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Design and optimization of efficient microfluidic platforms for particle manipulation and cell stimulation in systems biologyPaul, Alison Marie 25 August 2011 (has links)
The overall goal of this research was to develop an efficient microfluidic system to study signal transduction in stimulation dynamics. This research applied reactive transport fundamentals in concert with biological systems knowledge to completely understand diffusion of soluble signals, fluid and particle flow properties, and dynamics of cellular responses. First, a device capable of parallel multi-time-point cell stimulation and lysis on-chip was developed in collaboration. Second, to understand flow of cells through complex 3-D flow schemes, a Single-field Three-dimensional Epifluorescence Particle (STEP) imaging technique was developed. Using the STEP imaging technique, we were able to determine particle distributions and track individual particles in complex flow geometries. Third, during the design of the stimulation device it was observed that the cells do not distribute across the channel in the same way as the fluids. Based on the observation that geometry and particle size were most influential factors on particle distribution, it was hypothesized that our earlier observation and all observed phenomena in our experimental range were due to the volume exclusion of particles of finite size near the wall of the complex flow geometry. Overall, this work contributed to the realization of microfluidic platforms as powerful tools for probing areas of biology and medicine that are difficult with existing technology. The high-throughput format enabled simple and fast generation of large sets of quantitative data, with consistent sample handling. We demonstrated the necessary first steps to designing efficient unit operations on cells in microfluidic devices. The model can be used for informed design of unit operations in many applications in the future.
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Mesures expérimentales et modélisation du remaniement sédimentaire dans le bassin d’ArcachonBernard, Guillaume 10 July 2013 (has links)
Le remaniement sédimentaire, défini comme l’ensemble des mouvements de particules sédimentaires induits par les organismes benthiques, est l’une des deux composantes du phénomène de bioturbation. Il constitue un processus clé du fonctionnement des écosystèmes côtiers. Ce manuscrit présente une étude intégrée de ce depuis l’échelle de la simple particule sédimentaire jusqu’à celle de la communauté benthique in toto.Le développement d’une nouvelle approche expérimentale basée sur l’acquisition à haute fréquence et l’analyse de séries temporelles d’images de mouvements de luminophores le long de la paroi d’aquariums plats a permis de mesurer directement les mouvements élémentaires de particules de sédiment effectués par le bivalve A. alba. Cette approche a ainsi conduit à la première détermination expérimentale d’ « empreintes » du remaniement sédimentaire d’un invertébré marin, d’après le formalisme du modèle CTRW (Continuous Time Random Walk).Dans un second temps, le déploiement de cette nouvelle approche a permis d’évaluer, de manière dynamique (i.e. pendant des expériences de 48h) et sur l’ensemble de la partie de la colonne sédimentaire affectée par ce bivalve, le contrôle exercé par la température et par la disponibilité de matière organique fraîche sur les caractéristiques du processus de remaniement sédimentaire effectué par A. alba.Enfin, l’intensité du remaniement sédimentaire effectué par l’ensemble de la communauté benthique a été mesurée in-situ dans le Bassin d’Arcachon, à la fois dans un herbier à Zostera noltii et dans une zone de vase nue d’où celui-ci a disparu. Ceci a permis de déterminer les effets limitant de la présence d’herbier et de certaines espèces benthiques clés, sur le remaniement sédimentaire. / Sediment particle mixing, defined as the movements of sediment particles induced by benthic fauna, is one of the two components of bioturbation by benthic organisms. It is a key process of the ecological functioning in coastal areas. This manuscript presents an integrated study of sediment particle mixing process from the single sediment particle to the whole benthic community.The development of a new experimental approach, coupling high frequency acquisition of time series images of luminophores motions along thin aquaria glass walls, allowed for the direct measurement of elementary particle motions induced by the bivalve Abra alba. This constitutes the first experimental assessment of sediment particle mixing “fingerprints” in a marine invertebrate, according to the CTRW (Continuous Time Random Walk) model formulation.The deployment of this new approach also allowed for the determination of the control of water temperature and of fresh organic matter availability on sediment particle mixing induced by Abra alba. Moreover, the temporal (i.e., during 48h experiments) and spatial (i.e., over the whole section of the sediment column affected) dynamics of these effects were considered.At last, sediment particle mixing intensities induced by the whole benthic community were assessed in-situ in Arcachon Bay, within both a Zostera noltii meadow and a bare sediment mudflat where phanerogams were previously present. These results highlighted the restrictive effect of phanerogams themselves and of a restricted number of key benthic species, on sediment particle mixing.
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