A Na, K-ATPase (NKA) é uma proteína de membrana encontrada em organismos eucariotos multicelulares cuja atividade e funções já são amplamente discutidas na literatura. Sua unidade funcional corresponde a um heterodímero formado por duas subunidades , com regiões transmembrana. Espécies multiméricas como dímeros e tetrâmeros dessa enzima também são conhecidos por exercer atividade enzimática. As interações lipídio-proteína são intrínsecas para a NKA, por tal motivo, proteolipossomos constituídos de DPPC e DPPC:DPPE foram preparados por co-solubilização. Como controle, lipossomos de mesma composição foram produzidos por extrusão e/ou sonicação. Para as imagens de AFM, as amostras foram fixadas com glutaraldeído, para proteção mecânica e contra desidratação das vesículas. Para lipossomos de DPPC as imagens topográficas de AFM das vesículas apresentaram formato oval, superfície perfeitamente lisa e diâmetro médio de 151 + 46 nm, enquanto as vesículas de composição DPPC:DPPE, apesar de lisas, tiveram cantos pontiagudos e diâmetro médio de 98 + 28 nm. Imagens de fase de ambas as composições não apresentaram qualquer indicativo de diferenças na composição química, provavelmente devido à natureza de carga neutra dos dois fosfolipídios. As imagens de fase por AFM para os proteolipossomos tanto de DPPC-NKA, quanto DPPC:DPPE-NKA, revelaram resultados inéditos na literatura, onde a inserção da NKA aparece como nítidas regiões transições de fase de composição química distinta quando comparadas com os lipossomos. No entanto, as mudanças de fase são diferentes entre as composições estudadas, aparecendo como manchas escuras circulares para DPPC-NKA e mais visíveis como interstícios brilhantes para composição de DPPC:DPPE-NKA. As vesículas de DPPC-NKA apresentaram diâmetro médio de 390 + 326 nm e, nas imagens de topografia tridimensionais, protusões de 38 a 115 nm correspondentes às regiões de mudanças de fase, que, indicaram o diâmetro dos microdomínios relacionados à proteína. Já nas imagens para DPPC:DPPE-NKA o diâmetro médio dos proteolipossomos foi de 189 + 156 nm, e as protusões apareceram entre os interstícios, variando de 20 a 66 nm. O estudo de DSC dos lipossomos revelou que a concentração de glutaraldeído nas condições das análises de AFM, em torno de 5% (v/v), afetam as características físico-químicas para as composições com DPPE. A AFM foi eficiente para confirmar a reinserção da NKA em proteolipossomos pelas imagens de fase, e, para medir o diâmetro dos microdomínios pelas imagens de topografia. / Na, K-ATPase (NKA) is a membrane protein present in eukaryotic multicellular organisms. Its functions and activity are already widely described in the literature. Its minimal functional structure is a heterodimer of two main subunits , with transmembrane domains. However, dimers and tetramers of the enzyme are also known to have enzymatic activity. Since there are intrinsic lipid-protein interactions, NKA proteoliposomes composed of DPPC and DPPC:DPPE (1:1 molar ratio) were prepared by the co-solubilization method and liposomes of the same compositions were obtained by extrusion and/or sonication to be used as control. The samples to the AFM study were prepared using glutaraldehyde to protect the vesicles from mechanical shocks and dehydration. Liposomes composed of DPPC and DPPC:DPPE (1:1 molar ratio) were prepared by extrusion and sonication, respectively, as control. The topographical images for DPPC liposomes showed vesicles with an oval shape and smoothed surfaces with a mean diameter of 151 + 46 nm. DPPC:DPPE vesicles also presented smoothed surfaces, but with pointed corners and mean diameter of 98 + 28 nm. Phase images for both lipid compositions showed no differences in chemical composition. For DPPC:DPPE samples, this can be explained by the neutral net charge of both lipids. The proteoliposomes observed in the AFM phase images showed darker and large circular spots in the vesicles. These spots represent delays in the phase oscillation of the AFM probe and are associated with different chemical composition. The phase changes showed the reconstitution of the NKA in the proteoliposomes. When compared with topographical images, this spots matched protrusions. The mean diameter of DPPC-NKA proteoliposomes determined by AFM was 390 + 326 nm. In the three-dimensional topographical images of composition, protrusions from 38 to 115 nm near the areas of different phases indicate the diameters of the NKA microdomains. The phase changes for DPPC:DPPE-NKA appeared as bright interstices with the protrusions of the topographical images in between them. The size of these protrusions ranged from 20 to 66 nm and the mean diameter of the proteoliposomes was 189 + 156 nm. The DSC liposomes data showed that the glutaraldehyde concentration used in the AFM analysis affect the physical chemistry properties of the samples with DPPE. AFM proved to be an efficient method to confirm the reconstitution of into proteoliposomes with phase images and to determine the diameter of the protein microdomains with the topographical images.

A Na, K-ATPase (NKA) é uma proteína de membrana encontrada em organismos eucariotos multicelulares cuja atividade e funções já são amplamente discutidas na literatura. Sua unidade funcional corresponde a um heterodímero formado por duas subunidades , com regiões transmembrana. Espécies multiméricas como dímeros e tetrâmeros dessa enzima também são conhecidos por exercer atividade enzimática. As interações lipídio-proteína são intrínsecas para a NKA, por tal motivo, proteolipossomos constituídos de DPPC e DPPC:DPPE foram preparados por co-solubilização. Como controle, lipossomos de mesma composição foram produzidos por extrusão e/ou sonicação. Para as imagens de AFM, as amostras foram fixadas com glutaraldeído, para proteção mecânica e contra desidratação das vesículas. Para lipossomos de DPPC as imagens topográficas de AFM das vesículas apresentaram formato oval, superfície perfeitamente lisa e diâmetro médio de 151 + 46 nm, enquanto as vesículas de composição DPPC:DPPE, apesar de lisas, tiveram cantos pontiagudos e diâmetro médio de 98 + 28 nm. Imagens de fase de ambas as composições não apresentaram qualquer indicativo de diferenças na composição química, provavelmente devido à natureza de carga neutra dos dois fosfolipídios. As imagens de fase por AFM para os proteolipossomos tanto de DPPC-NKA, quanto DPPC:DPPE-NKA, revelaram resultados inéditos na literatura, onde a inserção da NKA aparece como nítidas regiões transições de fase de composição química distinta quando comparadas com os lipossomos. No entanto, as mudanças de fase são diferentes entre as composições estudadas, aparecendo como manchas escuras circulares para DPPC-NKA e mais visíveis como interstícios brilhantes para composição de DPPC:DPPE-NKA. As vesículas de DPPC-NKA apresentaram diâmetro médio de 390 + 326 nm e, nas imagens de topografia tridimensionais, protusões de 38 a 115 nm correspondentes às regiões de mudanças de fase, que, indicaram o diâmetro dos microdomínios relacionados à proteína. Já nas imagens para DPPC:DPPE-NKA o diâmetro médio dos proteolipossomos foi de 189 + 156 nm, e as protusões apareceram entre os interstícios, variando de 20 a 66 nm. O estudo de DSC dos lipossomos revelou que a concentração de glutaraldeído nas condições das análises de AFM, em torno de 5% (v/v), afetam as características físico-químicas para as composições com DPPE. A AFM foi eficiente para confirmar a reinserção da NKA em proteolipossomos pelas imagens de fase, e, para medir o diâmetro dos microdomínios pelas imagens de topografia. / Na, K-ATPase (NKA) is a membrane protein present in eukaryotic multicellular organisms. Its functions and activity are already widely described in the literature. Its minimal functional structure is a heterodimer of two main subunits , with transmembrane domains. However, dimers and tetramers of the enzyme are also known to have enzymatic activity. Since there are intrinsic lipid-protein interactions, NKA proteoliposomes composed of DPPC and DPPC:DPPE (1:1 molar ratio) were prepared by the co-solubilization method and liposomes of the same compositions were obtained by extrusion and/or sonication to be used as control. The samples to the AFM study were prepared using glutaraldehyde to protect the vesicles from mechanical shocks and dehydration. Liposomes composed of DPPC and DPPC:DPPE (1:1 molar ratio) were prepared by extrusion and sonication, respectively, as control. The topographical images for DPPC liposomes showed vesicles with an oval shape and smoothed surfaces with a mean diameter of 151 + 46 nm. DPPC:DPPE vesicles also presented smoothed surfaces, but with pointed corners and mean diameter of 98 + 28 nm. Phase images for both lipid compositions showed no differences in chemical composition. For DPPC:DPPE samples, this can be explained by the neutral net charge of both lipids. The proteoliposomes observed in the AFM phase images showed darker and large circular spots in the vesicles. These spots represent delays in the phase oscillation of the AFM probe and are associated with different chemical composition. The phase changes showed the reconstitution of the NKA in the proteoliposomes. When compared with topographical images, this spots matched protrusions. The mean diameter of DPPC-NKA proteoliposomes determined by AFM was 390 + 326 nm. In the three-dimensional topographical images of composition, protrusions from 38 to 115 nm near the areas of different phases indicate the diameters of the NKA microdomains. The phase changes for DPPC:DPPE-NKA appeared as bright interstices with the protrusions of the topographical images in between them. The size of these protrusions ranged from 20 to 66 nm and the mean diameter of the proteoliposomes was 189 + 156 nm. The DSC liposomes data showed that the glutaraldehyde concentration used in the AFM analysis affect the physical chemistry properties of the samples with DPPE. AFM proved to be an efficient method to confirm the reconstitution of into proteoliposomes with phase images and to determine the diameter of the protein microdomains with the topographical images.

A mathematical model is developed to investigate the grazing dynamics of tapping mode atomic force microscopes (AFM) subjected to a base harmonic excitation. The nonlinear dynamics of the AFM microcantilever are studied in both of the monostable and bistable phases with the microcantilever tip being, respectively, located in the monostable and bistable regions of the static bifurcation diagram in the reference configuration. Free-vibration responses of the AFM probes, including the microcantilever natural frequencies and mode shapes, are determined. It is found that, for the parameters used in a practical operation of an AFM, the natural frequencies and mode shapes of the AFM microcantilever are almost the same as those of a free-end microcantilever with the same geometry and made of an identical material. A multimode Galerkin approximation is utilized to discretize the nonlinear partial-differential equation of motion and associated boundary conditions governing the cantilever response and obtain a set of nonlinearly coupled ordinary-differential equations (ODE) governing the time evolution of the system dynamics. The corresponding nonlinear ODE set is then solved using numerical integration schemes. A comprehensive numerical analysis is performed for a wide range of the excitation amplitude and frequency. The tip oscillations are examined using nonlinear dynamic tools through several examples. The non-smoothness in the tip/sample interaction model is treated rigorously. A higher-mode Galerkin analysis indicates that period doubling bifurcations and chaotic vibrations are possible in tapping mode microscopy for certain operating parameters. It is also found that a single-mode Galerkin approximation, which accurately predicts the tip nonlinear responses far from the sample, is not adequate for predicting all of the nonlinear phenomena exhibited by an AFM, such as grazing bifurcations, and leads to both quantitative and qualitative errors. A point-mass model is also developed based on the single-mode Galerkin procedure to compare with the present distributed-parameter model. In addition, a reduced-order model based on a differential quadrature method (DQM) is employed to explore the dynamics of the AFM probe in the bistable phase where the multimode Galerkin procedure is computationally expensive. We found that the DQM with a few grid points accurately predicts the static bifurcation diagram. Moreover, we found that the DQM is capable of precise prediction of the lowest natural frequencies of the microcantilever with only a few grid points. For the higher natural frequencies, however, a large number of grid points is required. We also found that the natural frequencies and mode shapes of the microcantilever about non-contact equilibrium positions are almost the same as those of the free-end microcantilever. On the other hand, free-vibration responses of the microcantilever about contact equilibrium positions are quite different from those of the free-end microcantilever. Moreover, we used the DQM to discretize the partial-differential equation governing the microcantilever motion and a finite-difference method (FDM) to calculate limit-cycle responses of the AFM tip. It is shown that a combination of the DQM and FDM applied, respectively, to discretize the spatial and temporal derivatives provides an efficient, accurate procedure to address the complicated dynamic behavior exhibited by the AFM probe. The procedure was, therefore, utilized to study the response of the microcantilever to a base harmonic excitation through several numerical examples. We found that the dynamics of the AFM probe in the bistable region is totally different from those in the monostable region. / Ph. D.

This research concerns the development and characterization of methylene bis (p-cyclohexyl isocyanate/butanediol) (HMDI/BD) based polyurethanes used in connection with surface-active anti-microbial polyurethanes. Previously studied polyurethanes having an isophorone diisocyanate/butanediol (IPDI/BD) hard block contaminated water during dynamic contact angle (DCA) analyses. This contamination by unknown species confounds results from biocidal studies and jeopardizes the use of the polyurethane as a matrix polyurethane. By contrast, polyurethanes with methylene bis (p-cyclohexyl isocyanate)/butanediol hard block showed no contamination during DCA analysis. For this reason, further study of HMDI/BD/PTMO polyurethanes was conducted. HMDI/BD polyurethanes were synthesized with 15-50wt% hard block and a soft block of PTMO-2000 or PTMO-1000 where PTMO-2000 is poly (tetramethylene oxide) with a molecular weight of 2000g/mol and PTMO-1000 has a molecular weight of 1000g/mol. Characterization was performed with FT-IR and 1H NMR spectroscopy to verify polyurethane composition as well as hard block percentage. Thermal characterization was performed with modulated differential scanning calorimetry (MDSC). From MDSC, the glass transition temperatures of the soft and hard block for polyurethanes with PTMO-2000 as the soft block were -80°C and 86°C, respectively. For corresponding polyurethanes containing PTMO-1000 as the soft block, the measured Tgs for the soft and hard segments were -55°C and 65°C, respectively. The disparity between the respective soft and hard segment Tgs of these polyurethanes of differing soft block molecular weights is due to increased phase mixing that causes an increase in soft block Tg and a decrease in hard block Tg for the PTMO-1000 polyurethanes. From dynamic contact angle analyses of HMDI/BD/PTMO polyurethanes, the advancing and receding contact angles gradually decreased with each cycle but approached 80° and 60°, respectively. Results from force-distance curves with flamed glass slides obtained before and after immersion of the polyurethane coatings indicated that no water contamination occurred. Tensile tests demonstrated that hard block percentage, soft block molecular weight, and the amount of chain extender influences mechanical properties. For example, increasing hard block weight percentage increases the modulus. HMDI/BD(30)/PTMO-2000 (PU-1), HMDI/BD(25)/PTMO-2000, (PU-2) and HMDI/BD(35)/PTMO-2000 (PU-10) exhibited the best elastomeric properties. As the final outcome, lack of contamination and good mechanical properties made PU-2 and PU-9 (HMDI/BD(50)/PTMO-1000) suitable candidates as polyurethane matrices for polymer surface modifier evaluation.

<p>The work presented in this thesis concernes two sorts of nanostructures: energetic-ion-impact-induced surface tracks and gas-deposited WO₃ nanoparticles. Our aims to characterise these nanostuctures and understand the physical principles behind their formation are of general interests for basic science as well as of the field of nanotechnology.</p><p>AFM studies of irradiated organic surfaces showed that individual ion impacts generate craters, most often accompanied by raised plastically deformed regions. Crater sizes were measured as a function of ion stopping power and incidence angle on various surfaces. Observed crater volumes were converted into estimates of total sputtering yields, which in turn were correlated with data from collector experiments. The observations were compared to predictions of theoretical sputtering models. The observed plastic deformations above grazing-incidence-ion penetration paths agree with predictions of the pressure pulse model. However, closer to the ion track, evaporative sputtering can occur.</p><p>AFM images of gas-deposited WO₃ nanoparticle-films indicated the formation of agglomerates. The size distribution of the agglomerates was measured to be log-normal, <i>i.e.</i> similar to the size distribution of the gas-phase nanoparticles forming the deposit. By simulations we could relatively well reproduce this observation. The agglomerates exhibited high thermal stability below 250°C when considering their size, implying that these porous films can be useful in applications involving elevated temperatures in the 250°C range. The appearance of the nanoparticles in the tapping-mode AFM images was sensitive to the free amplitude of the oscillating tip. We could show by model calculations that the high adhesion between the tip and the sample could account for some of these observations.</p>

The work presented in this thesis concernes two sorts of nanostructures: energetic-ion-impact-induced surface tracks and gas-deposited WO3 nanoparticles. Our aims to characterise these nanostuctures and understand the physical principles behind their formation are of general interests for basic science as well as of the field of nanotechnology. AFM studies of irradiated organic surfaces showed that individual ion impacts generate craters, most often accompanied by raised plastically deformed regions. Crater sizes were measured as a function of ion stopping power and incidence angle on various surfaces. Observed crater volumes were converted into estimates of total sputtering yields, which in turn were correlated with data from collector experiments. The observations were compared to predictions of theoretical sputtering models. The observed plastic deformations above grazing-incidence-ion penetration paths agree with predictions of the pressure pulse model. However, closer to the ion track, evaporative sputtering can occur. AFM images of gas-deposited WO3 nanoparticle-films indicated the formation of agglomerates. The size distribution of the agglomerates was measured to be log-normal, i.e. similar to the size distribution of the gas-phase nanoparticles forming the deposit. By simulations we could relatively well reproduce this observation. The agglomerates exhibited high thermal stability below 250°C when considering their size, implying that these porous films can be useful in applications involving elevated temperatures in the 250°C range. The appearance of the nanoparticles in the tapping-mode AFM images was sensitive to the free amplitude of the oscillating tip. We could show by model calculations that the high adhesion between the tip and the sample could account for some of these observations.

This dissertation focuses on quantitative subsurface and mechanical properties imaging potential of AFM probes. Extensive modeling of AFM probes are presented for thorough understanding of capabilities and limitations of current techniques, these models are verified by various experiments, and different methods are developed by utilizing force-sensing integrated read-out active tip (FIRAT), which is an active AFM probe with broad bandwidth. For quantitative subsurface imaging, a 3-D FEA model of AFM tip-sample contact is developed and this model can simulate AFM tip scan on nanoscale-sized buried structures. FIRAT probe, which is active and broadband, is utilized for interaction forces imaging during intermittent contact mode and mechanical characterization capability of this probe is investigated. It is shown that probe dynamics, stiffness, stiffness ambiguity, assumed contact mechanics, and noise are important parameters for the accuracy of mechanical properties imaging. An active tip control mechanism is introduced to limit contact forces during intermittent contact mode. In addition to these, a combined ultrasonic AFM and interaction forces imaging method is developed and modeled to solve the reduced elasticity measurement sensitivity on composite materials. This method is capable of imaging a broader range of elasticity on combination samples such as metal nanoparticles in polymers at nanoscale.

In der vorliegenden Arbeit wird die Entwicklung von ausschließlich elektrostatisch arbeitenden Sensor-Aktor-Arrays zur Oberflächenprofilbestimmung an Mikroteilen beschrieben. Ein wesentliches Merkmal der Strukturen ist ihr großer Eigenzustellbereich von bis zu 20 Mikrometer. Die Auswertung atomarer Kräfte ermöglicht Wegauflösungen im Nanometerbereich. Auf Grund der geringen Abmessungen durch die mikromechanische Fertigung des Sensorelements und der integrierten Sensor-Aktorfunktion sind Anordnungen als Zeilenarray möglich. Die Entwicklung richtet sich auf Strukturen, welche in klassischer Oberflächentechnologie gefertigt werden können. Durchgeführte experimentelle Tests wurden mit Sensoren in Silizium-bulk-Mikromechanik (SCREAM) realisiert. Der Schwerpunkt der Arbeit behandelt die Charakterisierung der Sensorelemente und damit verbundene Layoutverbesserungen, wie das Einbringen von Feldstoppern und die Nutzbarkeit des Sensors zur Profilbestimmung von Oberflächen unter Beachtung industrieller Anforderungen. Vorteile des Einsatzes eines solchen Sensor-Aktor-Arrays liegen in der Miniaturisierung und dem vergleichsweise großen Eigenzustellbereich jedes einzelnen Sensors. Dadurch ist es möglich, technische Oberflächen, welche im Eigenzustellbereich des Sensorarrays liegen, ohne das Nachregeln einer übergeordneten Positioniereinheit im Profil zu bestimmen. Es wird gezeigt, wie die angewandte kapazitive Wirkungsweise des Sensors mit den sehr kleinen Nutzkapazitäten im Beisein von großen Parasitärkapazitäten zur Signalauswertung genutzt werden kann.

In this thesis work several manufacturing methods for graphene is discussed followed by an indepth study of graphene grown by a high temperature sublimation method (sublimation of siliconcarbide). The graphene surfaces studied have been grown by Graphensic AB, both graphenegrown on the Si-face and the C-face of the silicon carbide were studied. Six graphene samplesgrown 4H-SiC substrates were examined for homogeneity and surface morphology as well assome surface roughness parameters using Atomic Force Microscopy (AFM). The graphene wasstudied to get a better understanding of the surfaces and the growth mechanisms to improvemanufacturing parameters while also being informative for graphene sample customers. Anadditional graphene sample grown on 6H-SiC epitaxial layer was also studied to get a betterunderstanding of the sublimation mechanism. If graphene could be manufactured in a cheaprepeatable way the applications are endless and a new era of technology could emerge muchlike the silicon era that began several decades ago. In this thesis work the results are presentedas topography images as well as tables and histograms in the results section. The growth onthe Si-face is found to be well ordered when compared to the C-face which shows signs of alargely complex growth. The graphene on the Si-face lies on top of silicon carbide steps like acarpet with a buer layer interface against the silicon carbide. On the C-face this buer layeris not present but the graphene is deformed by buckling which is suspected to originate fromdierences in thermal properties between the graphene and the C-face. The in uence of AFMsettings for characterization of graphene while using intermittent mode have been evaluated andrecommendations are given. Finally a method for evaluating the homogeneity of the graphenelm is proposed but is in need of further verication.

In der vorliegenden Arbeit wird die Entwicklung von ausschließlich elektrostatisch arbeitenden Sensor-Aktor-Arrays zur Oberflächenprofilbestimmung an Mikroteilen beschrieben. Ein wesentliches Merkmal der Strukturen ist ihr großer Eigenzustellbereich von bis zu 20 Mikrometer. Die Auswertung atomarer Kräfte ermöglicht Wegauflösungen im Nanometerbereich. Auf Grund der geringen Abmessungen durch die mikromechanische Fertigung des Sensorelements und der integrierten Sensor-Aktorfunktion sind Anordnungen als Zeilenarray möglich. Die Entwicklung richtet sich auf Strukturen, welche in klassischer Oberflächentechnologie gefertigt werden können. Durchgeführte experimentelle Tests wurden mit Sensoren in Silizium-bulk-Mikromechanik (SCREAM) realisiert. Der Schwerpunkt der Arbeit behandelt die Charakterisierung der Sensorelemente und damit verbundene Layoutverbesserungen, wie das Einbringen von Feldstoppern und die Nutzbarkeit des Sensors zur Profilbestimmung von Oberflächen unter Beachtung industrieller Anforderungen. Vorteile des Einsatzes eines solchen Sensor-Aktor-Arrays liegen in der Miniaturisierung und dem vergleichsweise großen Eigenzustellbereich jedes einzelnen Sensors. Dadurch ist es möglich, technische Oberflächen, welche im Eigenzustellbereich des Sensorarrays liegen, ohne das Nachregeln einer übergeordneten Positioniereinheit im Profil zu bestimmen. Es wird gezeigt, wie die angewandte kapazitive Wirkungsweise des Sensors mit den sehr kleinen Nutzkapazitäten im Beisein von großen Parasitärkapazitäten zur Signalauswertung genutzt werden kann.