPA (aminomethylphosphonic acid). In a first moment, the work presents the development of a methodology in which the use capacitively coupled contactless conductivity detection (C4D) for determination of analytes is employed, coupled with electrophoretic separation in PT microchips. Several parameters that govern a good analytical reliability and with the intuit of optimizing the sensitivity of the system were evaluated. <br />In a second time and still allied to the PT microchips, the technique of chronoamperometry was used for detection. Electrochemical studies were initially conducted in conventional electrodes of gold and copper, looking for determine which metal is more sensitive to detect the herbicide glyphosate. Having the best choice planar copper electrodes were constructed from a combination of technologies for the production of toner masks, and printed circuit boards. <br />In both, C4D and chronoamperometric detection methodologies, well-resolved peaks were found for the glyphosate and AMPA. Time analysis of less than 80s were found including the separation and detection processes, and a good analytical repeatability was also found. The limits of detection (LOD) using C4D were 60,8 and 74,8 &micro;mol L-1 respectively for glyphosate and AMPA. As expected, lower LOD were obtained in the amperometric detection methodology, 1.9 &micro;mol L-1 for glyphosate, and 16.5 &micro;mol L-1 for AMPA. The applicability of the methods was tested by analyzing the herbicide and its metabolite in fortified water samples. Steps of derivatization and preconcentration off-line were not applied in this work, so the two methods showed as main advantages very low cost and time analysis, and simplicity of application.

Este trabalho descreve o desenvolvimento de um biossensor contendo transdução condutométrica sem contato (C4D, capacitively coupled contactless conductivity detection) e ácido fólico (FA) como biorreceptor em microchip, uma nova alternativa que poderá ser utilizada na determinação do biomarcador tumoral FR-&alpha;. Essa espécie exibe interações com FA altamente específicas, com constantes de formação da ordem de 109-1010. Os dispositivos microfluídicos, os quais consistiram de uma lâmina de vidro (integrando os eletrodos), dielétrico (contendo a fase biossensora) e substrato de poli(dimetilsiloxano) (PDMS, incorporando os microcanais), foram fabricados utilizando-se processos de fotolitografia e deposição de filmes finos em fase vapor. Objetivando melhorias nos níveis de detecção da C4D, estudos de sensibilidade com base em parâmetros da curva analítica foram conduzidos alterando-se a natureza do dielétrico e a configuração dos eletrodos. Posteriormente, estudos de caracterização foram realizados para as superfícies modificadas com os intermediários de imobilização; condições reacionais distintas (reagente, concentração, solvente e tempo) foram consideradas. As técnicas de microscopia eletrônica de varredura e espectroscopia de fotoelétrons excitados por raios-X foram usadas, respectivamente, a fim de se verificar a possível formação de aglomerados e permitir determinações qualitativas e quantitativas sobre as composições químicas das superfícies. Como resultado dos experimentos de sensibilidade e caracterização de superfície, adotamos os parâmetros seguintes para os ensaios de interações biomoleculares posteriores: filme de SiO2 como dielétrico, eletrodos seletivos à C4D com formato retangular e orientação antiparalela e monocamadas automontadas do reagente 3-aminopropil(trietoxisilano) como intermediário de imobilização de FA. As duas etapas finais do trabalho foram: otimização do tempo de funcionalização com FA (3, 5 e 7 h) e caracterização da fase biossensora, realizada a partir de medidas de C4D e microscopia de força atômica (AFM). Para o primeiro caso, os microchips foram aplicados a um padrão de anticorpo monoclonal específico a FA (&alpha;-FA). Os ensaios biomoleculares indicaram uma adsorção efetiva de FA junto à superfície de SiO2 silanizada, sem a ocorrência (ao menos em níveis significativos) de impedimentos estéricos de sua espécie bioativa. Dentre os tempos de funcionalização investigados, 3 h foi aquele que resultou em uma maior sensibilidade do método. Em termos da etapa de caracterização eletroquímica da fase biossensora, seus resultados mostraram haver correlação entre a resposta analítica e as interações FA/&alpha;-FA. Em adição, conforme indicaram as medidas de AFM, não houve alterações drásticas na morfologia do substrato (SiO2) em função dos processos de modificação química de superfície. Por fim, o uso da C4D como uma técnica de transdução em biossensores mostrou-se uma alternativa promissora para a análise do biomarcador tumoral FR-&alpha;. Dentre outros aspectos, essa plataforma analítica requer uma instrumentação simples, barata e portátil, não apresenta inconvenientes relacionados ao contato eletrodo/solução, dispensa o uso de mediadores redox e permite a determinação simultânea de multianalitos. Neste ínterim, alterações no transdutor devem ser implementadas visando um aumento na sensibilidade do método, o qual representa seu fator limitante principal. / This work describes the development of a biosensor containing capacitively coupled contactless conductivity transduction (C4D) and folic acid (FA) as bioreceptor in microchip, a new alternative that can be used in FR-&alpha; tumor biomarker analysis. FR-&alpha; exhibits highly specific interactions with FA, showing formation constants of the order of 109-1010. The microfluidic devices consisted of a glass layer (integrating the electrodes), dielectric (containing the biosensor phase), and poly(dimetilsiloxane) substrate (PDMS, incorporating microchannel). The microfabrication stage evolved photolithography processes, metal adsorption via sputtering, and plasma-enhanced vapor film deposition. In order to improve detection levels of C4D, sensitivity studies were conducted by changing the dielectric nature and electrode configuration. Through flow analysis with given electrolyte standards, the limits of detection and quantification were calculated based on analytical curve parameters. Subsequently, researches were performed to characterize the modified surfaces with immobilization intermediate considering reaction conditions distinct (reagent, concentration, solvent, and time). The techniques of scanning electron microscopy and X-ray photoelectron spectroscopy were employed, respectively, aiming to verify the clusters formation and allow qualitative and quantitative determinations about the surfaces chemical composition. From the results of sensitivity experiments and surface characterization, we adopt the following parameters for the biomolecular interactions assays: SiO2 film as dielectric, C4D selective electrodes with rectangular shape and antiparallel orientation, and self-assembled monolayers of 3-aminopropyl(triethoxysilane) as intermediary for immobilization of FA. The two final steps of the work were: optimizing the FA functionalization time (3, 5, and 7 h) and phase biosensor characterization, made from measures of C4D and atomic force microscopy (AFM). For the first case, due to the absence of FR-&alpha; standard for purchase, the microchips were applied to FA specific monoclonal antibody (&alpha;-FA). The biomolecular assay indicated effective adsorption of FA, without occurrence (at least in significant levels) of steric hindrance of its bioactive specie. Among the investigated times of functionalization, 3 h resulted in a higher sensitivity of the method. In terms of biosensor phase electrochemical characterization stage, their results evidenced correlation between analytical response and FA/&alpha;-FA interactions. Additionally, as the AFM measurements showed, drastic changes in the morphology of the substrate (SiO2) with the surface modification processes did not occur. Finally, the use of the C4D as transduction technical in biosensors proved to be a promissory alternative for FR-&alpha; tumor biomarker analysis. Among other features, this platform has not drawbacks related to the electrode/solution contact, dispenses the use of redox mediators, allows the simultaneous determination of multianalytes, and employs an instrumentation that is simple, cheap, and portable. Nevertheless, changes in the transducer should be implemented to increase the method sensitivity, which represents its main limiting factor.