L'enregistrement extracellulaire d'activité électrique est une technique très répandue en neurosciences, mais son application aux îlots pancréatiques et cellules bêta n'est que très récente. La facilité d'utilisation des MEAs (Microelectrode Arrays, Matrices de Microélectrodes) a ouvert de nouvelles perspectives à l'électrophysiologie des cellules bêta, dont des méthodes de criblage en clinique ou des approches biocapteur pour le pancréas artificiel. Cette thèse contribue à la conception et la caractérisation d'un biocapteur hybride composé de cellules pancréatiques cultivées sur un MEA et d'un système électronique de traitement du signal. Le système ainsi réalisé est le fruit de collaborations et projets pluridisciplinaires menés à l'IMS (groupe bioélectronique), en partenariat avec le CBMN (biologie cellulaire et biocapteurs), tous deux au sein de l'Université de Bordeaux. Les projets faisaient également appel au service d'endocrinologie des Hôpitaux Universitaires de Bordeaux, Montpellier, Grenoble et Genève.Les projets en question incluent:- ISLET-CHIP (ANR 2013-PRTS-0017), qui explore une méthode pour évaluer la qualité d'un greffon pancréatique destiné à des patients diabétiques de type I.- BIODIA (EU FEDER), qui caractérise la réponse électrique d'îlots à des stimuli de glucose, hormones et médicaments pour des applications de criblage, différentiation cellulaire, et en boucle-fermée.- DIAGLYC (Bourse régionale 2017 1R30 226), qui étudie l'utilisation du biocapteur hybride comme un capteur pour le pancréas artificiel.La thèse aborde le biocapteur dans ses aspects à la fois électronique et biologique, son intégration dans des expériences appliquées, et ses résultats expérimentaux. Elle s'intéresse également à la modélisation d'une boucle de régulation chez le patient diabétique de type I.D'abord, le système d'analyse électronique est décrit. Issue de l'équipe Elibio, elle réalise acquisition multicanaux et traitement du signal numérique. Elle est construite autour d'un FPGA (Field Programmable Gate Array) qui rend son architecture de calcul polyvalente et évolutive. Elle est capable de mesurer, afficher, et enregistrer des données multicanaux. Le calcul embarqué est optimisé pour de faibles latences de calcul, compatibles avec des applications en boucle fermée. La thèse décrit ses algorithmes de traitement et son architecture.Des résultats expérimentaux utilisant le système et ses algorithmes sont ensuite montrés pour illustrer la réponse des îlots à des stimuli de glucose, médicaments et hormones. L'activité des îlots est quantifiée en analysant leurs APs (Action Potentials, Potentiels d'Action), et plus notoirement leurs SPs (Slow Potentials, Potentiels Lents), une nouvelle signature électrique exclusivement mesurée sur les îlots pancréatiques. Ces mesures, en régimes statique et dynamique, contribuent non seulement à caractériser la réponse du biocapteur, mais aussi à mettre en évidence les algorithmes internes des îlots de Langerhans.Enfin, des approches pour l'intégration du biocapteur dans un pancréas artificiel sont étudiées. Les réponses au glucose et aux hormones sont modélisées et simulées dans un modèle des interactions glucose-insuline dans le corps entier. Ce concept est novateur dans le sens où le capteur en charge de mesurer le besoin d'insuline n'est pas seulement sensible au glucose, mais aussi aux hormones présentes dans le sang. / Extracellular recording of electrical activity is a widespread technique in neurosciences, but only recently has it been applied to pancreatic islets and beta cells. The ease of use of Microelectrode Arrays (MEAs) has opened new perspectives for the electrophysiology of pancreatic cells, including screening methods for clinics and biosensor approaches for the artificial pancreas. This thesis is a contribution to the design and characterization of a hybrid biosensor composed of pancreatic cells cultured on an MEA and dedicated processing electronics. This device is the product of multi-disciplinary projects conducted at IMS (Bioelectronics group), partnered with CBMN (Cell biology and Biosensors team), both at the University of Bordeaux. Projects also involved the endocrinology service of university hospitals in Bordeaux, Montpellier, Grenoble, and Geneva.The covered projects include:- ISLET-CHIP (French ANR 2013-PRTS-0017), investigating a method of evaluating the quality of a preparation prior to its transplantation in type-I diabetic patients.- BIODIA (EU FEDER), characterizing islet electrical response to glucose, hormone, and drug stimuli for screening, cell differentiation, and closed-loop approaches.- DIAGLYC (French regional grant 2017 1R30 226), investigating the use of the hybrid biosensor as an artificial pancreas front-end sensor.The thesis tackles the biosensor in both its electronic and biological aspects, its integration in applicative experimental setups, and experimental results. It also addresses the modeling of a closed regulation loop for type-I diabetic patients.First, the electronic processing platform is described. It is a custom board performing multichannel acquisition and digital signal processing. It is built around an FPGA (Field Programmable Gate Array) that makes its processing architecture versatile and evolutive. It is capable of measuring, displaying and storing multichannel data. Computation was optimized for low-processing latencies compatible with closed-loop configurations. Both its processing algorithms and architecture are detailed.Then, experimental results using this system and its algorithms are shown to illustrate islet response to glucose, drug, and hormone stimuli. Islet activity is quantified by analyzing Action Potentials (APs), and more importantly Slow Potentials (SPs), a novel electrical signature exclusively measured on pancreatic islets. These measurements in both steady state and dynamic regime help characterize the biosensor response, but also shed light on the endogenous algorithms of islets of Langerhans.Finally, approaches for integrating the biosensor in an artificial pancreas are investigated. The measured glucose and hormone responses are modeled and simulated in a full-body glucose-insulin system. This concept is novel in that the sensor in charge of measuring insulin demand in the body is not only sensitive to glucose, but also to blood hormones.

La bioélectronique utilise des signaux électriques pour interagir avec des systèmes biologiques. Les capteurs qui permettent la lecture électrique de marqueurs de maladies importantes et les implants/stimulateurs utilisés pour la détection et le traitement d'activité cellulaire pathologique ne sont que quelques exemples de ce que cette technologie peut offrir. Du fait de leurs propriétés électro-actives et mécaniques fascinantes, l'électronique organique ou les matériaux conjugués π ont été largement exploités dans le domaine de la bioélectronique. Le mélange intéressant entre conductivité électronique et ionique de ces polymères conducteurs permet le couplage entre les charges électroniques présentent dans le volume des films organiques avec les flux ioniques du milieu biologique. Le matériau prototypique de la bioélectronique organique est le polymère conducteur poly(3,4-éthylènedioxythiophène) (PEDOT) dopé avec du polystyrène sulfonate (PSS). Dans ce rapport, nous étudierons une approche pour moduler les propriétés mécaniques, électriques et électrochimiques du PEDOT: PSS et étudier leur impact sur la performance des transistors électrochimiques organiques. Par ailleurs, nous évaluerons l'effet de la micro-structuration et du nano-patterning sur l'impédance électrochimique des électrodes en or recouvertes de PEDOT: PSS utiles pour de futurs enregistrements et stimulations neurales. Enfin, nous démontrerons l'utilisation du PEDOT:PSS à micro-motifs pour l'adhésion et la migration de cellules. / Bioelectronics uses electrical signals to interact with biological systems. Sensors that allow for electrical read-out of important disease markers, and implants/stimulators used for the detection and treatment of pathological cellular activity are only a few examples of what this technology can offer. Due to their intriguing electroactive and mechanical properties, organic electronics or π-conjugated materials have been extensively explored regarding their use in bioelectronics applications. The attractive mixed electronic/ionic conductivity feature of conducting polymers enables coupling between the electronic charges in the bulk of the organic films with ion fluxes in biological medium. The prototypical material of organic bioelectronics is the conducting polymer poly(3,4-ethylenedioxythiophene) (PEDOT) doped with polystyrene sulfonate (PSS). PEDOT:PSS is commercially available, water-dispersible conjugated polymer complex that can be cast into films of high hole and cation conductivity, good charge storage capacity, biocompatibility, and chemical stability. In the present work we investigate an approach to tailor the mechanical, electrical, and electrochemical properties of PEDOT:PSS and study their impact on the performance of organic electrochemical transistors. In addition, we study the effect of micro-structuring and nano-patterning on the electrochemical impedance of PEDOT:PSS- coated gold electrodes for future neural recordings and stimulation. Moreover we demonstrate the use of micro-patterned PEDOT:PSS in cell adhesion and migration.

No description available.

In the 1970s it was discovered that organic polymers, a class of materials otherwise best know as insulating plastics, could be made electronically conductive. As an alternative to silicon semiconductors, organic polymers offer many novel features, characteristics, and opportunities, such as producing electronics at low costs using printing techniques, using organic chemistry to tune optical and electronic properties, and mechanical flexibility. The conducting organic polymers have been used in a vast array of devices, exemplified by organic transistors, light-emitting diodes, and solar cells. Due to their softness, biocompatibility, and combined electronic and ionic transport, organic electronic materials are also well suited as the active material in bioelectronic applications, a scientific and engineering area in which electronics interface with biology. The coupling of ions and electrons is especially interesting, as ions serve as signal carriers in all living organisms, thus offering a direct translation of electronic and ionic signals. To further enable complex control of ionic fluxes, organic electronic materials can be integrated with various ionic components, such as ion-conducting diodes and transistors. This thesis reports a background to the field of organic bioelectronic and ionic devices, and also presents the integration of ionic functions into organic bioelectronic devices. First, an electrophoretic drug delivery device is presented, capable of delivering ions at high spatiotemporal resolution. The device, called the organic electronic ion pump, is used to electronically control amyloid-like aggregation kinetics and morphology of peptides, and offers an interesting method for studying amyloids in vitro. Second, various ion-conducting diodes based on bipolar membranes are described. These diodes show high rectification ratio, i.e. conduct ions better for positive than for negative applied voltage. Simple ion diode based circuits, such as an AND gate and a full-wave rectifier, are also reported. The AND gate is intended as an addressable pH pixel to regulate for example amyloid aggregation, while the full-wave rectifier decouples the electrochemical capacity of an electrode from the amount of ionic charge it can generate. Third, an ion transistor, also based on bipolar membranes, is presented. This transistor can amplify and control ionic currents, and is suitable for building complex ionic logic circuits. Together, these results provide a basic toolbox of ionic components that is suitable for building more complex and/or implantable organic bioelectronic devices.

The following dissertation addresses a novel low cost process developed to fabricate a Vertical Organic Field Effect Transistor (VOFET). The solution processable VOFET is designed, fabricated and tested in the context of bioengineering domains. The scope of distinct biomedical applications has also been explored. Organic thin-film transistors are gathering industrial attention as a potential candidate for future electronics analogous to silicon technology. Low fabrication cost, structural miniaturization and low operational voltage are the challenges for fabricating an Organic Field Effect Transistor (OFET). To create these devices, OFETs require new design paradigms and wet processing routes. However, conventional lateral OFET geometry cannot satisfy these demands because of process complexities and the high cost to achieve sub-micron channel length. Despite these barriers, solvent sensitivity towards organic semiconductors, electrode patterning and masking make this process more challenging than are associated with current technologies. Therefore, the need for production of a low cost high efficiency OFET is of high importance. The soluble organic semiconductor exhibits promising device properties. The growing demand of organic electronics poses great difficulty in adapting standard photolithography patterning for fabrication. The main issue is incompatibility in handling organic materials. To circumvent these challenges, a novel fabrication process has been developed to build OFETs in vertical geometry. The novelty of this process allows for creation of sub-micron channel devices at very low cost. Solution processed VOFET devices are fabricated using a 13,6-N-sulfinylacetamidopentacene (NSFAAP) precursor. Low cost fabrication techniques such as spin coating and drop casting are employed for achieving submicron channel length. Nanoscale devices, i.e. channel lengths, L=265nm, 300nm and 535nm, are respectively fabricated using the spin coating technique. Output characteristics are recorded at an operational voltage of 1volt. Short channel effects dominate the device performance, resulting in a linearity effect in I-V characteristics. Strategies, such as perforated source electrode design and drop casting techniques, are evolved and employed to minimize the short channel effects. Space Charge Limited Current (SCLC) effects, better known as short channel effects, are observed during I-V characterizations at high longitudinal fields. The drop casting technique is used over Patterned Electrode (PE) for reducing these SCLC effects. Thick channel devices, i.e. L=2µm, are fabricated to minimize the SCLC effects. Low cost polyimide 3M kapton tape is used as masking material in between the stacked layers. Time-temperature balance is optimized during the precursor to pentacene growth process. Metrological characterizations such as TEM, SEM, AFM, Raman Spectroscopy and X-RD are performed to confirm the precursor to pentacene conversion. AFM scanning illustrates dendritic pentacene molecular growth at 170°C annealing. Consequently, the conversion temperature is optimized around 200°C. In life sciences, there is always striving for translational technology development that can mimic, integrate and manipulate the biological system. Electrical signals enhance the capabilities of electronics to interact and understand the signaling pathways in a biological system. Keeping this in view, the potential applications into biomedical areas, such as flexible sensors and biomedical imagers, are proposed. VOFET has been proposed as a mainstay for flexible cardiac sensors and as imagers. OFET sensors could be designed to cover highly stretchy and arbitrary cardiac tissue. Sensor web integration with pacemakers and Implantable Cardioverter Defibrillator (ICD) device systems has been proposed. The OFET imaging sensor holds potential for early detection of cancer by detecting nuclear level changes in breast cancer images. Nuclear pleomorphic (shape and size distortion of cancerous nuclei) feature detection and analysis could be a step forward in the direction of digital pathology. The conventional analysis approach is time-consuming and error prone as it depends on visual inspection by pathologists. The proposed approach is parallel in nature and supports the existing method of cancer detection.

<p>One of the fields where image processing finds its application but that remains as anunexplored territory is the analysis of cell morphology. This master thesis proposes a systemto carry out this research and sets the necessary technical basis to make it feasible, rangingfrom the processing of time-lapse sequences using image segmentation to the representation,description and classification of cells in terms of morphology.</p><p>Due to the highly variability of cell morphological characteristics several segmentationmethods have been implemented to face each of the problems encountered: Edge-detection,region-growing and marked watershed were found to be successful processing algorithms.This variability inherent to cells and the fact that human eye has a natural disposition to solvesegmentation problems finally lead to the development of a user-friendly interactiveapplication, the <em>Time Lapse Sequence Processor</em> (TLSP). Although it was initially consideredas a mere interface to perform cell segmentation, TLSP concept has evolved into theconstruction of a complete multifunction tool to perform cell morphology analysis:segmentation, morphological data extraction, analysis and management, cell tracking andrecognition system, etc. In its last version, TLSP v0.2 Alpha contains several segmentationtools, improved user interface and, data extraction and management capabilities.</p><p>Finally, a wide set of recommendations and improvements have been discussed, pointing the path for future development in this area.</p>

One of the fields where image processing finds its application but that remains as anunexplored territory is the analysis of cell morphology. This master thesis proposes a systemto carry out this research and sets the necessary technical basis to make it feasible, rangingfrom the processing of time-lapse sequences using image segmentation to the representation,description and classification of cells in terms of morphology. Due to the highly variability of cell morphological characteristics several segmentationmethods have been implemented to face each of the problems encountered: Edge-detection,region-growing and marked watershed were found to be successful processing algorithms.This variability inherent to cells and the fact that human eye has a natural disposition to solvesegmentation problems finally lead to the development of a user-friendly interactiveapplication, the Time Lapse Sequence Processor (TLSP). Although it was initially consideredas a mere interface to perform cell segmentation, TLSP concept has evolved into theconstruction of a complete multifunction tool to perform cell morphology analysis:segmentation, morphological data extraction, analysis and management, cell tracking andrecognition system, etc. In its last version, TLSP v0.2 Alpha contains several segmentationtools, improved user interface and, data extraction and management capabilities. Finally, a wide set of recommendations and improvements have been discussed, pointing the path for future development in this area.

Many organic electronic and bioelectronics devices rely on mixed (electronic and ionic) transport within a single organic layer. Although electronic transport in these materials is relatively well understood, a fundamental understanding of ion transport is missing. I developed a simple analytical model that describes ion transport in a planar junction between an electrolyte and a conducting polymer film. The model leads to predictions of the temporal evolution of drift length of ions and current. These predictions are validated by numerical simulations and by using realistic parameters, I show that the analytical model can be used to obtain the ion mobility in the film. Furthermore, I developed an experimental method which allows the application of the analytical model and leads to a straightforward estimation of the ion drift mobilities in conducting polymers. PEDOT:PSS was found to support efficient transport of common ions, consistent with extensive swelling of the film in water. Crosslinking the film decreased its swelling and the ion mobility. Understanding the high correlation of hydration and ionic conductivity enables us to engineer materials with high and defined ion mobilities. As an example tuning of ion mobility by adjusting the relative ratio of the hydroscopic phase to PEDOT:TOS is presented. Finally I performed electrochemical impedance spectroscopy during a moving front experiment, in order to give a physical interpretation of the impedance spectra at a conducting polymer/electrolyte junction.

Le diamant dopé bore (BDD) est un matériau extrêmement prometteur pour applications biomédicales par son unique combinaison de propriétés. Cette thèse a visé le développement de nouvelles structures de micro-électrodes en BDD et l'étude de leur intérêt et leurs performances pour des applications électroanalytiques et électrophysiologiques. En dépit de leurs propriétés électroanalytiques très supérieures à d'autres matériaux d'électrodes plus conventionnels, les électrodes BDD sont sujettes au «fouling», i.e. l'apparition d'un film à la surface du diamant qui réduit la réactivité électrochimique. Ceci est très compromettant dans des milieux complexes comme l'urine, les eaux stagnantes, des boissons, le plasma sanguin etc. Ici, un nouveau traitement d'activation a été développé pour nettoyer la surface des électrodes et recouvrir leur réactivité initiale, donc il permet leur usage pour de longues périodes d'enregistrement sans dégradation du signal. Ceci permet l'usage de ce type d'électrodes, pour des domaines d'applications, pour le suivi continu d'analytes, sans entretien spécifique, en solutions complexes. La grande originalité de ces techniques d'activation est qu'elle peut être menée directement dans l'analyte lui-même. En comparaison avec leurs équivalents en macro-électrodes, les microélectrodes permettent d'obtenir de plus grandes sensibilités, des courants résiduels moindre, des pertes ohmiques moindres, et donc des rapports signal à bruit meilleurs. Un procédé robuste et fiable a été optimisé pour la fabrication de réseaux de microélectrodes (MEA MicroElectrode Arrays) et d'ultra micro-électrodes, permettant par lithographie sur 4 pouces d'offrir une large flexibilité de fabrication. Par exemple, parmi d'autres prototypes, des microélectrodes BDD ont été utilisées pour applications de biocapteurs pour quantifier l'acide urique en temps quasi-réel. Bien que le diamant possède une très bonne biocompatibilité et des propriétés électrochimiques excellentes, la faible relative capacité de double couche limite leur application pour des applications électrophysiologiques. Des procédés de nanostructuration ont ainsi été mis au point pour accroitre les limites d'injection de charge. Parmi les approches, des procédés hybrides à base de polypyrrole se sont révélés prometteurs, de même que des procédés de gravure pour former de la «nano-herbe» diamant, très intéressantes pour la fabrication de MEAs en BDD. Ces matériaux à fort rapport d'aspect apparaissent comme d'excellents candidats pour applications d'interfaces neuronales et notamment pour la fabrication d'implants rétiniens.STAR / Boron doped diamond (BDD) electrodes are extremely promising in the field of biomedical applications as they exhibit a unique combination of properties. The thesis aims at developing new types of BDD microelectrodes and exploring their interests for electro-analytical and electrophysiological applications. Despite their superior electro-analytical properties, BDD electrodes are prone to fouling, which leads to a loss of electrode reactivity when used in biological fluids such as urine, waste waters, drinks, blood plasma, etc. A novel electrochemical treatment was developed to clean the electrode surface and to retrieve the initial reactivity, thereby enabling the use of BDD electrodes to long periods of measurements without degradation of the signal, thus significantly extending the field of monitoring and surveying applications up to domains where continuous analysis is required. The real novelty of the technique is that it does not require the use of a specific media and thus can be directly performed in the probed (bio-)fluid. Microelectrodes in comparison with macro-electrodes offer higher sensitivity, lower background current, lower ohmic losses and higher signal-to-noise ratio. A robust, high-yield, reliable, and reproducible process for fabricating a thin-film BDD micro and ultra-microelectrode arrays (MEA) was developed using a novel lithographic technique, based on clean room processing on 4 inch substrates, thus offering wide flexibility. For example, among other prototypes, BDD microelectrodes were developed as biosensors to quantify uric acid in human urine in quasi-real time. Although diamond film possesses good biocompatibility and excellent electrochemical properties, the low double-layer capacitance limits its application in electrophysiological applications. Increasing the charge injection limit was investigated by surface modification and nano-structuring. These include the synthesis of hybrid diamond-polypyrrole electrodes and nanograss BDD MEAs. Such high aspect ratio materials appear as excellent candidates for neurointerfacing applications such as for retinal implants.

De nombreux dispositifs pour l’électronique organique et la bioélectronique reposent sur le transport mixte (électronique et ionique).Le transport électronique dans les matériaux organique est relativement bien compris, mais une compréhension fondamentale du transport des ions est manquante. J'ai développé un modèle analytique qui décrit le transport d'ions dans une jonction planaire entre un électrolyte et un film de polymère conducteur.Le modèle permet des prédictions de l'évolution temporelle du courant et du drift length des ions.Ces prédictions sont validées par des simulations numériques et en utilisant des paramètres réalistes, je montre que le modèle analytique peut être utilisé pour obtenir la mobilité des ions dans le film. De plus, j'ai développé une méthode expérimentale qui permet l'application du modèle analytique et conduit à une estimation de la mobilité des ions dans les polymères conducteurs. Le PEDOT:PSS offre un transport efficace pour les ions, qui peut être mis en relation avec le gonflement important du film dans l'eau. Je montre que la réticulation du film diminue son gonflement ainsi que la mobilité des ions. Comprendre la forte corrélation entre l'hydratation et la conductivité ionique nous permet de développer des matériaux à mobilité ionique définie et importante. A titre d'exemple, le réglage de la mobilité ionique du PEDOT:TOS est présenté en ajustant le rapport relatif de la phase hygroscopique. Pour finir, j'ai effectué des mesures de spectroscopie d'impédance électrochimique au cours d'une expérience de moving front, afin de proposer une interprétation physique des spectres d'impédance mesurés à une jonction polymère conducteur/électrolyte / Many organic electronic and bioelectronics devices rely on mixed (electronic and ionic) transport within a single organic layer. Although electronic transport in these materials is relatively well understood, a fundamental understanding of ion transport is missing. I developed a simple analytical model that describes ion transport in a planar junction between an electrolyte and a conducting polymer film. The model leads to predictions of the temporal evolution of drift length of ions and current. These predictions are validated by numerical simulations and by using realistic parameters, I show that the analytical model can be used to obtain the ion mobility in the film. Furthermore, I developed an experimental method which allows the application of the analytical model and leads to a straightforward estimation of the ion drift mobilities in conducting polymers. PEDOT:PSS was found to support efficient transport of common ions, consistent with extensive swelling of the film in water. Crosslinking the film decreased its swelling and the ion mobility. Understanding the high correlation of hydration and ionic conductivity enables us to engineer materials with high and defined ion mobilities. As an example tuning of ion mobility by adjusting the relative ratio of the hydroscopic phase to PEDOT:TOS is presented. Finally I performed electrochemical impedance spectroscopy during a moving front experiment, in order to give a physical interpretation of the impedance spectra at a conducting polymer/electrolyte junction.