"Bacteria during the course of their life undergo a lot of developments on their surface. The changes that occur inside a cell result in the production of a variety of biopolymers on the cell surface. These polysaccharides have been found to play a major role in deciding the adhesive or repulsive nature of a bacterial cell. Based on the application the adhesive nature of a cell sometimes needs to be manipulated such that bacteria are required to have higher adhesions for bioremediation applications and in the case of bioreactors bacteria must not stick to walls to avoid fouling. In order to control adhesions of a cell to a variety of substrates, knowledge of the polysaccharides present on its surface is needed. Therefore the goal of the present study is to detect the sugars present on the surface of Pseudomonas putida KT2442 using Atomic force microscopy and to relate properties of the polysaccharides to bacterial adhesion. Previous experiments suggested that cellulose and other sugars were produced by Pseudomonas putida KT2442. Thus the cells were grown to late exponential phase and treated with cellulase to degrade any cellulose, if present, on the surface of the cells. Control experiments were done on untreated cells and cells that were not treated with cellulase but were centrifuged, since centrifugation is a part of the cellulase treatment and may also affect the bacterial surface. An appropriate (Steric) fitting model for the atomic force microscope (AFM) approach curves was applied to calculate the height and density of the polymer brush layer present on the cell surface. There was a decrease in the density of the polymer brush and increase in the height of the brush upon treatment with cellulase. Centrifugation alone did not affect the approach curves. From looking at the retraction curves it verified the results got from the approach curves and indicated stretching out of the polymer brush to greater distances after the treatment with cellulase. Another batch of cells was treated with dextranase to check for the presence of dextran on the cell surface. Dextranase treated cells behaved identical to the control cells, suggesting that dextran is not one of the polysaccharides present on the bacterial surface. No change was observed in retraction curves data for dextranase treated and untreated cells."

O objetivo deste estudo foi avaliar o pH e a liberação de íons cálcio dos cimentos AH Plus, MTA Fillapex e Sealapex, em diferentes períodos, quando empregados em obturações de canais radiculares, bem como, a qualidade das obturações, após 1 ano de realizadas. O pH e a liberação de íons cálcio foram avaliados por meio de um peagômetro e espectrofotômetro de absorção atômica, respectivamente. A qualidade das obturações foi avaliada por meio de um estereomicroscópio. Cento e vinte e quatro pré-molares inferiores humanos extraídos, unirradiculados, foram divididos em 01 grupo controle e 02 grupos experimentais, de acordo com a padronização do diâmetro do forame apical. Em seguida, os 02 grupos experimentais foram divididos em 4 subgrupos, considerando as técnicas de instrumentação e obturação empregadas. Após o preparo dos canais radiculares cada subgrupo teve o número de dentes divididos por dois, formando novos subgrupos, de acordo com os cimentos obturadores utilizados. Imediatamente após a obturação, a raiz de cada dente foi imersa em 15 mL de água ultrapura para avaliação do pH e liberação de íons cálcio nos seguintes períodos: 1 h, 1, 3, 7, 30, 180 e 365 dias, após a obturação. Após o período de 1 ano, os dentes foram seccionados transversalmente a 2, 4, 6 mm do forame apical para análise da qualidade das obturações, analisando-se a porcentagem de cimento, guta-percha e espaços vazios na massa obturadora. Os cimentos apresentaram pH neutro, porém, aos 180 dias, os valores mostraram um ligeiro aumento quando utilizados em canais com forames de diâmetro 0,25 mm. Quando utilizados em canais com forames de diâmetro 0,45 mm, o pH mostrou-se com valores maiores para todos os períodos sem distinção do cimento. Todos os cimentos liberaram íons cálcio, com menores valores para o AH Plus. Na estereomicroscopia, observaram-se variações nas porcentagens de cimento e guta-percha, considerando-se os diâmetros foraminais e técnicas obturadoras, e porcentagem insignificante de espaços vazios. / The objective of this study was to evaluate the pH and calcium ion release of AH Plus, MTA Fillapex and Sealapex cements at different time periods when used in root canal obturations, as well as the quality of the obturations one year after being performed. The pH and calcium ion release were evaluated by means of a pH meter and an atomic absorption spectrophotometer, respectively. The quality of the obturations was evaluated using a stereomicroscope. One hundred and twenty-four extracted human uniradicular mandibular premolars were divided into 1 control and 2 experimental groups, according to the standardization of the apical foramen diameter. Next, the 2 experimental groups were divided into 4 subgroups, considering the instrumentation techniques and obturations employed. After the preparation of the root canals, each subgroup had the number of teeth divided by two, forming new subgroups, according to the sealers used. Immediately after the obturation, the root of each tooth was immersed in 15 mL of ultrapure water for the evaluation of the pH and calcium ion release in the following time periods: 1 hr, 1, 3, 7, 30, 180 and 365 days after the obturation. After the period of one year, the teeth were transversely sectioned at 2, 4 and 6 mm from the root apex to examine the quality of the obturations, analyzing the percentage of cement, gutta-percha and voids in the obturator mass. The cements presented a neutral pH, which showed slightly increase as of 180 days when used in canals with foramens of 0.25 mm diameters. When used in canals with foramens with diameters of 0.45 mm, the pH presented higher values in all periods for all cements. All cements released calcium ions, with lower values for the AH Plus. In the stereomicroscopy, there were variations in the percentages of cement and gutta-percha, considering the foraminal diameters and obturation techniques and an insignificant percentage of voids.

Algumas indicações morfológicas sugerem que as superfícies de paisagem tropicais onde encontram-se os duricrusts ferruginosos, podem apresentar idades que variam desde o Quaternário até o Cretáceo, e que a laterização sobre estas superfícies pode ter sido iniciada simultaneamente ou não, estabelecendo-se uma sequência cronológica de formação escalonada de acordo com a elevação, em função da evolução geomorfológica das superfícies. Sabendo-se que no Planalto de Diamantina na Serra do Espinhaço Meridional (SdEM) são reconhecidos três níveis geomorfológicos correlatos aos ciclos de aplainamento da plataforma Sul-americana, o objetivo deste trabalho foi estudar as variações mineralógicas, geoquímicas e morfológicas de duricrusts ferruginosos da Serra do Espinhaço Meridional buscando entender os processos genéticos envolvidos e as implicações disto sobre a distribuição e evolução da paisagem regional e suas relações com os ciclos de aplainamento do continente Sul-americano. Para atingir estes objetivos, foram realizadas análises de difratometria de raios X, suscetibilidade magnética, microscopia eletrônica de varredura com sistema de energia dispersiva acoplado e análise elementar a partir da dissolução total dos minerais constituintes. Os resultados mostraram que no Planalto de Diamantina na SdEM coexistem níveis de ferricretes e lateritas na superfície correspondente ao ciclo erosivo Pós-Gondwanico. As superfícies lateríticas são provavelmente as formações supérgenas mais antigas da paisagem regional, originadas durante os processos denudacionais que ocorreram ao longo do ciclo erosivo Pós-Gondwanico. A erosão parcial de seu perfil laterítico constituíram o material fonte de Fe e Al para a gênese dos ferricretes distribuídos em superfícies elevadas (> 1200 m), especialmente aqueles em superfícies em torno de 1400 m de altitude. As variações paleoclimáticas do ciclo Pós-Gondwanico também proporcionaram ciclos erosivos alternados que resultaram no subescalonamento desta superfície, criando condições geomorfológicas favoráveis à gênese de ferricretes mais recentes (1200 - 1400 m). Além disso, os duricrusts ferruginosos do Planalto Diamantina (SdEM) sob influência do maciço quartzítico do Supergrupo Espinhaço e situados superfícies erosivas elevadas (> 1200 m), além de policíclicos, também podem apresentar características poligenéticas. / Some morphological indications suggest that the tropical landscape surfaces where the iron-rich duricrust are present may have ages varying from the Quaternary to the Cretaceous, and that the laterization on these surfaces may have been initiated simultaneously or not, establishing a sequence chronological formation according to the elevation, according to the geomorphological evolution of the surfaces. Three geomorphological levels correlated to the South American platform planing cycles are known on the Diamantina Plateau, for this the objective of this work was to study the mineralogical, geochemical and morphological variations of the iron-rich duricrust of the SdEM, trying to understand the genetic processes involved and the implications of this on the distribution and evolution of the regional landscape and its relations with the planning cycles of the South American continent. In order to reach these objectives, X-ray diffraction, magnetic susceptibility, scanning electron microscopy with dispersive energy system coupled and elemental analysis carried out from the total dissolution of the constituent minerals. The results showed that in the Diamantina Plateau at the SdEM, coexist levels of ferricretes and laterites on the surface corresponding to the Post-Gondwanic erosive cycle. The laterite surfaces are probably the oldest surviving formations of the regional landscape, originated during the denudation processes that occurred along the Post-Gondwanic erosive cycle. Partial erosion of its lateritic profile was source material of Fe and Al for the genesis of ferricretes distributed on high surfaces (> 1200 m), especially those on surfaces at 1400 m of altitude. The paleoclimatic variations of the Post-Gondwanic cycle also provided alternating erosive cycles that resulted in sub-scaling of this surface, creating geomorphological conditions favorable for genesis of earlier ferricretes (1200 - 1400 m). In addition, the ferricretes of the Diamantina Plateau (SdEM) under influence of the Quartzite massif of the Espinhaço Supergroup and located in elevated erosive surfaces (> 1200 m), besides polycyclics, may also present polygenetic characteristics.

Submitted by Cristiane Chim (cristiane.chim@ucpel.edu.br) on 2018-08-30T12:05:38Z No. of bitstreams: 1 Lucas Soares da Costa.pdf: 6533559 bytes, checksum: 540c854aef17b6fb97cad699d5be5f0a (MD5) / Made available in DSpace on 2018-08-30T12:05:38Z (GMT). No. of bitstreams: 1 Lucas Soares da Costa.pdf: 6533559 bytes, checksum: 540c854aef17b6fb97cad699d5be5f0a (MD5) Previous issue date: 2017-03-28 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPES / From the perspective of systems biology, the cell is the main element of information integration. Therefore, understanding the cell in its context is the key to unraveling many of the still unknown mechanisms of life and certain pathologies. Despite increasing automation in clinical analysis laboratories, many of them still employ optical microscopes for some specific analyzes. When working with microorganisms, it is most often desired to determine the cell concentration of the prepared suspension. One of the most common ways of obtaining this estimate is by counting under the microscope, using a Neubauer Chamber, also known as a Hemacytometer or Count Chamber. Each microscope image can contain hundreds of cells to be counted. Performing the counting visually entails a very large inaccuracy and requires high time, which also increases the time to obtain the final estimate. The current state of scientific research in Brazil and other third world countries reflects the need for a device that is easily accessible and inexpensive for researchers. Thus, in view of the aforementioned reasons, the development of a mechanism that matches these expectations becomes indispensable. In view of this perspective, the objective of this study was consolidated in the development of an algorithm for cellular counting through the capture of images from the Neubauer chamber. This algorithm results in counting accuracy greater than 90 % when compared to manual counting, as well as reducing the time from approximately two minutes to less than one second of the algorithm. / Na perspectiva de biologia de sistemas, a célula é o elemento principal da integração de informações. Portanto, compreender a célula no seu contexto é a chave para desvendar muitos dos mecanismos ainda desconhecidos da vida e de certas patologias. Apesar da crescente automatização nos laboratórios de análises clínicas, muitos deles ainda empregam microscópios óticos para algumas análises específicas. Quando se trabalha com microorganismos, na maioria das vezes deseja-se determinar a concentração de células da suspensão preparada. Uma das formas mais comuns de se obter esta estimativa é através da contagem ao microscópio, utilizando-se uma Câmara de Neubauer, também conhecida como Hemacitômetro ou Câmara de Contagem. Cada imagem de microscópio pode conter centenas de células a serem contadas. Realizar a contagem de forma visual acarreta em uma imprecisão muito grande e requer tempo elevado, o que também aumenta o tempo para obter a estimativa final. A situação atual da pesquisa científica no Brasil e em outros países de terceiro mundo reflete a necessidade de um dispositivo de fácil acesso e baixo custo aos pesquisadores. Assim, diante dos motivos supracitados, o desenvolvimento de um mecanismo que corresponda essas expectativas, faz-se indispensável. Frente a essa perspectiva, o objetivo desse estudo consolidou-se no desenvolvimento de um algoritmo para contagem celular através da captura de imagens da câmara de Neubauer. Este algoritmo resulta em uma precisão na contagem superior a 90% quando comparado a contagem manual, como também, reduzir o tempo de aproximadamente dois minutos para menos de um segundo do algoritmo.

La combinaison de plusieurs procédés d’imagerie et de mesure permet d’obtenir des ensembles de données complémentaires et parfois uniques. A l’aide d’une technique hybride de microscopie présentant des modalités de mesure différentes et des enregistrements synchrones, on peut recueillir des informations complémentaires sur des échantillons à l’échelle nanométrique. De plus, l’intégration de procédés nanorobotiques et de logiciels open-source permet une approche technologique pour la recherche sur les semi-conducteurs et les sciences des matériaux. Ce travail démontre le potentiel d’une telle technologie. Ce démonstrateur fonctionne dans la chambre d‘un MEB et sert de plateforme technologique dans laquelle sont intégrés différentes modalités, technologies et procédés. Un AFM basé sur un interféromètre optique compact permet l’imagerie de la topographie de surface tandis qu’un microscope à micro-ondes à balayage enregistre les caractéristiques électromagnétiques dans la gamme de fréquence des micro-ondes, le tout opérant dans le même MEB. L’engin est contrôlé par un ensemble de logiciels qui est optimisé pour la nanorobotique basée sur l‘imagerie. Ce démonstrateur technologique permet d’observer en direct la région d’intérêt à l’aide du microscope électronique tandis qu’est effectuée en champ proche la caractérisation de la surface de l’échantillon par intermédiaire des micro-ondes évanescentes et des forces intermoléculaires. Ensuite, est présenté un standard multimodal de test et qui valide la fonctionnalité de l’instrument démonstrateur. Le présent travail est complété par une analyse électrique de capacités MOS ainsi que leur approximation destinée au calibrage. / Various disciplines of micro- and nanotechnology requires combinatorial tools for the investigation, manipulation and transport of materials in the submicrometer range. The coupling of multiple sensing and imaging techniques allows for obtaining complementary and often unique datasets of samples under test. By means of an integrated microscopy technique with different modalities, it is possible to gain multiple information about nanoscale samples by recording at the same time. The expansion with nanorobotics and an open-source software framework, leads to a technology approach for semiconductor research and material science. This work shows the potential of such a multimodal technology approach by focusing on a demonstrator setup. It operates under high-vacuum conditions inside the chamber of a Scanning Electron Microscope and serves as a technology platform by fusing various microscopy modalities, techniques and processes. An Atomic Force Microscope based on a compact, optical interferometer performs imaging of surface topography, and a Scanning Microwave Microscope records electromagnetic properties in the microwave frequency domain, both operating inside an SEM. A software framework controls the instrument. The setup allows for observing with SEM, while imaging and characterizing with interacting evanescent microwaves and intermolecular forces simultaneously. In addition, a multimodal test standard is introduced and subsequently confirms the functionality of the demonstrator. Within this context, the work also includes an electrical analysis of micro-scale MOS capacitors, including an approximation for use in the calibration.

L'imagerie plénoptique est souvent présentée comme une révolution par rapport à l'imagerie standard. En effet, elle apporte plus de contrôle à l'utilisateur sur l'image finale puisque les dimensions spatiales et angulaires du champ de lumière offrent la possibilité de changer le point de vue ou de refaire la mise au point après coup ainsi que de calculer la carte de profondeur de la scène. Cependant, cela complique le travail du concepteur optique du système pour deux raisons. La première est qu'il existe une multitude d'appareils de capture plénoptique différents, chacun avec sa propre spécificité. La deuxième est qu'il n'existe pas de modèle qui relie le design de la caméra à ses propriétés optiques d'acquisition et qui puisse guider le concepteur dans sa tâche. Cette thèse répond à ces observations en proposant un modèle optique du premier ordre pour représenter n'importe quel appareil d'acquisition plénoptique. Ce modèle abstrait une caméra plénoptique par un réseau équivalent de caméras virtuelles existant en espace objet et qui effectue un échantillonnage identique de la scène. Ce modèle est utilisé pour étudier et comparer plusieurs caméras plénoptiques ainsi qu'un microscope plénoptique monté en laboratoire, ce qui révèle des lignes directrices pour la conception de systèmes plénoptiques. Les simulations du modèle sont aussi validées par l'expérimentation avec une caméra et le microscope plénoptique. / Light field imaging is often presented as a revolution of standard imaging. Indeed, it does bring more control to the user over the final image as the spatio-angular dimensions of the light field offer the possibility to change the viewpoint and refocus after the shot and compute the scene depth map.However, it complicates the work of the optical designer of the system for two reasons. The first is that there exist a multitude of different light field acquisition devices, each with its own specific design. The second is that there is no model that relates the camera design to its optical properties of acquisition and that would guide the designer in his task. This thesis addresses these observations by proposing a first-order optical model to represent any light field acquisition device. This model abstracts a light field camera as en equivalent array of virtual cameras that exists in object space and that performs the same sampling of the scene. The model is used to study and compare several light field cameras as well as a light field microscope setup which reveals guidelines for the conception of light field optical systems. The simulations of the model are also validated through experimentation with a light field camera and a light field microscope that was constructed in our laboratory.

[pt] Nesta tese, apresentamos um procedimento utilizado para a calibração do fotodetector e dos cantileveres utilizados em nosso AFM para a medida de força lateral. Desenvolvemos um código em Matlab para o controle do microscópio que permitiu a realização do estudo da influência da força normal na fricção. Também foi desenvolvido um segundo código em Matlab para a medida automatizada da adesão. Apresentamos e discutimos a influência da energia livre superficial na fricção e adesão de várias superfícies. Neste trabalho um estudo da influência da condensação capilar na forca lateral foi estudado para superfícies hidrofílicas, e hidrofóbicas. Encontramos que as nano asperezas podem realizar contatos singulares descritos pelo modelo de Hertz ou múltiplos contatos de acordo com o modelo de Greenwood. O tipo de contato entre as nano asperezas pode ser controlado através da hidrofobicidade e da umidade relativa no ambiente de medida. É verificado que os meniscos formados entre ponta e superfície influenciam a força lateral, através do aumento da força normal e também através da energia gasta pela ponta para arrastar ou deformar o capilar durante seu deslocamento sobre a superfície. O efeito da cinética de condensação capilar da água sobre a fricção foi também estudado. É mostrado que a molhabilidade é determinante para a definição dos mecanismos da dissipação de energia entre as nanoasperezas. Apresentamos também a influência da hidrofobicidade superficial no coeficiente de atrito. A correlação observada entre o ângulo de contato e o coeficiente de atrito reforça a importância da cinética da condensação capilar nos processos de fricção que ocorre na escala de nanômetros. / [en] In this work, the procedures developed to the calibration of the AFM photodetector and cantilevers for lateral force measurements in our AFM is presented. A Matlab code that controls the microscope allows the study of the influence of the normal force on the lateral one. A second Matlab code was developed in order to study the adhesion forces in an automated way. We present and discuss the influence of the surface free energy on the friction and adhesion forces. In this work, the lateral forces were measured at hydrophilic and hydrophobic surfaces. It was observed that the nano asperities may form single asperity contacts described by the Hertz model as well as multi-asperity type of contacts described by the Greenwood model. The nanoasperity contact may be controlled by the wettability and ambient relative humidity. It is seen that the capillar formed between the tip and the surface influences the tip-surface normal force and the friction forces due to the dissipation of energy caused by the drag or brake of the capillar meniscous. The effect of capillary condensation kinetics was studied as well. It is shown that the surface wettability is determinant to the energy dissipation mechanism in nanoscale. The influence of the surface wettability on the friction coefficient is presented. The observed correlation between the friction coefficient and contact angle enhances the influence of the surface wettability and its kinetics in the friction forces at nanoscale.

Dans ce travail de thèse, nous avons cherché d'une part, à répondre à la question du dimensionnement des nanosystèmes électromécaniques (NEMS) à base de nanotube de carbone (NTC) multiparois, à la fois théoriquement et expérimentalement, et d'autre part, à appliquer le savoir-faire développé à la réalisation de composants de types interrupteurs.<br /> Nous avons développé un modèle théorique pour décrire la déflexion d'un nanotube de carbone suspendu soumis à une force d'attraction électrostatique. Notre modèle montre qu'il existe une loi d'échelle reliant la déflexion électrostatique aux paramètres géométriques, électriques, et physiques des NEMS à base de NTC. Ce résultat constitue, en soi, un outil de dimensionnement pour la conception de ces dispositifs, car il permet de prédire leur comportement électromécanique sur une « large » gamme de paramètres opérationnels.<br /> Parallèlement, nous avons mis au point des procédés de fabrication pour réaliser des nanostructures intégrant un NTC suspendu actionnable électrostatiquement. Parmi ces différentes structures, la plus simple a été utilisée pour sonder les propriétés électromécaniques des NTC multiparois. Nous avons développé une méthode basée sur l'utilisation d'un microscope à force atomique pour mesurer la déflexion en fonction de la tension électrique d'actionnement. Les résultats de ces mesures, pour différents NTC (différents diamètre et longueur), montrent clairement, et sans paramètre ajustable, l'existence de la loi d'échelle prédite par notre modèle théorique. À partir de ces mesures, nous avons extrait le module d'Young des NTC. Pour des diamètres inférieurs à 30 nm, celui-ci est constant et vaut en moyenne 400 GPa. Au-delà, nous observons une forte diminution qui pourrait s'expliquer par l'entrée dans un régime de déformation non-linéaire.<br /> Enfin, nous montrons la réalisation d'un interrupteur électromécanique à base de NTC qui présente de bonnes caractéristiques de commutation.

Avec l'intégration de circuits intégrés de plus en plus denses, le besoin d'outils de caractérisation adaptés à ces échelles se fait ressentir. Identifier et analyser les problèmes de fiabilité survenant dans ces structures à des dimensions inférieures à 100 nm demande la mise au point d'instruments innovants. Ce travail de thèse a consisté dans un premier temps à développer un appareil à champs proches sensible aux propriétés mécaniques de surface, et dans un second temps à analyser les résultats expérimentaux en s'appuyant sur des approches analytiques et/ou numériques. Désigné sous le nom de microscope à force atomique à résonance de contact (CR-AFM), cet appareil est sensible à la rigidité effective de films minces sur substrat, ce qui lui permet de cartographier la rigidité mécanique de films minces. Nous avons mené un important travail de développement instrumental afin d'obtenir des résultats expérimentaux répétables et fiables, condition indispensable à une analyse quantitative. Puis nous avons utilisé le CR-AFM sur divers échantillons : empilements modèles (films de silice sur silicium, avec épaisseurs variables de silice), films de silice avec porosité variable, structures damascènes d'interconnexion cuivre,... Des images traduisant les variations d'élasticité de surface ont ainsi pu être construites. Pour quantifier ces variations, nous avons analysé nos résultats à l'aide de différents modèles (approches analytiques et numériques). Des simulations par éléments finis ont été réalisées pour étayer ces résultats.

Surface characteristics such as topography and critical dimensions serve as important indicators of product quality and manufacturing process performance especially at the micrometer and the nanometer scales. This paper first reviews different technologies used for obtaining high precision 3-D images of surfaces, along with some selected applications. Atomic force microscopy (AFM) is one of such methods. These images are commonly distorted by convolution effects, which become more prominent when the sample surface contains high aspect ratio features. In addition, data artifacts can result from poor dynamic response of the instrument used. In order to achieve reliable data at high throughput, dynamic interactions between the instrument's components need to be well understood and controlled, and novel image deconvolution schemes need to be developed. Our work aims at mitigating these distortions and achieving reliable data to recover metrology soundness. A summary of our findings will be presented. / Singapore-MIT Alliance (SMA)