Development of a microfluidic device based on Deterministic Lateral Displacement (DLD) for biological sample preparation, towards the extraction of extracellular vesicles

Abstract : Over the past decades, Extracellular Vesicles (EVs) have demonstrated strong potential as new biomarkers for liquid biopsy. Indeed, since EVs are fingerprints of parent cells, they can be exploited as early diagnostic tools. However, owing to their small size and high heterogeneity, EVs are challenging to extract from biofluids. In particular, reproducible and standardized protocols are required to perform fast, efficient, and cost-effective preparation of undamaged EV subpopulations from limited sample volumes. Deterministic Lateral Displacement (DLD) appears to be a promising microfluidic technology for this preparation by means of passive and label-free separation. DLD performs size-based separation of particles around a critical diameter that can be fine-tuned according to design parameters in an array of micropillars. Across the numerous biotechnological applications of DLD, none has yet successfully performed the complete extraction of EVs from unprocessed biofluids. This is the underlying motivation of this thesis, which outlines technological enhancements that make DLD separation more predictable, efficient, and easy-to-integrate. Based on both numerical and experimental developments, predictive models are proposed in order to anticipate particle behavior and to help in the design of efficient DLD devices. In addition to the optimization of single DLD devices, this thesis also addresses the issue of system integration. An innovative approach of serial connection between DLD modules is proposed to address the sequential sorting of particles from a complex biofluid and ensure that there is no loss of function of individual DLD devices when operated alone or in series. Two biological applications illustrate the potential of DLD-based sample preparation systems: the isolation of E. coli bacteria from human blood samples for sepsis diagnostics and the extraction of EVs from cell culture media with the perspective of liquid biopsy applications. And as sample preparation cannot be dissociated from detection or characterization, this thesis moreover highlights the potential integration of DLD in an all-in-one microfluidic device for both sample preparation and analysis of extracted EVs. Such a portable and autonomous device could overcome some of the current limitations with regard to the clinical use of EVs.
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Eloïse Pariset. Development of a microfluidic device based on Deterministic Lateral Displacement (DLD) for biological sample preparation, towards the extraction of extracellular vesicles. Biological Physics [physics.bio-ph]. Université Grenoble Alpes, 2018. English. ⟨NNT : 2018GREAY027⟩. ⟨tel-01978865⟩

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