This thesis is dealing with application of Correlative Probe and Electron Microscopy. All measurements were carried out by atomic force microscope LiteScope which is designed especially to be combined with electron microscopes. Advantages of Correlative AFM/SEM Microscopy are demonstrated on selected samples from field of nanotechnology and material science. Application of the correlative imaging was proposed and then realized particularly in case of low-dimensional structures and thin films. Further, this thesis deals with the possibility of combining Correlative AFM/SEM Microscopy with other integrated techniques of an electron microscope such as Focused Ion Beam and Energy Dispersive X-rays Spectroscopy.

In this thesis, Atomic Force Microscopy (AFM) is used to characterize Micro Fibrillated Cellulose (MFC) produced by two different methods according to their size and shape. For one of these MFC-types, their interaction with the humidity in the atmosphere is investigated and their swelling is calculated. MFC is a relatively new material based on cellulose fibres extracted from wood. This study is performed in co-operation with Stora Enso research centre. Stora Enso is a renewable material company which uses mostly wood based raw materials in their production. The measured swelling is ~ 5 % and depends on the number of elementary fibrils included in the fibre. / Atomkraftsmikroskopi (AFM) har använts för att karakterisera Mikro Fibrillerad Cellulosa (MFC), som är producerad med två olika produktionsmetoder, med avseende på deras storlek och form. För en av dessa MFC-typer så är deras påverkan av en varierande fuktig atmosfär undersökt och deras svällning är uträknad. MFC är ett relativt nytt material baserat på cellulosafibrer från trä. Detta examensarbete är utfört i samarbete med Stora Enso forskningscentrum. Stora Enso är ett företag fokuserat på förnyelsebara material som använder mestadels träbaserade råmaterial i deras prodiktion. Den uppmätta svällningen är ~ 5 % och beror av antalet elementära fibriller som ingår i fibern.

Retirement is an unavoidable and necessary phase in a person’s life. It is also one of the biggest transitions in the life cycle of a human being. Emeritus of the Apostolic Faith Mission of S.A. (AFM) have positive and negative feelings regarding their retirement. In the light of this fact the research question was posed: How can the AFM, from a pastoral perspective, understand the situation of, prepare and assist its emeritus in coping effectively with retirement - in the process addressing their holistic needs - maintaining a meaningful life in the service of God as integral members of His community? With reference to the abovementioned question it is the overall aim of this study to research the holistic needs of the pastors of the AFM, with the objective to create a pastoral model for retirement preparation. In so doing they can remain integral members of God`s community whose holistic needs are taken care of, in a Christian way. The study was done by using the practical-theological model of Osmer as starting point which comprises the following: * the normative task, where Scriptural perspectives on the topic have been researched; * the interpretative task, according to which material from other disciplines are studied; * an empirical research by way of interviews with emeritus; * and a practice theory to design a model for pre-retirement planning for pastors of the AFM. The normative task part of the research indicates that although the Bible does not say much regarding emeritus, definite direction is found regarding old age. God speaks through, and gives direction in his Word concerning the elderly. In the interpretative task part of the study very little literature is found regarding the retirement of clergy. Information exists, however, on the subject of the elderly. Numerous guidelines are also found in Scripture a propos the pastors as a holistic person in connection with body, soul and spirit. Research indicated that preparation for retirement is of cardinal importance. The results which surfaced from the empirical study, confirmed the normative and interpretative theoretical research. Subsequently the practice theory has been formulated from the hermeneutical interaction of the different elements of the research in the format of a model for pre-retirement planning. A model where the strategy for planning has been explained has been compiled finally. Matters related to the problems and fears of emeritus were addressed in creating a model. The intention of the pre-retirement education is to reduce the effect of negative stereotypes of retirement. It will further assist pastors to develop realistic goals for their retirement. With reference to the research question, central theoretical argument and the research reflected in chapters 2 to 5, the final conclusion states that pre-retirement planning for pastors of the AFM is very important. / PhD (Pastoral studies), North-West University, Potchefstroom Campus, 2014

In this thesis, atomic force microscopy (AFM), transmission electron microscopy (TEM) and optical microscopy techniques were used to investigate systematically the self-assembled nanostructure behaviour of two different types of spin-cast polymer thin films: poly(isoprene-b-ethylene oxide), PI-b-PEO diblock copolymers and [poly(9,9-dioctylfluorene-co-benzothiadiazole)]:poly[9,9- dioctyfluorene-co-N-(4-butylphenyl)-diphenylamine], F8BT:TFB conjugated polymer blends. In the particular case of the polymer blend thin films, the morphology of their composites with cadmium selenide (CdSe) quantum dot (QD) nanoparticles was also investigated. For the diblock copolymer thin films, the behaviour of the nanostructures formed and the wetting behaviour on mica, varying the volume fraction of the PEO block (fPEO) and the average film thickness was explored. For the polymer blend films, the effect of the F8BT/TFB blend ratio (per weight), spin-coating parameters and solution concentration on the phase-separated nanodomains was investigated. The influence of the quantum dots on the phase separation when these were embedded in the F8BT:TFB thin films was also examined. It was found that in the case of PI-b-PEO copolymer thin films, robust nanostructures, which remained unchanged after heating/annealing and/or ageing, were obtained immediately after spin coating on hydrophilic mica substrates from aqueous solutions. The competition and coupling of the PEO crystallisation and the phase separation between the PEO and PI blocks determined the ultimate morphology of the thin films. Due to the great biocompatible properties of the PEO block (protein resistance), robust PEO-based nanostructures find important applications in the development of micro/nano patterns for biological and biomedical applications. It was also found that sub-micrometre length-scale phase-separated domains were formed in F8BT:TFB spin cast thin films. The nanophase-separated domains of F8BT-rich and TFB-rich areas were close to one order of magnitude smaller (in the lateral direction) than those reported in the literature. When the quantum dot nanoparticles were added to the blend thin films, it was found that the QDs prefer to lie in the F8BT areas alone. Furthermore, adding quantum dots to the system, purer F8BT and TFB nano-phase separated domains were obtained. Conjugated polymer blend thin films are excellent candidates for alternatives to the inorganic semiconductor materials for use in applications such as light emitting diodes and photovoltaic cells, mainly due to the ease of processing, low-cost fabrication and mechanical flexibility. The rather limited optoelectronic efficiency of the organic thin films can be significantly improved by adding inorganic semiconducting nanoparticles.

Addition of nanopacticles in a base solvent leads to suspensions with enhanced physiochemical properties, compared to base solvent. This new type of suspensions is called nanofluids, with applications ranging from biomedicine to automotives. As a consequence extensive research is being conducted in the field, in particular, on the evaporation of these fluids as it leads to well-defined and highly ordered coffee-rings. However, the exact physics governing this phenomenon remain elusive. The goal of this experimental investigation is to elucidate how various parameters affect the progression of nanofluid coffee-stain formation. Examination of the coffee-ring structuring, produced by the free evaporation of sessile droplets containing nanoparticles, revealed an unexpected, disordered region at the exterior edge of the ring. A self-assembly mechanism with two components, particle velocity and wedge constraints, was proposed to describe the deposition of particles at contact lines of evaporating drops. Environmental pressure was used as a method to control particle crystallinity in the coffee-rings. Essentially, evaporation rate and pressure were found to be inversely proportional. Macroscopically, lowering pressure led to a transition from “stick-slip” to constant pinning. Nanoscopically, lowering pressure promoted crystallinity. Findings supported the proposed, in this thesis, particle self-assembly mechanism. Particle aspect ratio and flexibility were subsequently examined. Pinning strength was found to be a function of particle aspect ratio and rigidity, leading to constant pinning. The proposed, in this thesis, particle self-assembly mechanism was found to be applicable to a variety of aspect ratios and flexibilities. Lastly, particulate crystals grew following different pathways depending on particle flexibility.

This thesis uses the techniques of atomic force microscope (AFM) and conductive AFM (C-AFM) to study the conduction properties of n-type GaN films. A total of 16 samples were examined and grouped according to their surface morphologies and conduction behaviors. The most common type of surface morpliology was that of Ga-rich samples having undulating "hillocks" with interspersed holes. Although most of the samples had this common morphology, their local conduction behaviors were not all similar. Local I-V spectra of the tip-sample Schottky contact could be grouped according to three major types: low leakage, high leakage, and "p-type". The highest quality samples with low leakage were usually grown at moderate temperatures (~650°C). For such samples, localized leakage only occurred at screw dislocations located at small pits terminating surface hillocks. I-V spectra taken on and off such hillocks were fit in forward bias to determine whether field emission or Frenkel-Poole conduction were dominant. Although field emission is a good fit compared to Frenkel-Poole, yielding reasonable values for the barrier height, the results are not yet conclusive without variable temperature studies.

This study uses atomic force microscopy (AFM) to investigate the oxygen etching behavior of the following silicon surface orientations: (001), (111), (113), (5 5 12) and (112). Most etching was performed at sample temperatures between 650 °C and 800 °C, at pressures of 3.3×10-7 and 1.5×10-7 Torr, and at an exposure of 200 L. Surface orientation strongly influences the morphology resulting from extended etching. The surface orientations that are stable against etching and remain flat include Si(001), Si(111), and Si(113). Such surfaces also include island structures, which result from etching around oxide-induced pinning sites. The density of these islands increases at lower temperatures and higher pressures. The surface orientations that are unstable against oxygen etching and facet to other orientations include Si(5 5 12) and Si(112). These surfaces form sawtooth facets that are primarily composed of more stable (111) and (113) planes. By controlling the temperature and exposure during oxygen etching, it is therefore possible to form a variety of surface morphologies.

Master of Science / Department of Physics / Robert Szoszkiewicz / Nanoscale modifications of polymer surfaces by scratching them with sharp tips with curvature radii of tens of nanometers and at variable temperatures are expected to provide wealth of information characterizing wear response of these polymers. Such studies are important in the light of understanding the nanoscale behavior of matter for future applications in advanced polymer coatings. This thesis describes how Atomic Force Microscopy (AFM) and hot-tip AFM (HT-AFM) methods were used to characterize thermal and mechanical properties of a 30 nm thick film of poly(styrene-block-ethylene oxide), PS-b-PEO, and modify its lamellar surface patterns. Additionally, it is revealed how contact AFM and HT-AFM methods can efficiently characterize the wear response of two popular polymer surfaces, poly(methyl methacrylate), PMMA, and polystyrene, PS. The AFM and HT-AFM studies on PS-b-PEO copolymer were aimed at producing spatial alignment of respective PS and PEO parts. Instead, however, surface ripples were obtained. These measurements are explained using mode I crack propagation model and stick-and-slip behavior of an AFM tip. In addition, HT-AFM studies allowed extraction of several thermo-physical properties of a PS-b-PEO film at local volumes containing about 30 attograms of a polymer. These thermo-physical quantities are: PEO melting enthalpy of, 111 ± 88 J g[superscript]-1, PS-b-PEO local specific heat of 3.6 ± 2.7 J g[superscript]-1K[superscript]-1, and molecular free energy of Helmholtz of 10[superscript]-20 J nm[superscript]-2 for the PEO within PS-b-PEO. Utilizing a spiral scan pattern at constant angular speed and at various temperatures at the AFM tip-polymer interfaces, the wear response of PS and PMMA polymers was characterized. Cross-sections along the obtained spiral wear patterns provided plots of polymer corrugation as a function of scanning speed. From these studies it was found that the corrugation of the modified polymer surface decays exponentially with linear velocity of the scanning tip.

In this work, we used atomic force spectroscopy to obtain the elastic modulus of melanoma and keratinocytes fixed cells, with the purpose to determine the initial conditions for studies of confluente cultures of these cells in the future. The cell lines used were HaCaT cells and WM1366 melanoma cell, the last one is derived from a radial growth melanoma and both were analyzed, parental WM1366 cells (WM1366 shSCR cells) and galectin-3 silenced WM1366 cells (WM1366 shGal3). Cells were located and images of them were obtained by AFM contact mode under liquid conditions. Single force curves acquired in the central region of cells were used to determine the elastic modulus by the Hertzian contact model for the pyramidal tip, allowing to establish a comparison patter between cancer and normal cells. It was found that the melanoma cell (21.8 ± 0.5 kPa) exhibit smaller elastic modulus than keratinocytes cells (31.9 ± 0.4 kPa). For WM1366 shGal3 was found a elastic modulus of 16.1 ± 0.6 kPa, therefore, we found that for large indentation depth it is possible to distinguish between the same melanoma cell line, which represents general alterations in the organization of the cytoskeleton induced by the presence or absence of the galectin-3 protein. On the other hand, to detect local elastic modulus variations along the cell and to identify subcellular regions characterized by specific stiffness associated with local structures, we took elasticity maps in which a single force curve is acquired in each probe position. In order to interpret these maps, the cell was sliced into several different heights, curves of each height section were analyzed and represented in histograms, adjusted by the binomial distribution function. It was observed that the gradient of elastic modulus in cells from the nuclear region towards the cell periphery is more pronounced in cells devoid of galectin-3 than parental cells. The increased elastic modulus in the pericellular region of cells devoid of galectin-3 suggests that the organization of the extracellular matrix in these areas is different than those observed around HaCaT and shSCR WM1366 cells. / Neste trabalho, utilizamos espectroscopia de força atômica para obtenção do módulo elástico de células fixadas de melanoma e queratinócitos, com o objetivo de determinar as condições iniciais de estudos a serem realizados futuramente de culturas de células confluentes do mesmo tipo. As linhagem celulares utilizadas foram as células HaCaT e as células de melanoma WM1366, sendo a última derivada de um melanoma de crescimento radial sendo analisadas tanto as células parentais (células WM1366 shSCR) e as células WM1366 silenciadas com galectina-3 (WM1366 shGal3). As células foram localizadas e imageadas no modo AFM contato em meio líquido. Curvas de força adquiridas na região central das células foram utilizadas para determinar o módulo elástico, a partir do modelo de contato hertziano por uma ponta piramidal, permitindo estabelecer um padrão para comparação entre células normais e cancerígenas. Verificou-se que a célula de melanoma exibe menor módulo de elasticidade (21.8 ± 0.5 kPa) do que as células de queratinócitos (31.9 ± 0.4 kPa). Para as células WM1366 shGal3 foi encontrado um módulo elástico de 16.1 ± 0.6 kPa. Portanto, verificou-se que, para grandes profundidades de indentação, é possível distinguir entre a mesma linhagem de melanoma, células que apresentam alterações gerais na organização do citoesqueleto induzidas pela presença ou ausência da proteína galectina-3. Por outro lado, para detectar variações locais do módulo elástico ao longo da célula e identificar regiões subcelulares, caracterizadas por rigidez específica associada a estruturas locais, foram obtidos mapas de elasticidade nos quais uma única curva de força é adquirida em cada posição da sonda. Para interpretar estes mapas, a célula foi dividida em regiões de diferentes alturas e curvas de cada seção de altura foram analisadas e representadas em histogramas, ajustadas pela função de distribuição binomial. Observou-se que o gradiente de módulo de elasticidade em células da região nuclear em direção à periferia celular é mais acentuado em células desprovidas de galectina-3 do que em células parentais. O aumento do módulo de elasticidade na região pericelular das células desprovidas de galectina-3 sugere que a organização da matriz extracelular nestas áreas é diferente das observadas em torno das células HaCaT e shSCR WM1366.

Electron tomography allows visualization of objects in a form of reconstructed 3D virtual volumes with resolution power of electron microscopy. The thesis is focused primarily on biological applications of electron tomography applied on tilt series images acquired in transmission electron microscope at room temperature. Specifically, the interaction of tick-borne encephalitis virus with neural cells and 3D ultrastructure of the central electron-dense part of the flagellum 9 + 1 (Caryophyllaeides fennica) were investigated. Finally, electron tomography was combined and correlated with atomic force microscopy to allow repetitive examination of ultrathin sections on electron microscopy grids.