Les hydrogénases sont des métalloenzymes qui catalysent la conversion réversible du dihydrogène en protons et en électrons. Durant ma thèse, je me suis focalisé sur certains aspects du fonctionnement de l’hydrogénase à [NiFe] hétérodimérique de Desulfovibrio fructosovorans. Nous avons montré que contrairement au mécanisme communément admis d’inactivation aérobie de l’enzyme, l’O2 n’est pas incorporé en tant que ligand au niveau du site actif mais agit plutôt comme un simple oxydant. Ce résultat remet en question le mécanisme proposé pour expliquer la tolérance naturelle à l’O2 de certaines hydrogénases. L’analyse à l’aide d’un modèle cinétique des voltamogrammes cycliques complexes obtenus avec 16 variants a montré que les vitesses d’(in)activation en conditions anaérobies peuvent être accélérées de plusieurs ordres de grandeurs. Nous avons aussi montré et expliqué la corrélation entre ces vitesses et la tolérance à l’O2. Nous avons étudié une série de mutants qui produisent H2 beaucoup plus lentement que l’enzyme sauvage. Nous avons montré que la vitesse de cette réaction est déterminée par celle de l’étape de diffusion du H2, qui est lente dans les mutants. Finalement, nous nous sommes intéressés à une thréonine appartenant à la voie putative de transfert des protons. Nous avons démontré que cet acide aminé est effectivement impliqué dans le transport des protons. Il joue aussi un rôle crucial dans la stabilisation des intermédiaires formés au cours du cycle catalytique et probablement dans la détermination des vitesses de transfert électronique et de diffusion à travers le canal. / Hydrogenases are metalloenzymes which catalyse the reversible conversion of dihydrogen into protons and electrons. In my work, I focused on some aspects of the catalytic mechanism of the heterodimeric NiFe hydrogenase from Desulfovibrio fructosovorans. We demonstrated that, contrary to the commonly accepted mechanism of aerobic inactivation, the attacking O2 is not incorporated as an active site ligand but rather acts as an electron acceptor. This finding calls for a re-examination of the mechanism for O2 tolerance of the natural O2 tolerant NiFe hydrogenases. We also described a simple analytical model that we used to analyse the complex voltammetric signals of 16 mutants obtained by substituting an amino acid near the active site. We demonstrated that this substitution can accelerate anaerobic inactivation and reactivation by up to three orders of magnitude. We also demonstrated and explained the correlation between these rates and O2-tolerance. We studied mutants whose H2-production activity is impaired. We found that the rate limiting step of this reaction is the diffusion of hydrogen out of the enzyme, through the hydrophobic channel. Finally, we focused on a threonine belonging to the putative proton transfer pathway. We demonstrated that this amino acid is indeed implicated in proton transport. It may also play a crucial role in the stabilization of intermediates formed during the catalytic cycle, and probably also in determining the rate of electron transfer and diffusion along the gas channel.

Na literatura, a análise e a interpretação das linhas de emissão de H2 em nebulosas planetárias são feitas, em geral, considerando que a molécula somente seja produzida em ambientes neutros, como as regiões de fotodissociação ou com choques. No entanto, existem fortes evidências observacionais de que ao menos parte da emissão seja proveniente da região ionizada desses objetos. Em trabalhos anteriores mostramos que quantidades significativas de H2 podem sobreviver dentro dessa região hostil. No presente trabalho nosso objetivo é calcular e estudar a emissão de H2 em linhas no infravermelho produzidas na região ionizada de nebulosas planetárias, utilizando o código de fotoionização unidimensional Aangaba. Para isso, desenvolvemos diversas sub-rotinas que determinam o povoamento em níveis de energia da molécula e calculam a intensidade das linhas de emissão de H2. Obtivemos a intensidade de diversas linhas produzidas pela molécula H2 em nebulosas planetárias cujos parâmetros característicos (temperatura e luminosidade da estrela central, densidade do gás, tipo e tamanho dos grãos, etc.) estão na faixa de valores conhecidos para esses objetos. Como resultado de nosso trabalho, mostramos que a contribuição da região ionizada para a emissão de H2 de nebulosas planetárias pode ser significativa em diversas situações, particularmente quando a temperatura da estrela central é alta. Esse resultado pode explicar porque a detecção de linhas de H2 é mais provável em nebulosas planetárias bipolares, que têm estrelas tipicamente mais quentes. Além disso, verificamos que na região ionizada a excitação e a desexcitação colisional são mecanismos importantes de povoamento de todos os níveis rovibracionais do estado fundamental eletrônico de H2. Os mecanismos radiativos são também importantes, particularmente para os níveis de energia excitados. Os mecanismos de formação em estados excitados podem ter alguma influência no espectro de linhas produzidas pela desexcitação de níveis rotacionais bastante elevados, principalmente em ambientes densos. Em nossos modelos incluímos o efeito da molécula H2 no equilíbrio térmico do gás, verificando que a molécula H2 somente tem influência significativa na temperatura do gás em casos de temperatura da estrela central muito alta ou grande quantidade de grãos, principalmente através da desexcitação colisional. / The analysis and the interpretation of the H2 line emission of planetary nebulae have been done in the literature assuming that the molecule survives only in neutral environments, as in photodissociation or shocked regions. However, there is strong evidence that at least part of the H2 emission is produced inside the ionized region of such objects. In previous work we showed that significant amounts of H2 can survive inside the ionized region of planetary nebulae. The aim of the present work is to calculate and study the infrared line emission of H2 produced inside the ionized region of planetary nebulae using the one-dimensional photoionization code Aangaba. For this, we developed several numerical subroutines in order to calculate the statistical population of the H2 energy levels, as well as the intensity of the H2 infrared emission lines in physical conditions typical of planetary nebulae. We show that the contribution of the ionized region for the H2 emission can be significant, particularly in the case of nebulae with high temperature central stars. This result explains why H2 emission is more frequently observed in bipolar planetary nebulae (Gatley\'s rule), since this kind of object typically has hotter stars. We show that collisional excitation plays an important role on the H2 population of the rovibrational levels of the electronic ground state. Radiative mechanisms are also important, particularly for upper levels. Formation pumping may have some effects on the line intensities produced by de-excitation from very high rotational levels, specially in dense environments. We include the effects of H2 in the thermal equilibrium of the gas, concluding that H2 only contributes to the thermal equilibrium in the case of very high temperature of the central star or high grain to gas ratio, mainly through collisional de-excitation.

This thesis deals with the MIMO system analysis, with a comparison of a different frequency charactersitics, and including singular values characteristic. The H2 and H norms are defined for signals and a MIMO systems. A MIMO optimal control using an LQG, an H2 and an H control is defined and described. Their general advantages, disadvantages and similarities are summed up. An H2 and an H state controller and state observer synthesis is implemented in a MATLAB using linear matrix inequalities. Control of an inverted pendulum is designed using the H optimal cascade control, the H2 state control and the H state control. The results are compared with each other. The theoretical possibility of same H2 and H optimal state controller design is presented.

Le travail que je présente dans cette thèse est un travail principalement expérimental effectué au sein du laboratoire LAMAp de l'Université de Cergy-Pontoise, à l'aide du dispositif expérimental appelé FORMOLISM. Les principaux composants de ce dispositif sont: l'ultravide (10-10 mbar), l'ultra-froid (~5 K), des jets atomiques, la spectrométrie de masse TPD (Thermally Programmed Desorption) et la spectroscopie laser dans l'UV mettant en oeuvre des longueurs d'onde autour de ~200 nm. Grâce à la technique REMPI-TOF (Resonantly Enhanced Multi-Photon Ionization – Time Of Flight), nous avons étudié d'une part, la conversion de spin nucléaire de la molécule d'hydrogène H2 sur une surface de glace d'eau amorphe poreuse et d'autre part les processus de formation de cette molécule, qui est la plus abondante dans le milieu interstellaire, sur des surfaces simulant des grains de poussière interstellaire. Les résultats de cette étude présentent un intérêt capital en astrophysique. En effet cette formation ne peut se produire en phase gaz mais peut être expliquée par la rencontre de deux atomes d'hydrogène sur un grain de poussière du milieu interstellaire qui joue le rôle de catalyseur. Différents processus sont impliqués dans la formation de H2 qu'il s'agit d'identifier. Dans ce but, je présente des résultats entièrement nouveaux sur la formation de H2 moléculaire sur une surface de silicate amorphe. Je montre que la molécule peut se former dans un état rovibrationnellement excité de son niveau fondamental jusqu'à une température d'environ 70K et qu'elle est relâchée dans la phase gaz immédiatement après sa formation. Ces résultats démontrent la compétition entre deux mécanismes de formation à très basses température (<18 K) tandis qu'un autre mécanisme prend le relais jusqu'à 70K. / The work that I present in this thesis is primarily an experimental work carried out in the LAMAp laboratory at the University of Cergy-Pontoise, using the experimental device called FORMOLISM. The main components of this device are: ultra-high vacuum (10-10 mbar), ultra-low temperature (~5 K), atomic jets, TPD mass spectrometry (Thermally Programmed Desorption) and laser spectroscopy using ultraviolet wavelengths around ~200 nm. Thanks to the REMPI-TOF (Resonantly Enhanced Multi-Photon Ionization – Time Of Flight) technique, we have studied i) the nuclear spin conversion of the hydrogen molecule H2 on a porous amorphous water ice surface and ii) the processes of formation of this molecule, which is the most abundant in the interstellar medium, on surfaces simulating interstellar dust grains. The results of this study are of capital interest in astrophysics. Actually, this formation cannot occur in the gas phase but can only be explained by the encounter of two hydrogen atoms on a dust grain in the interstellar medium, the latter playing the role of a catalyst. Different processes are involved in the formation of H2 which require to be identified. With this aim, I present entirely new results on molecular H2 formation on an amorphous silicate surface. I show that the molecule can form in a rovibrationnally excited state of its ground state up to a temperature of about 70 K and that the molecule is released into the gas phase immediately after its formation. These results demonstrate the competition between two mechanisms of formation at very low temperature (<18 K) while another mechanism takes over up to 70 K.

Le travail présenté dans cette thèse s'inscrit dans le cadre de l'étude de l'efficacité de la commande CRONE avec effet anticipatif robuste dans les problèmes de poursuite et de régulation pour les systèmes monovariables et multivariables. L'objectif de l'anticipation est de concevoir des algorithmes de commande capables de minimiser un critère quadratique basé sur l'erreur entre la sortie du système et le signal de référence avec modération du niveau du signal de commande. La solution proposée repose sur l'association d'une commande robuste de type feedback à une commande anticipative de type feedforward. L'action feedforward utilise un filtre anticipatif synthétisé dans le domaine fréquentiel en utilisant des critères d'optimalité et de contraintes de type $mathcal{H}_2$ et $mathcal{H}_infty$. La réduction du nombre de paramètres du filtre anticipatif en utilisant une période d'échantillonnage plus lente a été traitée. Des exemples académiques illustrent les développements théoriques. Une application pratique sur un système de régulation hydraulique a montré la pertinence de cette nouvelle approche. / The work presented in this thesis is part of the study of the effectiveness of CRONE control with anticipative effect in the problems of tracking and control for monovariable and multivariable systems. The anticipation objective is to design a control algorithm that minimizes a quadratic error between the reference and the output of the system and at the same time achieve a good level of the control signal. The proposed solution combines a robust feedback control with a feedforward control. The feedforward action uses optimality criteria and an anticipative filter which is designed in the frequency domain using a mix of $mathcal{H}_2$ et $mathcal{H}_infty$ constraints. The reduction of the number of parameters of the anticipative filter by using a slower sampling period for the anticipative filter is treated. Academic examples highlight the theoretical developments. A practical application on a hydraulic control system has shown the efficiency of the developed approach.

A produ&ccedil;&atilde;o de energia &eacute; um dos grandes desafios deste s&eacute;culo, principalmente com a necessidade no desenvolvimento de processos que utilizem preferencialmente fontes renov&aacute;veis. Nesse contexto, &eacute; claro o interesse por pesquisas baseada no hidrog&ecirc;nio. Por&eacute;m, os ganhos ambientais efetivos est&atilde;o associados tamb&eacute;m &agrave; mat&eacute;ria-prima usada no processo de gera&ccedil;&atilde;o do hidrog&ecirc;nio, senso assim mais significativo quando do uso de fontes renov&aacute;veis. No presente trabalho foi estudado o efeito da adi&ccedil;&atilde;o de diferentes teores de CeO2 em catalisadores de NiZnO, preparados pelo m&eacute;todo da co-precipita&ccedil;&atilde;o. Os catalisadores foram estudados frente &agrave; rea&ccedil;&atilde;o de reforma a vapor do etanol (RVE) para produ&ccedil;&atilde;o de H2, e no decorrer do trabalho foi pertinente incluir o estudo da reforma a vapor da acetona (RVA), como complemento dos estudos da RVE. Al&eacute;m disso, esta &eacute; uma mol&eacute;cula modelo para reforma a vapor do bio-&oacute;leo. Nos sistemas catal&iacute;ticos sintetizados houve a forma&ccedil;&atilde;o da liga NiZn3, o que aparenta proporcionar um efeito sin&eacute;rgico entre esses elementos. Observou-se um efeito altamente promotor do CeO2 com rela&ccedil;&atilde;o a diminui&ccedil;&atilde;o do coque formado, devido a uma menor da forma&ccedil;&atilde;o da acetona, durante as etapas de reforma do etanol. O que consequentemente interferiu na produ&ccedil;&atilde;o de H2. O catalisador contendo 20% de CeO2 denominado NiZn20Ce apresentou um desempenho altamente promissor, pois de acordo com as an&aacute;lises de DRIFT, a presen&ccedil;a do CeO2 levou a forma&ccedil;&atilde;o de esp&eacute;cies de formiato, o que consequentemente interferiu em uma menor forma&ccedil;&atilde;o de coque e maior produ&ccedil;&atilde;o de H2. Foi comprovado tamb&eacute;m que a natureza dos dep&oacute;sitos carbon&aacute;ceos depende tanto do substrato utilizado quanto das etapas precursoras inicias que levar&atilde;o a forma&ccedil;&atilde;o desse coque, sendo o ponto chave para um melhor desempenho do catalisador. / Energy production is one of the great challenges of this century, especially with the need to develop processes that use preferentially renewable sources. In this context, it&acute;s evident the interest in research based on hydrogen. However, the environmental benefits are also associated with the raw material used on the hydrogen generation process, being more significant when used renewable sources. In the present work, it was studied the effect of CeO2 addition in NiZnO catalysts, prepared by coprecipitation method. The catalysts were studied front of the ethanol steam reforming reaction (ESR) to produce H2, and in the course of this study was relevant include study of acetone steam reforming (ASR), as a complement of ESR studies. Moreover, acetone is a molecule model for steam reforming of bio-oil. In the catalyst systems there has been the NiZn3 alloy formation, which provides a synergistic effect between these elements. It was observed a high CeO2 promoter effect regarding reduction of the formed coke, due to lower formation of acetone in the ethanol reforming steps, and consequently interfere in the H2 production. The catalyst denominated NiZn20Ce showed a high performance, according to DRIFT analyzes, the presence of the CeO2 led to the formation of species formate, which in turn interfere through a lower coke formation and higher H2 production. It was also established that the nature of the carbonaceous deposits depends as the substrate used as the precursors initial steps that led the formation of this coke, and it is the key to a better performance of the catalyst.

Made available in DSpace on 2017-07-24T19:38:04Z (GMT). No. of bitstreams: 1 Juliane Nadal Dias.pdf: 2277080 bytes, checksum: cfb6cfa9f705c82f0634bb16f025fd4f (MD5) Previous issue date: 2010-09-03 / Fundação Araucária de Apoio ao Desenvolvimento Científico e Tecnológico do Paraná / Photodynamic therapy (PDT) is a kind of treatment minimally invasive which depends on two components: photosensitizer and light. It consists of the administration of a drug which after reaching its action site is activated by light, whose wavelength is absorbed and catalyses the production of free radicals oxygen, and singlet oxygen, which trigger the cellular death of the unhealthy tissue, either by necrosis or apoptosis. As this technique appears as very promising in the treatment of bacteria infections and cancers (mainly skin cancer of the non-melanoma), several studies seek new photosensitizers compounds which are efficient and present little collateral effect. The aim of this study was to investigate the photodynamic efficiency of [5, 10, 15, 20-tetrakis(4-sulfonatophenyl) porphyrinato Iron (III)], chloride, [Fe(TPPS4)]Cl, and 5, 10, 15, 20-tetrakis(N-methyl-4-pyridyl) porphyrin p-toluenesulfonate, [H2(TMPyP)], compounds, through spectroscopic and cytotoxic analysis, using LED irradiation system, for later use in photodynamic therapy. The uric acid method was employed in order to determine the photodynamic activity, analyse the spectroscopic behavior regarding alterations in the environment pH, with and without CTAB and interpret photobleaching reactions. Toxicity was evaluated through in vitro experiments with microorganisms (Escherichia coli, Staphylococcus aureus e Candida albicans) and Artêmia salina. Results presented in this work with [H2(TMPyP)] porphyrin, using LED irradiation, demonstrated that the porphyrin:LED system can be used in PDT, once it presented satisfactory photodynamic activity, and significant inhibition of CFUs (Colony forming units), regarding the micro organisms under study, mainly when the red LED was used. Metalporphyrin [Fe(TPPS4)]Cl presented lower photodynamic activity values in both AU and microorganisms tests under the experimental conditions employed when compared to [H2(TMPyP)]. / A Terapia Fotodinâmica (TFD) é uma modalidade de tratamento minimamente invasiva que depende de dois componentes: fotossensibilizador (FS) e luz. Consiste na administração de um fármaco que, ao atingir o seu sítio de ação, é irradiado por luz, cujo comprimento de onda é absorvido, e catalisa a produção de radicais de oxigênio, e de oxigênio singlete, que desencadeiam a morte celular do tecido atingido, por necrose ou apoptose. Por ser promissora no tratamento em infecções bacterianas e em cânceres (principalmente em câncer de pele do tipo não melanoma), muitos estudos buscam novos compostos fotossenssibilizadores que sejam eficazes e apresentem poucos efeitos colaterais. O objetivo desse trabalho foi investigar a eficiência fotodinâmica dos compostos cloreto de [5, 10, 15, 20-tetrakis(4-sulfonatofenil)porfirinato ferro(III)], [Fe(TPPS4)]Cl, e p–toluenosulfonato de 5, 10, 15, 20-tetrakis(N-metil-4-piridil)porfirina, [H2(TMPyP)], através de ensaios espectroscópicos e citotóxicos utilizando sistema de LED para irradiação, visando posterior utilização em Terapia Fotodinâmica. Utilizou-se o método do ácido úrico para determinação da atividade fotodinâmica, a análise do comportamento espectroscópico a partir de alterações do pH do meio, na ausência e presença de CTAB, interpretações de reações de fotobranqueamento, e avaliou-se a atividade fotodinâmica através de ensaios in vitro com micro-organismos (Escherichia coli, Staphylococcus aureus e Candida albicans) e Artêmia salina. Os resultados apresentados neste trabalho com a porfirina [H2(TMPyP)] utilizando-se LED na irradiação, demonstraram que o sistema porfirina/LED tem potencial para ser utilizado em TFD, uma vez que apresentou atividade fotodinâmica satisfatória, e uma inibição significativa nas UFCs dos micro-organismos estudados, principalmente quando se fez uso de LED vermelho. A metaloporfirina [Fe(TPPS4)]Cl apresentou menores valores de atividade fotodinâmica no teste de AU e nos testes com micro-organismos, nas condições experimentais utilizadas, quando comparada com [H2(TMPyP)].

Submitted by Kamila Costa (kamilavasconceloscosta@gmail.com) on 2015-06-15T20:50:21Z No. of bitstreams: 1 Dissertacao-Adriano B dos S Frutuoso.pdf: 1799620 bytes, checksum: acda5b957dcaee1690413438f4a99e8e (MD5) / Approved for entry into archive by Divisão de Documentação/BC Biblioteca Central (ddbc@ufam.edu.br) on 2015-06-16T14:58:57Z (GMT) No. of bitstreams: 1 Dissertacao-Adriano B dos S Frutuoso.pdf: 1799620 bytes, checksum: acda5b957dcaee1690413438f4a99e8e (MD5) / Approved for entry into archive by Divisão de Documentação/BC Biblioteca Central (ddbc@ufam.edu.br) on 2015-06-16T15:00:11Z (GMT) No. of bitstreams: 1 Dissertacao-Adriano B dos S Frutuoso.pdf: 1799620 bytes, checksum: acda5b957dcaee1690413438f4a99e8e (MD5) / Made available in DSpace on 2015-06-16T15:00:11Z (GMT). No. of bitstreams: 1 Dissertacao-Adriano B dos S Frutuoso.pdf: 1799620 bytes, checksum: acda5b957dcaee1690413438f4a99e8e (MD5) Previous issue date: 2015-02-02 / FAPEAM - Fundação de Amparo à Pesquisa do Estado do Amazonas / This work approaches the angles stability control of a quadrotor. The vehicle parts (mechanical structure and electronic devices), dynamic modeling, controllers design methodology and the experimental results are also presented. Attitude controllers were designed using a PD/H2 discrete control structure, which the gain of H2 part was performed by solving a convex optimization problem, described in linear matrix inequalities form. The experiments indicates that angles dynamic responses can be changed by setting the gain of H2, without modifying the gains of the proportional-derivative part. The results of PD/H2 controllers were compared with a PD controller, in order to evaluate the overshoot and settling time. / Neste trabalho é abordado o controle de estabilidade dos ângulos de atitude de um MAV (Micro Aerial Vehicle) do tipo quadrirrotor. São apresentadas as partes constituintes do veículo (estrutura mecânica e dispositivos eletrônicos), a modelagem dinâmica dos ângulos de atitude, a metodologia de projeto dos controladores de atitude e os resultados experimentais. Em relação ao projeto dos controladores de atitude, estes foram projetados usando estrutura de controle discreta combinada PD/H2, cuja sintonia do controlador H2 por realimentação de estados foi realizada a partir da resolução de um problema de otimização convexo descrito na forma de desigualdades matriciais lineares. Os experimentos mostraram que as respostas dinâmicas dos ângulos de atitude poderiam ser alteradas com o ajuste dos controladores H2, sem modificar os ganhos da parte proporcional-derivativa. Os resultados obtidos para os controladores PD/H2 foram comparados com os controladores PD, de modo a avaliar os seus desempenhos com relação a tempo de acomodação e overshoot.

À ce jour, l’humanité est confrontée simultanément à une crise énergétique et environnementale due principalement à deux facteurs: la croissance démographique et la dépendance aux combustibles fossiles. C'est pourquoi l'urgence d'utiliser des sources d'énergie renouvelables, comme l'énergie solaire est une, solution potentielle. A ce titre, la production d'H2 décarboné par dissociation solaire de l'eau est une voie prometteuse. Néanmoins, pour atteindre l'objectif mentionné, il faut trouver un système photocatalytique (semi-conducteurs, SCs) idéal, qui nécessite quatre caractéristiques majeures: (1) une bonne capacité d'absorption de la lumière visible (2) des positions adéquates de BV et BC des SCs par rapport aux potentiels d’oxydation de l’eau et de réduction du proton (3) une utilisation efficace des photons absorbés et charges générées et (4) une bonne stabilité dans le temps. À cette fin, cette thèse contribue à la conception et à l’optimisation de trois matériaux innovants: les composites Au/g-C3N4, Au/TiO2 (P25)-gC3N4, et Au/TiO2 (NTs)-gC3N4 dont l’activité photocatalytique a été corrélé avec les propriétés physico-chimiques pour comprendre leurs performances photocatalytiques de production d'H2 sous irradiation solaire et visible. De manière annexe, certains de ces matériaux se sont également montrés performants pour les conversions du CO. Pour mettre en évidence l'efficacité des composites préparés, des études comparatives ont été testées en utilisant des références commerciales, pertinentes et les mélanges physiques correspondant. / To date, mankind is facing an energy and environmental crisis simultaneously due to mainly two factors: growth population and the dependency on fossil fuels. For this reason, the urgency of using renewables sources, e.g., solar energy, is a potential solution. For example, non-carbon based H2 production from solar light driven water photodissociation is a promising approach. Nevertheless, to target the mentioned objective, an ideal photocatalytic system (semiconductors, SCs) has to meet four main features: (1) capacity of absorption of visible-light (2) suitable VB and CB positions of SCs to undergo the two half reactions of water splitting (3) efficient use of absorbed photons and generated charges and (4) good stability over time. For this purpose, this thesis contributes to the design and optimization of three innovative materials: Au/g-C3N4, Au/TiO2 (P25)-gC3N4, and Au/TiO2 (NTs)-gC3N4 composites. Their photocatalytic activities were correlated with their physico-chemical properties. In addition some of these composites also exhibited interesting CO conversion yields. To highlight the efficiency on the as-prepared composites, comparative studies were tested using commercial, pertinent references, and physical mixtures homologs.

L'objectif du projet est la mesure du rapport de la masse de l'électron à celle du proton par spectroscopie vibrationnelle à deux photons sans effet Doppler de H2+. Le refroidissement des ions H2+ est nécessaire et ne peut être fait que par refroidissement sympathique par des ions Be+ refroidis par laser. La première partie présente la conception et la réalisation d'un piège linaire adapté au confinement des ions H2+ et Be+ et permettant les accès optiques nécessaires pour les lasers et l'imagerie. La seconde partie décrit les sources laser à 626 (à base de diodes laser ou de lasers à fibres) ainsi que la cavité de doublage de fréquence utilisés pour obtenir le faisceau à 313 nm pour refroidir les ions Be+. La dernière partie expose les premiers résultats obtenus qui montrent que le montage est opérationnel. Elle présente l'observation et la caractérisation de cristaux de Coulomb de Be+ ainsi que des cristaux de Coulomb mixtes Be+/H2+ où les ions H2+ sont créés par impact électronique à partir du gaz résiduel. Ces résultats montrent que le montage est prêt pour réaliser la spectroscopie de H2+. / The objective of the project is to perform high-resolution Doppler-free two-photon vibrational spectroscopy of the H2+ molecular ion for accurate electron to proton mass ratio determination. Trapping and laser-cooling of the H2+ ions is necessary. Since H2+ molecular ions cannot be directly laser cooled, a solution is the implementation of sympathetic cooling via a laser cooled Be+ ions. In the first part of this thesis the design and implementation of a linear Paul trap is presented. This trap is suitable to confine H2+ and Be+ ions while allowing good optical access for multiple laser sources and the imaging system. The second part describes a low power, diode based laser system to provide narrow linewidth 626nm light, and a high power, fiber laser based laser system. Subsequently, a bow-tie cavity is presented to frequency double the 626nm light to obtain 313nm light for Be+ Doppler-cooling. In the last part the operational readiness of the trap and the fiber laser based 313nm laser source is demonstrated by the observation of Be+ Coulomb crystals. Electron impact ionization of H2 present in the residual gas allows to observe sympathetic cooling and shows that the developed setup can provide the experimental conditions for precision spectroscopy of H2+.