abstract: In this work, atomic force microscopy (AFM) and time resolved confocal fluorescence microscopy are combined to create a microscopy technique which allows for nanometer resolution topographic and fluorescence imaging. This technique can be applied to any sample which can be immobilized on a surface and which can be observed by fluorescence microscopy. Biological problems include small molecular systems, such as membrane receptor clusters, where very high optical resolutions need to be achieved. In materials science, fluorescent nanoparticles or other optically active nanostructures can be investigated using this technique. In the past decades, multiple techniques have been developed that yield high resolution optical images. Multiple far-field techniques have overcome the diffraction limit and allow fluorescence imaging with resolutions of few tens of nanometers. On the other hand, near-field microscopy, that makes use of optically active structures much smaller than the diffraction limit can give resolutions around ten nanometers with the possibility to collect topographic information from flat samples. The technique presented in this work reaches resolutions in the nanometer range along with topographic information from the sample. DNA origami with fluorophores attached to it was used to show this high resolution. The fluorophores with 21 nm distance could be resolved and their position on the origami determined within 10 nm. Not only did this work reach a new record in optical resolution in near-field microscopy (5 nm resolution in air and in water), it also gave an insight into the physics that happens between a fluorescent molecule and a dielectric nanostructure, which the AFM tip is. The experiments with silicon tips made a detailed comparison with models possible on the single molecule level, highly resolved in space and time. On the other hand, using silicon nitride and quartz as tip materials showed that effects beyond the established models play a role when the molecule is directly under the AFM tip, where quenching of up to 5 times more efficient than predicted by the model was found. / Dissertation/Thesis / Ph.D. Physics 2012

COSTA, Erivelton Façanha da. Caracterização de Células Vermelhas por Microscopia de Força Atômica. 2006. 146 f. Dissertação (Mestrado em Física) - Programa de Pós-Graduação em Física, Departamento de Física, Centro de Ciências, Universidade Federal do Ceará, Fortaleza, 2006. / Submitted by Edvander Pires (edvanderpires@gmail.com) on 2015-05-05T20:59:55Z No. of bitstreams: 1 2006_dis_efcosta.pdf: 10007653 bytes, checksum: e0be816e57ec642d42d294c9688b5c9f (MD5) / Approved for entry into archive by Edvander Pires(edvanderpires@gmail.com) on 2015-05-07T16:55:52Z (GMT) No. of bitstreams: 1 2006_dis_efcosta.pdf: 10007653 bytes, checksum: e0be816e57ec642d42d294c9688b5c9f (MD5) / Made available in DSpace on 2015-05-07T16:55:53Z (GMT). No. of bitstreams: 1 2006_dis_efcosta.pdf: 10007653 bytes, checksum: e0be816e57ec642d42d294c9688b5c9f (MD5) Previous issue date: 2006 / The optical microscopy is the most employed technique used for visualize red blood cells (RBCs). But, due to its resolution limit, it is necessary to use other complementary techniques to study the cells, such as: the scanning and transmission electron microscopy, and the scanning probing microscopy. The Atomic Force Microscopy (AFM) is included in the last group. This work refers to the possibilities of using AFM in life science, focusing on the erythrocytes characterization. Five experiments involving red blood cells and AFM were carried out: AB+ and O+ blood types differentiation; RBCs membrane study of donors and patients with MDS (Myelodysplastic Syndrome); suitable preparation of red blood cells for AFM analysis; volume study of erythrocytes; and finally aging process observation of RBC in air. The first experiment determined the cell membrane roughness for AB+ and O+ groups, which were different. For the second one, depressions were found on the cell surface of both MSD patients and healthy people. These "holes" were deeper in the first group. The third experiment showed the importance of sample (RBCs) preparation for each AFM specific analysis. The fourth experimental procedure showed the AFM technique capability for providing volume information, which was also used in the last experiment to monitor the aging process of RBCs in air. / A maneira mais difundida na observação de células sanguineas (hemácias) é aquela que utiliza microscopia ótica convencional. Devido ao limite de resolução dos instrumentos óticos, novas técnicas de microscopia colocam-se como alternativas para o estudo de células, tais como: a microscopia eletrônica (de varredura e transmissão) e as técnicas de varredura por sonda. Inclui-se neste último grupo a microscopia de força atômica (AFM). Este trabalho discute as possibilidades de uso da Microscopia de Força Atômica ($emph{Atomic Force Microscopy}$ - AFM) em ciências da vida, para ser mais específico, na caracterização de eritrócitos. Cinco experimentos envolvendo hemácias e AFM estão aqui descritos: diferenciação entre os grupos sanguíneos AB+ e O+; análise do perfil da membrana eritrocitária de indivíduos sadios e portadores de SMD; preparação de células vermelhas para análise em microscopia de força atômica; análise volumétrica de células vermelhas; e monitoramento do envelhecimento de um eritrócito ao ar usando o AFM. No primeiro experimento, a rugosidade das membranas celulares dos grupos AB+ e O+ mostraram-se diferentes. Já no segundo experimento, depressões foram encontradas sobre a membrana de pacientes com SMD e indivíduos sadios, contudo, aparentemente mais profundas no primeiro grupo. O terceiro estudo trouxe à tona a importância da preparação adequada dos eritrócitos para medidas específicas em AFM. A quarta experiência comprovou a capacidade da técnica AFM de fornecer informação de volume, o que também foi utilizado no último experimento para monitorar o envelhecimento de uma hemácia ao ar.

Made available in DSpace on 2016-06-02T19:12:02Z (GMT). No. of bitstreams: 1 3197.pdf: 6315859 bytes, checksum: 36b44ae7699dfee5945b8beb022fe8eb (MD5) Previous issue date: 2010-05-24 / Financiadora de Estudos e Projetos / The porcelain stoneware tile is a ceramic product that has presented a great increase on production, since it had excellent technical properties and raised aesthetic performance. To improve its aesthetic characteristics, it is submitted to a polishing process that influences a lot of interested properties of the products, due mainly to changes observed on the superficial roughness. In this way, the roughness was investigated throughout the superficial polishing process of the glazed and unglazed (technical) porcelain stoneware tile and it was correlated with the gloss. For this, the Atomic Force Microscope (AFM) was used, being ideal to measure the roughness in nanometer scale and to visualize the nanotexture of many types of surface materials. The specular part of the gloss was measured through the equipment known as glossmeter. The results achieved shown that the technical porcelain stoneware tile in the first stage is more rough than the glazed. On the technical porcelain stoneware tile the roughness reduction was more accented between the grit 80 and 1000 stages, and the gloss was practically double between the first and the last (POLIDO) stages. The surface covering with the polymeric resin reduced significantly the product gloss, without it causes brusque alterations in superficial roughness. On the glazed porcelain stoneware tile the roughness reduction and the gloss increased occurred mainly between the grit 600 and 1800 stages. Between the grit 1800 and 8000 stages had not observed great changes on the roughness or gloss, showing that the polishing becomes unnecessary from the grit 1800. In general way, the gloss reduction occurred more visibly in stages of polishing with abrasives of larger grains, while the abrasives of smaller grains had given the final gloss, intervene poorly in roughness. The results interpretation had supplied subsidies to a better understanding and control of the process responsible for the final characteristics between the glazed and technical products, and the differences between these typologies. / O porcelanato é um produto cerâmico que apresenta grande expansão na produção, já que possui excelentes propriedades técnicas e um elevado desempenho estético. Para melhorar suas características estéticas, é submetido a um processo de polimento que influencia diversas propriedades de interesse do produto, devido principalmente às mudanças verificadas na rugosidade superficial. Desta forma, a rugosidade foi investigada ao longo do processo de polimento da superfície de porcelanatos esmaltados e não esmaltados (técnicos) e correlacionada com o brilho obtido. Para isso, foi utilizado o Microscópio de Força Atômica (AFM), ideal para medir a rugosidade em escala nanométrica e visualizar a nanotextura de muitos tipos de superfícies de materiais. A parte especular do brilho foi medida por meio do equipamento conhecido por brilhômetro. Os resultados obtidos mostraram que o porcelanato técnico no primeiro estágio analisado é mais rugoso que o esmaltado. No porcelanato técnico a diminuição da rugosidade foi mais acentuada entre os estágios de grit 80 e 1000 e o brilho do produto praticamente dobrou entre o primeiro e último estágio (POLIDO). A aplicação de resina impermeabilizante causou uma diminuição brusca do brilho, sem alterar significativamente a rugosidade. No porcelanato esmaltado a diminuição da rugosidade e o aumento do brilho ocorreram principalmente entre os estágios de grit 600 e 1800. Entre os estágios de grit 1800 e 8000 não foram observadas grandes mudanças na rugosidade, nem tampouco no brilho, mostrando que o polimento torna-se desnecessário a partir do grit 1800. De modo geral, a diminuição da rugosidade ocorreu mais visivelmente nas etapas de polimento com abrasivos de grãos maiores, enquanto os abrasivos de grãos menores deram o brilho final das peças, interferindo pouco na rugosidade. A interpretação dos resultados forneceu subsídios para um melhor entendimento e controle do processo responsável pelas características finais dos produtos esmaltados e técnicos e as diferenças entre estas duas tipologias.

The possibilities of novel catalytic scanning probe lithography (cSPL) on starch using α-amylase was investigated. For this thin homogeneous layers of starch with good coverage were prepared by spin coating a starch solution on a silicon base. Amylase immobilized to an atomic force microscopy (AFM) cantilever tip were prepared and dragged along a spin coated starch surface. This after verifying the enzyme immobilization method using (3-Aminopropyl)triethoxysilane (APTES) on a silicon surface. In addition an unmodified cantilever tip were dipped in amylase solution and were dragged along a starch surface to investigate possibilities of dip-pen nanolithography (DPN). The preliminary experiments with AFM based enzymatic lithography were promising but non-conclusive. There are still many parameters not fully explored such as water availability, activity and reach of the amylase, speed of the enzymatic process and difference in structure between the starch and the shorter saccharides that are left after the hydrolysis

Du fait de leur faible coût de production et de leur intégration possible sur substrat flexible, les cellules photovoltaïques organiques sont prometteuses pour répondre aux besoins futurs en énergie. Leurs performances reposent sur l'architecture de la cellule et sur la nature des matériaux choisis. Par conséquent, le contrôle à l'échelle nanométrique de la couche active (formation de nanodomaines purs, organisation moléculaire...), ainsi que le développement de nouvelles molécules aux propriétés électroniques et structurales optimisées apparaissent comme des paramètres clés. Dans ce contexte, les travaux présentés dans cette thèse visent à étudier le lien entre la morphologie des films minces et les propriétés de transport à l'échelle nanométrique en fonction des matériaux actifs utilisés. Pour cela deux voies ont été explorées. La première voie repose sur l'utilisation des propriétés d'auto-organisation des cristaux liquides pour améliorer la formation et l'organisation de nanodomaines. Pour cette étude, nous avons choisi d'associer un donneur d'électron classique, le poly-3-hexylthiophène (P3HT), et de le mélanger avec un complexe de nickel [Ni(4dopedt)2] possédant des propriétés cristallines liquides colonnaires. L'étude par Microscopie à Force Atomique (AFM), Conductive-AFM (C-AFM), absorption UV-visible et spectrométrie Raman des films de mélanges démontre l'effet structurant du cristal liquide sur les chaînes de P3HT, en fonction de l'épaisseur de la couche et des traitements thermiques effectués. La deuxième voie explorée repose sur l'ingénierie moléculaire. Suite à une étude bibliographique, nous avons conçu puis synthétisé différentes petites molécules fluorées capables d'agir comme accepteurs d'électrons. Leur synthèse est réalisée en plusieurs étapes, privilégiant une méthode de couplage innovante, l'hétéroarylation directe. Les molécules obtenues ont été caractérisées par les techniques analytiques classiques, puis soumises à une étude de relation structures/propriétés. D'une part, les analyses optiques, électrochimique et thermique ont révélé leur grande stabilité et leur intérêt potentiel pour l'application visée. D'autre part, leur étude en film mince, par spectroscopie d'absorption UV-visible, de fluorescence et par AFM, révèle l'influence de la substitution du squelette conjugué par des atomes de fluor ainsi que de la modification des chaînes alkyles des groupements terminaux sur les propriétés optoélectroniques et structurelles des molécules. / Organic solar cells appear as a promising technology to meet future energy requirements, owing to their low production costs, their great flexibility and their ability to be integrated into light devices. Their performances rely on their architecture and the nature of the chosen materials. As a consequence, two of the key parameters for their development are the control the active layer at a nanometric scale (molecular organisation and the formation of pure compound nanodomains) and the development of new small molecules with optimized electronic and structural properties. This work comes in that aim : the study of the relation between thin film morphology and transport properties at the nanometric scale as function of the chosen materials. Two ways have been explored. The first way relied on self-organisation properties of a liquid crystal for improving the formation and organisation of nanodomains. In this purpose, we have associated a well-known electron donor, the poly-3-hexylthiophene (P3HT), with a complex of nickel, named as [Ni(4dopedt)2], exhibiting columnar liquid crystal properties. Atomic Force Microscopy (AFM), Conductive-AFM (C-AFM), UV-visible absorption and Raman spectroscopy on bulk films have shown the structuring effect of the liquid crystal on the P3HT chains as a function of the films thicknesses and thermal annealing. The second way was based on molecular engineering. Following a bibliographic study, we have designed and synthetized different fluorinated small molecules with electron acceptor capability. For this, a more economical and cleaner synthesis technique has been employed: the direct arylation. These molecules have been characterized by classical analytic technics, and a study of the relation between structure and properties has been carried out. On the one hand, optical, electrochemical and thermal analyses have shown their good stability and their potential for the aimed application. On the other hand, their study in thin film by UV-visible absorption, fluorescence and AFM have shown the influence of backbone substitution by fluorine atoms as well as the impact of the nature of alkyl end chains on the optoelectronic and structural properties of these molecules.

L’étude porte sur la propagation lente de fissure à température ambiante dans deux classes de polymères à l’état vitreux : le PMMA et la résine époxy-amine stœchiométrique DGEBA-IPD comme prototypes respectifs de thermoplastique (TP) et de thermodurcissable (TD). Des mélanges TD/TP présentant une séparation de phase de nano-domaines de thermoplastique dans une phase dominante de thermodurcissable ont été aussi explorés en vue de leur intérêt en tant que matrice à morphologie complexe utilisée dans les composites à renfort de fibre de carbone destinés à l’aéronautique. Un montage expérimental original a été développé pour le suivi in situ de propagation lente de fissures, combinant une caméra optique, un microscope à force atomique (AFM) et un échantillon sollicité en géométrie Double Cleavage Drilled Compression (DCDC). La possibilité d’accéder ainsi à une gamme de vitesses de propagation entre le pm/s et le nm/s a permis de mettre en évidence pour la toute première fois un régime de propagation stationnaire dans les thermodurcissables (résines pures et mélanges TD/TP) analogue au comportement établi pour les thermoplastiques, et de le caractériser en termes d’une loi cinétique reliant la vitesse de propagation au facteur d’intensité des contraintes (SIF). L’analyse in situ AFM a de plus permis de caractériser les différentes modalités de déformation à l’échelle de la zone de process en combinant l’imagerie topographique et des techniques de corrélation d’images numériques (DIC). Un modèle de prédiction des énergies de rupture en propagation stationnaire ainsi qu’en régime de stick-slip a été développé, en intégrant les champs de déformation visco-plastiques mesurés dans la zone de process, la vitesse de déformation locale, et la loi de comportement des matériaux mesurée à l’échelle macroscopique. / This work deals with the slow crack growth in glass polymer materials at room temperature, namely PMMA and stoechiometric epoxy-amine DGEBA-IPD resin as archetypes of thermoplastic (TP) and thermoset (TS) respectively. TS/TP blends developing phases separation in nano-domains of thermoplastic during the cure process have also been studied since they are used as matrix with a complex morphology in carbon fibers reinforced composites materials for aircrafts. An innovative experimental setup has been developed for in situ investigation of slow crack growth by combining optical measurements, in situ atomic force microscopy (AFM) and a Double Cleavage Drilled Compression (DCDC) sample. By this way, a steady state crack propagation regime has been highlighted for a first time in thermosets (pure and TS/TP blends) with crack speed ranging from pm/s to nm/s. Thus crack speed and stress intensity factor (SIF) diagrams were established for each material. Furthermore, in situ AFM images allowed characterizing deformations at the process zone scale by combining topographic imaging with Digital Images Correlation (DIC) technique. Finally, we developed a model for predicting fracture energy for steady state propagation and stick-slip regime, based on the visco-plastic strain fields measured round the process zone, the local strain rate, and the constitutive laws of materials measured at the macroscopic scale.

The human body consists of a number of very complex, highly specialised organs which perform a variety of functions that are essential to life and health. One of the main functions of the skin, the largest of the human organs, is to maintain the integrity of the body. It does this by acting as a physical barrier, preventing micro-organisms and other potentially harmful substances from entering the body. When the integrity of the skin is damaged through injury, a self-protective mechanism is triggered and the reparative wound healing process begins. Under normal circumstances the wound healing process culminates in the skin recuperating its normal characteristics and functions at the site of the injury, with only a small visible mark being left behind. However, in some cases the wound healing process may become altered leading to the production of abnormal scars, such as keloids. Keloid scars are formed from scar tissue at the site of an injury, as a result of excessive tissue repair that extends beyond the boundaries of the original wound. These scars are characterised by excess collagen deposition produced during the wound healing process. It is estimated that as many as 20% of the black and Hispanic population are affected by keloid scarring. In addition to the aesthetic aspect, keloid scars can also be painful, itchy and prone to become infected. Keloid scar formation can be triggered by skin injuries caused by, for example, acne, wounds, shaving, burns, and surgical incisions. The mechanism by which keloid scars form is currently not well understood and consequently no effective treatments exist to date.This thesis describes an investigation into the mechanical properties of single keloid and normal skin fibroblast cells for the purpose of establishing if there is a quantitative difference between the two types of cells. This information will be of benefit to researchers looking for a better understanding of the keloid formation mechanism and for those seeking improved treatments. An atomic force microscope (AFM) was employed to indent single Keloid and normal skin fibroblast cells taken from five patients. Values for the apparent Young's modulus of the cells were then calculated by fitting the experimental data using Hertz's model. Apparent Young's modulus values were then compared. The findings of the analysis indicate that statistically, there is a significant difference in the Young's modulus values of normal and keloid cells, with keloid cells exhibiting substantially greater stiffness than normal skin fibroblast cells. To enable the keloid and normal skin fibroblast cells to be studied in as close to their natural, physiological environment as is possible the AFM experiments described in this thesis were undertaken in a phosphate buffered saline (PBS) solution. In such cases the use of a fluid medium presents additional complexities, not least of which is the introduction of a hydrodynamic drag force due to viscous friction of the cantilever with the liquid which can affect the experimental data obtained and consequently any material properties calculated as a result of using these data. In order to investigate the effect of dragging force on the experimental data obtained from the AFM a novel integrated finite element based model was developed. The model, described in this thesis, provides quantification of the drag force in AFM measurements of soft specimens in fluids, consequently enabling more accurate interpretation of the data obtained from AFM experimentation. The model is validated using extensive data obtained from AFM experimentation undertaken in a number of fluids of different viscosities, at a variety of tip velocities and platform-tip separations and by comparison with an existing analytical model. The novel model is shown to accurately account for drag forces in AFM in fluid media without the need for extrapolation of experimental data and can be employed over the range of tip geometries and velocities typically utilised in AFM experimentation.The work described in this thesis demonstrates that the AFM is a valuable tool that can be used to successfully investigate the mechanical properties of biological samples in fluids. It was shown that increased accuracy in the interpretation of data obtained from AFM experimentation can be obtained by taking into account the hydrodynamic drag force due to viscous friction of the cantilever with the liquid. The investigation into the mechanical behaviour of keloid cells described in this thesis significantly adds to the yet small body of research undertaken on keloid cells to date. The findings of the investigation will provide valuable information that will be of benefit in the future to researchers looking to develop effective treatments for the prevention, reduction or removal of keloid scars.

The barrier properties of polymers are a significant factor in determining the shelf or device lifetime in polymer packaging. Nanocomposites developed from the dispersion of nanometer thick platelets into a host polymer matrix have shown much promise. The magnitude of the benefit on permeability has been different depending on the polymer investigated or the degree of dispersion of the platelet in the polymer. In this dissertation, the effect of density changes in the bulk and at the polymer-platelet interface on permeability of polymer nanocomposites is investigated. Nanocomposites of nylon, PET, and PEN were processed by extrusion. Montmorillonite layered silicate (MLS) in a range of concentrations from 1 to 5% was blended with all three resins. Dispersion of the MLS in the matrix was investigated by using one or a combination of X-ray diffraction (XRD), scanning electron microscopy (SEM) and transmission electron microscopy (TEM). Variation in bulk density via crystallization was analyzed using differential scanning calorimetry (DSC) and polarized optical microscopy. Interfacial densification was investigated using force modulation atomic force microscopy (AFM) and ellipsometry. Mechanical properties are reported. Permeability of all films was measured in an in-house built permeability measurement system. The effect of polymer orientation and induced defects on permeability was investigated using biaxially stretched, small and large cycle fatigue samples of PET and nylon nanocomposites. The effect of annealing in nylon and nanocomposites was also investigated. The measured permeability was compared to predicted permeability by considering the MLS as an ideal dispersion and the matrix as a system with concentration dependent crystallinity.

A variety of approaches were conducted to obtain a comprehensive understanding of the adsorption of Natural Organic Matter (NOM) isolates on metal oxides (MeO). Adsorption experiments with a series of small molecular weight (MW), oxygenated, aromatic organic acids were performed with Aluminum oxide (Al2O3), Titanium oxide (TiO2), and Zirconium oxide (ZrO2) surface. The experiments were conducted in batch mode at pH 4.2 and 7.6. The adsorption of simple organic acids was described by Langmuir model, and exhibited strong dependence on the relative abundance of carboxyl group, aliphaticity/aromaticity, length of alkyl chain, and the presence of hydroxyl group. The adsorption of the model compounds was high at low pH and decreased with increasing the pH. Isolated NOM fraction of strong humic character, i.e., hydrophobic (HPO) (high in MW, aromaticity, and acidity), i.e., Suwannee River fulvic acid (SRW HPO), showed strong adsorption on all MeO. However, fractions with similar acidic character, and lower MW exerted weak adsorption. NOM fraction that incorporated polysaccharides and proteins like structures (i.e., biopolymers) was not significantly adsorbed compared to HPO fractions. Interestingly, biopolymer adsorption on Heated Aluminum oxide particles (HAOP) was higher than that on Al2O3, TiO2, and ZrO2. These different adsorption profiles were related to their physicochemical characteristics of NOM and MeO, and thus, showed different interacting mechanisms and were studied by Atomic Force Microscopy (AFM). Hydrogen bonding was suggested as the main mechanism between NOM of strong hydrophilic character (i.e., biopolymers) and Al2O3, TiO2 and ZrO2 coated wafers. The strength of the hydrogen bonding was influenced by the hydrophilicity degree of MeO surface, ionic strength, and cation type. NOM fractions with strong humic character showed repulsive forces that are electrostatic in nature with MeO of high negative charge density. Hydrogen bonding and ligand exchange mechanisms are proposed to control the adsorption mechanism at high ionic strength with less negatively-charged MeO surface. Strong interactions forces was recorded between NOM molecules with different properties, more specifically with high MW humic and non humic fractions. These forces are controlled by cation type, and NOM chemical structure.

A conductive atomic force microscopy (c-AFM) investigation of GaN nanostructures is reported for strain engineering optoelectronic and piezotronic devices. The use of AFM enables the simultaneous correlation between the surface morphology and charge carrier transport through the nanostructures. The samples under investigation are molecular beam epitaxy (MBE) grown InGaN/GaN nanowires on Ti coated Mo substrate and GaN nanowires on ITO. The metal-semiconductor interface between the metallic substrates and the GaN nanostructures form the bottom contact. A Pt-Ir coated AFM probe is used to create a Schottky top nano-contact. The two interfaces form a metal-semiconductor-metal (MSM) structure. Force and temperature-dependent IV curves are obtained and analyzed, and the MSM structure parameters are extracted. Modulation of both the conductivity and Schottky barrier height (SBH) is revealed. Drastic reduction of the barrier is observed to drive the junctions to ideal MSM under a combination of force and temperature, revealing a dynamic and controlled two-way switching of the devices from rectifying to ideal linear IV properties. Through compressive force modulation by AFM tip, a symmetric 80 meV reduction in SBH at ±0.7 V is realized for the sample grown on Mo. By a combination of temperature and force modulation, a 40 meV increase in SBH is achieved at 0.53 V for the sample on ITO. These results show that the formed structure is ideal for applications in optoelectronics, sensing, piezotronic, piezo-phototronic, and nano-energy harvesting devices.