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Theses

Développement d'un système de microfluidique de gouttes pour l'analyse d'échantillons complexes

Abstract : Droplet microfluidics is a technology with a huge potential for miniaturization and automation of conventional methods in bioanalysis. Indeed, droplet microfluidics has many functionalities, such as merging, sorting and cell encapsulation, that can reproduce manipulations in standard protocols in bioanalysis on very small volumes with advantages such as a low samples and reagents consumption and reduction of analysis duration. During this thesis, we developed protocols for two types of biological analysis, using magnetic particles manipulation in 100 nL droplets.The first project is about single cell multiomics analysis. We implemented a droplet microfluidics protocol for the separation of mRNA and DNA from complex samples: from a mix of pre-purified RNA and DNA to a suspension of less than ten cells. At each steps, the droplet system performances were evaluated and the results were compared to extractions made in tubes in a non-microfluidics way. The results are very promising since they demonstrate for the first time in such a system the sequential separation of DNA and RNA from cell lysate or a few cells encapsulated and lysed in droplets.The second project is about the sample preparation in droplet for MALDI-TOF mass spectrometry analysis. This project was built in collaboration researchers from CEA Saclay. They have shown that it is very interesting to deposit samples in droplets on MALDI plates to increase the detection sensitivity thanks to a focusing effect. We have implemented on an original integrated droplet microfluidic approach for protein analysis by MALDI-TOF MS: from in drop sample preparation by supported enzymatic digestion to on plate deposition and peptides local pre-concentration. Development was done with lysozyme as a model protein and thanks to our in-drop digestion and deposition protocol we identified this protein by mass fingerprint from 200 fmol protein with a good reliability.
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Submitted on : Tuesday, September 14, 2021 - 4:09:09 PM
Last modification on : Tuesday, November 16, 2021 - 4:01:43 AM

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Mathilde Richerd. Développement d'un système de microfluidique de gouttes pour l'analyse d'échantillons complexes. Chimie analytique. Université Paris sciences et lettres, 2021. Français. ⟨NNT : 2021UPSLS051⟩. ⟨tel-03344129⟩

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