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Adhésion et mécanique standardisées de lymphocytes T : rôles dans l'activation par anticorps et cellules présentatrices d'antigène, sous force / Standardized adhesion and mechanics of T lymphocytes : roles in activation by antibodies and antigen-presenting cells, under forceSadoun, Anaïs 05 December 2018 (has links)
Les événements biochimiques de l'activation T ont été décrits depuis longtemps et sont bien connus. A l'échelle moléculaire la liaison du Récepteur des Cellules T (TCR) présent à la surface du lymphocyte T avec un peptide (du soi ou non soi), ce dernier étant chargé sur le Complexe Majeur d'Histocompatibilité (MHC), présent à la surface des Cellules Présentatrices d'Antigène (CPA) conduit à l'initiation de la réponse immunitaire. La réponse des lymphocytes T est extrêmement spécifique, sensible, robuste et semble présenter des caractéristiques de mécano-transduction. En effet, il a été démontré récemment que le TCR agirait comme un mécanosenseur alors que le lymphocyte T peut sentir la mécanique de son environnement à une échelle cellulaire. Cependant, la majorité de ces études ont été réalisées en opposant un lymphocyte T à un substrat inerte limitant la compréhension sur la contribution éventuelle de chaque partenaire cellulaire car la présence de l'APC peut induire des changements dans l'organisation du lymphocyte T.Le but de cette thèse a été de mettre en place un suivi de l’activation des lymphocytes T pat une APC modèle, sous force grâce à l’utilisation de la microscopie à force atomique. Il a mené à plusieurs développements méthodologiques originaux validés de manière expérimentale avec un système cellulaire modèle (hybridomes murins vs. Cellules COS modifiées pour être des APC pouvant être modifiées à souhait grâce à l’expression d’une grande variété de molécules impliquées dans la réponse immunitaire). / The biochemical events of T activation have been described for a long time and are well known. At the molecular level, the binding of the T-cell receptor (TCR) present on the surface of the T-cell with a peptide (self or non-self) loaded on the Major Histocompatibility Complex (MHC), present on the surface of the Antigen Presenting Cells (APC), leads to the initiation of the immune response. In addition, the T cell response is extremely specific, sensitive, robust and appears to have mechano-transduction characteristics. Indeed, it has recently been demonstrated that the TCR would act as a mechanosensor while the T lymphocyte can feel the mechanics of its environment on a cellular scale. However, the majority of these studies were performed by opposing/facing a T cell to an inert substrate (e.g., bead, functionnlized glass slides molecularly decorated or not), limiting the understanding of the possible contribution of each cell partner because the presence of APC can induce changes in the organization of the T cell. The aim of this thesis was to set up a follow-up of the activation, under force, between a model APC and a T lymphocyte. It has led to several original methodological developments experimentally validated with a model cell system (mouse hybridomas vs. COS cells modified to be complexifiable APCs).
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Influência da fase de crescimento celular na ação fotodinâmica: avaliação morfológica, mecânica e bioquímica, em células de Candida albicans / Influence of the cell growth phase on photodynamic action: morphological, mechanical and biochemical evaluation in cells of Candida albicansBaptista, Alessandra 24 November 2015 (has links)
Estudos têm demonstrado o potencial da terapia fotodinâmica antimicrobiana (aPDT) na inativação de diferentes células microbianas. No geral, são três as fases de crescimento dos microrganismos: fase lag, exponencial e estacionária. Os objetivos deste estudo foram avaliar a susceptibilidade de células de Candida albicans em diferentes fases de crescimento, submetidas à aPDT, associando azul de metileno (50 μM) e luz de emissão vermelha (λ= 660 nm) e investigar alterações morfológicas, mecânicas e bioquímicas, antes e depois da aPDT, por microscopia eletrônica de varredura, de força atômica e por espectroscopia no infravermelho por transformada de Fourier. Os resultados obtidos sugerem que, em parâmetros letais, células em fase estacionária de crescimento (48 h) são menos susceptíveis à aPDT, quando comparadas àquelas em fases lag (6 h) e ex-ponencial (24 h) de crescimento. Entretanto, em parâmetros subletais, células de 6 h e 48 h mostraram a mesma susceptibilidade à aPDT. Em sequência, os experimentos foram realizados em parâmetros considerados subletais para células crescidas por 6 e 48 h. A avaliação morfológica mostrou menor quantidade de matriz extracelular em células de 6 h comparada àquelas de 48 h. A espectroscopia de força atômica mostrou que células em fase lag perderam a rigidez após a aPDT, enquanto que células em fase estacionária mostraram comportamento in-verso. Ainda, células de 48 h diminuíram sua adesividade após a aPDT, enquanto que células de 6 h e 24 h tornaram-se mais adesivas. Os resultados bioquímicos revelaram que as diferenças mais significativas entre as células fúngicas de 6 h e 48 h ocorreram na região de DNA e carboidratos. A aPDT promoveu mais alterações bioquímicas na região de DNA e carboidratos em células de 6 h e em lipídios e ácidos graxos em células de 48 h. Nossos resultados indicam que a fase de crescimento celular desempenha papel importante no sítio de ação da aPDT em células de C. albicans. / Studies have demonstrated the potential of antimicrobial photodynamic therapy (aPDT) on the inactivation of different microbial cells. Overall, there are three phases of cell growth: lag phase, exponential phase and stationary phase. The objectives of this study were to evaluate the susceptibility of Candida albicans in different growth stages submitted to aPDT, with methylene blue (50μM) and red light (λ = 660 nm) and to investigate morphological, mechanical and biochemical changes before and after aPDT, by scanning electron microscopy, atomic force microscopy and by Fourier transform infrared spectroscopy. The results suggested that with lethal parameters, cells in stationary phase (48 h) are less susceptible to aPDT, compared to those in lag phase (6 h) and exponential phase (24 h). However, in sub-lethal parameters 6 h and 48 h cells showed the same susceptibility to aPDT. The following results were obtained in sub-lethal parameters. The morphological evaluation showed lower amount of extra-cellular matrix at 6 h compared to cells growth for 48 h. The atomic force spectroscopy showed that cells in lag phase lost cell wall rigidity after aPDT, while cells in stationary phase showed a reverse behavior. Furthermore, 48 h cells presented a decrease in their adhesiveness after aPDT, whereas cells growth for 6 h and 24 h become more adhesive. The biochemical evaluation showed that the most significant differences among the fungal cells growth for 6 h and 48 h in DNA and carbohydrates. The aPDT caused more expressive alterations on DNA and carbohydrates in cells growth for 6 h, while cells growth for 48 h presented significant alterations on lipids and fatty acids. Our results indicate that cell growth phase play an important role on the target sites affected by aPDT in C. albicans cells.
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Medidas de expoentes críticos de filmes de diamante por meio de microscopia de força atômica / Measures of critical exponents of diamond films using atomic force microscopySilveira, Marcilei Aparecida Guazzelli da 28 May 1999 (has links)
Neste trabalho investigamos a dinâmica de crescimento de filmes de diamante sintetizados por meio de deposição química a vapor ativada por plasma de microondas (CVD). A caracterização foi feita utilizando, fundamentalmente, microscopia de força atômica (AFM). Analisamos o comportamento da rugosidade dos filmes como função da escala de observação e do tempo de deposição. Dessa maneira verificamos a existência de leis de potência para o crescimento e determinamos os expoentes críticos associados a essas leis. Os resultados obtidos estão em bom acordo com o processo de crescimento descrito pela equação estocástica KPZ. Os mecanismos principais são a deposição aleatória de partículas na superfície, o crescimento lateral e a dessorção. / Diamond films have been grown by Microwave Plasma assisted Chemical Vapor Deposition (CVD). The characterization has been made mainly by Atomic Force Microscopy (AFM). We could analyze the roughness behavior with the scale of observation and with the deposition time. We could determine the critical exponents associated with these laws. The results suggest that the growth process is in good agreement with the stochastic growth equation known as KPZ. The most important mechanisms are the random deposition, the lateral growth and the desorption.
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Sulfide and UV/ozone treatments on III-V semiconductors =: 用硫及紫外光/臭氧處理III-V 族半導體. / 用硫及紫外光/臭氧處理III-V 族半導體 / Sulfide and UV/ozone treatments on III-V semiconductors =: Yong liu ji zi wai guang/xiu yang chu li III-V zu ban dao ti. / Yong liu ji zi wai guang/xiu yang chu li III-V zu ban dao tiJanuary 1998 (has links)
by Choy Wing Hong. / Thesis (M.Phil.)--Chinese University of Hong Kong, 1998. / Includes bibliographical references (leaves 95-102). / Text in English; abstract also in Chinese. / by Choy Wing Hong. / ABSTRACT --- p.vi / ACKNOWLEDGEMENTS --- p.x / LIST OF FIGURES --- p.xi / LIST OF TABLES --- p.xiii / Chapter Chapter 1 --- Introduction --- p.1 / Chapter 1.1 --- Background --- p.1 / Chapter 1.2 --- Surface passivation techniques --- p.2 / Chapter 1.2.1 --- Sulfide solution passivation --- p.2 / Chapter 1.2.2 --- Gas-phase sulfide passivation --- p.3 / Chapter 1.2.3 --- Ultra-violet and ozone exposure --- p.4 / Chapter 1.3 --- Surface structure of sulfide-passivated surface --- p.5 / Chapter 1.4 --- Surface structure of ultra-violet/ozone oxidation --- p.8 / Chapter 1.5 --- Objectives of present study --- p.10 / Chapter Chapter 2 --- Instrumentation --- p.12 / Chapter 2.1 --- Introduction --- p.12 / Chapter 2.2 --- Atomic force microscopy (AFM) --- p.12 / Chapter 2.2.1 --- The development of AFM --- p.12 / Chapter 2.2.2 --- Basic principles of AFM --- p.12 / Chapter 2.2.3 --- Forces and their relevance to atomic force microscopy --- p.13 / Chapter 2.2.3.1 --- Van Der Waals forces --- p.15 / Chapter 2.2.3.2 --- Repulsive forces --- p.15 / Chapter 2.2.3.3 --- Capillary forces --- p.15 / Chapter 2.2.4 --- Displacement sensor of AFM --- p.15 / Chapter 2.2.4.1 --- Electron tunneling --- p.16 / Chapter 2.2.4.2 --- Optical interference --- p.16 / Chapter 2.2.4.3 --- Laser beam deflection --- p.16 / Chapter 2.2.5 --- Instrument specification --- p.17 / Chapter 2.2.5.1 --- Contact mode AFM --- p.17 / Chapter 2.3 --- X-ray photoelectron spectroscopy --- p.19 / Chapter 2.3.1 --- The development of XPS --- p.19 / Chapter 2.3.2 --- Basic principles of XPS --- p.19 / Chapter 2.3.3 --- XPS experiments --- p.23 / Chapter 2.3.4 --- Quantitative analysis --- p.26 / Chapter 2.3.4.1 --- Atomic concentration of a homogenous materials --- p.26 / Chapter 2.3.4.2 --- Layer structure --- p.27 / Chapter 2.4 --- Rutherford backscattering spectrometry (RBS) --- p.29 / Chapter 2.4.1 --- Basic principles --- p.29 / Chapter 2.4.2 --- Kinematics --- p.29 / Chapter 2.4.3 --- Channeling --- p.31 / Chapter Chapter 3 --- Surface treatments --- p.32 / Chapter 3.1 --- Semiconductor wafer --- p.32 / Chapter 3.2 --- Cleaning procedures --- p.32 / Chapter 3.3 --- Polysulfide passivation --- p.34 / Chapter 3.4 --- UV/Ozone oxidation --- p.39 / Chapter Chapter 4 --- Surface roughness and oxide contents of sulfide passivation --- p.41 / Chapter 4.1 --- Introduction --- p.41 / Chapter 4.2 --- Experimental methodology --- p.42 / Chapter 4.3 --- Etching --- p.44 / Chapter 4.3.1 --- Etching effect of polysulfide solution --- p.45 / Chapter 4.3.2 --- Possible consequences of the etching effect --- p.45 / Chapter 4.4 --- Oxide contents --- p.47 / Chapter 4.4.1 --- Oxide gained during polysulfide solution treatment --- p.47 / Chapter 4.4.2 --- Oxide gained after polysulfide passivation --- p.47 / Chapter 4.5 --- Surface roughness --- p.49 / Chapter 4.5.1 --- Surface roughness after different passivation methods --- p.49 / Chapter 4.5.2 --- The sticking probability after different passivations --- p.51 / Chapter 4.6 --- The spiral ladder of solution-phase passivation --- p.55 / Chapter 4.7 --- Conclusions --- p.58 / Chapter Chapter 5 --- Sulfide on Ge/GaAs heterojunction --- p.59 / Chapter 5.1 --- Introduction --- p.59 / Chapter 5.1.1 --- Band structure of Ge/GaAs heteroj unction --- p.59 / Chapter 5.1.2 --- Lattice match of Ge/GaAs heteroj unction --- p.60 / Chapter 5.1.3 --- The growth of Ge on GaAs using molecular beam epitaxy --- p.62 / Chapter 5.2 --- The growth of Ge on GaAs using thermal pulse annealing --- p.63 / Chapter 5.3 --- Sulfide as an atomic interdiffusion barrier --- p.65 / Chapter 5.3.1 --- Experimental methodology --- p.65 / Chapter 5.3.2 --- Crystallinity of Ge --- p.67 / Chapter 5.3.3 --- Results and discussions --- p.67 / Chapter 5.3.3.1 --- RBS and XPS results --- p.67 / Chapter 5.3.3.2 --- AFM and I-V results --- p.71 / Chapter 5.4 --- Conclusions --- p.71 / Chapter Chapter 6 --- UV/03 on Ge/GaAs heterojunction --- p.72 / Chapter 6.1 --- Introduction of UV/o3 oxidation --- p.72 / Chapter 6.2 --- UV/o3 oxidation on GaAs --- p.74 / Chapter 6.3 --- Ge on UV/o3 treated GaAs --- p.76 / Chapter 6.3.1 --- Experimental methodology --- p.76 / Chapter 6.3.2 --- Crystallinity of Ge --- p.77 / Chapter 6.3.3 --- AFM results --- p.77 / Chapter 6.3.4 --- RBS results --- p.80 / Chapter 6.4 --- Diodes --- p.82 / Chapter 6.4.1 --- Fabrication of diode --- p.82 / Chapter 6.4.2 --- Diode characteristics --- p.84 / Chapter 6.4.3 --- I-V characteristics --- p.90 / Chapter 6.5 --- Conclusions --- p.90 / Chapter Chapter 7 --- Conclusion and future work --- p.93 / Chapter 7.1 --- Conclusions --- p.93 / Chapter 7.2 --- Future works --- p.94 / Reference --- p.95
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The Effect of Biopolymer Properties on Bacterial Adhesion: an Atomic Force Microscopy (AFM) StudyAbu-Lail, Nehal Ibrahim 18 September 2003 (has links)
"The effect of bacterial surface biopolymers on bacterial adhesion to surfaces was studied through experiments and modeling. Atomic Force Microscopy (AFM) provided the tool to measure the interaction forces between different bacterial cells and silicon nitride tips under different chemical conditions at a nanoscopic level. Two bacterial strains were considered: Pseudomonas putida KT2442 and Escherichia coli K-12 JM109. This study addressed the following issues: 1) the effect of solution ionic strength and solvent polarity on adhesion between Pseudomonas putida KT2442 and the silicon nitride AFM tip, 2) role of heterogeneity of bacterial surface biopolymers on bacterial adhesion, 3) role of lipopolysaccharides (LPS) on adhesion at three different scales: continuous, batch, and nanoscale, and 4) nature of interactions between E. coli JM109 and a model surface (silicon nitride tip). To address the first issue, formamide, water, and methanol were used to investigate the effect of polarity on surface characteristics of biopolymers on the bacterial surface while a range of salt concentrations between that of water to 1 M KCl were used to study the effect of ionic strength. The adhesion increased with decreasing polarity of the solvent, indicating that the polymers on the bacterial surface are hydrophilic in nature. The adhesion was slightly affected by ionic strength variations up to a concentration of 0.1 M KCl; this may have been due to the fact that the ionic concentration in the solution did not counterbalance the ionic concentration in the biopolymer brush on the bacterial surface. However, a dramatic increase in the adhesion magnitude was observed when the salt concentration increased above 0.1 M KCl. This transition in adhesion with ionic strength from a low to high value induced a transition in the elasticity of the bacterial surface biopolymers. The biopolymer brush layer did change from rigid to soft with increasing the ionic strength. The elasticity was quantified mainly by the use of the freely jointed chain (FJC) model. Our interest in investigating the role of heterogeneity on adhesion developed from the results of the first study. The bacterial surface polymers were thought to be different in their chemical and physical nature since they were found to span a range of segment lengths. Analyzing the adhesion forces for P. putida KT2442 showed that the bacterial surface is heterogeneous. The heterogeneity was evident on the same cell surface and between different cells from the same population. To resolve the third issue, approximately, 80% of the surface LPS of E. coli K-12 JM109 were removed by treating the cells with 100 mM ethylenediaminetetraacetic acid (EDTA). The effect of LPS removal on the adhesion of the cells to the silicon nitride tip was studied in water and phosphate buffered silane (PBS). The adhesion results from the AFM experiments were compared to batch retention experiments with glass as the substratum and column attachment experiments with columns packed with quartz sand. LPS controlled bacterial adhesion to the different surfaces in the study at three scales: batch, continuous, and nano-scale. Finally, the nature of interactions between E. coli JM109 and a model surface (silicon nitride tip) were investigated in solvents of varying polarity (formamide, water, and methanol). The Young’s modulus of elasticity for the bacterial surface was estimated by fitting of the Hertzian model to the force-indentation curves. Young’s modulus values increased as the solvent polarity decreased, indicating a stiffer bacterial surface in lower polarity solvents. The average adhesion force in each solvent was negatively correlated with the dielectric constant of the solvent, suggesting hydrophilic biopolymers. Specific and non-specific interaction forces between the AFM tip and the biopolymers were further characterized by applying a Poisson statistical analysis to the discrete adhesion data. The specific and non-specific interaction forces were the highest in methanol (-4 and -1.48 nN respectively). These values are in accordance with the high adhesion magnitude values measured with AFM in methanol. The results of my different studies emphasized the important role of AFM in studying biological interactions to different surfaces and in characterizing bacterial surface biopolymers."
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Investigation of Measurement Artifacts Introduced By Horizontal Scanning Surface Profiling InstrumentsBergstrom, Torbjorn S 08 January 2002 (has links)
Horizontal scanning instruments, such as, atomic force microscopes and scanning laser microscopes, acquire three-dimensional topographic maps of surfaces, at scales ranging from tenths of nanometers to hundreds of millimeters, by measuring elevations along a series of traces scanning a region of the surface. Random and systematic errors may influence parameters calculated from these topographic maps. This work investigates anisotropic artifacts in atomic force microscope and a scanning laser microscope measurements by looking at difference between parameters calculated in the tracing and scanning directions. It is found that horizontal scanning profiling instruments systematically introduce anisotropic measurement artifacts when measuring both isotropic and anisotropic surfaces.
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Rhéologie et tribologie aux nanoéchelles / Rheology and tribology at the nanoscaleComtet, Jean 03 July 2018 (has links)
Dans ce manuscrit, nous mesurons la réponse mécanique à l’échelle nanométrique de divers systèmes issus de la matière molle en utilisant un microscope à force atomique basé sur un diapason à quartz. Utilisé comme un nano-rhéomètre, cet instrument permet une mesure quantitative des propriétés viscoélastiques des matériaux et des processus frictionnels et dissipatifs aux nanoéchelles. Nous montrons d’abord que les liquides ioniques confinés aux nanoéchelles peuvent subir un changement dramatique de leurs propriétés mécaniques, suggérant une solidification capillaire. Cette transition est favorisée par la nature métallique des interfaces confinantes, montrant la présence d’effets électrostatiques subtils dans ces électrolytes denses. Nous étudions ensuite les mécanismes de plasticité à l’échelle atomique en mesurant la réponse viscoélastique de jonctions d’or de quelques atomes de diamètre. Nous mettons en évidence une transition sous cisaillement entre un régime élastique, puis plastique, jusqu’à la liquéfaction complète de la jonction. Nous caractérisons ainsi de manière fine les mécanismes de plasticité dans ces systèmes moléculaires. Finalement, nous montrons les effets profonds que les interactions à l’échelle nanométrique peuvent avoir sur le comportement macroscopique de la matière molle. Nous mesurons le profil frictionnel entre paires de particules de suspensions de PVC et de maïzena. Nos mesures mettent en lumière le rôle dominant des interactions locales entre particules dans la rhéologie non-newtonienne des suspensions. / In this manuscript, we use a tuning fork based atomic force microscope to measure the mechanical response of various soft matter systems at the nanoscale. This instrument is used as a nano-rheometer, allowing quantitative measurements of viscoelastic material properties, and unprecedented characterization of friction and dissipation at the nanoscale. First, we show that ionic liquids can undergo a dramatic change in their mechanical properties when confined at the nanoscale, pointing to a capillary freezing transition. This transition is favored by the metallic nature of the confining substrates, suggesting the occurrence of subtle electrostatic effects in those dense electrolytes. Second, we probe plasticity at the individual atomic level, by measuring the viscoelastic rheological response of gold junctions of few atoms diameter. For increasing shear, we uncover a transition from a purely elastic regime to a plastic flow regime, up to the complete shear-induced melting of the junction. Our measurements give unprecedented insights on the plastic mechanisms at play in those molecular systems. Finally, we show that nanoscale interactions can have profound effects on the macroscopic behavior of soft materials. Focusing on the nonnewtonian flow behavior of concentrated suspensions of particles, we measure the nanoscale frictional force profile between pairs of particles of PVC and cornstarch suspensions. Our measurements highlight the dominant role of local interparticle interactions on the macroscale rheology of suspensions.
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Desenvolvimento do biofilme bacteriano em superfícies de metais puros / Bacterial biofilm development onto pure metals surfaceMendes, Natália Helena 29 September 2015 (has links)
Biofilmes são aglomerados complexos de células microbianas que crescem na superfície de um material sólido, como um metal. As superfícies metálicas são amplamente utilizadas em dispositivos biomédicos cirúrgicos e em superfícies de mobiliário intra-hospitalares as quais podem ser infectadas por bactérias epidemiologicamente importantes e permitir o desenvolvimento de um biofilme. O objetivo desse trabalho foi avaliar a superfície topográfica dos metais puros, incluindo: chumbo, cromo, estanho, ferro e níquel, avaliar a aderência bacteriana nestas superfícies, com a consequente formação de biofilme e a potencial citotoxicidade dos metais por meio de microscopia de força atômica (MFA), microscopia óptica e microscopia eletrônica de varredura (MEV). A metodologia constou de observações microscópicas e procedimentos bacteriológicos. A aderência bacteriana foi verificada por meio de MEV e a viabilidade celular bacteriana por contagem de Unidades Formadoras de Colônia (UFC). A citotoxicidade dos metais foi avaliada frente a células CHO-K1 por ensaio XTT. As bactérias selecionadas foram: Escherichia coli, Pseudomonas aeruginosa (gram-negativos); Staphylococcus epidermidis e Staphylococcus aureus (gram-positivos) Para realizar o estudo, foram preparados corpos de prova dos metais puros e colocados em contato com cada uma das bactérias (da ordem 108 UFC/mL). Os resultados mostraram a formação de biofilme em cada um dos corpos de prova. A contagem das células viáveis demonstrou a recuperação de 105 UFC/mL para Escherichia coli, Pseudomonas aeruginosa e Staphylococcus aureus após contato com os metais chumbo, cromo, estanho, ferro e níquel, e para Staphylococcus epidermidis após contato com chumbo, níquel e cromo houve uma redução bacteriana de 103 UFC/mL. Para Staphylococcus epidermidis após contato com estanho foram recuperadas 1,14 x 102 UFC/mL e para o ferro, houve recuperação de 1,3 x 103 UFC/mL. A MEV demonstrou nestas superfícies metálicas, os bacilos e cocos aderidos e agrupados em uma massa amorfa formando biofilme. Os resultados da rugosidade (Ra) de cada uma destas superfícies obtidos por MFA em varredura em 2 microns da superfície, mostram que o Ra do chumbo foi de 38.258 μm; estanho 13.481 μm; níquel 3.929 μm; ferro 3.689 μm e cromo 2.097 μm. Os resultados do teste XTT, após contato com as células CHO-K1, mostram que o ferro foi citotóxico para estas células (p<0,05) diferença estatisticamente significante em relação ao controle negativo. Os metais puros avaliados nas condições experimentais do estudo mostram que as superfícies dos metais puros não impedem a aderência bacteriana e formação de biofilme das bactérias selecionadas. / Biofilms are complex microbial cell clusters that are growing on the solid material surface, as a metal. The metalic surfaces are widely used in surgical biomedical devices and hospital furniture surfaces which can be infected by important bacteria and to allow biofilm development. The aim of this work was to evaluate the topographic surface of the lead, chromium, tin, iron and nickel pure metals, evaluate bacterial adhesion in these surfaces with subsequent biofilm formation and the potential cytotoxicity of metals through force microscopy atomic (AFM), optical microscopy and scanning electron microscopy (SEM). The methodology consisted of microscopic observations and bacteriological procedures. Bacterial adherence was observed by SEM and the bacterial cell viability by colony forming units colony counts (CFU). The cytotoxicity of metals was evaluated using CHO-K1 cells by XTT assay.The isolated bacteria were: Escherichia coli, Pseudomonas aeruginosa (gram-negative), and Staphylococcus epidermidis and Staphylococcus aureus (gram positive). In order to realize this study, samples of pure metals were prepared and put in touch with each bacteria (in 108 CFU/mL). The results show biofilm formation in each of the specimens. The counting of viable cells demonstrated a recovery 105 CFU/mL for Escherichia coli, Pseudomonas aeruginosa and Staphylococcus aureus after contact with lead, chromium, tin, iron and nickel metals and Staphylococcus epidermidis after contact with lead, nickel and chromium there was a bacterial reduction of 103 CFU/mL. For Staphylococcus epidermidis after contact with tin were recovered 1.14 x 102 CFU / mL and the iron, there was a recovery of 1.3 x103 CFU/mL. The data show that there was a bacterial reduction of 103 CFU/mL viable cells. Staphylococcus epidermidis on contact with tin were recovered 1,14 x 102 and the iron recovery was 1,3 x 103 UFC/mL. SEM showed the metal surfaces, bacilli and coccus adhered and grouped in an amorphous mass forming biofilm. The results of the roughness (Ra) of each of the surfaces obtained by AFM scan of 2 microns from the surface, show that the RA lead was 38,258 uM; Tin 13,481 uM; Nickel 3,929 uM; iron 3,689 μm and chrome 2,097 μm. The results of XTT, after contact with the CHO-K1 cells, show that iron was cytotoxic to these cells (p <0.05) statistically significant difference compared to the negative control. Pure metals evaluated in the experimental conditions of the study show that the surfaces of pure metals do not prevent bacterial adherence and biofilm formation of the bacteria selected.
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Sistema de análise de imagens SEBS por microscopia de força atômica / Image analysis system SEBS by atomic force microscopyValencia, Carolina Elisa Guillen 04 April 2014 (has links)
Neste trabalho, se pretende caracterizar a morfologia de filmes finos poliméricos por meio de técnicas de processamento de imagens, utilizando principalmente a geometria computacional e técnicas de classificação de padrões. Os objetivos principais foram quantificar as grandezas geométricas das estruturas observadas nos filmes finos e descrever padrões de superfície formados nestes filmes. Foram estudadas imagens obtidas por microscopia de força atômica (AFM) de amostras de filmes finos SEBS [poliestireno-poli(etileno-co-butileno)-poliestireno], depositados sobre um substrato de mica por técnicas de imersão. Os filmes finos SEBS são considerados de grande interesse devido à formação de estruturas auto-organizadas na escala nanométrica. A caracterização e a obtenção da morfometria dos filmes são de relevância neste trabalho, pois contribuem para o entendimento da dinâmica de formação destes padrões nas nanoestruturas estudadas. Foram analisadas diferentes morfologias, como forma de gotículas com anéis concêntricos e forma de tiras e pontos regularmente espaçados. Os resultados obtidos permitem caracterizar os padrões observados. / In this work, we intend to characterize the morphology of polymer thin films by techniques of image processing, mainly using computational geometry and pattern classification. The main objectives were to quantify the geometrical structures observed in thin films and describe surface patterns formed in these films. Were studied images obtained by atomic force microscopy (AFM) of SEBS [polystyrene-poly(ethylene-co-butylene)-polystyrene] thin films samples, deposited on a mica substrate by dip-coating technique . SEBS thin film polymers have great interest due to the formation of self-organized structures on the nanometer scale. The characterization and obtaining measurements of the morphology of the thin films are of relevance in this work, because they contribute to the understanding of the formation dynamics of these patterns in nanostructures studied. We analyzed different morphologies, such as droplets form with concentric rings and stripe and regularly spaced points forms. The results allow to characterize the observed patterns.
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Influência de agentes clareadores na rugosidade de compósitos dentários / Influence of bleaching gels on roughness of dental compositesEduardo Varanda 18 December 2009 (has links)
O clareamento dental é o meio disponível mais simples, comum e conservador para o cirurgião-dentista proporcionar aos pacientes o padrão de cor de seus dentes mais desejado. Em alguns casos, os dentes que vão ser clareados podem apresentar restaurações realizadas com compósitos dentais, que são mais suscetíveis a alterações químicas, quando comparados a outros materiais restauradores. Alguns estudos mostraram que diferentes concentrações de agentes clareadores levaram a um aumento significativo da rugosidade superficial e das porosidades em compósitos dentais. Este estudo avaliou o efeito de dois agentes clareadores (Whiteness HP Blue 20%, Whiteness HP Max) sobre a rugosidade superficial de dois compósitos dentais, um micro-híbrido (Esthet X, Denstply) e outro nanoparticulado (Z 350, 3M ESPE). Um total de oito corpos de prova (9 x 2 mm) foram confeccionados com auxílio de uma matriz de teflon, sendo divididos em 4 grupos (Esthet X + Whiteness HP Blue 20%; Esthet X + Whiteness HP Max; Z 350 + Whiteness HP Blue 20%; Z 350 + Whiteness HP Max), sendo n=2. Os corpos de prova foram armazenados em saliva artificial neutra e, após 24 horas, foram polidos com discos de óxido de Alumínio (Sof-lex, 3M ESPE). Após sete dias de imersão salivar, cada corpo de prova foi levado a um microscópio de força atômica para obtenção do valor inicial de rugosidade superficial (Ra em nm). Em seguida, sem remover o corpo de prova do microscópio, o agente clareador foi aplicado sobre a superfície do corpo de prova, segundo as instruções do fabricante, de modo que permitisse uma posterior observação da mesma área do corpo de prova observada inicialmente, para obtenção do valor final de rugosidade superficial. Imagens em duas e três dimensões foram obtidas de cada corpo de prova para observação de alterações da topografia. Os resultados foram tratados estatisticamente por ANOVA e pelo teste de contraste Student-Newman-Keuls (p < 0,05). Não houve alteração significativa na rugosidade superficial (Ra) dos corpos da prova de compósitos micro-híbrido e nanoparticulado, submetidos aos agentes clareadores Whiteness HP Blue 20% e Whiteness HP Maxx. No entanto, independente do agente clareador utilizado, foram observadas maiores alterações topográficas nas imagens de microscopia de força atômica da superfície do compósito micro-híbrido do que nas imagens do nanoparticulado. / Dental bleaching is the simplest, the most common and conservative available way for dentists to offer their patients the standard color of teeth more desired. In some cases, the teeth that will be bleached may include restorations made of resin composites, which are more prone to chemical alteration compared with other restoration materials. Some studies have showed that different concentrations of bleaching gels might lead to a significant increase in surface roughness and amount of porosities in composite resins. This study evaluated the effect of two bleaching gels (Whiteness HP Blue 20%, Whiteness HP Max) on surface roughness of two resin composites, micro-hybrid (Esthet X, Denstply) and nanoparticled (Z350, 3M ESPE). A total of eight specimens (9 x 2 mm) were produced with a Teflon mold help, and were divided into four groups (Esthet X + Whiteness HP Blue 20%; Esthet X + Whiteness HP Max; Z 350 + Whiteness HP Blue 20%; Z 350 + Whiteness HP Max), with n=2. They were stored in neutral artificial saliva and, after 24 hours, were polished with aluminium oxide discs (Sof-lex, 3M ESPE). After seven days of salivary immersion, each specimen was taken to an atomic force microscope for recording initial value of surface roughness (Ra in nm). After that procedure, without removing the specimen from microscope, the bleaching gel was applied on the surface of the specimen, following manufacturers instructions, in order to allow posterior observation of the same area of the specimen initially observed, for recording the final value of surface roughness. Two-dimensional and 3D images were taken from each specimen group for observation of surface alterations. The results were statistically analyzed by ANOVA and the SNK test (p< 0.05). There were no statistically significant differences in the surface roughness (Ra) for specimens of micro-hybrid and nanoparticled composites, submitted to the bleaching agents Whiteness HP Blue 20% and Whiteness HP Maxx. However, there were noticed larger surface alterations in atomic force microscope images on the surface of micro-hybrid composites than on the nanoparticled, independently of the bleaching agent used.
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