The image analysis techniques presented in the présent thesis have been developed as part of a European projeet dedicated to the development of an automatic membrane protein crystallization pipeline. A large number of samples is simultaneously produced and assessed by transmission electron microscope (TEM) screening. Automating this fast step implicates an on-fine analysis of acquired images to assure the microscope control by selecting the regions to be observed at high magnification and identify the components for specimen characterization.The observation of the sample at medium magnification provides the information that is essential to characterize the success of the 2D crystallization. The resulting objects, and especially the artificial membranes, are identifiable at this scale. These latter present only a few characteristic signatures, appearing in an extremely noisy context with gray-level fluctuations. Moreover they are practically transparent to electrons yielding low contrast. This thesis presents an ensemble of image processing techniques to analyze medium magnification images (5-15 nm/pixel). The original contribution of this work lies in: i) a statistical evaluation of contours by measuring the correlation between gray-levels of neighbouring pixels to the contour and a gradient signal for over-segmentation reduction, ii) the recognition of foreground entities of the image and iii) an initial study for their classification. This chain has been already tested on-line on a prototype and is currently evaluated.

In most of the tribological experiments studying friction and wear behaviour, the contact interface is hidden. The present work attempts to overcome this hidden-interface problem by carrying out real-time tribological experiments inside Transmission Electron Microscope (TEM). This is achieved by developing an in situ TEM triboprobe which can carry out nanoscale indentation, sliding and reciprocating tests on an electron transparent sample inside TEM. A novel in situ TEM triboprobe is developed by characterising the individual components involved in the development. Coarse positioning of a sharp probe is achieved using inertial sliders. Fine motion of the probe is controlled using a 4-quadrant tube piezoceramic. This triboprobe is capable of carrying out high stiffness tribological experiments inside TEM. The interface is viewed at high resolutions in real time during the experiments using a movie rate CCD camera. In indentation experiments a sharp probe is brought into contact with the sample surface. During indentation of Aluminium alloy tribolayer, it has been observed that the cracks originate from subsurface and propagate to the surface causing delamination-like material removal. Indentation experiments on protruding silicon particle in Aluminium-Silicon (Al-Si) alloy shows that initial deformation is elastic. Once the load is increased, the particle starts indenting the soft aluminum matrix, and results in sinking of the particle into the aluminium matrix. Once the particle starts sinking, the increase in the displacement causes the generation of a crack and the propagation of this crack results in the fracture of the particle. The sliding experiments inside TEM allowed the direct visualization of asperity level interaction during sliding. The preliminary experimental results of nanoscale sliding experiments carried out using an AFM tip as the sample. The adhesive instability is observed as snap-in and snap-out events. The snap-out distance seems to depend on the local geometry of the contact. To simulate reciprocating wear, a sharp diamond probe is brought into contact with Al-Si alloy and reciprocated sinusoidally at 0.5Hz. At lower loads no wear is observed. However, when the normal load is increased, material starts getting removed in thin slivers, and most of the wear debris generated get swept away from the track. Some wear debris get entrapped in between the sliding surfaces; subsequently they join to form larger wear particles. The trapped particles generated during the test act like rollers and a significant increase in the stroke-length is observed accompanying the rolling action of the particle. The phenomena like agglomeration and dissociation of the wear particles has also been observed. Repeated deformation of the trapped particles leads to the formation of tiny liquid drop on some of the wear debris. The liquid consists of gallium which comes from the sample preparation technique. The interaction between the liquid droplets has been studied by carrying out liquid-bridge pulling experiments. Liquid gallium gets cooled with time during tensile pulling of the droplets. A nano-filament is formed between the droplets during pulling. After some time, the droplet gets solidified and coalescence of droplets does not take place. Further frictional heating was necessary to form the bridge again. The in situ TEM triboprobe, which allow the tribological processes to be observed dynamically under high resolutions, is a power full tool in detecting fundamental tribological interactions.

This diploma thesis deals with the structural analysis of semiconductor nanowires by transmission electron microscopy. The construction of microscope is introduced together with its basic imaging modes and with the function of each construction element in these modes. In the experimental part the results of analysis of several germnaium nanowires are discussed, with emphasis on their crystallographic structure and orientation.

Masters of Science / The infants of women who smoke during pregnancy have a lower birth mass than those born of women who abstain. Animal studies reveal that reduced growth due to maternal nicotine exposure during gestation is accompanied by lung hypoplasia. Biochemical analysis suggests that these lungs contain more cells which implies that lung damage occurs. In this study we examined the in vivo effects of maternal nicotine exposure (lmg/Kg/day), the equivalent of 32 cigarettes per day, on the following parameters of fetal and neonatal Wistar rat lung:(i) the content and distribution of glycogen in fetal and neonatal lung (ii) the status of connective tissue in neonatal lung (iii) the cell composition of the alveoli in neonatal lung. Fetal rat lungs of ages 17, 18, 19 and 20 days and neonatal lungs of 1, 7, 14 and 21 day old pups were used. Light microscope techniques and special stains were used to investigate glycogen, connective tissue, macrophage numbers and morphological status of the lungs. Fetal rat lungs of ages 17, 18, 19 and 20 days and neonatal lungs of 1, 7, 14 and 21 day old pups were used. Light microscope techniques and special stains were used to investigate glycogen, connective tissue, macrophage numbers and morphological status of the lungs. Transmission electron microscope (TEM) techniques were employed to investigate the characteristics and composition of the alveolus The results show clearly that maternal nicotine exposure elevates pulmonary alveolar macrophage numbers'(PAM's) and lung glycogen levels. The quantity of elastic fibres in 1 day old neonates was significantly reduced but no changes in the quantity of reticulin and collagen fibres was observed. As a result of this change in connective tissue status, emphysema-like lesions and alveolar collapse was evident in the lungs of nicotine-exposed pups. TEM investigations revealed that changes to the composition of alveoli occurred. These included increased numbers of type II pneumocytes with high numbers of lamellar bodies with degenerative changes. Thickening of the blood-air barrier was also observed. The effect of maternal nicotine exposure has been documented in this study. However, it has not been possible to pinpoint the mechanisms involved but explanations have been proposed. Further research is required to elucidate the mechanisms by which nicotine produces these effects. Information thus obtained could help prevent the harmful effects to the fetus and neonate caused by smoking during pregnancy.

Les techniques d'analyse des images présentées dans cette thèse sont élaborées dans le cadre du Projet Européen dédié au développement d'une plateforme automatique pour l'évaluation de la cristallisation des protéines membranaires. Un grand nombre d'échantillons est simultanément produit et évalué par microscopie électronique en transmission (MET). Pour rendre cette tache automatique, une analyse en ligne des images acquises est indispensable ; des régions d'intérêt essentielles pour le pilotage du microscope sont progressivement sélectionnées afin d'évaluer les cristaux de protéines a fort grossissement.L'observation de l'échantillon à moyen grossissement fournit des informations nécessaires pour la caractérisation du succès de la cristallisation 2D. Les objets résultants, et en particulier le gris. De plus, ces membranes sont pratiquement transparentes aux électrons et donc, apparaissent faiblement contrastées.Cette thèse présente un ensemble de techniques de traitement d'images pour leur analyse à moyen grossissement (5-15 nm/pixel). La contribution originale de ce travail est située sur i) une évaluation statistique des contours en mesurant la corrélation entre les niveaux de gris proche du contour et un filtre de gradient pour réduire le sur segmentation, ii) la reconnaissance des objets de l'image, iii) une étude préliminaire de classification. Cette chaîne est en cours de validation sur un prototype. / The image analysis techniques presented in the présent thesis have been developed as part of a European projeet dedicated to the development of an automatic membrane protein crystallization pipeline. A large number of samples is simultaneously produced and assessed by transmission electron microscope (TEM) screening. Automating this fast step implicates an on-fine analysis of acquired images to assure the microscope control by selecting the regions to be observed at high magnification and identify the components for specimen characterization.The observation of the sample at medium magnification provides the information that is essential to characterize the success of the 2D crystallization. The resulting objects, and especially the artificial membranes, are identifiable at this scale. These latter present only a few characteristic signatures, appearing in an extremely noisy context with gray-level fluctuations. Moreover they are practically transparent to electrons yielding low contrast. This thesis presents an ensemble of image processing techniques to analyze medium magnification images (5-15 nm/pixel). The original contribution of this work lies in: i) a statistical evaluation of contours by measuring the correlation between gray-levels of neighbouring pixels to the contour and a gradient signal for over-segmentation reduction, ii) the recognition of foreground entities of the image and iii) an initial study for their classification. This chain has been already tested on-line on a prototype and is currently evaluated.

The topic of this master’s thesis is the design of a sample manipulator for TEM. Regarding which the segment of the sample manipulator used to move the sample in the direction of the x-axis (so-called x-stage) was designed. The introductory theoretical part is concerned with a brief description of transmission electron microscopy focused on the imaging methods used in TEM and the procedures of preparing and loading TEM samples. The theoretical part follows with the design of precise mechanisms, especially with respect to the inaccuracies of their operation and the technological aspect of the design. In the practical part, a detailed analysis of the design of the x-stage is presented. Kinematics and dynamics of the mechanism is analyzed in depth, following with the description of the overall design of the mechanism and some of its parts in detail.

The objective of my work was to develop HDPE clay nanocomposites for packaging with superior barrier (gas and water) properties by economical processing technique. This work also represents a comparative study of thermoplastic nanocomposites for packaging based on linear low density polyethylene (LLDPE), high density polyethylene (HDPE) and Nylon12. In this study properties and processing of a series of linear low density polyethylene (LLDPE), high density polyethylene (HDPE) and Nylon 12 nanocomposites based on Na-MMT clay and two different aspect ratio grades of kaolinite clay are discussed.

With the advent of microelectronics and micromechanical systems, the benefits of miniaturized technology became evident. With the discovery of carbon nanotubes by Iijima in 1991, a material has been found that offers superior porperties such as high tensile strength, excellent electrical and heat conductivity while being lightweight, flexible and tunable by the specific atomic arrangement in its structure. The first part of this thesis deals with a new synthesis approach, which combines the known routes of chemical vapour deposition and laser ablation. The results concerning diameter and yield fit well within an established model for the nucleation and growth of carbon nanotubes and extend it by considering a larger parameter space. Furthermore, conventional laser ablation has been used to synthesize C-13 augmented carbon nanotubes, whose diameters depend among the usual synthesis parameters also on the C-13 content, an influence which is in line with the changed thermal conductivities of isotope mixtures. Manipulation of carbon nanotubes inside a transmission electron microscope forms the second part of this thesis. With the help of an in-situ nanomanipulator, several experiments involving the mechanical and electrical properties of carbon nanotubes have been performed. Two-probe resistances of individual nanotubes have been measured and the observation of individual shell failures allowed for the determination of current limits per carbon shell. With the help of electrical current, a nanotube was modified in its electrical characteristics by reshaping its structure. By application of DC-currents or square current pulses, the filling of iron- or cementite-filled multi-wall carbon nanotubes has been found to move in a polarity-defined direction guided by the nanotube walls. Depending on the current, nanotube shape, and composition of the filling different regimes of material transport have been identified, including the reworking of the inner nanotube shells. The application of a high driving current leads to a complete reworking of the host nanotube and the current-induced growth of carbonaceous nanostructures of changed morphology. Utilizing the obtained results, a transport mechanism involving momentum transfer from the electron wind to the filling atoms and a solid filling core during transport is developed and discussed. Finally, measurements of mechanical properties using electrically induced resonant or non-resonant vibrations inside the transmission electron microscope have been observed and important mechanical parameters have been determined with the help of a modified Euler-Bernoulli-beam approach. / Mit dem Aufkommen von Mikroelektronik und mikromechanischen Systemen wurden die Vorteile miniaturisierter Geräte augenscheinlich. Mit der Entdeckung von Kohlenstoff-Nanoröhren durch Iijima 1991 wurde ein Material gefunden, welches überlegene Eigenschaften wie hohe Festigkeit, exzellente elektrische und Wärmeleitfähigkeit zeigt, während es zeitgleich leicht und flexibel ist. Diese Eigentschaften können durch eine Änderung der spezifischen atomaren Anordnung in der Nanoröhrenhülle beeinflusst werden. Der erste Teil dieser Dissertationsschrift behandelt einen neuartigen Syntheseansatz, welche die bekannten Syntheserouten der chemischen Gasphasenabscheidung und Laserablation kombiniert. Die Ergebnisse bezüglich des Durchmessers und der Ausbeute lassen sich gut mit einem etablierten Modell der Nukleation und des Wachstums von Kohlenstoff-Nanoröhren beschreiben - sie erweitern es, indem sie einen größeren Parameterraum berücksichtigen. Des Weiteren wurde konventionelle Laserablation benutzt, um C-13 angereicherte Kohlenstoff-Nanoröhren herzustellen, deren Durchmesser nicht nur von den üblichen Parametern, sondern auch vom C-13 Anteil abhängt. Diese Abhängigkeit geht mit der veränderten thermischen Leitfähigkeit von Isotopenmischungen einher. Die Manipulation von Kohlenstoff-Nanoröhren in einem Transmission-Elektronenmikroskop formt den zweiten Teil der Dissertationschrift. Mit Hilfe eines in-situ Manipulators wurden vielfältige Experimente durchgeführt, um die mechanischen und elektrischen Eigenschaften der Kohlenstoff-Nanoröhren zu bestimmen. Zweipunktmessungen des Widerstands einzelner Nanoröhren und die Beobachtung des Versagens einzelner Kohlenstoffschichten erlaubte die Bestimmung der Stromtragfähigkeit einzelner Hüllen. Mit Hilfe eines elektrischen Stromes konnte eine Nanoröhre durch die veränderung der Struktur in ihren elektrischen Eigenschaften verändert werden. Unter Verwendung dauerhaften oder gepulsten Gleichstroms konnte die Eisen- oder Zementit-Füllung der Kohlenstoff-Nanoröhren in eine polaritätsabhängige Richtung bewegt werden. Die Füllung wurde dabei durch die Wände der Nanoröhre geführt. Abhängig von Strom, Form der Nanoröhre und Zusammensetzung der Füllung ließen sich verschiedene Bereiche des Materialtransports identifizieren, u.a. das Umarbeiten einiger innerer Kohlenstoffschichten. Ein hoher Strom hingegen bewirkt eine Umarbeitung der kompletten Nanoröhre und strominduziertes Wachstum von Kohlenstoff-Nanostrukturen mit veränderter Morphologie. Mit Hilfe der gewonnenen Resultate wurde ein Transportmodell entwickelt, welches den Impulstransfer von Elektronen an Füllungsatome sowie einen festen Füllungskern während des Transports diskutiert. Messungen der mechanischen Eigenschaften, welche mit Hilfe von resonanter oder nicht-resonanter elektrischer Anregung von Schwingungen im Transmissions-Elektronenmikroskop durchgeführt wurden bilden den Abschluss der Arbeit. Durch die Beobachtungen konnten mit einem modifizierten Euler-Bernoulli-Balkenmodell wichtige mechanische Eigenschaften bestimmt werden.