In both, cell secretome analysis and bacteria evolution, controlled handling of particles with a few to sub-micrometers in size and media exchange are inevitable in order to investigate body fluid’s proteins or change the surrounding culture conditions for pivoted evolution. Typically, nanofiltration and ultra-centrifugation are employed which can lead to cell damage, need large sample volumes and have a high sample loss. Using contactless and label-free acoustic cell manipulation, disadvantages of other magnetic, dielectric or hydrodynamic methods can be avoided. Here, a novel design using acoustic forces for small particle trapping and media exchange is thoroughly numerically investigated including first- and second-order acoustic effects. The device comprises parallel aligned medium and air channels separated by a thin wall. Particle trapping occurs at this thin wall. The medium channel dimensions (height and width) and thin wall thickness are optimized with respect to trapping forces. Thinnest walls are preferable and an aspect ratio of 0.8. First preliminary experimental variation with polystyrene particles showed good agreement with the simulations. Thereby the particle trapping efficiency is evaluated under quiescent flow conditions. For particle trapping, a device with a channel height of 290μm and an aspect ratio of 0.7 is superior which supports the numerical results. Finally, medium exchange of E. coli bacteria is demonstrated with best results for a device with a channel height of 450μm and an aspect ratio of 0.8 showing that 13.4% of the initial bacteria were released after medium exchange which can be used for further processing.

The development of techniques targeting the manipulation of particles of different<p>sizes - mostly in the nano to millimeter scale - when dispersed in a carrier medium, is an increasingly important topic in many fields such as biotechnology,nanotechnology, medicine, biophysics and environmental monitoring and remediation. The underlying rationale for using such techniques stands in the sometimes compelling requirements of avoiding clogging as in micro/nano channel flows, of limiting sedimentation and wall interactions in particle/cell counting, of enhancing particle-surface interaction as in bio-sensing or of facilitating characterization and sorting as in bio-physical applications. Being developed in the frame of a Belgian national project devoted to the characterization and counting of pollutant in water media by digital holographic microscopy, this thesis tackles a peculiar class of particle manipulation techniques, commonly known as Focusing. The main goal of focusing is to avoid at best wall particle interactions and sedimentation, prevalent issues for dispersions owing in micro/mini-channels especially for applications such as optical characterization and counting.<p><p>The main attention was given to two flow focusing techniques - Hydrodynamic and Acoustic Focusing - for their wide range applicability and cost effectiveness. Hydrodynamic Focusing consists in controlling the position and spreading of the sample under investigation by means of a so-called sheath flow. A low-cost, nevertheless effective, prototype has been conceived, designed, manufactured and tested. It allowed for controlling the spreading of the sample stream and achieving a focusing ratio accounting for only 4% of the original stream width.<p><p>Acoustic Focusing takes advantage of the time-averaged pressure fields induced by the creation of standing waves in channels to manipulate and focus the dispersed particles. In the frame of this thesis, several devices have been developed using square cross section glass mini-channels. Aside from the cost-effectiveness, particles where focused in a somehow unexpected but high reproducible 3D matrix-like structure. A novel numerical model has also been implemented in order to study the conditions leading to the 3D structure formation. A good agreement between experimental and numerical results was found./Ce projet de thèse portant sur la manipulation de micro-particules dans des minicanaux s'inscrit dans le développement de cellules de flux pour des applications biologiques, qui est l'une des problématiques du projet HOLOFLOW, soutenu par<p>la région de Bruxelles Capitale. Les cellules de flux doivent permettre l'observation et la reconnaissance des micro-organismes vivants dans une large gamme de dimensions (de quelques microns à 1mm) avec la microscopie holographie digitale.<p>La problématique d'observation et de manipulation des microorganismes en flux est liée au clogging (bouchage) et à la sédimentation qui limitent la durée de vie des cellules d'observation. Ce projet de thèse s'inscrit dans cette problématique et propose deux axes d'étude pour limiter l'interaction entre organismes et canaux, la focalisation hydrodynamique, basée sur le guidage de flux, et la focalisation acoustique, basée sur la manipulation des particules.<p><p>La focalisation hydrodynamique est une technique basée sur l'injection différentiée de l'échantillon à observer et d'un fluide support. La différence des vitesses d'injection des flux permet de contrôler la dispersion des particules afin d'optimiser leur observation. Dans le cadre de cette thèse, un prototype à bas-coût a été développé et construit, permettant de focaliser les particules dans un faisceau jusqu'à 4% de leur faisceau incident.<p><p>La focalisation acoustique utilise la création d'une onde acoustique stationnaire afin de regrouper les particules en suspension au centre du canal. Au cours de cette thèse, plusieurs prototypes ont été réalisés, mettant en évidence la formation de motifs tridimensionnaux. Un model numérique a été spécialement développé afin d'étudier les conditions de génération de ces motifs, et de nombreuses expériences ont été menées afin de s'assurer de leur reproductibilité. Une bonne adéquation entre la position des particules mesurée et calculée numériquement a été démontrée. / Doctorat en Sciences de l'ingénieur / info:eu-repo/semantics/nonPublished