The biosynthesis of proteins occurs at the ribosomes, where amino acids are linked together into linear chains. Nascent protein chains may undergo several different processes during their synthesis. Some proteins begin to fold, while others interact with chaperones, targeting factors or processing enzymes. Nascent membrane proteins are targeted to the cell membrane for integration, which involves the translocation of periplasmic domains and the insertion of membrane-embedded parts. The aim of this thesis was to gain insights about the dynamics of nascent peptide chains undergoing folding, membrane translocation and integration. To this end, we explored the use of arrest peptides (APs) as force sensors. APs stall ribosomes when translated unless there is tension in the nascent peptide chain: the higher the tension, the more full-length protein can be detected. By using APs, we could show that a transmembrane helix is strongly ‘pulled’ twice on its way into the membrane and that strong electric forces act on negatively charged peptide segments translocating through the membrane. Furthermore, we discovered that APs could be used to detect protein folding and made the surprising discovery that a small protein domain folded well inside the ribosomal tunnel. Finally, we explored the arrest-stability of a large set of AP variants and found two extremely stable APs.

No description available.

Ces dernières décennies, l'électronique moléculaire a suscité un intérêt croissant. La construction de jonctions métal / molécules / métal est une étape fondamentale dans la compréhension de ce domaine. Nous avons été témoins d’avancées importantes concernant les jonctions moléculaires tant sur le plan théorique que sur le plan expérimental. Cette thèse se concentre principalement sur l'étude du transport de charge à travers les jonctions moléculaires. Des polymères conducteurs et des filaments de cuivre ont été déposés, par électrochimie avec un microscope électrochimique à balayage (SECM), entre une pointe et une électrode substrat. Ainsi, nous avons développé une nouvelle façon de réaliser des contacts atomiques et des jonctions moléculaires permettant de contrôler, d’activer et de protéger ces systèmes.La fabrication de jonctions à grille redox de polymères conducteurs, tel que le PEDOT et le PBT, a été effectuée dans l’intervalle micrométrique séparant les deux électrodes du SECM. Ces nano-jonctions, hautement stables et réversibles, ont montré des conductances de 10-7-10-8 S dans leur état conducteur. Ces résultats, liés à la croissance du polymère, donnent à penser que la conductance de l'ensemble de la jonction est régie par 20 à 100 oligomères.Afin d’obtenir des nano-jonctions de manière contrôlée, une méthode combinant la stratégie dite « Break Junction » (BJ) et le SECM a été mise en place. Une nano-jonction peut être obtenue en éloignant la pointe de sa position initiale. Les variations de conductance obtenues ont montré que des jonctions moléculaires au PEDOT peuvent être brisées par paliers. Des paliers de conductance ont été mesurés par SECM-BJ, et sont comparables à ceux observés par des approches STM-BJ classiques. La technique SECM-BJ s’est avérée efficace pour la fabrication et l’étude de jonctions moléculaires de polymères à grille redox. Le SECM permet également de réaliser des nano-jonctions en utilisant une stratégie d'auto-terminaison. La croissance du polymère peut être arrêtée dès que quelques brins de polymère relient les deux électrodes initialement séparées. La taille de la jonction peut donc être contrôlée par cette méthode. Les jonctions au PTFQ et PFETQ ont montré des propriétés de transport ambipolaires. Lorsque les jonctions sont constituées de plusieurs fibres, un déséquilibre dans le transport est observé entre canaux de type p- et n-. Au contraire, un équilibre est mis en évidence lorsque les jonctions atteignent une taille nanométrique. Nous attribuons cet effet à un mécanisme de transport qui passe d’un régime diffusif (loi d’Ohm) à un régime balistique (quantique) lorsque les dimensions du dispositif deviennent nanométriques.Par ailleurs, le comportement d’électrodes d’ITO avec des nanoparticules d’or (Au NPs/ITO) dénote la présence de plasmons localisés de surface (LSP). Ces substrats ont été utilisés, sous irradiation lumineuse, pour activer la jonction démontrant ainsi que la résonance plasmon peut induire une réduction électrochimique. La diminution de conductance observée peut être attribuée à des électrons chauds générés par les plasmons sur les nanoparticules d’Au piégées dans la jonction de PEDOT, réduisant celui-ci en un état isolant.Enfin, des nano-fils de cuivre ont été élaborés par SECM en utilisant un procédé électrochimique. L’étude du transport a permis de suivre la formation de ces fils entre des électrodes asymétriques. Une étude similaire a été conduite sur une électrode constituée d’un film de silice mésoporeuse sur ITO. Les films ont une épaisseur de 115 nm et les filaments de cuivre sont protégés par encapsulation dans des canaux poreux verticaux d’environ 3 nm de diamètre. / Molecular electronics has attracted increasing interest in the past decades. Constructing metal/molecules/metal junctions is a basic step towards the investigation of molecular electronics. We have witnessed significant development in both experiment and theory in molecular junctions. This thesis focuses mainly on the study of charge transport through molecular junctions. Conducting polymers and copper filaments were electrochemically deposited with a scanning electrochemical microscope (SECM) configuration between a tip and a substrate electrode. In doing so, we have developed a new way to fabricate atomic contact and molecular junctions, and we have explored the possibility to control, protect and switch these systems.Firstly, SECM, where two microelectrodes are located face-to-face separated by a micrometric gap, has been successfully used for the fabrication of redox-gated conducting polymers junctions, such as PEDOT and PBT. Highly stable and reversible redox-gated nano-junctions were obtained with conductance in the 10-7-10-8 S range in their conducting states. These results, associated with the wire-like growth of the polymer, suggest that the conductance of the entire junction in the conductive state is governed by less than 20 to 100 oligomers.Secondly, to obtain the nano-junctions in a controllable way, a break junction strategy combined with the SECM set up is adopted. A nano-junction could be acquired by pulling the tip away from its initial position. And conductance traces showed that PEDOT junctions can be broken step by step before complete breakdown. Similarly as STM-BJ conductance steps were observed on a PEDOT molecular junction before break down by using SECM-BJ. SECM break junction technique proved to be an efficient way of molecular junction fabrication studies, especially for redox gated polymer molecular junctions. Moreover, a self-terminated strategy is found to be another way to obtain nano-junctions. An external resistance connected to the electrode plays an important role in controlling the size of conducting polymer junctions.PFTQ and PFETQ molecular junctions exhibit well-defined ambipolar transport properties. However, an unbalanced charge transport properties in n- and p- channel for these two polymer junctions was observed when the junctions are in the fiber device scale. In contrast, when molecular junction changes into nano-junction, a balanced n- and p-channel transport property is acquired. We propose that such effect is due to charge transport mechanism changing from diffusive (ohm’s law) to ballistic (quantum theory) when the junction size is reduced from fiber devices to nanodevices.High stable Au NPs/ITO electrodes exhibit a well localized surface plasmon (LSP) behavior. These plasmonic substrates have been successfully used to trigger switching of molecular junctions under light irradiation, demonstrating that surface plasmon resonance can induce electrochemical reduction. Such conductance reduction can be attributed to the hot electrons plasmonically generated from gold nanoparticles trapped into the PEDOT junction, resulting in PEDOT being reduced and changed to an insulating state.Finally, copper metallic nanowires were generated using an electrochemical self-terminated method based on SECM configuration. The presence of a few atoms that control the electron transport highlights the formation of metallic nanowires between the asymmetric electrodes. Furthermore, a similar study was performed on mesoporous silica film on ITO used as a substrate electrode. The mesoporous silica films have vertically aligned channels with a diameter of about 3 nm and a thickness of 115 nm, which play a crucial role in protecting the copper filament.

Esta tese apresenta os achados e avanços obtidos na fabricação, caracterização e aplicação de pontas de prova eletroquímicas multifuncionais para a obtenção de informações eletroquímicas resolvidas no espaço em diversas superfícies/interfaces por meio de técnicas de microscopia de varredura de ponta de prova eletroquímica (SEPM, em inglês). Diferentes designs de pontas de prova multifuncionais foram investigados e, devido a natureza não convencional destas, novos métodos para fabricação e caracterização foram desenvolvidos. Os benefícios da utilização de pontas de prova multifuncionais para a obtenção de informações eletroquímicas resolvidas no espaço ficaram evidente durante a realização de experimentos \"prova de conceito\", onde a maior densidade de informação obtida permitiu o estudo de sistemas mais complexos e a aquisição de informações eletroquímicas livre de interferência topográfica mesmo em superfícies não planas. A hibridização de diferentes técnicas de microscopia de varredura de ponta de prova eletroquímica em uma única ponta de prova também foi investigada o que se provou extremamente útil para a aquisição de imagens eletroquímicas de alta resolução, livres de influências topográficas, quando utilizada a técnica de microscopia de condutividade iônica (SICM, em inglês) como sensor de topografia do substrato investigado. Por ultimo, uma nova técnica, baseada na microscopia de condutividade iônica, que se utiliza de pontas de prova eletroquímicas multifuncionais fabricadas a partir de uma nanopipeta de um único canal, foi desenvolvida. Esta nova técnica se mostrou extremamente ponderosa, capaz de obter informações a respeito da topografia e mapear sítios ativos sobre um substrato utilizando uma nanopipeta de um único canal com alta resolução especial e temporal a uma taxa de aproximadamente 4000 pixels por Segundo. / This thesis presents the findings and advances made on fabrication, characterization and application of multifunctional electrochemical probes to acquire space resolved electrochemical information on diverse surfaces/interfaces employing Scanning Electrochemical Probe Microscopy (SEPM) techniques. Different multifunctional probes designs were investigated and new and innovative methods for fabrication and characterization of those probes were developed, which was necessary due to the unconventional nature of most of the probes studied. The benefits of using multifunctional probes for space resolved electrochemical measurements was clear during \"proof-of-concept\" experiments, where the increased density of information allowed the study of complex systems and the acquisition of topography-free electrochemical information of rough surfaces. The hybridization of different SEPM techniques in a single probe tip was also investigated, and this was found to be extremely beneficial, especially for acquiring high-resolution, topography-free, electrochemical images employing Scanning Ion Conductance Microscopy as a topography feedback. Finally, a new SICM technique, based on the use of a multifunctional probe tip fabricated from a single barrel nanopipette, was developed. This new technique was found to be extremely powerful, capable of acquiring information on topography and map active sites over substrates using a single barrel pipette with high spatial and temporal resolution at a rate of approx. 4000 pixels per second.

O trabalho descrito nesta tese mostra de forma detalhada a fabricação e caracterização de diferentes microssensores eletroquímicos os quais têm sido recentemente utilizados como sondas em grupo de técnicas conhecida como Scanning Electrochemical Probe Microscopy (SEPM). Desta forma, a caracterização de superfícies pode ser feita explorando diferentes fenômenos interfaciais relevantes à Ciência. Neste sentido, as interfaces de materiais cristalinos como hidroxiapatita (materiais dentários) e calcita foram o foco de estudo neste trabalho. Assim, diferentes técnicas SEPM foram exploradas no sentido de se obter informações relevantes relacionadas aos processos dentários, como a erosão ácida e hipersensibilidade. Inicialmente, microeletrodos de platina foram desenvolvidos empregando uma metodologia convencional na qual são utilizados microfibras encapsuladas em capilares de vidro. Scanning Electrochemical Microscopy (SECM) no modo amperométrico foi utilizada para obtenção de informações com relação às alterações de topografia do esmalte dentário causadas pelo contato com substâncias ácidas. Adicionalmente, SECM foi empregada no estudo do transporte de espécies eletroativas em amostras de dentina e investigações relacionadas ao bloqueio dos túbulos empregando-se cremes dentais comerciais foram realizadas. A permeação de peróxido de hidrogênio pela dentina também foi estudada. Os resultados de SECM foram comparados com imagens SEM obtidas nas mesmas condições experimentais. Microeletrodos de membrana ionófora íon-seletiva (Ion Selective Microelectrodes-ISMEs) sensíveis a íons cálcio também foram desenvolvidos e caracterizados, com subsequente aplicação em SECM no modo potenciométrico. A dissolução ácida de esmalte bovino (erosão dentária) foi investigada em diferentes valores de pH (2,5; 4,5; 6,8). Além disso, o transporte de íons cálcio através de membranas porosas sintéticas foi avaliado a uma distância tip/substrato de 300µm. Alterações no fluxo de íons cálcio foram correlacionadas em experimentos realizados na presença e ausência de campos magnéticos gerados por nanopartículas de magnetita incorporadas à membrana porosa. Microcristais de calcita facilmente sintetizados pelo método de precipitação foram utilizados como superfície modelo para investigações interfaciais, cujos resultados podem ser correlacionados aos materiais dentários. Desta forma, nanopipetas de vidro preenchidas com eletrólito suporte foram fabricadas e utilizadas como sonda em Scanning Ion Conductance Microscopy (SICM). O mapeamento topográfico de alta resolução espacial da superfície de um microcristal de calcita foi obtido utilizando o modo de varredura hopping mode. Adicionalmente, sondas multifuncionais ISME-SICM também foram desenvolvidas e caracterizadas para investigações simultâneas com relação às alterações topográficas e quantificação de íons cálcio sobre a superfície de um microcristal de calcita. A adição de reagentes ácidos no canal SICM promoveu a dissolução da superfície do microcristal, sendo obtidos dados cinéticos de dissolução. Investigações em meio neutro também foram realizadas utilizando a sonda ISME-SICM. Os resultados experimentais obtidos também foram comparados com aqueles oriundos de simulação computacional. / The study reported in this thesis shows in details the fabrication and characterization of different electrochemical microsensors which have been employed as probes in SEPM. Thus, the characterization of surfaces can be performed by exploiting different interfacial phenomena that are relevant to life sciences. In this sense, the interfaces of crystalline materials such as hydroxyapatite (dental materials) and calcite were the focus of this study. Thus, different SEPM techniques were explored in order to obtain relevant information related to dental materials processes such as acid erosion and hypersensitivity. Initially, platinum microelectrodes were developed employing conventional methodology that utilizes microfibers encapsulated in glass capillaries. Amperometric SECM mode was used to obtain information regarding the topography changes of tooth enamel caused by contact with acid chemicals. In addition, SECM was used to study the transport of electroactive species in dentin samples. Investigations related to the treatment of dental hypersensitivity and dental whitening were also evaluated. SECM results were compared with SEM images obtained under the same experimental conditions. Ion-selective microelectrode (ISME) based on the ionophore membrane sensitive to calcium ions were also developed and characterized followed by application in SECM potentiometric mode. The acid dissolution of bovine enamel (dental erosion) was investigated at different pH values (2.5; 4.5; 6.8). In addition, the transport of calcium ions through synthetic porous membranes was evaluated at a tip/substrate distance of 300µm. Changes in calcium ion flux were studied in the presence and absence of magnetic fields generated by magnetite nanoparticles incorporated into the porous membrane. Calcite microcrystals easily synthesized by precipitation method were used as a model of an interfacial surface for investigations which can be correlated to the dental materials. Thus, glass nanopipette filled with supporting electrolyte was fabricated and used as SICM probe. The high resolution topographic mapping of the calcite microcrystal was obtained using hopping mode. Additionally, ISME-SICM multifunction probes were developed and characterized for simultaneous investigations related to the topographical changes and quantification of local calcium ions on the surface of a calcite microcrystal. The addition of acidic reagents in the SICM channel promoted the dissolution of the microcrystal surface and dissolution kinetic data were obtained. Investigations in neutral medium were also studied using the ISME-SICM multifunctional probe. The experimental results were also compared with those obtained by computer simulation.

O trabalho descrito nesta tese mostra de forma detalhada a fabricação e caracterização de diferentes microssensores eletroquímicos os quais têm sido recentemente utilizados como sondas em grupo de técnicas conhecida como Scanning Electrochemical Probe Microscopy (SEPM). Desta forma, a caracterização de superfícies pode ser feita explorando diferentes fenômenos interfaciais relevantes à Ciência. Neste sentido, as interfaces de materiais cristalinos como hidroxiapatita (materiais dentários) e calcita foram o foco de estudo neste trabalho. Assim, diferentes técnicas SEPM foram exploradas no sentido de se obter informações relevantes relacionadas aos processos dentários, como a erosão ácida e hipersensibilidade. Inicialmente, microeletrodos de platina foram desenvolvidos empregando uma metodologia convencional na qual são utilizados microfibras encapsuladas em capilares de vidro. Scanning Electrochemical Microscopy (SECM) no modo amperométrico foi utilizada para obtenção de informações com relação às alterações de topografia do esmalte dentário causadas pelo contato com substâncias ácidas. Adicionalmente, SECM foi empregada no estudo do transporte de espécies eletroativas em amostras de dentina e investigações relacionadas ao bloqueio dos túbulos empregando-se cremes dentais comerciais foram realizadas. A permeação de peróxido de hidrogênio pela dentina também foi estudada. Os resultados de SECM foram comparados com imagens SEM obtidas nas mesmas condições experimentais. Microeletrodos de membrana ionófora íon-seletiva (Ion Selective Microelectrodes-ISMEs) sensíveis a íons cálcio também foram desenvolvidos e caracterizados, com subsequente aplicação em SECM no modo potenciométrico. A dissolução ácida de esmalte bovino (erosão dentária) foi investigada em diferentes valores de pH (2,5; 4,5; 6,8). Além disso, o transporte de íons cálcio através de membranas porosas sintéticas foi avaliado a uma distância tip/substrato de 300µm. Alterações no fluxo de íons cálcio foram correlacionadas em experimentos realizados na presença e ausência de campos magnéticos gerados por nanopartículas de magnetita incorporadas à membrana porosa. Microcristais de calcita facilmente sintetizados pelo método de precipitação foram utilizados como superfície modelo para investigações interfaciais, cujos resultados podem ser correlacionados aos materiais dentários. Desta forma, nanopipetas de vidro preenchidas com eletrólito suporte foram fabricadas e utilizadas como sonda em Scanning Ion Conductance Microscopy (SICM). O mapeamento topográfico de alta resolução espacial da superfície de um microcristal de calcita foi obtido utilizando o modo de varredura hopping mode. Adicionalmente, sondas multifuncionais ISME-SICM também foram desenvolvidas e caracterizadas para investigações simultâneas com relação às alterações topográficas e quantificação de íons cálcio sobre a superfície de um microcristal de calcita. A adição de reagentes ácidos no canal SICM promoveu a dissolução da superfície do microcristal, sendo obtidos dados cinéticos de dissolução. Investigações em meio neutro também foram realizadas utilizando a sonda ISME-SICM. Os resultados experimentais obtidos também foram comparados com aqueles oriundos de simulação computacional. / The study reported in this thesis shows in details the fabrication and characterization of different electrochemical microsensors which have been employed as probes in SEPM. Thus, the characterization of surfaces can be performed by exploiting different interfacial phenomena that are relevant to life sciences. In this sense, the interfaces of crystalline materials such as hydroxyapatite (dental materials) and calcite were the focus of this study. Thus, different SEPM techniques were explored in order to obtain relevant information related to dental materials processes such as acid erosion and hypersensitivity. Initially, platinum microelectrodes were developed employing conventional methodology that utilizes microfibers encapsulated in glass capillaries. Amperometric SECM mode was used to obtain information regarding the topography changes of tooth enamel caused by contact with acid chemicals. In addition, SECM was used to study the transport of electroactive species in dentin samples. Investigations related to the treatment of dental hypersensitivity and dental whitening were also evaluated. SECM results were compared with SEM images obtained under the same experimental conditions. Ion-selective microelectrode (ISME) based on the ionophore membrane sensitive to calcium ions were also developed and characterized followed by application in SECM potentiometric mode. The acid dissolution of bovine enamel (dental erosion) was investigated at different pH values (2.5; 4.5; 6.8). In addition, the transport of calcium ions through synthetic porous membranes was evaluated at a tip/substrate distance of 300µm. Changes in calcium ion flux were studied in the presence and absence of magnetic fields generated by magnetite nanoparticles incorporated into the porous membrane. Calcite microcrystals easily synthesized by precipitation method were used as a model of an interfacial surface for investigations which can be correlated to the dental materials. Thus, glass nanopipette filled with supporting electrolyte was fabricated and used as SICM probe. The high resolution topographic mapping of the calcite microcrystal was obtained using hopping mode. Additionally, ISME-SICM multifunction probes were developed and characterized for simultaneous investigations related to the topographical changes and quantification of local calcium ions on the surface of a calcite microcrystal. The addition of acidic reagents in the SICM channel promoted the dissolution of the microcrystal surface and dissolution kinetic data were obtained. Investigations in neutral medium were also studied using the ISME-SICM multifunctional probe. The experimental results were also compared with those obtained by computer simulation.

Esta tese apresenta os achados e avanços obtidos na fabricação, caracterização e aplicação de pontas de prova eletroquímicas multifuncionais para a obtenção de informações eletroquímicas resolvidas no espaço em diversas superfícies/interfaces por meio de técnicas de microscopia de varredura de ponta de prova eletroquímica (SEPM, em inglês). Diferentes designs de pontas de prova multifuncionais foram investigados e, devido a natureza não convencional destas, novos métodos para fabricação e caracterização foram desenvolvidos. Os benefícios da utilização de pontas de prova multifuncionais para a obtenção de informações eletroquímicas resolvidas no espaço ficaram evidente durante a realização de experimentos \"prova de conceito\", onde a maior densidade de informação obtida permitiu o estudo de sistemas mais complexos e a aquisição de informações eletroquímicas livre de interferência topográfica mesmo em superfícies não planas. A hibridização de diferentes técnicas de microscopia de varredura de ponta de prova eletroquímica em uma única ponta de prova também foi investigada o que se provou extremamente útil para a aquisição de imagens eletroquímicas de alta resolução, livres de influências topográficas, quando utilizada a técnica de microscopia de condutividade iônica (SICM, em inglês) como sensor de topografia do substrato investigado. Por ultimo, uma nova técnica, baseada na microscopia de condutividade iônica, que se utiliza de pontas de prova eletroquímicas multifuncionais fabricadas a partir de uma nanopipeta de um único canal, foi desenvolvida. Esta nova técnica se mostrou extremamente ponderosa, capaz de obter informações a respeito da topografia e mapear sítios ativos sobre um substrato utilizando uma nanopipeta de um único canal com alta resolução especial e temporal a uma taxa de aproximadamente 4000 pixels por Segundo. / This thesis presents the findings and advances made on fabrication, characterization and application of multifunctional electrochemical probes to acquire space resolved electrochemical information on diverse surfaces/interfaces employing Scanning Electrochemical Probe Microscopy (SEPM) techniques. Different multifunctional probes designs were investigated and new and innovative methods for fabrication and characterization of those probes were developed, which was necessary due to the unconventional nature of most of the probes studied. The benefits of using multifunctional probes for space resolved electrochemical measurements was clear during \"proof-of-concept\" experiments, where the increased density of information allowed the study of complex systems and the acquisition of topography-free electrochemical information of rough surfaces. The hybridization of different SEPM techniques in a single probe tip was also investigated, and this was found to be extremely beneficial, especially for acquiring high-resolution, topography-free, electrochemical images employing Scanning Ion Conductance Microscopy as a topography feedback. Finally, a new SICM technique, based on the use of a multifunctional probe tip fabricated from a single barrel nanopipette, was developed. This new technique was found to be extremely powerful, capable of acquiring information on topography and map active sites over substrates using a single barrel pipette with high spatial and temporal resolution at a rate of approx. 4000 pixels per second.

La structuration et la fonctionnalisation de réseaux de fibres optiques ont été utilisées afin de réaliser différents capteurs électrochimiques et luminescents. Ce type de support permet de concevoir des capteurs capables de détecter à distance dans un milieu confiné, difficilement accessible ou dangereux. Deux capteurs pour la détection du mercure cationique (Hg2+) sont décrits dans cette thèse. Le premier utilise un dérivé de la rhodamine, qui est lié de façon covalente à la surface en verre du réseau, par silanisation. Le signal fluorescent de cette sonde est augmenté en présence de mercure ce qui permet de le quantifier. Le second capteur combine l’électrochimie sur ces réseaux de fibres optiques, grâce à une fine couche conductrice déposée à sa surface. Le réseau est ensuite modifié avec un complexe de Ruthénium(II), qui peut être électropolymérisé par l’intermédiaire de la fonction cyclopentadithiophène (CPDT). La détection du mercure est alors réalisée par électrochimiluminescence (ECL), qui est collectée à distance, grâce aux propriétés électro-modulables du film polymère. Les réseaux de fibres optiques nanostructurés ont également été utilisés pour la réalisation d’une sonde SECM, présentant un réseau dense de nanoélectrodes collectives. La réalisation d’un tel outil utilise le positionnement basé sur les forces de cisaillement et peut être utilisé pour la structuration de surfaces conductrices ou isolantes par SECM. / The structuration and the functionalisation of optical fiber bundles have been used to design different optical and electrochemical sensors. The use of these tools allows the realisation of sensors for remote detection in a confined environment, which may be dangerous or not easily accessible. Two sensors for the detection of inorganic mercury (Hg2+) are described in this thesis. The first sensor uses a rhodamine derivative, which is covalently functionalized on the surface of the glass optical fibers by silanisation. The fluorescent signal of this probe increases in presence of mercury, which allows its quantification. The second sensor uses the combination of electrochemistry on the optical fiber bundle, thanks to a thin conductive layer deposited on its surface. The bundle is then modified by electropolymerisation of a Ruthenium (II) complex through the cyclopentadithiophene (CPDT) moiety. The detection of mercury is then realised by elecrogenerated chemiluminescence (ECL), which is collected through the optical fiber bundle, thanks to the optical properties of the polymer film. The nanostructured array of optical fibers has also been used to create a new SECM probe composed of a dense nanoelectrode array. Such a tool is obtained through Shearforce positioning and can be used for the structuration of conductive or insulating surfaces by SECM.

La présence de matière biologique (biofilms) dans les sites de stockage géologique profond, d'éléments toxiques ou encore de l'eau potable des aquifères est maintenant clairement démontrée. Cette biomasse est à l'origine de processus physiques et chimiques qui modifient considérablement la durabilité et la pérennité des sites concernés. Ces processus, principalement de type oxydo-réductif, sont encore mal compris. Ceci est principalement dû aux méthodes d'investigation, principalement macroscopiques, loin de l'échelle micrométrique caractéristique des bactéries. Seules des études, basées sur des méthodes d'investigation locale, peuvent apporter les informations requises. Ainsi, nous avons développé un dispositif expérimental basé sur l'utilisation combinée de la microscopie optique (en transmission), la microscopie à force atomique (AFM) et la microscopie AFM en mode électrique et électrochimique (EC_AFM) afin d'obtenir des informations simultanées sur la topographie de l'échantillon et sur les processus électrochimiques à l'échelle des bactéries. La première étape sensible consistait à utiliser l'AFM sur des échantillons biologiques en milieu liquide: nous présentons ici les résultats de l'imagerie AFM en milieu liquide de plusieurs types de bactéries dans leurs conditions physiologiques naturelles (conditions in vivo). Aucun protocole d'immobilisation, ni chimique ni mécanique, n'a été nécessaire; et pour la première fois, les mouvements de reptation de cyanobactéries Nostoc ont été étudiés par l'AFM. Les études AFM ont permis d'acquérir des données topographiques mais aussi mécaniques : nous avons pu ainsi mesurer le module d'Young, la pression de turgescence de différentes souches bactériennes (Anabaenopsis circularis, Rhodococcus wratislaviensis). Cette étude complète, a révélé que l'imagerie AFM est donc possible sur des espèces vivantes en mouvement. Ces résultats ouvrent une grande fenêtre sur de nouvelles études d'intérêts tels que la formation de biofilms et les propriétés dynamiques de bactéries dans des conditions physiologiques réelles. La deuxième étape délicate était de combiner l'AFM aux mesures optiques et électriques. Nous avons développé un nouveau dispositif expérimental permettant (i) le suivi de l'évolution de la croissance bactérienne par la mesure des propriétés optiques comme la densité optique DO (pour le développement bactérien en volume – milieu planctonique) , ou l'analyse de l'image du substrat par comptage du nombre de bactéries sur la surface de l'échantillon (biofilm), et (ii) les mesures électriques et électrochimiques. L'ensemble de ces résultats sera prochainement appliqué au développement de nouveaux outils de surveillance d'une biodépollution de terrain contaminé par les hydrocarbures, par le suivi in situ et en temps réel de l'activité de bactéries dépolluantes (ECOTECH_BIOPHY ANR). / The presence of biological matter (biofilms) in deep geological sites for storage of, for instance, toxic elements or groundwater in aquifers was clearly proved. That biomass triggers physical and chemical processes which greatly modify the durability and the sustainability of the sites. These processes, mainly from oxidative/reductive reactions, are poorly understood. This is mainly due to the fact that former studies were done at the macroscopic level far away from the micrometric scale where relevant processes induced by bacteria take place. Investigations at microscopic level are needed. Thus, we developed an experimental set-up based on the combined use of optical microscopy (transmission), atomic force microscopy (AFM) and electrical and electro-chemical AFM microscopy (EC_AFM) in order to get simultaneous information on topographic and electro-chemical processes.The first highly sensitive step was to use AFM with biological samples in liquid environment: we present here a study about AFM imaging of living, moving or self-immobilized bacteria, in their genuine physiological liquid medium and in true in vivo conditions. No external immobilization protocol, neither chemical nor mechanical was needed. For the first time, the native gliding movements of Gram negative Nostoc cyanobacteria upon the surface were studied by AFM. AFM height and mechanical stiffness data were simultaneously acquired. From these, mechanical parameters, inner turgor pressure and Young modulus, were derived for different bacterial species (Anabaenopsis circularis, Rhodococcus wratislaviensis). Our study revealed that AFM imaging is thus possible on moving living species. These results open a large window on new studies of both dynamical phenomena of practical and fundamental interests such as the formation of biofilms and dynamic properties of bacteria in real physiological conditions.The second delicate step was to combine AFM and optical measurements with electrical ones. We mounted a new experimental set-up coupling real-time (i) monitoring of optical properties as the optical density (OD) evolution related to bulk bacterial growth in liquid or as the counting of number of bacteria adhering on the surface of the sample as well and (ii) electrical and electrochemical measurements. Furthermore, these results will shortly be applied to the optimized monitoring of the in-situ activity of bacteria consuming oil pollutants, following this way, in real-time, the bioremediation of an oil-contaminated soil (ANR ECOTECH_BIOPHY program).

La corrosion localisée des alliages d'aluminium a pu être reliée qualitativement au rôle de la microstructure, des phases intermétalliques (IM) en particulier, qui provoquent la sensibilisation à la corrosion localisée des alliages d’aluminium contenant par exemple du cuivre. Mais un effort de modélisation est nécessaire si on veut aller vers de nouvelles méthodes d’essais.L’objectif de ce travail est d'étudier et de modéliser, sur des systèmes métallurgiques très simplifiés, les effets de couplage entre réactions électrochimiques et chimiques qui ne sont pas pris en compte dans l'analyse des essais conventionnels.L’endommagement lors de l’amorçage de la corrosion microstructurale a pu être simulé numériquement par résolution, par la méthode des éléments finis, de l’équation de Nernst-Planck décrivant les phénomènes de transport et les réactions des espèces chimiques en solution.Le comportement en milieu non chloruré de trois combinaisons où le couple IM (cathode)/matrice Al (anode) peut être assimilé à un couple Cuivre/Aluminium a été étudié, ceci de façon à rendre compte de la dissolution sélective qui fait que les phases S ou θ, présentes dans les alliages au cuivre, subissent un enrichissement en cuivre en surface. Pour valider le modèle « AlOH3 » on s’est donc appuyé sur la mesure de la vitesse de corrosion de l’anode pour trois combinaisons testées appelées systèmes modèles.En terme de mécanismes, la confrontation de l’ensemble de ces expériences aux résultats de la simulation numérique démontre que l’amorçage de la dissolution de l’anode nécessite un changement de pH local, et donc que la dépassivation locale de la surface d’aluminium par un effet chimique est l’élément déclenchant du couplage électrique. Pour aller vers une application industrielle et simuler successivement, l’amorçage et la propagation de la corrosion intergranulaire, il reste pour l’amorçage à intégrer les effets d’interaction entre phases et pour la propagation à construire à partir de ce modèle numérique, un modèle valable en milieu confiné et désaéré qui devra satisfaire aux règles physiques imposées par la dimension du système que constitue le joint de grain / In literature, localized corrosion of aluminum alloys has been qualitatively related to the role of microstructure, intermetallic phases (IM) in particular, which induces sensitization to localized corrosion of aluminum alloys containing copper, for example. But modeling effort is needed to go further in the development of new testing methods.The objective of this work is to study and model on very simplified metallurgical systems, the effects of coupling between chemical and electrochemical reactions that are not taken into account in the analysis of conventional tests.The damage induced by the microstructural corrosion initiation has been numerically simulated by resolution, by the finite element method, of the Nernst-Planck equation describing the mass transport phenomena and reactions of chemical species in solution.Behavior in non-chlorinated medium of three combinations where the couple IM (cathode) / Al matrix (anode) has been assumed to be close from a Copper / Aluminium couple has been studied, mimicking the selective dissolution of S or θ phases present in copper alloys leading to a surface copper enrichment. Validation of the model "AlOH3" defined in this study, was based on the measurement of the corrosion rate of the anode for the three model systems which were tested.In terms of mechanisms, the comparison of the experimental results of the numerical simulation clearly highlights that the initiation of the dissolution of the anode requires a change in the local pH, and therefore the local depassivation of the surface aluminum by a chemical effect is the triggering element of the electrical coupling.To transfer this numerical approach to an industrial application and simulate successively, the initiation and the propagation of intergranular corrosion, it remains, for initiation to introduce the effects of interaction between phases and for propagation to build, from this numerical model, a robust model in confined and deaerated environment which must comply with the rules imposed by the physical size of the system as defined by the grain boundary