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  • About
  • The Global ETD Search service is a free service for researchers to find electronic theses and dissertations. This service is provided by the Networked Digital Library of Theses and Dissertations.
    Our metadata is collected from universities around the world. If you manage a university/consortium/country archive and want to be added, details can be found on the NDLTD website.
101

Investigations into Hot-Tip Scanning Electrochemical Microscopy and Redox Flow Battery Applications

Zhao, Zhiling 29 August 2019 (has links)
No description available.
102

Electrochemical Studies of Nickel/Sulfuric Acid Oscillating Systems and the Preparation and Testing of Copper Coupled Microelectrode Array Sensors

Clark, David Quentin 12 August 2016 (has links)
The electrochemical behavior of nickel (Ni) in different concentrations of sulfuric acid (H2SO4) was studied via cyclic voltammetry (CV) over a range of potentials (0.0 V– 3.0 V) at room temperature. The presented work displays novel experiments where external forcing by a platinum (Pt) electrode changed the proton concentration at a Ni electrode surface in order to control the frequency and magnitude of periodic oscillations produced. When studying unique phenomena such as the Ni phenomena in this thesis, efficient, durable, and inexpensive technology is always beneficial. A coupled microelectrode array sensor or CMAS which has been used for over four decades to study pitting corrosion, crevice corrosion, intergranular corrosion, galvanic corrosion, and other heterogeneous electrochemical processes were fabricated in a novel, systematic, inexpensive, and time efficient process. The presented work shows how to make the CMAS and proved that they functioned properly.
103

Effects of Hyperosmotic Medium on Hepatocyte Volume, Transmembrane Potential and Intracellular K<sup>+</sup> Activity

Wang, Kening, Wondergem, Robert 04 November 1991 (has links)
Hepatocyte transmembrane potential (Vm) behaves as an osmometer and varies with changes in extracellular osmotic pressure created by altering the NaCl concentration in the external medium (Howard, L.D. and Wondergem, R. (1987) J. Membr. Biol. 100, 53). We now have demonstrated similar effects on Vm by increasing external osmolality with added sucrose and not altering ionic strength. We also have demonstrated that hyperosmotic stress-induced depolarization of Vm results from changes in membrane K+ conductance, gK, rather than from changes in the K+ equilibrium potential. Vm and aki of hepatocytes in liver slices were measured by conventional and ion-sensitive microelectrodes, respectively. Cell water vols. were estimated by differences in wet and dry weights of liver slices after 10-min incubations. Effect of hyperosmotic medium on membrane transference number for K+, tk, was measured by effects on Vm of step-changes in external [K+]. Hepatocyte Vm decreased 34, 52 and 54% when tissue was superfused with medium made hyperosmotic with added sucrose (50, 100 and 150 mM). Correspondingly, aKi increased 10, 18 and 29% with this hyperosmotic stress of added sucrose. Tissue water of 2.92 ± 0.10 kg H2O/kg dry weight in control solution decreased to 2.60 ± 0.05, 2.25 ± 0.06 and 2.22 ± 0.05 kg H2O/kg dry weight with additions to medium of 50, 100 and 150 mM sucrose, respectively. Adding 50 mM sucrose to medium decreased tK from 0.20 ± 0.01 to 0.05 ± 0.01. Depolarization by 50% with hyperosmotic stress (100 mM sucrose) also occurred in Cl-free medium where Cl- was substituted with gluconate. We conclude that hepatocytes shrink during hyperosmotic stress, and the aKi increases. The accompanying decrease in Vm is opposite to that expected by an increase in aKi, and at least in part results from a concomitant decrease in gK. Changes in membrane Cl- conductance most likely do not contribute to osmotic stress-induced depolarization, since equivalent decreases in Vm occurred with added sucrose in cells depleted of Cl- by superfusing tissue with Cl-free medium.
104

Electrochemical studies of external forcing of periodic oscillating systems and fabrication of coupled microelectrode array sensors

Clark, David 01 May 2020 (has links)
This dissertation describes the electrochemical behavior of nickel and iron that was studied in different acid solutions via linear sweep voltammetry, cyclic voltammetry, and potentiostatic measurements over a range of temperatures at specific potential ranges. The presented work displays novel experiments where a nickel electrode was heated locally with an inductive heating system, and a platinum (Pt) electrode was used to change the proton concentration at iron and nickel electrode surfaces to control the periodic oscillations (frequency and amplitude) produced and to gain a greater understanding of the systems (kinetics), oscillatory processes, and corrosion processes. Temperature pulse voltammetry, linear sweep voltammetry, and cyclic voltammetry were used for temperature calibration at different heating conditions. Several other metal systems (bismuth, lead, zinc, and silver) also produce periodic oscillations as corrosion occurs; however, creating these with pure metal electrodes is very expensive. In this work, metal systems were created via electrodeposition by using inexpensive, efficient, coupled microelectrode array sensors (CMASs) as a substrate. CMASs are integrated devices with multiple electrodes that are connected externally in a circuit in which all of the electrodes have the same amount of potential applied or current passing through them. CMASs have been used for many years to study different forms of corrosion (crevice corrosion, pitting corrosion, intergranular corrosion, and galvanic corrosion), and they are beneficial because they can simulate single electrodes of the same size. The presented work also demonstrates how to construct CMASs and shows that the unique phenomena of periodic oscillations that can be created and studied by using coated and bare copper CMASs. Furthermore, these systems can be controlled by implementing external forcing with a Pt electrode at the CMAS surface. The data from the single Ni electrode experiments and CMAS experiments were analyzed by using the Nonlinear Time-Series Analysis approach.
105

FABRICATION AND STUDY OF AC ELECTRO-OSMOTIC MICROPUMPS

Guo, Xin 07 May 2013 (has links)
In this thesis, microelectrode arrays of micropumps have been designed, fabricated and characterized for transporting microfluid by AC electro-osmosis (ACEO). In particular, the 3D stepped electrode design which shows superior performance to others in literature is adopted for making micropumps, and the performance of such devices has been studied and explored. A novel fabrication process has also been developed in the work, realizing 3D stepped electrodes on a flexible substrate, which is suitable for biomedical use, for example glaucoma implant. There are three major contributions to ACEO pumping in the work. First, a novel design of 3D “T-shaped” discrete electrode arrays was made using PolyMUMPs® process. The breakthrough of this work was discretizing the continuous 3D stepped electrodes which were commonly seen in the past research. The “T-shaped” electrodes did not only create ACEO flows on the top surfaces of electrodes but also along the side walls between separated electrodes. Secondly, four 3D stepped electrode arrays were designed, fabricated and tested. It was found from the experiment that PolyMUMPs® ACEO electrodes usually required a higher driving voltage than gold electrodes for operation. It was also noticed that a simulation based on the modified model taking into account the surface oxide of electrodes showed a better agreement with the experimental results. It thus demonstrated the possibility that the surface oxide of electrodes had impact on fluidic pumping. This methodology could also be applied to metal electrodes with a native oxide layer such as titanium and aluminum. Thirdly, a prototype of the ACEO pump with 3D stepped electrode arrays was first time realized on a flexible substrate using Kapton polyimide sheets and packaged with PDMS encapsulants. Comprehensive experimental testing was also conducted to evaluate the mechanical properties as well as the pumping performance. The experimental findings indicated that this fabrication process was a promising method to create flexible ACEO pumps that can be used as medical implants and wearable devices. / Thesis (Ph.D, Mechanical and Materials Engineering) -- Queen's University, 2013-05-06 10:57:48.077
106

Structuration de surfaces organiques et inorganiques par lithographie électro-colloïdale : principe et applications / Structuration of organic and inorganic surfaces by electrocolloidal lithography : principle and applications

Bazin, Damien 05 December 2012 (has links)
De nombreuses techniques de lithographie sont proposées aujourd'hui pour structurer des surfaces à l'échelle micrométrique et nanométrique. Parmi elles, la lithographie colloïdale est intéressante en raison notamment du faible coût du procédé. Dans cette thèse, nous avons développé une nouvelle technique appelée « lithographie électro-colloïdale » qui est basée sur l'utilisation de particules colloïdales soumises à des champs électriques continus et alternatifs. Avec des temps de préparation courts et une instrumentation peu coûteuse, des surfaces structurées polymériques et métalliques ont été produites puis testées pour différentes applications (immobilisation de protéines, réseaux de microélectrodes, surfaces superhydrophobes). / Many lithography techniques have been developed to structure surfaces at the micrometer and sub-micrometer ranges. Among them, colloidal lithography is interesting because the process is inexpensive and does not require the use complex instruments. In this thesis, we have developed a new technique called « electro-colloidal lithography » which is based on the use of colloidal particles organized using alternating and direct electric fields. With short preparation times and inexpensive instruments, polymeric and metallic structured surfaces have been prepared and tested for different applications (protein immobilization, microelectrode arrays, superhydrophobic surfaces)
107

Desenvolvimento de sensor eletroquímico para monitoramento de chumbo em resíduos de disparos de armas de fogo / Development of an electrochemical sensor for lead analysis in gunshot residues

Salles, Maiara Oliveira 26 August 2011 (has links)
Duas metodologias distintas foram desenvolvidas para a determinação de chumbo em Resíduos de Disparos de Armas de Fogo (RDAF) utilizando microeletrodo de ouro. Na primeira delas, a superfície eletródica foi modificada com filme de bismuto em meio alcalino (pH = 13) e na outra as análises foram realizadas com microeletrodo não modificado em meio ácido com cloreto. Os RDAF foram coletados do alvo atingido pela bala e das mãos do atirador. Dos alvos, foi recortada uma região próxima ao orifício feito pela bala. Já para remoção dos RDAF das mãos do atirador, foi utilizado um algodão embebido em solução de EDTA 2% (m/v). Os resíduos foram submetidos a um processo de extração que consistiu em adicionar 10 mL de solução de HNO3 10% (m/m) ao frasco contendo o alvo ou o algodão com os RDAF. Em seguida, a mistura foi submetida ao ultrassom por 2 horas a uma temperatura de 60°C. As amostras provenientes dos alvos foram analisadas com o microeletrodo de ouro modificado com bismuto em meio alcalino por voltametria de redissolução anódica. Com essa metodologia, obteve-se um limite de detecção (3&#963;/sensibilidade) de 12,5 nmol L-1, uma faixa linear entre 40 e 6700 nmol L-1 e um desvio padrão relativo de 3,4% (n = 12). Foi possível analisar amostras provenientes de duas armas diferentes (pistola 9 mm e revólver 0.38 polegadas) e três munições distintas (normal, spl® e CleanRange®). Constatou-se um aumento da quantidade de chumbo presente nos resíduos à medida que o atirador se aproxima do alvo e que os valores dependem da arma e munição utilizadas. Com a outra metodologia as análises foram realizadas na própria solução extratora, com adição de cloreto, sem qualquer modificação da superfície eletródica. Neste caso, o limite de detecção (3&#963;/sensibilidade) encontrado foi de 1,7 nmol L-1, a faixa linear ficou entre 10 e 100 nmol L-1 e o desvio padrão relativo de 10 medidas foi 2,5%. Foram analisados resíduos provenientes das mãos dos atiradores de quatro diferentes armas (revólver 0.38 polegadas, espingardas calibre 12, 22 e 38) e de seis diferentes munições (CleanRange®, normal, semi-enjaquetada, 3T®, Eley® e CBC®). Os resultados obtidos com a metodologia proposta foram comparados com aqueles oriundos de análise por espectroscopia de absorção atômica e houve concordância em um nível de confiança de 95%. Foi também realizada uma análise quimiométrica das soluções contendo os RDAF. Os resultados mostraram a possibilidade de discriminação em relação à arma e munição utilizadas. Os filmes formados nas superfícies eletródicas foram estudados utilizando o Microscópio Eletroquímico de Varredura (SECM), a Microbalança Eletroquímica de Cristal de Quartzo (MECQ) e o Microscópio Eletrônico de Varredura (MEV). A deposição dos filmes de bismuto, chumbo e bismuto/chumbo em meio alcalino ocorre de forma heterogênea e esses filmes reagem com oxigênio e ferricianeto. Em meio ácido, a presença do cloreto na solução de depósito do filme de chumbo provoca aumento na rugosidade do filme, com consequente melhoria no sinal analítico. Também se observou que o cloreto facilita o processo de redissolução do filme de chumbo. / Two different methodologies were developed to analyze lead from gunshot residues (GSR) with a gold microelectrode: modification of the electrode surface with a bismuth film in alkaline solution (pH = 13) and use of an unmodified gold microelectrode in acidic media with chloride. GSR were collected from cloth targets and shooters\' hand after test firing. From the targets, a region close to the bullet hole was cutted off. GSR were removed from the shooters\' hand using a cotton swab embedded in a 2% (m/v) EDTA solution. The target and the cotton swab were placed in a steril vial containing 10 mL of a 10% (m/m) HNO3 solution to extract the GSR. Then, all vials were placed in the ultrasound for 2 hours at 60°C. Samples from the targets were analyzed with a gold microelectrode modified with a bismuth film in alkaline solution by anodic stripping voltammetry with a detection limit of 12.5 nmol L-1 (3&#963;/slope), linear range between 40 and 6700 nmol L-1 and a relative standard deviation of 3.4% (n = 12). Two different handguns (pistoll 9 mm and a revolver 0.38 inches) and tree different ammunition (normal, spl® and CleanRange®) were analyzed and the lead amount was dependent on the gun and ammunition used in the tests. An increase in the amount of lead from the GSR was observed as the distance between the shooter and the target decreased. Lead analyses were also performed in the extraction solution with bare gold microelectrodes in the presence of chloride ions. In this case, the detection limit was 1.7 nmol L-1 (3&#963;/slope), the linear range lied between 10 and 100 nmol L-1 and the relative standard deviation for 10 measurements was 2.5%. GSR residues form four different guns (revolver 0.38 inches, 12 caliber pump, a repeating rifle 0.38 and a semi automatic .22 caliber rifle) and six different ammunitions (CleanRange®, normal, semi jacketed, 3T®, Eley® and CBC®) were analyzed. The results obtained with the proposed methodology were compared with those from atomic absorption spectroscopy analysis and the paired t-test student indicated that there was no significant difference between the results obtained with both methods at a 95% confidence level. A chemometric analysis was also performed and the results showed the possibility to discriminate the guns and the ammunitions used. Metallic flms formed onto the electrode surface were studied by using Scanning Electrochemial Microscopy (SECM), Electrochemical Quartz Crystal Microbalance (EQCM) and Scanning Electronic Microscopy (SEM). The deposition of bismuth, lead and bismuth/lead films in alkaline solution is heterogeneous and the films react with oxygen and ferricyanide. In acidic media, the presence of chloride ions in the depositing solution causes an increase in the film roughness and facilitates the lead film dissolution
108

Estudos eletroquímicos envolvendo diferente complexos de amin-rutênio e o ligante NO. Desenvolvimento de eletrodo modificado para análise de óxido nítrico / Electrochemical studies involving different amin-ruthenium complexes and NO ligand. Development of modified electrode for nitric oxide analysis

Mori, Vânia 19 May 2003 (has links)
Neste trabalho foram realizados estudos eletroquímicos com complexos de rutênio do tipo trans-[Ru(NH3)4L1L2]n+, aos quais se coordenam ligantes apropriados visando à modelagem de estruturas capazes de atuar como \"captadoras\" de NO. A síntese e caracterização desses complexos foram feitas pelo grupo de pesquisa do Professor Dr. Douglas Wagner Franco, IQSC-USP. Estes complexos foram adequadamente imobilizados em eletrodos e microeletrodos de ouro com filme de óxido de molibdênio. Com isso, foi desenvolvido um sensor eletroquímico para NO em solução aquosa de tampão fosfato, simulando o pH fisiológico (pH &#732;7,4). Os eletrodos modificados foram empregados na quantificação de NO em amostras de albumina bovina com peroxidonitrito na presença de tempo, verificando-se a produção de NO nesse meio. Essas amostras foram fornecidas pela Professora Dra Ohara Augusto, IQ-USP São Paulo. Os estudos referentes os complexos de rutênio visaram principalmente à determinação da velocidade de liberação do NO desses complexos, através do mecanismo EC. Para isso, utilizaram-se duas técnicas eletroquímicas, cronoamperometria de duplo degrau de potencial e voltametria com eletrodo rotativo de disco-anel. Além dos complexos coordenados com o ligante NO foram estudados complexos coordenados com o ligante sulfato. Os resultados obtidos pelas duas técnicas eletroquímicas foram concordantes, e estavam de acordo com o esperado, mediante aos estudos prévios realizados pelo grupo do Professor Dr. Douglas Wagner Franco. As constantes de velocidades encontradas situaram-se - na faixa de 10 a 10-3 s-1. Essas variações estão relacionadas com as propriedades químicas de cada ligante. Outro estudo eletroquímico realizado com a molécula de óxido nítrico em solução de tampão fosfato (pH &#732;7,4) foi feito no intuito de determinar a concentração exata de uma solução saturada de NO. Para isso, utilizou-se a técnica de cronamperometria, com eletrodo de trabalho um microeletrodo de platina de raio igual a 25 &#181;m. A concentração da solução saturada de NO, foi de (2,1 &#177; 0,3) mmolL-1. Os resultados da concentração foram confirmados pelo método clássico titulação volumétrica, no qual obteve-se o valor da concentração de (1,95 &#177; 0,02) mmolL-1. Esses resultados estão de acordo com resultados descritos na literatura. / The trans-[Ru(NH3)4NOL]3+ (L = 4-NH2py, Him, L-hist, 4-pic, py, 4-Clpy, nia, isn, 4-CNpy and pz) complexes were eletrochemically investigated by using double potencial step chronoamperometry and rotating ring-disc electrode voltammetry. Values for the rate of NO substitution by water molecule ranged from 0.02 s-1 (4-pic) to 0.34 s-1 (pz) at 25ºC. Gold surfaces were modified with an electrochemically deposited layer of non-stoichiometric molybdenum oxides. At these surfaces, trans-[Ru(III)(NH3)4(4pic)SO4]+ complex was incorporated in a controlled way by cycling consecutively the potential in the range +0.50 to -0.25 V at pH &#732; 3. Very reproducible voltammetric curves corresponding to the electrochemical process of the ruthenium complex were obtained, confirming the immobilisation of the material into the molybdenum oxide film. The anodic oxidation of nitric oxide at physiological pH &#732;7.4 in phosphate buffer was investigated at the modified electrode containing the molybdenum oxide + trans-[Ru(III)(NH3)4(4pic)SO4]+ complex and an enhancement in the current response was observed compared to the signal at a bare electrode. The rate for NO electrochemical oxidation was dependent on the amount of catalytic ruthenium sites dispersed into the molybdenum oxide film, suggesting the participation of the metallic ion in an out-sphere mechanism. A linear relationship between current signals measured by square wave voltammetry and NO concentration was obtained in the 0 to 10 &#181;M range. The applicability of the modified electrode as a sensor for real-time NO monitoring was also demonstrated.
109

Desenvolvimento de sensor eletroquímico para monitoramento de chumbo em resíduos de disparos de armas de fogo / Development of an electrochemical sensor for lead analysis in gunshot residues

Maiara Oliveira Salles 26 August 2011 (has links)
Duas metodologias distintas foram desenvolvidas para a determinação de chumbo em Resíduos de Disparos de Armas de Fogo (RDAF) utilizando microeletrodo de ouro. Na primeira delas, a superfície eletródica foi modificada com filme de bismuto em meio alcalino (pH = 13) e na outra as análises foram realizadas com microeletrodo não modificado em meio ácido com cloreto. Os RDAF foram coletados do alvo atingido pela bala e das mãos do atirador. Dos alvos, foi recortada uma região próxima ao orifício feito pela bala. Já para remoção dos RDAF das mãos do atirador, foi utilizado um algodão embebido em solução de EDTA 2% (m/v). Os resíduos foram submetidos a um processo de extração que consistiu em adicionar 10 mL de solução de HNO3 10% (m/m) ao frasco contendo o alvo ou o algodão com os RDAF. Em seguida, a mistura foi submetida ao ultrassom por 2 horas a uma temperatura de 60°C. As amostras provenientes dos alvos foram analisadas com o microeletrodo de ouro modificado com bismuto em meio alcalino por voltametria de redissolução anódica. Com essa metodologia, obteve-se um limite de detecção (3&#963;/sensibilidade) de 12,5 nmol L-1, uma faixa linear entre 40 e 6700 nmol L-1 e um desvio padrão relativo de 3,4% (n = 12). Foi possível analisar amostras provenientes de duas armas diferentes (pistola 9 mm e revólver 0.38 polegadas) e três munições distintas (normal, spl® e CleanRange®). Constatou-se um aumento da quantidade de chumbo presente nos resíduos à medida que o atirador se aproxima do alvo e que os valores dependem da arma e munição utilizadas. Com a outra metodologia as análises foram realizadas na própria solução extratora, com adição de cloreto, sem qualquer modificação da superfície eletródica. Neste caso, o limite de detecção (3&#963;/sensibilidade) encontrado foi de 1,7 nmol L-1, a faixa linear ficou entre 10 e 100 nmol L-1 e o desvio padrão relativo de 10 medidas foi 2,5%. Foram analisados resíduos provenientes das mãos dos atiradores de quatro diferentes armas (revólver 0.38 polegadas, espingardas calibre 12, 22 e 38) e de seis diferentes munições (CleanRange®, normal, semi-enjaquetada, 3T®, Eley® e CBC®). Os resultados obtidos com a metodologia proposta foram comparados com aqueles oriundos de análise por espectroscopia de absorção atômica e houve concordância em um nível de confiança de 95%. Foi também realizada uma análise quimiométrica das soluções contendo os RDAF. Os resultados mostraram a possibilidade de discriminação em relação à arma e munição utilizadas. Os filmes formados nas superfícies eletródicas foram estudados utilizando o Microscópio Eletroquímico de Varredura (SECM), a Microbalança Eletroquímica de Cristal de Quartzo (MECQ) e o Microscópio Eletrônico de Varredura (MEV). A deposição dos filmes de bismuto, chumbo e bismuto/chumbo em meio alcalino ocorre de forma heterogênea e esses filmes reagem com oxigênio e ferricianeto. Em meio ácido, a presença do cloreto na solução de depósito do filme de chumbo provoca aumento na rugosidade do filme, com consequente melhoria no sinal analítico. Também se observou que o cloreto facilita o processo de redissolução do filme de chumbo. / Two different methodologies were developed to analyze lead from gunshot residues (GSR) with a gold microelectrode: modification of the electrode surface with a bismuth film in alkaline solution (pH = 13) and use of an unmodified gold microelectrode in acidic media with chloride. GSR were collected from cloth targets and shooters\' hand after test firing. From the targets, a region close to the bullet hole was cutted off. GSR were removed from the shooters\' hand using a cotton swab embedded in a 2% (m/v) EDTA solution. The target and the cotton swab were placed in a steril vial containing 10 mL of a 10% (m/m) HNO3 solution to extract the GSR. Then, all vials were placed in the ultrasound for 2 hours at 60°C. Samples from the targets were analyzed with a gold microelectrode modified with a bismuth film in alkaline solution by anodic stripping voltammetry with a detection limit of 12.5 nmol L-1 (3&#963;/slope), linear range between 40 and 6700 nmol L-1 and a relative standard deviation of 3.4% (n = 12). Two different handguns (pistoll 9 mm and a revolver 0.38 inches) and tree different ammunition (normal, spl® and CleanRange®) were analyzed and the lead amount was dependent on the gun and ammunition used in the tests. An increase in the amount of lead from the GSR was observed as the distance between the shooter and the target decreased. Lead analyses were also performed in the extraction solution with bare gold microelectrodes in the presence of chloride ions. In this case, the detection limit was 1.7 nmol L-1 (3&#963;/slope), the linear range lied between 10 and 100 nmol L-1 and the relative standard deviation for 10 measurements was 2.5%. GSR residues form four different guns (revolver 0.38 inches, 12 caliber pump, a repeating rifle 0.38 and a semi automatic .22 caliber rifle) and six different ammunitions (CleanRange®, normal, semi jacketed, 3T®, Eley® and CBC®) were analyzed. The results obtained with the proposed methodology were compared with those from atomic absorption spectroscopy analysis and the paired t-test student indicated that there was no significant difference between the results obtained with both methods at a 95% confidence level. A chemometric analysis was also performed and the results showed the possibility to discriminate the guns and the ammunitions used. Metallic flms formed onto the electrode surface were studied by using Scanning Electrochemial Microscopy (SECM), Electrochemical Quartz Crystal Microbalance (EQCM) and Scanning Electronic Microscopy (SEM). The deposition of bismuth, lead and bismuth/lead films in alkaline solution is heterogeneous and the films react with oxygen and ferricyanide. In acidic media, the presence of chloride ions in the depositing solution causes an increase in the film roughness and facilitates the lead film dissolution
110

Signaux électriques des îlots pancréatiques enregistrés sur matrices de microélectrodes : caractérisation et application au phénotypage d'animaux transgéniques / Electrical signals from pancreatic islets recorded on multielectrode arrays : characterization and application to the phenotyping of transgenic animals

Lebreton, Fanny 17 December 2014 (has links)
Les cellules β des îlots de Langerhans jouent un rôle central dans l’homéostasie glucidique car elles seules sécrètent l’insuline, unique hormone hypoglycémiante de l’organisme. La cellule β est un détecteur du glucose qui couple sa réponse sécrétoire et son expression génique aux niveaux ambiants de glucose. Le couplage entre le métabolisme du glucose et l’exocytose des granules d’insuline implique la génération d’une activité électrique. Son étude est importante pour déchiffrer la façon dont la cellule β encode la demande en insuline de l’organisme. Afin de contourner les limites des approches électrophysiologiques classiques incompatibles avec les études à long-terme, les enregistrements extracellulaires par matrice de microélectrodes (MEA) ont été mis en place.L’objectif de ma thèse était de mieux comprendre les signaux complexes enregistrés par MEAs. Cette étude a révélé l’existence d’une nouvelle signature électrique des cellules des îlots, les slow potentials (SP), qui reflète la fonction de couplage des cellules β. Les SP jouent un rôle important dans l’homéostasie du glucose et représentent un biomarqueur de la fonction normale des îlots. La réponse en hystérèse des îlots au glucose suggère l’existence d’un algorithme d’encodage de la demande en insuline intégrée au niveau du micro-organe. De plus, ce nouveau signal a été exploité pour le phénotypage d’îlots de souris invalidées pour le gène GluK2, que nous avons utilisées comme modèle d’interaction entre les cellules α et β. La caractérisation de ce nouveau type de signal constitue aussi une avancée importante pour le développement d’un biocapteur destiné à être intégré dans le futur à un pancréas artificiel. / Pancreatic β cells are central to glucose homeostasis because they are the only cell that secretes insulin, the sole hypoglycemic hormone in the organism. The β cell is a glucose sensor that regulates its secretory response and gene expression according to ambient glucose levels. The coupling between glucose metabolism and insulin granule exocytosis involves the generation of electrical activity. An investigation of this activity is important to decipher how β cells encode the organism’s insulin demand. In order to overcome the limits of classically used electrophysiological approaches that are not compatible with long-term studies, extracellular recordings using multielectrode arrays (MEA) have been set-up.My thesis aim was to better understand the complex signals recorded with MEA. This study revealed the existence of a new electrical signature of islet cells: slow potentials (SP) that reflect the coupling function of β cells. SP play an important role in glucose homeostasis and represent a biomarker of normal functioning of islets. The observed hysteretic response of islets to glucose suggests the existence of an algorithm encoding the insulin demand embedded at the microorgan level. Moreover, this new signal was used for the phenotyping of GluK2 deficient mouse islets that were employed as an α-to-β cell interaction model. The characterization of this new signal is an important progress in the development of a biosensor intended to be integrated in an artificial pancreas in the future.

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