Ce travail de thèse a été consacré à la mise au point d'une nouvelle microscopie électrochimique à force atomique (AFM-SECM) à haute résolution, totalement innovante, désignée par l'acronyme MT/AFM-SECM pour « Mediator Tethered–AFM/SECM », et pour laquelle le médiateur rédox ferrocène (Fc) est lié à la micro-électrode sonde combinée par le biais de chaînes flexibles polyéthylène glycol (PEG). Il a été établi que les têtes ferrocènes ainsi attachées à l'extrémité de la pointe-sonde AFM-SECM sont capables de sonder la réactivité locale d'un substrat. Il a de plus été démontré que les pointes AFM-SECM Fc-PEGylées peuvent être utilisées en imagerie en mode tapping, permettant l'acquisition simultanée d'un courant de feedback électrochimique et de la topographie du substrat, et ce avec une résolution latérale et verticale de l'ordre du nanomètre. Ce nouveau type de microscopie SECM, devrait trouver de nombreuses applications en partie parce qu'il permet d'obtenir une résolution SECM de l'ordre du nanomètre, dictée par la longueur des chaînes PEG, sans avoir à utiliser des sondes nanométriques. Cette nouvelle microscopie présente donc une résolution suffisante pour permettre de localiser des macromolécules individuelles, en particulier des molécules d'enzyme rédox, sur une surface. Mais de plus, parce qu'elle est libre des contraintes diffusionnelles de la SECM « classique », la microscopie MT/AFM-SECM devrait, en principe, permettre également de caractériser le fonctionnement cinétique de ces enzymes, molécule d'enzyme par molécule d'enzyme.

<p>In this diploma work I present the first experimental investigations</p><p>of carbon nanotubes at Karlstads University. Raw nanotube powder</p><p>of single walled carbon nanotubes have been dispersed primarily in</p><p>1,2-dichloroethane. The solutions have been spincoated on Au(111)</p><p>substrates. In order to determine the solubility of carbon nanotubes</p><p>in the solution the samples have been investigated in an atomic force</p><p>microscope.</p><p>Single walled carbon nanotubes deposited on a Au(111) substrate</p><p>have been investigated in a scanning tunneling microscope. Atomically</p><p>resolved STM images of single walled carbon nanotubes were obtained.</p><p>Scanning tunneling spectroscopy spectra was taken on a tube revealing</p><p>its chirality. The measured data from the nanotubes was compared to</p><p>calculations and confirmed their properties.</p><p>Dry direct contact transfers of individual single walled carbon nanotubes</p><p>have been done as a first step when trying to deposit carbon</p><p>nanotubes on reactive surfaces in ultra-high vacuum. Individual nanotubes</p><p>were found, confirming the success of dry direct contact transfer.</p>

This thesis research focuses on the characterization of thinfilms made of poly(vinylidene fluoride) (PVDF) using an atomic force microscope. Thinfilms of PVDF were fabricated by a spin coating method with different conditions and characterized using the Atomic Force Microscopy (AFM) for morphological changes. Phase and conformational changes of PVDF were investigated using both wide angle X-ray diffraction (WAXD) and Fourier Transform Infrared Spectroscopy (FTIR). From this analysis, in-situ corona poling with annealing of spin-cast PVDF enabled a phase change from α to the mixture of β and γ phases. This process can decrease the complexity of the conventional method which requires mechanical stretching before poling PVDF in addition to thermal annealing for β phase transformation. This thesis describes some materials and surface properties of solution-cast PVDF thinfilms with various conditions such as topography and phase image, adhesion force, friction force, and roughness. Through the AFM topography and phase images, polymeric behavior and spherulites are discussed in the later part of the thesis.

Les films minces de polymères font désormais partie intégrante de notre vie quotidienne sous forme par exemple d’adhésifs et de lubrifiants. Les macromolécules qui les composent présentent de nombreux paramètres modulables tels que leur nature chimique, leur masse moléculaire ou encore leur flexibilité, permettant d’atteindre des performances très contrastées et d’envisager un large domaine d’applications, notamment dans le secteur des nanotechnologies. Ces nouveaux développements nécessitent cependant de maîtriser la stabilité de ces couches minces de polymère. En effet, que ce soit dans le cadre de la tendance à la miniaturisation nécessitant de structurer les films à des échelles nanométriques ou dans le cadre d’applications tribologiques faisant appel aux propriétés mécaniques des polymères, leur stabilité est un critère essentiel qui conditionne leurs performances. De nombreuses propriétés des systèmes polymères sont directement déterminées par les interfaces du matériau dont l’étude est commodément réalisée par l’intermédiaire d’un confinement des chaînes. Dans ce contexte, nous étudions dans une première partie les conséquences de la nature chimique d’un polymère à l’état vitreux soumis à une contrainte mécanique appliquée en surface. Nous montrons tout d’abord que la cristallisation permet d’obtenir une structuration hiérarchique de la surface des films. Sur base de ces travaux, nous avons ensuite réalisé une structuration de surface par l’intermédiaire d’un microscope à force atomique (nanorubbing) qui permet un contrôle extrêmement précis des déformations imposées. Enfin, nous tâcherons d’apporter une contribution à la controverse sur la mobilité des chaînes polymères au sein de films minces. La seconde partie de notre travail s’intéresse au comportement rhéologique de longues chaînes de polymères confinées que nous étudions grâce au processus de démouillage de films minces, une conséquence directe de l’influence des interfaces sur la stabilité des films. Les propriétés des fluides viscoélastiques sont détaillées afin de mettre en évidence l’importance de paramètres physiques trop souvent négligés tels que le stress résiduel. Nous démontrons enfin que l’étude de l’interface associée aux dynamiques de retrait du film permet de caractériser la relaxation de chaînes confinées et d’en déduire la stabilité du système étudié.

Extracellular microenvironment, including its components and biophysical parameters such as matrix structure and stiffness, is a crucial determinant of cellular function. There exists interdependency between cellular behaviors and the extracellular matrix (ECM), whereby cells are constantly sensing and modifying their surroundings in response to physical stress or during processes like wound repair, cancer cell invasion, and morphogenesis, to create an environment which supports adaptation. To date, knowledge of the distinct regulatory mechanisms of this complex relationship is little, while the urge is evident as it plays a significant role in understanding tissue remodeling. Cells are observed to align with the parallel arrays of collagen fibrils found in tissues such as bone, tendon, and cornea, suggesting the importance of ordered matrices in defining cell functions. In this study, epitaxial growths of ordered two-dimensional collagen matrices were created, with parallelly aligned fibrils on muscovite mica, and novel triangular pattern matrix on phlogopite mica. Using Fluorescence and Atomic Force Microscopy, we were able to observe cell polarization along with stress fiber formation and matrix deformation at high resolution. Cells were observed to be able to penetrate between collagen fibrils and generate traction anisotropically to polarize. These ordered collagen matrices serve as an excellent model to study cellular remodeling of ECM in vitro, in which this fundamental apprehension of cell-matrix relationship is of crucial importance to manipulate the system and obtain desired cell functions.

In this diploma work I present the first experimental investigations of carbon nanotubes at Karlstads University. Raw nanotube powder of single walled carbon nanotubes have been dispersed primarily in 1,2-dichloroethane. The solutions have been spincoated on Au(111) substrates. In order to determine the solubility of carbon nanotubes in the solution the samples have been investigated in an atomic force microscope. Single walled carbon nanotubes deposited on a Au(111) substrate have been investigated in a scanning tunneling microscope. Atomically resolved STM images of single walled carbon nanotubes were obtained. Scanning tunneling spectroscopy spectra was taken on a tube revealing its chirality. The measured data from the nanotubes was compared to calculations and confirmed their properties. Dry direct contact transfers of individual single walled carbon nanotubes have been done as a first step when trying to deposit carbon nanotubes on reactive surfaces in ultra-high vacuum. Individual nanotubes were found, confirming the success of dry direct contact transfer.

One of the important challenges in surface bioengineering is the fabrication of robust and regular supramolecular structures, tuning the chemical and topographical properties of the surface, in order to include functional biomolecules, which preserve their activity and/ or induce the desired biological process.Laccases, are redox enzyme, that catalyse the oxidation of a broad range of polyphenols and aromatic substrates. Wide variety of application in the industry has been reported, and lately their use for biosensors development. Bacterial S-layers are very interesting systems since they self-assemble forming 2-D crystals (S-layers) on many type of surfaces, and they can be fused with other biomolecules maintaining their functionality.HegG2 cell line is an hepatoma cell line that has been used for cancer research. These cells maintain part of the normal metabolic capacity of hepatocytes, what make them a useful tool for high-throughput in vitro toxicity assays, as well as in the development of bioartificial livers. The objective of this work was to generate biocompatible surfaces to immobilise lacase, S-layers and HepG2 cells, on which they mantain they active. Different surfaces with defined functionalities have been constructed with synthetic polyelectrolytes, using the layer-by-layer technique and soft lithography. At the nanoscale, an enzyme (laccase) was covalently immobilised on a gold/polyethylenimineI/glutaraldhyde layer, preserving its activity. The immobilisation was studied with a quartz crystal microbalance with dissipation monitoring (QCM-D) and its activity assayed with a spectrophotometer.Besides, bacterial surface proteins (SbpA, SbpA-EGFP and SbpA-STV) were adsorbed on polyelectrolytes multilayers. The combination of soft-lithography with a protein resistant polyelectrolyte (PLL-g-PEG) led to the construction of micro-structured surfaces of functional bacterial proteins. Surface wetability, fluorescence and atomic force microscopy (AFM) were used to characterise those interfaces.Increasing the complexity, attachment of HepG2 cells on polyelectrolytes was studied. Cell adopted different morphologies depending on the hosting underlying polyelectrolyte as observed by transmission, scanning electron and atomic force microscopes. The adhesion and spreading of the cells that were monitored with QCM-D and transmission microscopy, and assayed with crystal violet, showed a higher affinity of the cells toward the adlayer formed on PEI, PAH and PLL in comparison with PSS and PLL-g-PEG. Force spectroscopy studies with AFM showed higher repulsion between PSS surfaces and the cell surface, and different local cell mechanical properties between cells attached to PEI and PSS. / La ingeniería de biomateriales busca obtener materiales biológicamente activos, ajustando las propiedades químico-físicas (y topográficas) de la superficie de interés. Las lacasas, son enzimas que catalizan la oxidación de un gran número de polifenoles con aplicación en la industria, así como en el desarrollo de biosensores. Las proteínas bacterianas S, son sistemas muy interesantes ya que se auto-ensamblan formando cristales en 2-D y se pueden fusionar con otras biomoléculas. La línea celular hepática cancerosaHepG2, se ha empleado en estudios relacionados con el cáncer, citotoxicidad , se ha incorporado en dispositivos extracorporales para suplir funciones hepáticas, ya que a pesar de su transformación mantienen ciertas funciones de los hepatocitos normales. El objetivo de este trabajo fue generar superficies biocompatibles en las que se inmovilizó lacasa y se adsorbieron proteínas (de fusión) bacterianas y células, comprobando su funcionalidad.El ajuste de la química superficial se realizó por adsorción capa tras capa de polielectrolitos y para su estructuración se utilizó litografía blanda ("micro-contact printing"). La lacasa se inmobilizó covalentemente sobre oro cubierto con PEI (polietilenimina) través de gluraldehído, para posteriormente recubrirla con otros polielectrolitos. Dicho proceso se monitoreó con QCM (microblanza de cuarzo) y espectrofotometría (actividad enzimática). Las proteínas bacterianas se adsobieron selectivamente sobre muliticapas de polielectrolitos previamente estructuradas. Su funcionalidad se comprobó usando AFM (microscopía de fuerza atómica) y microscopía de fluorescencia. Las células HepG2 se inmobilizaron sobre superficies homogéneas y estructuradas de con distintos polielectrolitos. Se comprobó su viabilidad por ensayos con MTT y se observaron con SEM (microscopía de escaneo de electrones). El proceso de adhesion de las células sobre las multicapas de polielectrolos se estudió en función del tiempo con QCM y microscopía de transmisión, y fue testado con cristal violeta. Se utilizó AFM para estudiar la interacción entre las células con los polielectrolitos.El protocolo de inmovilización usado es aplicable para la lacasa, la cual conserva su actividad catalítica en la presencia de ABTS. La enzima se recubrió con multicapas de polielectrolitos pero la determinación de su actividad se ve dificultada por la interferencia con el ABTS.Por primera vez se utilizó "micro-contact printing" para construir superficies estructuradas, en las que se adsorbieron proteínas (de fusión) S, generando superficies funcionales en escala nanométrica dentro de microestructuras. Cabe destacar que la posibilidad de manipular la distribución de la proteína en escala micro es un requerimiento básico para el desarrollo de biosensores. En lo que se refiere a la interacción célula/superficie, se encontró que las células HepG2 adoptan diferente morfología dependiendo del substrato al que se adhieren. Mientras polielectrolitos positivos (PEI, PAH) adsorben moléculas que inducen la expansión celular, PLL-g-PEG (interfase neutra e hidrofílica) y PSS (polielectrolito negativo) no favorecieron la expansión celular. El QCM-D revelan que las células no son detectadas cuando se depositan sobre PSS (lo opuesto sucede cuando se adhieren sobre PEI) aunque se mantiene adheridas. Las propiedades mecánicas de las células varían de acuerdo al sustrato al que se adhieren, más rígidas sobre PEI. La máxima adhesión de las puntas (funcionarizadas con PEI y PSS) del AFM a la superficie celular es independiente de la carga aplicada y de su recubrimiento, para un tiempo nulo de residencia de la punta sobre la superficie celular. La máxima adhesión observada fue de 750 pN para un tiempo de residencia de residencia de 3 s, cuando el tip fue funcionalizado con PEI.

No description available.

In this thesis, we study high quality InN films grown on sapphire (0001) by plasma-assisted molecular beam epitaxy (PAMBE). We used double layers methods to reduce lattice mismatch successfully. In this experiment, we have two series of samples, about series of A use low temperature GaN (LT-GaN) as the buffer layer as compared with series of B use high temperature AlN (HT-AlN) as the buffer layer. By in situ reflection high-energy electron diffraction (RHEED), we got film¡¦s surface situation. Surface morphology of the samples was observed by atomic force microscope (AFM). By high resolution X-ray diffraction (HR-XRD) methods was analyzed quality and composition of InN films. Van der Pauw method (Hall) was used to determine carrier concentration and mobility. The optical properties of InN films under different growth conditions were investigated by photoluminescence (PL). By changing growth temperature of these samples, we found the series of A having some fine characters as the InN(0002) rocking curve was 343 arcsec and InN(10-12) rocking curve was nearly 1000 arcsec. The mobility and carrier density of these samples were approximately 1000 cm2/Vs and 3 x 1018 cm-3 by Van der Pauw method.

Investigations of specific and nonspecific interactions of biomolecules at liquid/solid interfaces are presented. To investigate specific multivalent ligand-receptor interactions, bivalent antibodies and haptens bound to solid supported membrane were used as models for ligand-receptor coupling. Novel microfabrication strategies, which included spatially addressed bilayer arrays and heterogeneous microfluidic assays, in conjunction with total internal reflection microscopy, was employed to achieve this goal. These high throughput techniques allow thermodynamic data of binding interactions to be acquired with only a few microliters of analyte and superior signal to noise. The results yield both the first and second dissociation constant for bivalent IgG antibodies with membrane bound hapten molecules. Studies were conducted both as a function of hapten density and cholesterol content in the membrane. Another research area of this dissertation is the molecular level description of nonspecific adsorption and displacement of the model protein, fibrinogen, onto hydrophilic surfaces. Techniques such as atomic force microscopy, immunochemical assays, fluorescence microscopy, and vibrational sum frequency spectroscopy were employed to probe this system. The results demonstrate that the protein's &#945;C domains play the critical role. When fibrinogen is adsorbed to a hydrophilic surface via these moieties, its displacement rate in the presence of human plasma is approximately 170 times faster than when these domains are not in direct surface contact. Even more significantly, spectroscopic studies show evidence for highly aligned Arg and Lys residues interacting with the negatively charged substrate only when the &#945;C domains make direct surface contact. The interfacial ordering of these residues appears to be the hallmark of a weak and labile electrostatic attraction between the substrate and the adsorbed macromolecule.