No presente trabalho, microeletrodos de platina foram fabricados e empregados juntamente com a técnica de Microscopia Eletroquímica de Varredura (Scanning Electrochemical Microscopy - SECM) para a obtenção de informações a respeito da erosão ácida do esmalte dentário. Em uma etapa inicial do projeto, um procedimento de fabricação de microeletrodos utilizando o equipamento Micropipette Puller foi desenvolvido. A selagem e o polimento da área ativa dos dispositivos foram consideradas etapas críticas do processo de construção. Uma bomba a vácuo foi empregada durante a fabricação dos dispositivos, o que aumentou a eficiência da selagem do vidro sobre a microfibra. Os parâmetros do equipamento necessários à fabricação dos dispositivos foram otimizados e desta forma obteve-se uma metodologia reprodutível e eficiente. Assim, microeletrodos de diversos materiais e dimensões foram construídos e avaliados quanto à utilização em SECM. Uma metodologia alternativa para a redução no tamanho da ponta de prova foi proposta. Um polimento cuidadoso na ponta de prova foi feito e através de comparações entre Curvas de Aproximação teóricas e experimentais, chegou-se a microeletrodos com RG (razão raio do vidro e raio do microeletrodo) próximos de 15. Em uma segunda etapa do projeto, a erosão ácida do esmalte dentário foi investigada utilizando SECM. Empregando um microeletrodo de platina, foram obtidas imagens SECM que revelaram haver um intenso consumo de íons hidrogênio na interface esmalte dentário-solução ácida. Isso ocorre devido à reação de desmineralização da camada protetora do dente, sendo comprovado que este processo é relativamente rápido. Imagens SECM também mostraram diferenças topográficas do esmalte dentário e, desta forma, após um intenso processo de desmineralização ácida, a SECM foi utilizada para estimar a espessura da camada de esmalte erodida. Com o auxílio das Curvas de Aproximação concluiu-se que, após 48 horas de exposição, a camada de esmalte erodida foi de 18 µm e após 96 horas a espessura da camada erodida aumentou para 25 µm. Microscopia de Força Atômica também foi utilizada para visualização das mudanças morfológicas após o esmalte permanecer em contato com diferentes bebidas de elevada acidez. Desta forma, pode-se concluir que em longo prazo o esmalte dentário pode ser removido devido ao contato com substâncias ácidas, expondo a dentina ao ambiente bucal e gerando problemas de hipersensibilidade dentária. / In this study, platinum microelectrodes were fabricated with a Micropipette Puller and used in Scanning Electrochemical Microscopy (SECM) to obtain information on the acid erosion of tooth enamel. The sealing and polishing of the active area of the fabricated devices are considered critical steps in the construction process, hence a vacuum pump was used to enhance the efficiency of the seal of the glass microfiber. The parameters of the equipment were optimized allowing a reproducible and efficient method to be developed. Thus, microelectrodes of various materials and dimensions were constructed and evaluated for use in SECM. An alternative method for reducing the size of the probe was also proposed. A careful polishing of the probe was done and by comparing theoretical and experimental approach curves, microelectrodes with RG values near to 15 were obtained. The acid erosion of dental enamel was investigated using SECM and a platinum microelectrode as a probe to H+ ions. The obtained images revealed a significant and relatively fast consumption of H+ ions at the interface enamel-acid solution, as a consequence of the demineralization reaction of the protective layer. SECM images also showed topographical differences with respect to the tooth enamel and after an exhaustive process of acid demineralization, the thickness of the layer of eroded enamel was estimated. By using approach curves it was concluded that the thickness of the eroded enamel layer was around 18 µm after 48 hours of exposure, this value being enhanced to 25 µm after 96 hours. Atomic Force Microscopy was also used to confirm the morphological changes after exposing the enamel to different beverages with high acidity. Accordingly, one can conclude that the acid erosion of enamel exposes the dentin to the oral environment and creates problems associated with dental hypersensitivity.

No presente trabalho, microeletrodos de platina foram fabricados e empregados juntamente com a técnica de Microscopia Eletroquímica de Varredura (Scanning Electrochemical Microscopy - SECM) para a obtenção de informações a respeito da erosão ácida do esmalte dentário. Em uma etapa inicial do projeto, um procedimento de fabricação de microeletrodos utilizando o equipamento Micropipette Puller foi desenvolvido. A selagem e o polimento da área ativa dos dispositivos foram consideradas etapas críticas do processo de construção. Uma bomba a vácuo foi empregada durante a fabricação dos dispositivos, o que aumentou a eficiência da selagem do vidro sobre a microfibra. Os parâmetros do equipamento necessários à fabricação dos dispositivos foram otimizados e desta forma obteve-se uma metodologia reprodutível e eficiente. Assim, microeletrodos de diversos materiais e dimensões foram construídos e avaliados quanto à utilização em SECM. Uma metodologia alternativa para a redução no tamanho da ponta de prova foi proposta. Um polimento cuidadoso na ponta de prova foi feito e através de comparações entre Curvas de Aproximação teóricas e experimentais, chegou-se a microeletrodos com RG (razão raio do vidro e raio do microeletrodo) próximos de 15. Em uma segunda etapa do projeto, a erosão ácida do esmalte dentário foi investigada utilizando SECM. Empregando um microeletrodo de platina, foram obtidas imagens SECM que revelaram haver um intenso consumo de íons hidrogênio na interface esmalte dentário-solução ácida. Isso ocorre devido à reação de desmineralização da camada protetora do dente, sendo comprovado que este processo é relativamente rápido. Imagens SECM também mostraram diferenças topográficas do esmalte dentário e, desta forma, após um intenso processo de desmineralização ácida, a SECM foi utilizada para estimar a espessura da camada de esmalte erodida. Com o auxílio das Curvas de Aproximação concluiu-se que, após 48 horas de exposição, a camada de esmalte erodida foi de 18 µm e após 96 horas a espessura da camada erodida aumentou para 25 µm. Microscopia de Força Atômica também foi utilizada para visualização das mudanças morfológicas após o esmalte permanecer em contato com diferentes bebidas de elevada acidez. Desta forma, pode-se concluir que em longo prazo o esmalte dentário pode ser removido devido ao contato com substâncias ácidas, expondo a dentina ao ambiente bucal e gerando problemas de hipersensibilidade dentária. / In this study, platinum microelectrodes were fabricated with a Micropipette Puller and used in Scanning Electrochemical Microscopy (SECM) to obtain information on the acid erosion of tooth enamel. The sealing and polishing of the active area of the fabricated devices are considered critical steps in the construction process, hence a vacuum pump was used to enhance the efficiency of the seal of the glass microfiber. The parameters of the equipment were optimized allowing a reproducible and efficient method to be developed. Thus, microelectrodes of various materials and dimensions were constructed and evaluated for use in SECM. An alternative method for reducing the size of the probe was also proposed. A careful polishing of the probe was done and by comparing theoretical and experimental approach curves, microelectrodes with RG values near to 15 were obtained. The acid erosion of dental enamel was investigated using SECM and a platinum microelectrode as a probe to H+ ions. The obtained images revealed a significant and relatively fast consumption of H+ ions at the interface enamel-acid solution, as a consequence of the demineralization reaction of the protective layer. SECM images also showed topographical differences with respect to the tooth enamel and after an exhaustive process of acid demineralization, the thickness of the layer of eroded enamel was estimated. By using approach curves it was concluded that the thickness of the eroded enamel layer was around 18 µm after 48 hours of exposure, this value being enhanced to 25 µm after 96 hours. Atomic Force Microscopy was also used to confirm the morphological changes after exposing the enamel to different beverages with high acidity. Accordingly, one can conclude that the acid erosion of enamel exposes the dentin to the oral environment and creates problems associated with dental hypersensitivity.

Future nuclear fission reactors (GEN IV) are designed to include fast breeder reactor technologies, which can accept transuranics (elements heavier than uranium) as fuel. This has the potential of being more fuel efficient but requires the closing of the nuclear fuel cycle: full recycling of existing and newly generated nuclear waste to extract uranium and transuranic elements which can be reused as fuel. In the UK a system being investigated is electrochemical pyroprocessing which uses molten LiCl-KCl eutectic (LKE), which aims to recover uranium by electrodeposition on an inert (steel) electrode and the transuranics by electrodeposition as alloys with an active metal electrode (bismuth, cadmium or aluminium). Of the three active metal candidates, aluminium has the best separation efficiency of actinides and lanthanides, which is important as lanthanides are neutron poisons and so are not to be extracted. The development of pyroprocessing requires fundamental understandings of electrochemical alloy formation, as well as on-line monitoring tools to ensure the reprocessing occurs safely and efficiently. To that end, this thesis investigates cerium-aluminium alloying (a non-radioactive model system for plutonium-aluminium) on macro- and microelectrodes to understand the limiting factors during the alloying reaction at each electrode scale and also the circumstances under which the Ce3+ concentration can be reliably determined for on-line monitoring. On a bulk aluminium macroelectrode one cerium-aluminium alloying reaction was observed. This reaction was kinetically limited by the phase change from cerium insertion into the aluminium, and resulted in lattice expansion and progressive roughening of the electrode surface. These factors made it difficult to reliably calculate the Ce3+ concentration. Li+ from the solution was also able to reduce and form alloys with aluminium, approximately 0.3 V more negative than the first cerium-aluminium alloying peak. Since lithium atoms are smaller than cerium, and there is an abundance of Li+ in the salt, lithium-aluminium alloy was found to form preferentially to cerium-aluminium alloy at these more negative potentials. By co-depositing Al3+ and Ce3+ together on a tungsten electrode which is inert under these conditions (it does not alloy), the kinetic barrier to alloy formation by cerium insertion was decreased, which is beneficial to studying the thermodynamics of alloying. Studies of pure aluminium plating and pure cerium plating showed each individual reaction was diffusion limited, with an increased contribution of convection to the mass transport at slow scan rates. Co-deposition on macroelectrodes with a low ratio of [CeCl3]:[AlCl3] showed only one cerium-aluminium alloying peak. The co-deposition currents, and ratio of oxidation peaks charges, showed that co-deposition was occurring with both species under diffusion control, resulting in an amorphous alloy with a Ce:Al ratio that smoothly varied with the [CeCl3]:[AlCl3] ratio. This was in contrast to the alloying behaviour of cerium with liquid bismuth, in which co-deposition occurred at specific ratios determined by the crystal phases that could be formed at the applied potentials, with higher co-deposition ratios being achieved at more negative potentials. Co-deposition on macroelectrodes with a high ratio of [CeCl 3]:[AlCl3] could result in up to five cerium-aluminium alloy peaks, corresponding to all five CexAly crystalline phases predicted by the phase diagram. This phase change from amorphous to crystalline was promoted by the high Ce:Al ratio in the amorphous alloy resulting from the high [CeCl3]:[AlCl3] ratio and by plating pure cerium on the surface, which could then insert into the alloy. Charge analysis of these peaks confirmed the expected stoichiometries of the crystal phase from these in-situ measurements which is important for rapid analysis, whereas all previous literature has relied on ex-situ techniques which cooled the alloy, possibly changing its composition and structure. In all circumstances of alloy formation on macroelectrodes, the rate of reduction of Ce3+ was time dependent and sensitive to convection. This significantly complicated analysis of the electrochemical signal, making it very difficult to reliably calculate the concentration of Ce3+, which is required for on-line monitoring. Co-deposition on in-house microfabricated tungsten microelectrodes resulted in steady state currents for both pure aluminium deposition and cerium-aluminium co-deposition (up to the beginning of lithium-aluminium alloying). Thus, unlike on macroelectrodes, the deposition rate occurred at the flux ratio of each species from solution and only one oxidation peak was observed corresponding to the amorphous cerium-aluminium phase, even at high [CeCl3]:[AlCl3] ratios. The steady state alloying current meant that calculating the Ce3+ concentration was relatively simple from co-deposition on microelectrodes. Co-deposition was highly beneficial for studying alloying, however to avoid the addition of Al3+ to the molten salt, in-house microfabricated thin film aluminium microelectrodes were also used to study alloying. Alloying on microfabricated thin film aluminium microelectrodes was hampered by the formation of a native aluminium oxide layer, which prevented cerium insertion into the aluminium. The oxide layer could be disrupted by reduction of lithium, which showed steady state currents (albeit with significant capacitance) could be achieved for alloying by cerium insertion. However, the full surface area of the microelectrode could not be attained and all microelectrodes lost their aluminium layer after multiple lithiation/de-lithiation cycles. These devices need further development to overcome the oxide layer, or prevent its formation, in order to study alloying in greater detail with aluminium microelectrodes to fully realise their advantages for sensing and monitoring in pyroprocessing.

The work within this thesis focuses on the study of alloy formation using an active liquid metal electrode for fundamental analysis and for the extraction and separation of the lanthanides and actinides in a pyroprocessing system. The electrochemical work herein is performed in a molten salt of lithium chloride and potassium chloride at its eutectic point (LKE). This salt is a likely candidate for pyroprocessing due to its relatively low melting point and resistance to degradation on exposure to high levels of radiation. The active electrode material under examination is bismuth due to its propensity to alloy with other elements, its relatively low melting point, high density and non-toxicity. The alloying processes studied are those of bismuth-lithium and bismuth-cerium. Lithium is the limiting reduction reaction defining the negative solvent limit in LKE. As a result, understanding the processes that would occur if the electrode were to be pushed to such negative potentials is of significant importance. Cerium is a commonly-used surrogate for plutonium, which is an element of relatively high concentration in waste nuclear fuel and is of significant interest to the nuclear international community in waste fuel recycling. This work examines the alloying processes in terms of which intermetallic compounds are formed and by what mechanisms. This is achieved through the use of co-deposition on a macro tungsten rod, employing a number of electrochemical techniques to extract pertinent information. Lithium electrodeposition and alloying with bismuth (at the negative solvent limit) was found to form BiLim alloy with increasing m at more reducing potentials, followed by the deposition of near pure lithium. Mixing of these two then gave rise to specific bismuth-lithium alloys and the apparent ejection of a lithium metal fog into the molten salt, which resulted in the chemical reduction of Bi3+ and the loss of the bismuth electrodeposition current. When electrodepositing cerium on, and alloying with, bismuth, the formation of intermetallic compounds is governed by potential with a maximum BiCem stoichiometry of m = 1 with equimolar Bi3+ and Ce3+. However, at concentrations of cerium greater than that of bismuth, alloys much richer in cerium were also deposited at more negative potentials. There is evidence that deposited cerium may also escape into solution and chemically react with Bi3+. In-house microelectrodes are also developed and used for this purpose, both through co-deposition and direct alloy formation on a liquid bismuth thin-film microelectrode. This work demonstrates that these devices provide a richness of information due to their highly beneficial microelectrode properties. A means of controllably depositing bismuth from an aqueous plating bath, without dendrite formation, on both platinum and tungsten microelectrodes was devised. This was followed by electrodeposition of bismuth films on these devices in LKE. Platinum was found to be an active electrode material, alloying with bismuth, while tungsten remained inert. Nonetheless, both electrode types produced characteristic microelectrode behaviour, which was successfully used to determine the diffusion coefficient of bismuth in LKE. A comparison of bismuth-cerium and cerium alloying on a thin film liquid bismuth microelectrode found that the latter indicated the formation of BiCe2 where only BiCe had been seen previously during co-deposition in an equivalent salt. This is thought to be due to the thin film liquid bismuth microelectrode configuration with enhanced Ce3+ mass transport. This response was also used to calculate the diffusion coefficient of cerium inside the bismuth film, which was found to be slightly slower than for Ce3+ in LKE.

Neste trabalho são apresentados resultados sobre o uso da voltametria com microeletrodos para o estudo de sistemas micelares. Estes estudos se basearam na obtenção de coeficientes de difusão de micelas em solução aquosa. Para tanto, o ferroceno foi utilizado como sonda eletroativa, haja vista que esta molécula é pouco solúvel em água e possui grande afinidade pelo interior micelar. A partir de experimentos envolvendo o tensoativo brometo de hexadecil trimetil amônio (C16TABr) pôde-se calcular parâmetros como o coeficiente de difusão do agregado na concentração micelar crítica e o coeficiente de interação intermicelar com a aplicação dos dados de coeficiente de difusão obtidos pela teoria de interação linear. O coeficiente de interação intermicelar permitiu a estimativa da concentração de eletrólito necessária para a mudança morfológica da micela (micela esférica para micela alongada). O raio hidronâmico da micela foi o calculado a partir da relação Stokes-Einstein. Um estudo complementar envolveu a adição de n-decanol à solução de tensoativo, e comparações dos coeficientes de difusão obtidos na presença e na ausência do álcool foram efetuadas. Estudos subseqüentes foram realizados empregando outros tensoativos (C14TABr, C12TABr e C10TABr) com a finalidade de verificar a dependência do coeficiente de difusão micelar com o comprimento da cauda hidrofóbica da molécula do tensoativo. A influência de outros parâmetros como o raio hidrodinâmico e a concentração de eletrólito necessária para a transição morfológica com o comprimento da cauda hidrofóbica também foi estudada. Em experimentos voltamétricos envolvendo a oxidação eletroquímica do ferroceno em meio de C16TABr e de SDS (dodecil sulfato de sódio) foi verificado que o sinal anódico ocorre em potenciais menos positivos em soluções de SDS, fato justificado pela interação da micela de SDS, negativamente carregada, com a superfície do eletrodo positivamente carregado. A investigação da interação do produto de oxidação eletroquímica do ferroceno, o íon ferrocínio, com as micelas de C16TABr foi realizada em experimentos envolvendo uma microestrutura geradora-coletora. Esta estrutura consiste em dois eletrodos de área similar separados por um canal de espessura micrométrica. A utilização deste dispositivo possibilitou a obtenção do coeficiente de difusão do ferrocínio e, a partir deste dado, foi possível fazer inferências sobre a afinidade deste íon pelo interior micelar. / Voltammetry with platinum disc microelectrodes was employed to gain information on diffusion coefficients, D, of micelles in aqueous solution. Owing to the low solubility in water and high affinity for the micellar core, ferrocene was used as electroactive probe. Diffusion coefficients of hexadecyl trimethyl ammonium bromide (C16TABr) were determined in solutions containing the surfactant in different concentrations and the data were fitted according to the linear interaction theory. This procedure allowed the diffusion coefficient at the critical micelle concentration to be found, as well as the intermicellar interaction parameter. The latter was used to estimate the minimum concentration of electrolyte that promotes morphological changes in the micelles (sphere to rod transition). The micellar hydrodynamic radius was calculated by using the Stokes-Einstein equation. A complementary study involved the addition of n-decanol to surfactant solutions and the diffusion coefficient was compared with the one obtained in the absence of the fatty alcohol. Experiments were also performed in solutions containing other surfactants (C14TABr, C12TABr e C10TABr) to investigate the dependence of D on the length of the hydrophobic tail. The influence of the hydrodynamic radii and supporting electrolyte concentration for the sphere to rod transition on the surfactant tail was also studied. A brief study on the electrochemical oxidation of ferrocene at two different micellar media (C16TABr and SDS sodium dodecyl sulfate) was done. The voltammetric experiments showed that the anodic process occurs at less positive potentials in solutions containing SDS and a likely explanation involves the electrostatic interaction between the micelle and the electrode surface. Experiments with a homemade twin-electrode thin-layer cell (TETLC) were performed in order to evaluate the interaction of ferrocinium, the product of ferrocene oxidation, with C16TABr micelles. The diffusion coefficient of ferrocinium was obtained and the affinity of this generated electroactive species with the micellar core was discussed.

Neste trabalho são apresentados resultados sobre o uso da voltametria com microeletrodos para o estudo de sistemas micelares. Estes estudos se basearam na obtenção de coeficientes de difusão de micelas em solução aquosa. Para tanto, o ferroceno foi utilizado como sonda eletroativa, haja vista que esta molécula é pouco solúvel em água e possui grande afinidade pelo interior micelar. A partir de experimentos envolvendo o tensoativo brometo de hexadecil trimetil amônio (C16TABr) pôde-se calcular parâmetros como o coeficiente de difusão do agregado na concentração micelar crítica e o coeficiente de interação intermicelar com a aplicação dos dados de coeficiente de difusão obtidos pela teoria de interação linear. O coeficiente de interação intermicelar permitiu a estimativa da concentração de eletrólito necessária para a mudança morfológica da micela (micela esférica para micela alongada). O raio hidronâmico da micela foi o calculado a partir da relação Stokes-Einstein. Um estudo complementar envolveu a adição de n-decanol à solução de tensoativo, e comparações dos coeficientes de difusão obtidos na presença e na ausência do álcool foram efetuadas. Estudos subseqüentes foram realizados empregando outros tensoativos (C14TABr, C12TABr e C10TABr) com a finalidade de verificar a dependência do coeficiente de difusão micelar com o comprimento da cauda hidrofóbica da molécula do tensoativo. A influência de outros parâmetros como o raio hidrodinâmico e a concentração de eletrólito necessária para a transição morfológica com o comprimento da cauda hidrofóbica também foi estudada. Em experimentos voltamétricos envolvendo a oxidação eletroquímica do ferroceno em meio de C16TABr e de SDS (dodecil sulfato de sódio) foi verificado que o sinal anódico ocorre em potenciais menos positivos em soluções de SDS, fato justificado pela interação da micela de SDS, negativamente carregada, com a superfície do eletrodo positivamente carregado. A investigação da interação do produto de oxidação eletroquímica do ferroceno, o íon ferrocínio, com as micelas de C16TABr foi realizada em experimentos envolvendo uma microestrutura geradora-coletora. Esta estrutura consiste em dois eletrodos de área similar separados por um canal de espessura micrométrica. A utilização deste dispositivo possibilitou a obtenção do coeficiente de difusão do ferrocínio e, a partir deste dado, foi possível fazer inferências sobre a afinidade deste íon pelo interior micelar. / Voltammetry with platinum disc microelectrodes was employed to gain information on diffusion coefficients, D, of micelles in aqueous solution. Owing to the low solubility in water and high affinity for the micellar core, ferrocene was used as electroactive probe. Diffusion coefficients of hexadecyl trimethyl ammonium bromide (C16TABr) were determined in solutions containing the surfactant in different concentrations and the data were fitted according to the linear interaction theory. This procedure allowed the diffusion coefficient at the critical micelle concentration to be found, as well as the intermicellar interaction parameter. The latter was used to estimate the minimum concentration of electrolyte that promotes morphological changes in the micelles (sphere to rod transition). The micellar hydrodynamic radius was calculated by using the Stokes-Einstein equation. A complementary study involved the addition of n-decanol to surfactant solutions and the diffusion coefficient was compared with the one obtained in the absence of the fatty alcohol. Experiments were also performed in solutions containing other surfactants (C14TABr, C12TABr e C10TABr) to investigate the dependence of D on the length of the hydrophobic tail. The influence of the hydrodynamic radii and supporting electrolyte concentration for the sphere to rod transition on the surfactant tail was also studied. A brief study on the electrochemical oxidation of ferrocene at two different micellar media (C16TABr and SDS sodium dodecyl sulfate) was done. The voltammetric experiments showed that the anodic process occurs at less positive potentials in solutions containing SDS and a likely explanation involves the electrostatic interaction between the micelle and the electrode surface. Experiments with a homemade twin-electrode thin-layer cell (TETLC) were performed in order to evaluate the interaction of ferrocinium, the product of ferrocene oxidation, with C16TABr micelles. The diffusion coefficient of ferrocinium was obtained and the affinity of this generated electroactive species with the micellar core was discussed.

This research is dedicated to develop novel batch fabrication procedures for two distinct AFM (Atomic Force Microscope) probes integrated with electrodes enabling electrical sample characterization and electrochemical sample surface profiling respectively. These AFM probes allow for highly accurate control of the probe positioning, low contact force and sample surface imaging with high lateral resolution. As an electrical characterization tool, a nickel micro four-point probe integrated with solid nickel tips was developed. Low electrical resistance of the probe and contact resistance were achieved due to the solid nickel cantilever and tips. Low aspect ratio solid metal tips reduced contact resistance resulting in stable electrical measurement. Conductivity loss easily experienced while using metal coated AFM cantilevers was overcome by solid nickel tip integration to the electrically conductive AFM cantilevers. The fabrication method introduces selective conical nickel tip etching in silicon dioxide etching chambers. A novel batch fabrication method for SECM-AFM (Scanning Electrochemical Microscope-Atomic Force Microscope) tip integrated with a ring electrode was developed as a tool for electrochemical imaging as well as topological imaging. The electroactive area at an exactly defined distance above the apex of the AFM tip is fabricated using an inverse silicon mold technique. The electrode at a deliberately chosen distance from the end of a scanning probe tip allowing electrochemical sample imaging separated from sample topology imaging. The ring electrode coated with polymer entrapping enzymes enabled the probe to detect ATP from living epithelial cells.

Urine culture, the current gold standard for urinary tract infection (UTI) diagnosis, does not produce results in an acceptable length of time. An ultra-sensitive, cost-effective electrochemical biosensing platform with nanostructured microelectrodes was designed to address the need for a rapid, point-of-care (PoC) test that could achieve a sample-to-answer time in less than an hour. Printed circuit boards and metallized glass slides were processed using various techniques and then tested for their ability to form nanostructured microelectrodes. Peptide nucleic acid probes for the bacteria and yeast as well as ten probes for antibiotic resistance genes were designed and synthesized for use with the new platform. Validation of the sensor's specificity was performed using high concentrations (100nM) of synthetic DNA oligomers. Furthermore, a clinically relevant sensitivity of 103 cfu/mL was demonstrated by detecting 4 pathogen lysates (Staphylococcus saprophyticus, Pseudomonas aeruginosa, Enterococcus faecalis and Klebsiella pneumoniae) in a buffered solution.

Urine culture, the current gold standard for urinary tract infection (UTI) diagnosis, does not produce results in an acceptable length of time. An ultra-sensitive, cost-effective electrochemical biosensing platform with nanostructured microelectrodes was designed to address the need for a rapid, point-of-care (PoC) test that could achieve a sample-to-answer time in less than an hour. Printed circuit boards and metallized glass slides were processed using various techniques and then tested for their ability to form nanostructured microelectrodes. Peptide nucleic acid probes for the bacteria and yeast as well as ten probes for antibiotic resistance genes were designed and synthesized for use with the new platform. Validation of the sensor's specificity was performed using high concentrations (100nM) of synthetic DNA oligomers. Furthermore, a clinically relevant sensitivity of 103 cfu/mL was demonstrated by detecting 4 pathogen lysates (Staphylococcus saprophyticus, Pseudomonas aeruginosa, Enterococcus faecalis and Klebsiella pneumoniae) in a buffered solution.

Willfully controlling the focus of an extracellular stimulus remains a significant challenge in the development of neural prosthetics and therapeutic devices. In part, this challenge is due to the vast set of complex interactions between the electric fields induced by the microelectrodes and the complex morphologies and dynamics of the neural tissue. Overcoming such issues to produce methodologies for targeted neural stimulation requires a system that is capable of (1) delivering precise, localized stimuli a function of the stimulating electrodes and (2) recording the locations and magnitudes of the resulting evoked responses a function of the cell geometry and membrane dynamics. In order to improve stimulus delivery, we developed microfabrication technologies that could specify the electrode geometry and electrical properties. Specifically, we developed a closed-loop electroplating strategy to monitor and control the morphology of surface coatings during deposition, and we implemented pulse-plating techniques as a means to produce robust, resilient microelectrodes that could withstand rigorous handling and harsh environments. In order to evaluate the responses evoked by these stimulating electrodes, we developed microscopy techniques and signal processing algorithms that could automatically identify and evaluate the electrical response of each individual neuron. Finally, by applying this simultaneous stimulation and optical recording system to the study of dissociated cortical cultures in multielectode arrays, we could evaluate the efficacy of excitatory and inhibitory waveforms. Although we found that the proximity of the electrode is a poor predictor of individual neural excitation thresholds, we have shown that it is possible to use inhibitory waveforms to globally reduce excitability in the vicinity of the electrode. Thus, the developed system was able to provide very high resolution insight into the complex set of interactions between the stimulating electrodes and populations of individual neurons.