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Applications of microfluidics and optical manipulation for photoporation and imagingRendall, Helen A. January 2015 (has links)
Optical manipulation covers a wide range of techniques to guide and trap cells using only the forces exerted by light. Another optical tool is photoporation, the technique of injecting membrane-impermeable molecules using light, which has become an important alternative to other injection techniques. Together they provided sterile tools for manipulation and molecule delivery at the single-cell level. In this thesis, the properties of low Reynolds fluid flows are exploited to guide cells though a femtosecond Bessel beam. This design allows for high-throughput optical injection of cells without the need to individually target cells. A method of 'off-chip' hydrodynamic focusing was evaluated and was found to confine 95.6% of the sample within a region which would receive a femtosecond dose compared to 20% without any hydrodynamic focusing. The system was tested using two cell lines to optically inject the membrane-impermeable dye, propidium iodide. This resulted in an increase of throughput by an order of magnitude compared to the previous microfluidic design (to up to 10 cells per second). Next optical trapping and photoporation were combined to create a multimodal workstation. The system provides 3D beam control using spatial light modulators integrated into a custom user interface. The efficiency of optical injection of adherent cells and trapping capabilities were tested. The development of the system provides the groundwork for exploration of the parameters required for photoporation of non-adherent cells. Finally optical trapping is combined with temporally focused multiphoton illumination for scanless imaging. The axial resolution of the system was measured using different microscope objectives before imaging cells stained with calcein. Both single and a pair of recently trypsinised cells were optically trapped and imaged. The position of the trapped cells was manipulated using a spatial light modulator in order to obtain a z-stack of images without adjusting the objective position.
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Evaluation des performances d'un système de localisation de véhicules de transports guidés fondé sur l'association d'une technique radio ULB et d'une technique de retournement temporel. / Performance evaluation of a location system guided transport vehicles based on the combination of UWB radio technology and a time reversal techniqueFall, Bouna 14 November 2013 (has links)
En transports guidés, la localisation précise des trains s’avère vitale pour une exploitation nominale du système de transport. Dans un environnement de propagation tel que celui d’une emprise ferroviaire, un capteur de localisation efficace est complexe à concevoir et à valider lorsqu’il doit opérer en présence de nombreux obstacles fixes et mobiles constitués par l’infrastructure et les trains. Afin de concevoir un tel capteur, nous proposons dans ce travail de thèse l’emploi de techniques innovantes dites de diversité spectrale que l’on retrouve également sous la dénomination de radio Ultra Large Bande (ULB). Dans ce travail, cette dernière est également associée à la technique de Retournement Temporel (RT) afin de tirer partie de cet environnement de propagation complexe. L’objectif visé est d’obtenir une localisation fiable et robuste des véhicules ferroviaires par focalisation de signaux ULB en direction des antennes sol ou trains. Des études théoriques alliées à des simulations ont été effectuées portant sur les propriétés de focalisation d’énergie de la technique de retournement temporel en tenant compte de plusieurs paramètres liés aux configurations antennaires, aux canaux de propagation rencontrés et à l’électronique utilisée. L’apport du retournement temporel sur la précision du système de localisation Ultra Large Bande a été quantifié en comparant le système de localisation ULB conventionnel, sans retournement temporel puis, en associant le RT. Les résultats théoriques et de simulations de la solution proposée ont été validés par des expérimentations menées en chambre anéchoïque ainsi qu’en environnement indoor. / In guided transport, the precise location of trains is vital for nominal operation of the transport system. In a propagation environment such as a railway line, an effective location sensor is complex to design and validate since it must operate in the presence of many fixed and mobile obstacles constituted by the infrastructure and the trains. In this thesis, to design such a sensor, we propose the use of so-called spectral diversity techniques also found under the name of Ultra Wideband radio (UWB). In this work, the latter is also associated with the Time Reversal (TR) technique to take advantage of the complex propagation environment. The objective is to obtain a reliable and robust location of rail vehicles by focusing UWB signals to antennas or ground trains. Theoretical studies combined with simulations were performed on the properties of energy focusing of TR technique taking into account several parameters related to antennal configurations, propagation channels and the railway environment. The contribution of TR on the accuracy of the positioning system was quantified by comparing the conventional UWB positioning system alone and then, combining it with TR. The theoretical results and simulations of the proposed solution have been validated by experiments carried out in an anechoic chamber and in indoor environment.
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Inversion of Nonlinear Dispersive Wave and its Application in Determining Tsunami Wave SoureLi, Lieh-Yu 13 April 2011 (has links)
In this study, the method of deciding the water level of the initial tsunami is proposed by using spatial-temporal focusing (Coalescence) theory and waveform inversion reciprocal with Green function. Tsunami and earthquake are so closely bonded that the current tsunami numerical model is dependent on the parameters of the fault and the initial tsunami water level by calculating the theory of
half flexibility. But in fact, it is not easy to have the parameters of seabed fault so that the initial tsunami water level is very hard to get a accurate value. On the other hand, although the parameters of fault can be speculated by seismic waves, because ground is uneven medium, therefore, it is still a lot of improvement to get the parameters of fault by using seismic waves. For the tsunami simulation, if you have the value of the initial tsunami water level, the fault parameters can be estimated.Since the propagation of tsunami in the ocean is a linear behavior, the propagating process is affected by the topography of the ocean and the nonlinear effect
so minimal that it is to satisfy the linear shallow water equations and the requirement of reversibility;However, in fact, the values of the water level measured by the tide stations on the coast are influenced by the shoaling effect so that the reversibility of linear system can not be directly applied to Coastal areas.Therefore, the overall Inversion procedure on this study consists of two parts; the first one is that the usage of variable
coefficient Korteweg-de Vries (vKdV) equation and the Coalescence theory inverses the data gathered by the Coastal tide stations to the water level data where the depth is more than 50m on the linear region, and compares the above results with the stimulation and confirms the accuracy of the inversed waveform;The second one is that according to the reversibility of the linear system the use of least squares and least squares QR- decomposition (LSQR) method reproduce the initial tsunami wave source that compares with the initial tsunami wave source by stimulating and has a very good conformity. The seismic parameters can be easily decided by the above results.
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Kampinės dispersijos panaudojimas galingų ir ultratrumpų impulsinių šviesos pluoštų formavimui netiesinių sąveikų metu / The use of angular dispersion for formation of high peak power and ultrashort pulsed light beams in nonlinear interactionsZaukevičius, Audrius 27 December 2012 (has links)
Disertacija yra skirta išnagrinėti ir paaiškinti impulsinių šviesos pluoštų parametrinio stiprinimo metu vykstančius reiškinius. Ypatingas dėmesys yra skiriamas erdvinei-laikinei impulsinio šviesos pluošto dinamikai stiprinimo metu. Visi nagrinėjamų reiškinių tyrimai buvo atliekami taikant skaitmeninio modeliavimo metodus. Modelyje panaudotos lygtys buvo išvestos iš Maksvelo lygčių, išsamiai aptariant lygčių išvedimo metu daromas prielaidas. Naudojantis pateiktu modeliu buvo gauti skaitmeniniai rezultatai, kurie parodė, jog nekolineariame parametriniame stiprintuve, kuomet kaupinimo ir signalinio impulsų frontai nėra sutapatinti, signalinis impulsas tampa pakrypęs ir be kampinės dispersijos taip pat įgyja ir erdvinį čirpą. Nustatyta, kad indukuotų erdvinės ir kampinės dispersijų mažėjimo sparta, didinant signalinio impulso laikinį čirpą, yra skirtinga. Pagrindiniai šio teorinio tyrimo rezultatai buvo patvirtinti eksperimentiškai. Taip pat šioje disertacijoje yra pristatomas naujas ir universalus impulsinių kūginių bangų parametrinio stiprinimo metodas, kuriame sustiprinti impulsai patys susispaudžia laike ir erdvėje paprasčiausiai sklisdami laisvoje erdvėje. Šis metodas leidžia formuoti didelio smailinio intensyvumo invariantiškus bangų paketus, galinčius nusklisti daugelį Relėjaus nuotolių norimoje medžiagoje. Priešingai nei įprastame čirpuotų impulsų stiprinimo metode, šis metodas nereikalauja impulsų spaustuvo, o tai žymiai palengvina parametrinį kelių optinių ciklų... [toliau žr. visą tekstą] / This thesis is aimed to provide an extensive picture of the phenomena encountered in optical parametric amplification of pulsed light beams. The particular attention is paid for the spatio-temporal dynamics of the pulsed light beams being amplified. All the research has been done by means of numerical methods. The equations used in the model were derived from the Maxwell’s equations and the assumptions made along the derivation of equations were discussed. Using the presented model it has been numerically demonstrated, that in noncollinear optical parametric amplifier, when pulse fronts of pump and signal are not matched, the signal pulse becomes tilted and aside from angular dispersion acquires a spatial chirp. It has been shown, that the magnitudes of the induced spatial and angular dispersions decrease at different rates with the increase of the signal pulse temporal chirp. The main results of this study were verified experimentally. Additionally, in this thesis we have proposed a novel and versatile method for pulsed conical wave parametric amplification with subsequent spatio-temporal compression during propagation in free space. It allows to form ultra-intense and propagation invariant wave-packets capable to propagate over many Rayleigh range in the desired material. In contrast to ordinary chirped-pulse amplification technique it does not require a pulse compressor, thus greatly facilitates the parametric amplification of few-cycle light pulses.
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The use of angular dispersion for formation of high peak power and ultrashort pulsed light beams in nonlinear interactions / Kampinės dispersijos panaudojimas galingų ir ultratrumpų impulsinių šviesos pluoštų formavimui netiesinių sąveikų metuZaukevičius, Audrius 27 December 2012 (has links)
This thesis is aimed to provide an extensive picture of the phenomena encountered in optical parametric amplification of pulsed light beams. The particular attention is paid for the spatio-temporal dynamics of the pulsed light beams being amplified. All the research has been done by means of numerical methods. The equations used in the model were derived from the Maxwell’s equations and the assumptions made along the derivation of equations were discussed. Using the presented model it has been numerically demonstrated, that in noncollinear optical parametric amplifier, when pulse fronts of pump and signal are not matched, the signal pulse becomes tilted and aside from angular dispersion acquires a spatial chirp. It has been shown, that the magnitudes of the induced spatial and angular dispersions decrease at different rates with the increase of the signal pulse temporal chirp. The main results of this study were verified experimentally. Additionally, in this thesis we have proposed a novel and versatile method for pulsed conical wave parametric amplification with subsequent spatio-temporal compression during propagation in free space. It allows to form ultra-intense and propagation invariant wave-packets capable to propagate over many Rayleigh range in the desired material. In contrast to ordinary chirped-pulse amplification technique it does not require a pulse compressor, thus greatly facilitates the parametric amplification of few-cycle light pulses. / Disertacija yra skirta išnagrinėti ir paaiškinti impulsinių šviesos pluoštų parametrinio stiprinimo metu vykstančius reiškinius. Ypatingas dėmesys yra skiriamas erdvinei-laikinei impulsinio šviesos pluošto dinamikai stiprinimo metu. Visi nagrinėjamų reiškinių tyrimai buvo atliekami taikant skaitmeninio modeliavimo metodus. Modelyje panaudotos lygtys buvo išvestos iš Maksvelo lygčių, išsamiai aptariant lygčių išvedimo metu daromas prielaidas. Naudojantis pateiktu modeliu buvo gauti skaitmeniniai rezultatai, kurie parodė, jog nekolineariame parametriniame stiprintuve, kuomet kaupinimo ir signalinio impulsų frontai nėra sutapatinti, signalinis impulsas tampa pakrypęs ir be kampinės dispersijos taip pat įgyja ir erdvinį čirpą. Nustatyta, kad indukuotų erdvinės ir kampinės dispersijų mažėjimo sparta, didinant signalinio impulso laikinį čirpą, yra skirtinga. Pagrindiniai šio teorinio tyrimo rezultatai buvo patvirtinti eksperimentiškai. Taip pat šioje disertacijoje yra pristatomas naujas ir universalus impulsinių kūginių bangų parametrinio stiprinimo metodas, kuriame sustiprinti impulsai patys susispaudžia laike ir erdvėje paprasčiausiai sklisdami laisvoje erdvėje. Šis metodas leidžia formuoti didelio smailinio intensyvumo invariantiškus bangų paketus, galinčius nusklisti daugelį Relėjaus nuotolių norimoje medžiagoje. Priešingai nei įprastame čirpuotų impulsų stiprinimo metode, šis metodas nereikalauja impulsų spaustuvo, o tai žymiai palengvina parametrinį kelių optinių ciklų... [toliau žr. visą tekstą]
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Méthodes optiques innovantes pour le contrôle rapide et tridimensionnel de l’activité neuronale / Advanced optical methods for fast and three-dimensional control of neural activityHernández Cubero, Óscar Rubén 22 January 2016 (has links)
La révolution en cours des outils optogénétiques - des protéines photosensibles génétiquement induites qui peuvent activer, inhiber et enregistrer l'activité neuronale - a permis d'ouvrir une nouvelle voie pour relier l'activité neuronale et la cognition. Néanmoins, pour profiter au mieux de ces outils nous avons besoin de méthodes optiques qui peuvent projeter des schémas d'illumination complexes dans le cerveau. Pendant mon doctorat, j'ai travaillé sur deux nouveaux systèmes complémentaires pour la stimulation de l'activité neuronale. Le premier système combine des déflecteurs acousto-optiques et une illumination Gaussienne à faible ouverture numérique pour produire une photo activation rapide des outils optogénétiques. La capacité d'accès aléatoire du système permet de délivrer des séquences d'illumination spatialement et temporellement complexes qui simulent avec succès les schémas physiologiques de l'activité des fibres moussues dans des tranches de cerveaux. Ces résultats démontrent que les schémas de stimulation optogénétique peuvent être utilisés pour recréer l'activité en cours et étudier les microcircuits du cerveau dans un environnement physiologique. Alternativement, l'holographie générée par ordinateur (HGO) permet d'améliorer grandement les stimulations optogénétiques en répartissant efficacement la lumière sur plusieurs cibles cellulaires simultanément. Néanmoins, le confinement axial se dégrade pour des schémas d'illuminations larges. Afin de d'améliorer ce point, l’HGO peut être combinée avec une technique de focalisation temporelle qui confine axialement la fluorescence sans dépendre de l'allongement latéral. Les précédentes configurations maintiennent l'excitation non linéaire à un unique plan focal spatiotemporel. Dans cette thèse, je décris deux méthodes différentes qui permettent de dépasser ces limitations et de permettre la génération de schémas focalisés tridimensionnellement, à la fois spatialement et temporellement. / The ongoing revolution of optogenetic tools – genetically encoded light-sensitive proteins that can activate, silence and monitor neural activity – has opened a new pathway to bridge the gap between neuronal activity and cognition. However, to take full advantage of these tools we need optical methods that can deliver complex light patterns in the brain. During my doctorate, I worked on two novel and complementary optical systems for complex spatiotemporally neural activity stimulation. The first system combined acousto-optic deflectors and low numerical aperture Gaussian beam illumination for fast photoactivation of optogenetic tools. The random-access capabilities of the system allowed to deliver complex spatiotemporal illumination sequences that successfully emulated physiological patterns of cerebellar mossy fiber activity in acute slices. These results demonstrate that patterned optogenetic stimulation can be used to recreate ongoing activity and study brain microcircuits in a physiological activity context. Alternatively, Computer Generated Holography (CGH) can powerfully enhance optogenetic stimulation by efficiently shaping light onto multiple cellular targets simultaneously. Nonetheless, the axial confinement degrades for laterally extended illumination patterns. To address this issue, CGH can be combined with temporal focusing that axially confines fluorescence regardless of lateral extent. However, previous configurations restricted nonlinear excitation to a single spatiotemporal focal plane. In this thesis, I describe two alternative methods to overcome this limitation and enable three-dimensional spatiotemporal focused pattern generation.
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