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Title: Microfluidic droplets : cell-based and (bio-)chemical assays on the micro-scale
Author: Hübner, A.-M.
Awarding Body: University of Cambridge
Current Institution: University of Cambridge
Date of Award: 2009
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The first chapter of this thesis introduces the reader into the field of microdroplets. A brief literature review is given summarising the achievements in biochemistry and biology using microfluidic droplets. In Chapter 2 the techniques that were applied to fabricate devices and to generate droplets are highlighted. Chapter 3 demonstrates that fluorescently labelled single cells can be controllably compartmentalised within aqueous microdroplets. A high-throughput screening was performed by detecting the expression of the fluorescent protein in individual cells with simultaneous measurement of droplet size and cell occupancy. Chapter 4 comprises a cell-based enzymatic assay performed inside picolitre droplets. Comparing studies on enzymatic activity of wild type and a mutant enzyme successfully demonstrate the feasibility of using microfluidic droplets to provide time-resolved kinetic measurements. Chapter 5 describes the design, fabrication and use of a single layered poly(dimethylsiloxane) microfluidic structure for the entrapment and release of microdroplets in a plate reader format controlled entirely by liquid flow. Such an array-based approach is used to characterise droplet shrinkage, aggregation of encapsulated E. coli cells and enzymatic reactions. Chapter 6 and 7 widens the approach described in Chapter 5 by entrapping droplets next to each other. Those entrapped droplets could be fused by means such as lasers and electric fields. With such an approach the formation of a chemical complex was monitored with sub-millisecond time resolution. Furthermore, depending on the surfactant that was chosen leakage and cross-contamination of an encapsulated solute between two adjacent droplets could be investigated. Chapter 8 presents an approach of droplets incubation for several minutes up to almost an hour in a continuously operating device. Chapter 9 lists all methods that were applied to generate, manipulate and utilise the droplets in each individual experiment.
Supervisor: Not available Sponsor: Not available
Qualification Name: Thesis (Ph.D.) Qualification Level: Doctoral
EThOS ID:  DOI: Not available