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Microelectrophoresis System Utilizing Conductivity Detection Analyzing Biological Molecues, Michelle Galloway

LSU Doctoral Dissertations

Microfabrication technology has proven to be a valuable tool for creating polymer-based devices utilized in chemical and biochemical assays. Although, reducing the size of the device allows for short analysis times and reduces the reagent demand to ultrasmall volumes (< 1 nanoliter), a resulting consequence is the constraint placed on the limits of detection associated with the detector hardware required for readout. To overcome such constraints, laser-induced fluorescence (LIF) is often employed as a detection method as it provides low detection limits, which approach the single molecule level. Unfortunately, most LIF systems do not offer the benefits of miniaturization, with the detector components (i.e. laser, optics, filters) often times requiring a much larger footprint compared to the device. Another readout strategy that has shown promise for these devices is conductivity detection. Detection can be accomplished using either conventional-size or microfabricated electrodes, which can be integrated on the device. Although conductivity has been commonly used to detect inorganic or small organic species, the potential for detection of biological species has received little attention. In this work, an integrated conductivity detector was developed for the analysis of amino acids, peptides, proteins, and oligonucleotides (double-stranded DNA). Using the detector, mass detection sensitivities in the range of 10^-18 - 10^-21 moles were achieved. To increase the throughput of the system a state-of-the-art, multichannel device with a conductivity array detector was devised. This device, which consists of a 16-channel fluidic network and a printed circuit board, is geared toward automating three-processing steps onto a single fluidic platform including purification, preconcentration and detection for downstream parallel processing.