This work is a numerical and analytical study of wave motion through dynamic materials (DM). This work focuses on showing several results that greatly extend the applicability of the checkerboard focusing effect. First, it is shown that it is possible to simultaneously focus dilatation and shear waves propagating through a linear elastic checkerboard structure. Next, it is shown that the focusing effect found for the original €œperfect€� checkerboard extends to the case of the checkerboard with smooth transitions between materials, this is termed a functionally graded (FG) checkerboard. With the additional assumption of a linear transition region, it is shown that there is a region of existence for limit cycles that takes the shape of a parallelogram in (m,n)-space. Similar to the perfect case, this is termed a €œplateau€� region. This shows that the robustness of the characteristic focusing effect is preserved even when the interfaces between materials are relaxed. Lastly, by using finite volume methods with limiting and adaptive mesh refinement, it is shown that energy accumulation is present for the functionally graded checkerboard as well as for the checkerboard with non-matching wave impedances. The main contribution of this work was to show that the characteristic focusing effect is highly robust and exists even under much more general assumptions than originally made. Furthermore, it provides a tool to assist future material engineers in constructing such structures. To this effect, exact bounds are given regarding how much the original perfect checkerboard structure can be spoiled before losing the expected characteristic focusing behavior.

After nearly 100 years of particle acceleration, particle accelerator experiments continue providing results within the field of high energy physics. Particle acceleration is used worldwide in practical applications such as radiation therapy and materials science research. Unfortunately, these accelerators are large and expensive. Dielectric Laser Acceleration (DLA) is a promising technique for accelerating particles with high acceleration gradients, without requiring large-scale accelerators. DLA utilizes the electric field of a high energy laser to accelerate electrons in the proximity of a nanostructured dielectric surface.The aim of this project was limited to laser beam routing and imaging techniques for a DLA experiment. The goal was to design the laser beam pathway between the laser and the dielectric sample, and testing a proposed imaging system for aiming the laser. This goal was achieved in a test setup using a low-energy laser. In the main setup including a femtosecond laser, the result indicated lack of focus. For a full experimental setup, a correction of this focus is essential and the beam path would need to be combined with a Scanning Electron Microscope (SEM) as an electron source.

A espectrometria de emissão óptica com plasma induzido por laser (LIBS) constitui um método alternativo para a determinação simultânea de macro e micronutrientes em pastilhas de materiais vegetais. No entanto, pouca informação com relação às condições de fluência e de focalização do laser mais apropriadas para a análise de pastilhas de materiais vegetais são disponíveis na literatura. O objetivo deste trabalho foi avaliar os efeitos da fluência, do diâmetro de focalização do laser e do tamanho das partículas das amostras nas análises de pastilhas de tecidos vegetais por LIBS. O arranjo experimental foi composto por um laser pulsado de Nd:YAG a 1064 nm (pulsos de 5 n\'delta\', 360 mJ, 10 Hz) e os sinais de emissão foram coletados por um telescópio acoplado por fibra óptica ao espectrômetro Echelle com detector ICCD. Os parâmetros instrumentais foram ajustados em 20 pulsos acumulados, 2,0 \'mü\'s de atraso e 5,0 \'mü\'s de integração. Amostras laboratoriais consistiram de pastilhas preparadas com folhas moídas criogenicamente (90 % das partículas < 50 \'mü\'m). Maiores intensidades das linhas Ca I 442,554, Mg I 277,983, P I 213,618, Al I 309,271, B I 249,773, Cu I 324,755, Fe II 261,187, Mn II 257,610 e Zn II 206,200 nm foram obtidas com fluências entre 35 e 60 J cm-2 e diâmetros de focalização entre 600 e 1050 \'mü\'m. O uso de fluências e diâmetros de focalização maiores concorreu para melhorar os coeficientes de variação (CV) das medidas. Fixando-se o diâmetro de focalização em 750 \'mü\'m, observou-se que, para a maioria dos analíticos, o CV das medidas foi reduzido 2 vezes quando a fluência foi alterada de 20 para 50 J cm-2. Nestas condições, estes resultados foram atribuídos à maior massa removida e, conseqüentemente, à amostragem mais representativa. Os coeficientes angulares das curvas de calibração também aumentaram quando a fluência foi acrescida. No entanto, não foram verificadas diferenças significativas nos coeficientes de correlação das curvas de calibração e nos limites de detecção empregando-se 25 e 50 J cm-2. Embora as intensidades dos sinais de emissão aumentem com a fluência, a emissão de fundo também aumenta. Por outro lado, o tamanho das partículas das amostras afeta as eficiências de atomização e excitação no plasma induzido por laser. Quando partículas maiores são parcialmente vaporizadas, os sinais de emissão resultantes dos processos de atomização e excitação das espécies constituintes são menores do que os provenientes de partículas menores. Nesta dissertação, melhorias significativas da intensidade dos sinais de emissão e dos CVs das medidas foram observados quando pastilhas preparadas com partículas < 75 \'mü\'m foram analisadas. Não foram observadas diferenças significativas nas intensidades dos sinais de emissão e no CV das medidas nas análises de pastilhas preparadas com partículas entre 20 e 75 \'mü\'m. Pastilhas preparadas com partículas > 75 \'mü\'m apresentaram problemas de coesão e, após ablação, apresentaram crateras não uniformes. A análise destas pastilhas resultou em CV das medidas geralmente > 15 %. Os resultados obtidos neste trabalho confirmaram que é recomendável utilizar amostras de calibração e amostras-teste com distribuição do tamanho das partículas semelhante para evitar efeitos físicos de matriz, que alteram as interações laser-amostra / Most recently, it was demonstrated that laser induced breakdown spectrometry (LIBS) constitutes an alternative for the simultaneous determination of macro- and micronutrients in pellets of plant materials. However, there is no information regarding the most appropriate fluence and laser focusing for analysis of pellets of plant materials. The aim of this work was to evaluate the effect of laser focusing and fluence on LIBS analysis of plants. The effect of particle size on LIBS analysis of pellets of plant materials as well as sample preparation approaches were also discussed. The experimental setup was designed by using a Q-switch Nd:YAG laser (5 n\'delta\', 360 mJ, 10 Hz, _ \'lâmbda\' = 1064 nm) and the emission signals were collected by lenses into an optical fiber coupled to an echelle spectrometer equipped with a high-resolution ICCD. Instrumental parameters consisted of 20 accumulated laser pulses, 2.0 \'mü\'s delay time and 5.0 \'mü\'s integration time gate. Pellets prepared from cryogenically ground plant leaves (90 % particles < 50 \'mü\'m; median = 10 \'mü\'m) were used as laboratory samples and 6 laser spot diameters (from 180 to 1050 \'mü\'m) at different laser fluences were evaluated. Results indicate that elements emission intensities increased with both laser fluence and spot diameter. Higher intensities for Ca I 442.554, Mg I 277.983, P I 213.618, Al I 309.271, B I 249.773, Cu I 324.755, Fe II 261.187, Mn II 257.610, and Zn II 206.200 nm emission lines were observed with fluences in the 35 - 60 J cm-2 range and spot diameters between 600 and 1050 \'mü\'m. Higher spot diameters improved mass removal and minimized repeatability drawbacks. It was observed that the coefficients of variation (CV) of site-to-site measurements decreased by using higher laser focusing diameters. By fixing the laser spot diameter at 750 \'mü\'m, the CV of measurements improved at least 2-fold for all analytes when 50 J cm-2 was applied in comparison to 20 J cm-2. These results can be attributed to a larger mass removal obtained at higher laser fluences and better laser sampling representativeness. Sensitivity did also increase with laser fluence but no significant differences were observed in the detection limits using 25 and 50 J cm-2. Although emission intensities increased with laser fluence the background emission and noise increased as well. On the other hand, particle size distribution of powdered samples affects the atomization and excitation efficiencies by laser induced plasmas. When larger particles are incompletely vaporized the resulting emission signals were lower than those obtained from smaller particles. In this work, significant improvements on emission intensities and CV of measurements were observed when pellets made with particles < 75 \'mü\'m were analyzed. No significant differences were observed (emission intensities and CV of measurements) in the analysis of pellets prepared with particles in the 20-75 \'mü\'m range. Pellets prepared with particles > 75 \'mü\'m presented cohesion drawbacks and resulted non uniform craters after laser ablation. The CV of measurements for most analytes in these materials was > 15 %. Therefore, similar particle size distribution between calibration and test samples is recommended to avoid physical matrix effects on laser-sample interaction

Third-harmonic generation (THG) is a fundamental nonlinear optical process that has been used in different applications such as third-order nonlinear materials characterization and nonlinear microscopy. It is widely employed since the third-order nonlinearity is the most important in isotropic materials and THG occurs in all media regardless of symmetry. In the tightly focused laser beam condition THG is observed only at the materials interfaces, where the focal symmetry is broken due to the presence of two media with different refractive index and/or third-order susceptibilities. Measuring slabs of different types of optical glasses, using femtosecond laser pulses, we could explain the asymmetric THG intensity profile observed at the interfaces. The harmonic generated at the exit interface is systematically stronger than the one generated at the entrance and this phenomenon can be understood by taking into account the presence of self-focusing effects. Basically, the self-focusing reduces the beam waist radius at the exit interface, resulting in greater laser irradiance and, consequently, higher THG. This study was then extended to the interfaces of a cuvette filled with organic solvents. Such systems present four interfaces and a mixture of nonlinear processes contributions since the cuvette walls present only electronic nonlinearity and the solvents present both electronic and orientational ones. In this way, the solvents may present an additional self-focusing contribution and, due to the noninstantaneous nature of the orientational process, the self-focusing from the solvent may be influenced by the pulse duration. In this case, the THG, which is an instantaneous electronic phenomenon, can be indirectly affected by pulse duration by means of the self-focusing effect. Usually, the slow orientational contribution is not considered for materials characterization by THG which may lead to incorrect nonlinear coefficient values, that means our study is important from the fundamental physics point of view and also for applications such as materials characterization. Based on the application of THG in nonlinear microscopy, we also present a microscopy technique which makes use of spatial frequency-modulated imaging (SPIFI) with single element detection. The microscope was developed at Colorado School of Mines (CSM) during an internship. The system uses a spatial light modulator (SLM) to provide the spatial frequency modulation and permits enhanced resolution images. THG SPIFI images are shown for the first time and we also report images obtained by other nonlinear optical process. In summary, the studies presented in this PhD work are of great importance for THG fundamental understanding, materials characterization and nonlinear optical microscopy. / Geração de terceiro harmônico (GTH) é um processo óptico não linear fundamental que tem sido usado em diferentes aplicações, como em caracterização óptica não linear de materiais e microscopia não linear. Ele é amplamente empregado uma vez que a não linearidade de terceira ordem é a mais importante em materiais isotrópicos e GTH ocorre em todos os meios independente da simetria. Na condição de feixe fortemente focalizado a GTH é observada apenas nas interfaces do material, onde a simetria focal é quebrada devido à presença de dois meios com diferentes índices de refração e/ou susceptibilidades de terceira ordem. Medindo lâminas de diferentes tipos de vidros ópticos, com pulsos de laser de femtossegundos, nós explicamos o perfil assimétrico de intensidade de GTH observado nas interfaces. O harmônico gerado na interface de saída é sistematicamente mais intenso do que o gerado na entrada e este fenômeno pode ser entendido levando-se em conta a presença do efeito de autofocalização. Basicamente, a autofocalização reduz a cintura do feixe na interface de saída do material, resultando em uma maior irradiância e, consequentemente, maior GTH. Este estudo foi estendido para o caso de interfaces de uma cubeta preenchida com diferentes solventes orgânicos. Tais sistemas apresentam quatro interfaces e uma mistura na contribuição dos processos não lineares, dado que as paredes da cubeta apresentam apenas não linearidade eletrônica e os solventes podem apresentar não linearidades tanto eletrônicas quanto orientacionais. Neste sentido, os solventes podem apresentar uma contribuição adicional de autofocalização e, devido à natureza não instantânea do processo orientacional, a autofocalização proveniente do solvente pode ser influenciada pela duração do pulso. Neste caso, a GTH, que é um fenômeno eletrônico (instantâneo), pode ser indiretamente afetada pela duração do pulso por meio do efeito de autofocalização. Usualmente, a contribuição orientacional não é considerada na caracterização de materiais por GTH, o que pode levar à valores incorretos para os coeficientes não lineares, o que significa que nosso estudo é importante do ponto de vista de física fundamental como também em aplicações como caracterização de materiais. Por conta da aplicação da GTH em microscopia não linear, apresentamos também nesta tese uma técnica de microscopia, que baseia-se em uma modulação em frequência espacial para imageamento (SPIFI) com uso de um detector de elemento único. O microscópio foi desenvolvido na Colorado School of Mines (CSM) durante um período de estágio. O sistema utiliza um modulador espacial de luz (SLM) para produzir a modulação em frequência espacial e permite obtenção de imagens em alta resolução. Imagens por GTH SPIFI são mostradas pela primeira vez e também apresentamos imagens obtidas por outros processos ópticos não lineares. Em resumo, os estudos apresentados neste trabalho de doutorado são de grande importância para o entendimento fundamental do processo de GTH, caracterização de materiais e microscopia óptica não linear.

L’essor spectaculaire des systèmes de communications sans fil a entrainé une forte augmentation du trafic des données qui ne devrait cesser de croître au cours des prochaines années. La future génération de réseaux cellulaires (5G) doit être capable de supporter cette croissance du trafic, tout en présentant une consommation énergétique réduite par rapport aux réseaux existants. Parmi les différentes technologies étudiées, le retournement temporel (RT) se présente comme un sérieux candidat pour répondre à ces contraintes. En effet, les nombreuses études théoriques sur le sujet ont montré que la combinaison du RT et de l’OFDM possède des performances intéressantes, notamment grâce à ses propriétés de compression temporelle et de focalisation spatiale. Cependant, les hypothèses prises dans les études théoriques ne sont pas toujours compatibles avec la réalisation pratique d’un tel système. L’objectif de ces travaux de thèse est donc de proposer des solutions aux hypothèses non réalistes, afin de les mettre en oeuvre dans la réalisation d’un prototype combinant RT et OFDM dans un contexte MISO. Dans un premier temps, la mise en oeuvre de l’estimation du canal au niveau de l’émetteur et de la synchronisation du système MISO RT-OFDM est étudiée. Ensuite, une solution de calibration au niveau de l’émetteur est proposée pour compenser le caractère non réciproque du canal de propagation vu de la bande de base. L’ensemble de ces solutions ainsi que les propriétés de focalisation du RT sont validés à partir de mesures expérimentales sur le prototype réalisé. Enfin, la mise en oeuvre du premier prototype de modulation spatiale à la réception utilisant le RT est présentée. / The spectacular growth of wireless communications systems has led to a sharp increase in data traffic, which should continue to grow over the next few years. The future generation of cellular networks (5G) must be able to support this growth of traffic, while presenting reduced energy consumption compared to existing networks. Among the different studied technologies, time reversal (TR) is a serious candidate to meet these constraints. Indeed, the numerous theoretical studies on the subject have shown that the combination of TR and OFDM has interesting performance, specifically thanks to its temporal compression and spatial focusing properties. However, the assumptions made in the theoretical studies are not always compatible with the practical implementation of a real system. The objective of this thesis is to propose solutions to unrealistic theoritical hypotheses, in order to implement them in a real hardware prototype combining TR and OFDM in a MISO context. First, the implementation of the channel estimation at the transmitter side, and the synchronization of the MISO TR-OFDM system is studied. Then, a calibration solution at the transmitter is proposed to compensate for the non-reciprocal nature of the baseband propagation channel. All these solutions as well as the focusing properties of TR are validated by experimental measurements using the developed prototype. Finally, the implementation of the first TR prototype of spatial modulation at the receiver side is presented.

submitted by Liu Dan. / Thesis (M.Sc.)--Chinese University of Hong Kong, 1998. / Includes bibliographical references (leaves 66-80). / Abstract also in Chinese. / CONTENTS --- p.i / LIST OF TABLES --- p.iii / LIST OF FIGURES --- p.iv / LIST OF ABBREVIATIONS --- p.v / ACKNOWLEDGEMENTS --- p.vi / ABSTRACT --- p.vii / Chapter Chapter 1 --- INTRODUCTION --- p.1 / Chapter 1.1 --- History --- p.1 / Chapter 1.2 --- Immunoglobulins --- p.3 / Chapter 1.2.1 --- Structure --- p.3 / Chapter 1.2.2 --- Properties of immunoglobulins --- p.7 / Chapter 1.3 --- Monoclonal proteins and monoclonal gammopathies --- p.12 / Chapter 1.3.1 --- Monoclonal proteins --- p.12 / Chapter 1.3.2 --- Monoclonal gammopathies --- p.14 / Chapter 1.4 --- Laboratory investigation of monoclonal immunoglobulin --- p.17 / Chapter 1.4.1 --- The current procedure of investigation in laboratory --- p.17 / Chapter 1.4.2 --- Problems in identifying monoclonal immunolgobuin --- p.19 / Chapter 1.5 --- Comparison of different techniques --- p.20 / Chapter 1.5.1 --- Immunoelectrophoresis --- p.20 / Chapter 1.5.2 --- Immunofixation electrophoresis --- p.22 / Chapter 1.5.3 --- Isoelectric focusing and immunoisoelectric focusing --- p.24 / Chapter 1.6 --- Aim of the present study --- p.27 / Chapter 1.7 --- Design of experiment --- p.27 / Chapter Chapter 2 --- MATERIALS AND METHODS --- p.30 / Chapter 2.1 --- Study subjects --- p.30 / Chapter 2.2 --- Apparatus --- p.30 / Chapter 2.2 --- Apparatus --- p.30 / Chapter 2.3 --- Reagents and materials --- p.32 / Chapter 2.4 --- Preparation of gels --- p.35 / Chapter 2.5 --- Isoelectric focusing procedure --- p.36 / Chapter 2.6 --- Acid fixation and staining --- p.37 / Chapter 2.7 --- Technical factors affecting results --- p.38 / Chapter Chapter 3 --- RESULTS --- p.40 / Chapter 3.1 --- Interpretation of results in isoelectric focusing --- p.40 / Chapter 3.2 --- Affecting factors --- p.47 / Chapter 3.3 --- Comparison of the results between IEF and IFE --- p.53 / Chapter Chapter 4 --- DISCUSSION --- p.59 / Chapter Chapter 5 --- CONCLUSION --- p.65 / References --- p.66

Les mégacaryocytes sont des cellules de la moelle osseuse, à l’origine de la production des plaquettes sanguines. Quand elles arrivent à maturité, elles grossissent et émettent des prolongements de cytoplasme à travers la paroi des vaisseaux irriguant la moelle. Dans la circulation sanguine, ces prolongements, soumis aux forces de l’écoulement, s’allongent et se rompent pour former des plaquettes. Des techniques microfluidiques capables de produire des plaquettes in vitro existent et sont une alternative prometteuse au don. Mais le rendement reste à améliorer. Pour cela, il est nécessaire de mieux comprendre la fragmentation des mégacaryocytes en plaquettes. Ce travail de doctorat s’inscrit dans ce contexte et sera développé en deux axes principaux dans ce manuscrit. Dans une première partie nous développons une méthode pour trier des cellules en fonction de leur déformabilité, afin de savoir si les propriétés mécaniques d’un mégacaryocyte sont liées à leur stade de maturité. La méthode a d’abord été mise au point avec des microcapsules. Leurs propriétés mécaniques sont déterminées par analyse inverse à partir de la mesure de leur forme en écoulement dans des constrictions droites. Puis le dispositif utilisé a été miniaturisé pour s’adapter à la taille des cellules. Pour la caractérisation de leurs propriétés mécaniques, deux outils ont été utilisés: l’analyse inverse et la microscopie à force atomique sans pointe. Une deuxième partie porte sur l’étude de l’élongation et de la rupture de mégacaryocytes soumis écoulement. Nous avons quantifié les variations spatiotemporelles du taux d’élongation et développé un protocole d’ablation laser pour étudier les mécanismes de rupture de cellules en élongation. / When they mature in the bone marrow, the precursors of platelets, called megakaryocytes, grow and extend protrusions able to join blood circulation. There these protrusions elongate and break into platelets. Microfluidic techniques for in vitro platelet production represent a promising alternative to donation. In order to enhance platelet production and match the needs of clinical applications such as transfusion, we need to better understand the fragmentation of megakaryocytes into platelets. Our contribution will be described in this manuscript in two main axes. First, in order to know if mechanical properties of megakaryocytes can indicate their maturity stage, we develop a cell sorting method based on deformability. The method is first validated with microcapsules. Their mechanical properties are determined by inverse analysis from their shape under flow in straight microchannels. Then the device is downscaled. The characterization of cell mechanical properties are performed using inverse analysis and tipless atomic force microscopy. Second, we study megakaryocyte elongation and rupture in a microfluidic device. We quantify the spatial and temporal variations of the elongation rate and develop a laser ablation protocol to trigger and study the rupture of elongating cells.

The developing flow field in a parallel plate microchannel, induced by wall motion, has been modeled numerically. This type of flow simulates the physical driving mechanism that exists in electro-osmotically generated flow with large channel diameter-to-Debye length ratios (Z). The physics of the flow field were compared between the moving wall model (MWM) and electro-osmotic flow (EOF) at Reynolds numbers of 1 and 1800, and Z > 2500. Also, Z-values between 50 and 500 were studied to investigate the accuracy of the MWM. Results show that for Z-values greater than 100 the MWM shows good agreement with EOF. The dynamics of the developing flow field for the MWM were explored for channel length-to-hydraulic diameter ratios (aspect ratio) of 5, 10, 20 and 40 at ten Reynolds numbers, Re (based on the wall velocity), below Re < 2000. The results show that far from the inlet the maximum fluid velocity occurs at the walls, as is expected, and the minimum velocity occurs at the channel center. Near the channel inlet, however, the centerline velocity is not a minimum but reaches a local maximum due to a resulting pressure imbalance generated by the wall motion. As the aspect ratio increases, the centerline velocity tends to approach the wall velocity far downstream from the inlet. Increases in the Reynolds number have the opposite effect on the centerline velocity. The hydrodynamic developing region, defined by that section of the channel where the wall shear stress is changing, also depends on the channel aspect ratio and Re, and is greater than the developing region for classical pressure-driven flow of a parallel plate channel. Also, the flow field physics was analyzed for a process called electro-mobility focusing (EMF). EMF is a process that separates and detects species of like charge with the use of electro-phoresis and EOF utilizing a varying voltage gradient. The velocity distribution and the effective diffusion were solved for analytically, for both a linear and non-linear voltage gradient, using the MWM and the creeping flow approximations. The resulting equations aid in optimizing the detection system by forcing the lowest effective diffusion (uniform velocity profile) to the detection location.

L'objectif de cette thèse était d'explorer de nouvelles applications cardiaques pour l'histotripsie et de développer les outils permettant leur mise en place non-invasive. La thérapie ultrasonore cardiaque est en effet encore assez peu développée aujourd’hui, à cause de la difficulté à traiter un organe en mouvement permanent, et très bien protégé derrière la cage thoracique.Nous avons d'abord montré in vivo, sur un modèle ovin, que l’on pouvait sectionner les cordages mitraux de manière non-invasive ainsi que traiter la sténose aortique calcifiée. Engendrer de la cavitation sur les feuillets valvulaires permet effectivement d’agir à distance sur les calcifications, et de globalement assouplir la valve.Simultanément, nous avons développé un dispositif pour la thérapie cardiaque non invasive, fondé sur le concept de cavité à retournement temporel. Ce dispositif permet l'émission d'impulsions ultrasonores de haute intensité dans un très grand volume d’intérêt. L’on peut ainsi déplacer le point de thérapie en 3 dimensions de manière entièrement électronique, et sans déplacer mécaniquement l’appareil. Après optimisation, ce dispositif a permis de créer des lésions mécaniques bien contrôlées dans une région d'intérêt de 2 000 cm3.Pour faire face au défi que représente la cage thoracique, nous avons développé une méthode de focalisation adaptative et l'avons mise en œuvre dans un prototype 2D du dispositif. Avec cette méthode, nous pouvons non seulement construire un front d'onde ultrasonore adaptatif qui se propage de manière préférentielle à travers les espaces intercostaux, mais grâce aux propriétés des cavités à retournement temporel, nous pouvons également augmenter la pression focale obtenue sur la cible de thérapie.Enfin, pour approfondir ce travail sur la focalisation adaptative, et nous avons considéré le cas de l'imagerie transcrânienne. Pour cette application, nous avons choisi d’utiliser la focalisation par retournement temporel dans le bruit de speckle, pour corriger les aberrations induites par le crâne. En simulations numériques, nous avons pu calculer les modulations de phase et d'amplitude induites par les os et améliorer le contraste et la résolution d'une image B-mode. / The objective of this thesis was to explore new applications for cardiac histotripsy, and to develop the tools making it possible non-invasively. Cardiac ultrasound therapy indeed still remains limited due to the tremendous challenge of treating a constantly and rapidly moving organ, well protected behind the ribcage.We first showed in vivo, on a large animal model, that histotripsy could be used non-invasively to cut mitral chordae, and to treat calcified aortic stenosis in a beating heart. Cavitation on the valve leaflets can indeed locally and remotely act on the calcifications, and globally soften the valve. Simultaneously, we developed a therapeutic device allowing completely non-invasive cardiac shock-wave therapy based on the time reversal cavity concept. In particular, this device allows the emission of high intensity ultrasound pulses, and provides 3D electronical steering of the therapy focal spot in a large volume. After a thorough optimisation process, this device was capable of creating well controlled mechanical lesions over a 2 000 cm3 region of interest. To tackle the challenge of ultrasound propagation through the rib cage, we developed an adaptive focusing method (DORT method through a time reversal cavity), and implemented it in a 2D prototype of the device. With this method, we not only could build an adaptive ultrasonic wavefront propagating preferentially through the intercostal spaces, but due to time reversal cavities properties, we could also increase the peak pressure obtained on target.Finally, we pushed our work on adaptive focusing further, and considered the case of transcranial imaging. For this application, we chose to use the time reversal of speckle noise technique, to correct the aberrations induced by the skull. In numerical simulations, we were able to derive the phase and amplitude modulations induced by the bones, and could improve the contrast and resolution of a B-mode image.

Ein Signalweg während der neuronalen Entwicklung im adulten Gehirn ist der PI3K-PTEN-Akt Signalweg. Akt ist eine Kinase die drei verschiedene Isoformen besitzt, welche durch die Phosphorylierung von S473 und T308 aktiviert werden. KO Modelle der Isoformen haben gezeigt, dass nicht alle Funktionen von anderen Isoformen kompensiert werden können. Die genaue Rolle der einzelnen Isoformen in einem neuronalen Zusammenhang ist nur wenig untersucht. Ziel dieser Arbeit war, eine detaillierte Analyse der einzelnen Akt Isoformen nach der Aktivierung des PI3K-PTEN Signalweges. Dazu wurde im Labor eine neue Methode zur isoelektrischen Fokussierung etabliert., welche Proteine nach ihrer Ladung trennt und somit eine Analyse der Dynamik von Akt Phosphorylierungen in neuronalen Zellen erlaubt. Im Zuge dieser Arbeit konnten wir bisher unerkannte Merkmale der Akt Aktivierung und Phosphorylierung identifizieren. Wir konnten zeigen, dass die S473 und T308 Phosphorylierung in Neuroblastomazellen unabhängig voneinander auftreten kann und, dass verschiedene Akt1 Moleküle unterschiedlich auf die Inhibition von PI3K reagieren. Außerdem konnten wir Verschiebungen in der Aktivierung und in der Expression der unterschiedlichen Isoformen während der postnatalen Gehirnentwicklung der Ratte feststellen. Des Weiteren konnten wir zeigen, dass die Aktivierung von Akt von dem Signal und dem Alter der Neurone abhängig ist. Noch nicht vollständig differenzierte Neurone reagieren vor allem auf BDNF Stimulation, wohingegen adulte, differenzierte Neurone hauptsächlich auf EGF reagieren und dort explizit Akt2 über EGFR und PI3K-p110α Signale aktiviert wird. Im Gegensatz dazu führt der Verlust von PTEN zu einer Aktivierung von hauptsächlich Akt1. Zusammenfassend zeigt diese Arbeit einen komplexen Zusammenhang der Phosphorylierung von Akt auf, welcher Signal- und Entwicklungsabhängig ist bei dem unterschiedliche Akt Populationen auf Wachstumsfaktoren und auf PTEN Verlust reagieren. / A major pathway involved in neuronal development is the PI3K-PTEN-Akt pathway. Akt comprises three isoforms, which are activated by phosphorylation of the residues S473 and T308. KO animals for the isoforms have shown differential as well as redundant functions of the three isoforms. However, their individual role in neuronal signaling pathways has not yet been studied in great detail. The aim of this study was to obtain further insight into differential Akt isoform signaling in response to changes in the activity of PI3K and PTEN pathway. A new isoelectric focusing method was established, which allowed us to separate Akt proteins according to their charge, therefore, providing a refined read-out to study dynamics of Akt phosphorylation in a neuronal background. In the course of this project we were able to identify previously undescribed features of Akt phosphorylation and activation. First, we could provide evidence for an uncoupling of the two activating phosphorylation events at S473 and T308 in neuroblastoma cells and differential sensitivities of Akt1 forms towards PI3K inhibition. Secondly, we found a transient shift in Akt isoform activation and abundance during postnatal rat brain development. Thirdly, we were able to show that the activation of different Akt isoforms is dependent of the upstream signal as well as the age of the neuron. Immature neurons were found to be highly responsive to BDNF treatment, whereas mature neurons were most responsive to EGF stimulation leading exclusively to activation of Akt2 in an EGFR- and PI3K/p110α-dependent manner. Stimulation of Akt phosphorylation by the loss of PTEN led to an activation of mainly Akt1 forms, which suggests inherent differences in the Akt pools that are accessible to growth factors dependent PI3Ks as compared to the pools that are controlled by PTEN. In summary, this thesis demonstrates the presence of complex phosphorylation events of Akt in a developmental- and signal-dependent manner in neurons.