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Towards a Microfluidic Toolbox for Proteomics: Novel Sample Pre-processing and Separation Techniques

Microfluidics was introduced in the early 1990’s and was posited to usher in a new age of integrated analysis systems in the form of labs-on-a-chip. To date, numerous embodiments of microfluidic technologies including fully integrated analysis systems have been described for various applications. Microfluidics can be sub-divided into two paradigms based on fluid manipulation in streams or droplets. In the former, streams of fluids flow through micron-dimension channels, and typical volumes manipulated are in the pico-liter to nano-litre range. These devices are mainly employed for rapid, high efficiency chemical separations, among other applications. In the latter, droplets are manipulated on a dielectric-coated array of microelectrodes in a process called digital microfluidics (DMF). In DMF each droplet is individually addressable, giving superior spatial control over fluid droplets with volumes in the pico-liter to micro-litre range. Independently addressable droplets make DMF amenable to carrying out sequential reactions. This thesis presents methods towards the integration of these two microfluidic paradigms into “hybrid microfluidic” platforms. Hybrid devices contain a DMF array for sample preparation and a microfluidic channel network for on-line analysis by chemical separation. Sample transfer between the platforms is made by way of a digital-channel interface, which has been fabricated in two geometries: side-on and vertical. Chemical separations on hybrid devices are performed in various open-channel and chromatographic modes. In open-channel methods analytes are separated by microchannel zone electrophoresis (MZE) or micellar electrokinetic chromatography (MEKC). In chromatographic separations porous polymer monolithic (PPM) columns were created in situ by UV-initiated polymerization of acrylate monomers. Prior to integration into hybrid microfluidic devices PPMs were optimized for use in gradient elution microchannel electrochromatography (MEC) of peptides. It is anticipated that hybrid microfluidic devices will bridge a large bottleneck for myriad analyses by combining sample preparation with on-line analysis by chemical separation.

Identiferoai:union.ndltd.org:TORONTO/oai:tspace.library.utoronto.ca:1807/29976
Date15 September 2011
CreatorsWatson, Michael
ContributorsWheeler, Aaron
Source SetsUniversity of Toronto
Languageen_ca
Detected LanguageEnglish
TypeThesis

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