The potential application of a hydrogel-based strong anion-exchange (Q) membrane to purify plasmid DNAs was evaluated. The maximum binding capacity of plasmid DNA was estimated to be 12.4 mg/ml of membrane volume with a plasmid DNA recovery of ~ 90%, which is superior to other commercially available anion-exchange resins and membranes. The membrane was able to retain its structural integrity and performance after multiple cycles of usage (> 30 cycles). The inherent properties of plasmid DNA, membrane adsorbent, and the ionic environment on membrane performance were identified as the factors affecting membrane performance and their effects were systematically investigated. Plasmid DNAs with smaller tertiary structure have shorter dynamic radius and/or lowersurface charge densities, which tended to have a better adsorption and recovery than those with larger tertiary structure. Environmental Scanning Electron Microscopy (ESEM) revealed that the hydrogel structure is more porous on one side of membrane than the other, and higher plasmid DNA adsorption and recovery capacities were observed if the more porous side of the membrane was installed upward of flow in the chromatographic unit. ESEM also revealed improved pore distribution and increased membrane porosity if membrane was pre-equilibrated in the buffer solution for 16 hours. The development of better flow through channel in the hydrogel membrane upon extensive soaking further improved plasmid DNA adsorption and recovery capacities. The ionic environment affects the tertiary size of plasmid DNA; and the optimal operating pH of membrane chromatography was different for the plasmid DNAs investigated in this study. The relative contribution of these factors to improve membrane chromatography of plasmid DNAs was analyzed using statistical modeling. It was found that the adsorption of plasmid DNA was mainly affected by the available adsorptive area associated with membrane porosity, whereas the recovery of plasmid DNAs was mainly affected by the environmental pH. A novel, RNase-free, and potentially scalable bioprocess was synthesized using the hydrogel membrane as the technology platform for the manufacturing of pharmaceutical-grade plasmid DNA. High bioprocess recovery and product quality were primarily associated with the optimal integration of impurity removal by calcium chloride precipitation and anion-exchange membrane chromatography and the implementation of isopropanol precipitation as a coupling step between the two impurity-removing steps. Complete removal of total cellular RNA impurity was demonstrated without the use of animal-derived RNase. High-molecular-weight (HMW) RNA and genomic DNA (gDNA) were removed by selective precipitation using calcium chloride at an optimal concentration. Complete removal of the remaining low-molecular-weight (LMW) RNA was achieved by membrane chromatography using the high-capacity and high-productive hydrogel membrane. The simultaneous achievement of desalting, concentrating and buffer exchange by the coupling step of isopropanol precipitation and the high efficiency and resolution of DNA-RNA separation by anion-exchange membrane chromatography significantly reduced the operating complexity of the overall bioprocess, increased the overall recovery of plasmid DNA, and enhanced product quality by removing trace amounts of impurities of major concern for biomedical applications, such as gDNA, proteins, and endotoxin.

Le bois est un matériau complexe, hétérogène et anisotrope. Ses propriétés mécaniques varient fortement en fonction de l'essence et de l'échelle considérées. Pour mieux comprendre le comportement du bois en fonction de sa structure, nous proposons dans le cadre de cette thèse une étude expérimentale à l'échelle des tissus qui est réalisée sur plusieurs types de bois : bois de peuplier, y compris bois de tension et bois d'épicéa. Compte tenu des faibles dimensions des échantillons testés, les mesures de déformation sont effectuées " sans contact " afin de s'assurer de ne pas perturber l'échantillon en cours de l'essai.Les premières séries de mesures sont réalisées sur les trois zones (tendue, normale, opposée) et selon les 3 directions matérielles (longitudinale, radiale et tangentielle) d'une tige de peuplier inclinée. Les résultats obtenus permettent de comparer des comportements mécaniques du bois de peuplier dans toutes les zones considérées en fonction de ses propriétés microscopiques.Les deuxièmes séries de mesures sont réalisées en compression, grandes déformations, dans la chambre d'un ESEM (Microscope électronique à balayage environnemental) sur du bois feuillu (peuplier) et sur du bois résineux (épicéa). Grâce aux images obtenues en période d'essai, du domaine élastique jusqu'à la densification, la réponse de chaque type du bois à la même sollicitation est illustrée.Enfin, une partie de la modélisation numérique par la MPM (Materiel Point Method) est proposée dans notre projet envisagé comme prolongement de ce travail. Il permettra de simuler le comportement mécanique du bois en grande déformation.

The potential application of a hydrogel-based strong anion-exchange (Q) membrane to purify plasmid DNAs was evaluated. The maximum binding capacity of plasmid DNA was estimated to be 12.4 mg/ml of membrane volume with a plasmid DNA recovery of ~ 90%, which is superior to other commercially available anion-exchange resins and membranes. The membrane was able to retain its structural integrity and performance after multiple cycles of usage (> 30 cycles). The inherent properties of plasmid DNA, membrane adsorbent, and the ionic environment on membrane performance were identified as the factors affecting membrane performance and their effects were systematically investigated. Plasmid DNAs with smaller tertiary structure have shorter dynamic radius and/or lowersurface charge densities, which tended to have a better adsorption and recovery than those with larger tertiary structure. Environmental Scanning Electron Microscopy (ESEM) revealed that the hydrogel structure is more porous on one side of membrane than the other, and higher plasmid DNA adsorption and recovery capacities were observed if the more porous side of the membrane was installed upward of flow in the chromatographic unit. ESEM also revealed improved pore distribution and increased membrane porosity if membrane was pre-equilibrated in the buffer solution for 16 hours. The development of better flow through channel in the hydrogel membrane upon extensive soaking further improved plasmid DNA adsorption and recovery capacities. The ionic environment affects the tertiary size of plasmid DNA; and the optimal operating pH of membrane chromatography was different for the plasmid DNAs investigated in this study. The relative contribution of these factors to improve membrane chromatography of plasmid DNAs was analyzed using statistical modeling. It was found that the adsorption of plasmid DNA was mainly affected by the available adsorptive area associated with membrane porosity, whereas the recovery of plasmid DNAs was mainly affected by the environmental pH. A novel, RNase-free, and potentially scalable bioprocess was synthesized using the hydrogel membrane as the technology platform for the manufacturing of pharmaceutical-grade plasmid DNA. High bioprocess recovery and product quality were primarily associated with the optimal integration of impurity removal by calcium chloride precipitation and anion-exchange membrane chromatography and the implementation of isopropanol precipitation as a coupling step between the two impurity-removing steps. Complete removal of total cellular RNA impurity was demonstrated without the use of animal-derived RNase. High-molecular-weight (HMW) RNA and genomic DNA (gDNA) were removed by selective precipitation using calcium chloride at an optimal concentration. Complete removal of the remaining low-molecular-weight (LMW) RNA was achieved by membrane chromatography using the high-capacity and high-productive hydrogel membrane. The simultaneous achievement of desalting, concentrating and buffer exchange by the coupling step of isopropanol precipitation and the high efficiency and resolution of DNA-RNA separation by anion-exchange membrane chromatography significantly reduced the operating complexity of the overall bioprocess, increased the overall recovery of plasmid DNA, and enhanced product quality by removing trace amounts of impurities of major concern for biomedical applications, such as gDNA, proteins, and endotoxin.

This research presents the results on an experimental investigation to identify the significant factors influencing horizontal cracking in continuously reinforced concrete pavements (CRCP). An in-depth analysis of the microstructure, morphological characteristics of the interfacial transition zone (ITZ) and the observation of cracking using the environmental scanning electron microscope (ESEM) was done. Characterization of oxides using Fourier transform infrared spectroscopy (FTIR) and electron dispersive x-ray spectroscopy (EDS) was also performed. Water to cement ratio (w/c) and rebar temperature had a significant influence on the rebar-concrete bond strength. The 28-day shear strength measurements showed an increase in rebar-concrete bond strength as the water to cement ratio (w/c) was reduced from 0.50 to 0.40. There was a reduction in the peak pullout load as the temperature increased from 14oF to 252oF for the corroded and non-corroded rebar experiments. The corroded rebar pullout test results showed a 20-50 % reduction in bond strength compared to the non-corroded rebars. FTIR measurements indicated a presence of lepidocrocrite (γ -FeOOH) and maghemite (γ -Fe2O3) on the ITZ. ESEM images showed the existence of microcracks as early as three days after casting with the bridging of these cracks between coarse aggregate locations in the interfacial zone propagating through the mortar.

The first part of the thesis deals with electron microscopy and physical model fluid flow. In the next part, a three-dimensional model is created in programme SolidWorks. It is the model of scintilating detector with aperture or screen with three hundred little apertures. These models are submitted to analysis focusing on investigation of the gas flow at different pressure in chamber of specimen ESEM using Cosmos FloWorks system. The results of the analysis of both apertures variants are compared with the request to minimize the pressure in the trajectory of the secondary electrons as well as in the chamber of specimen.

The thesis is focused on the design of a scintillation detector with one screen for environmental scanning electron microscope. At the beginning of the work is an introduction to microscopy. Further work is focused on the description of ESEM microscope and a scintillation detector. The following chapter is devoted to the dynamics of flow. They list the types of flow and mathematical description of equations. The next chapter is about using the software and the way of solution of flow. In the following chapter describes a proposal of the diaphragm and extraction method of the detector area. They are described some of type aperture. The last chapter is the conclusion, where the work is summarized.

The thesis is concerned with the design of the shape of suction canals used for pumping in the scintillation detector. The goal is to perform analysis of the model of the detector according to the current conception and then perform an optimalisation of the suction canals as per demands to their function. In the beginning of the thesis the history of the microscope and electron microscopy is described. The next chapter is focused on a more detailed description of ESEM. The following charter is dedicated to describing the scintillation detector.The thesis recounts the kinds of liquid flow. In conclusion the programs SolidWorks and Ansys Fleunt are described. Next part of this thesis describes in detail parameters‘ settings of simulation calculation. Following chapter introduces designed changes of suction channels shape and its results are shown and described.

This thesis deals with scanning electron microscope working at higher pressure in the specimen chamber. The main goal was to study the voltage contrast on the PN junction of the transistor under suitable working conditions for using environmental scanning microscope. The observation of sample was enabled by a scintillation detector designed for observation of high pressure.

La problématique énoncée par l’utilisation de matériaux composites C/C denses implique la connaissance et la maîtrise des processus de dégradation auxquels ils sont soumis. L’utilisation de moyens de caractérisation in-situ de ces voies de dégradation constitue alors un atout considérable pour leur anticipation. Ainsi, l’utilisation de la MEBE en Température associée à une caractérisation cristallographique par MET et une confrontation ex-situ par Analyse thermogravimétrique a abouti à l’obtention de lois cinétiques caractérisant la propagation de l’oxydation dans toutes les directions de l’espace. A la suite de cette étape expérimentale, une approche numérique basée sur l’utilisation d’algorithmes de Monte-Carlo Cinétique, a alors été mise en place pour modéliser ces observations tant sur le plan atomique avec la modélisation de la loi cinétique d’oxydation linéique, que meso et macroscopique par la simulation de la loi cinétique de perte de masse dans le cas particulier du HOPG. / The problem stated by the use of composites C / C dense implies knowledge and control of degradation processes to which they are subjected. The use of in-situ characterization of these means of degradation pathways then is a considerable asset for their advance. Thus, the use of ESEM in temperature associated with a crystallographic characterization by TEM and ex situ confrontation by thermogravimetric analysis resulted in obtaining kinetic laws characterizing the propagation of oxidation in all directions. Following this experimental stage, a numerical approach based on the use of algorithms Kinetic Monte-Carlo, was then introduced to model these observations both at the atomic level with the modeling of the oxidation kinetics law linear, as meso-and macro-simulation by the kinetic law of mass loss in the case of HOPG.

En vue de la réduction des problèmes engendrés par les phénomènes de retrait-gonflement des sols dans la dépression de la Lama au Sud du Bénin, nous avons entrepris des recherches dans le but de maîtriser le comportement de ces sols. Six échantillons de sols fins dont quatre argileux de cette région ont été prélevés. Différents essais ont été réalisés : une caractérisation macroscopique concernant les propriétés mécaniques et géotechniques et les essais microscopiques prenant en compte les analyses chimiques, thermiques, minéralogiques et microstructurales. Les sols étudiés sont constitués de différents minéraux argileux. Celle à structure fibreuse est à l'origine d’un comportement macroscopique particulier décelé par l’essai de gonflement libre. Les essais ont confirmé le fort potentiel de gonflement et de retrait de certains sols qui sont donc à l'origine des fortes dégradations rencontrées.Les résultats montrent que les sols traités se distinguent par les performances mécaniques acquises et peuvent être utilisés à différents niveaux de la chaussée selon leur teneur en liant.Les différentes méthodes utilisées ont révélé des facettes particulières de l'étude de traitement des sols fins. Principalement, nous avons noté que les informations apportées par les observations au Microscope Electronique à Balayage, les analyses thermiques et les analyses de Diffraction aux Rayons X ont permis de compléter l’analyse des résultats géotechniques. En somme, les méthodes de caractérisation géotechnique permettent d’avoir une idée générale du potentiel de gonflement, mais pour les études d’envergure, la réalisation d’études microscopiques est primordiale. / To help reduce the problems caused by phenomena due to the shrink-swell soil in the depression of the Lama in southern Benin, we have undertaken research in order to control the behavior of these soils. Six soil samples, including four clay from this area were taken. Different types of tests were performed: a macroscopic characterization concerning mainly mechanical and geotechnical properties and microscopic tests taking into account the chemical, thermal, microstructural and mineralogical analysis. The soils studied consist of different clay minerals. The fibrous structure is at the origin of a particular macroscopic behavior detected by the free swell test. The tests confirmed the potential for swelling and shrinking of some soils that are causing heavy damage.Three soils treated with different contents at Binder Hydraulic Road, were subjected to several tests. The results show that these soils are distinguished by the acquired mechanical performance and may be used at different levels of the floor according to binder content.Also, the different methods revealed specific facets of fine soil treatment study. Mainly, we noted that the information provided by the observations scanning electron microscope, thermal analysis and the analysis of diffraction X-Ray are very important and helped complete the analysis of geotechnical results for soil treatment. In sum, geotechnical characterization methods used to get a general idea of the swelling potential, but for large-scale studies, conducting microscopic studies is paramount.