University Ph.D. Oral Examination
Title: On-Chip Isotachophoresis Assays for High Sensitivity Electrokinetic Preconcentration, Separation and Indirect Fluorescence Detection
Candidate: Tarun Khurana
Advisor: Prof. Juan G. Santiago
Department of Mechanical Engineering
Time: Wednesday, July 2nd 2008, 2:00 pm
refreshments served at 1:45 pm
Location: McCullough building, room 122 (map attached)
Abstract:
Microfluidic devices have been particularly attractive for separation based chemical and biological analysis since the small length scales bring fundamental improvements in reagent volume, analysis time, resolution and separation efficiency. However, smaller length scales and volumes are also associated with lower detection sensitivity and therefore, microchip electrophoresis analysis is often less sensitive and is more commonly used for fluorescent analytes since fluorescence detection platform offers higher sensitivity. This presentation will focus on leveraging an electrophoresis technique termed isotachophoresis (ITP) for improving the detection sensitivity of on-chip electrophoresis assays and extending its scope to non-fluorescent analytes.
ITP is a robust sample preconcentration technique focuses analytes into zones that are ~10 µm wide. Such extreme compression of analytes results in drastic improvement in the detection sensitivity and resolution of electrophoretic separation system. We present a theoretical and experimental study of dynamics of ITP preconcentration that helps identify and optimize experiment parameters to achieve high sample preconcentration We have also demonstrated an indirect detection technique based on ITP to detect non fluorescent analytes on a standard fluorescence detection platforms. We leverage ITP to preconcentrate and separate analytes into distinct analyte zones and use a set of fluorescent species with different electrophoretic mobilities to demarcate the boundaries of these analyte zones and thereby, indirectly detect the non-fluorescent analytes present. We obtain ~1 µM detection sensitivity with this assay with high repeatability and have demonstrated indirect detection of a variety of analytes such as amino acids, organic acids and environmental toxins such as phenols and cresol.
Title: On-Chip Isotachophoresis Assays for High Sensitivity Electrokinetic Preconcentration, Separation and Indirect Fluorescence Detection
Candidate: Tarun Khurana
Advisor: Prof. Juan G. Santiago
Department of Mechanical Engineering
Time: Wednesday, July 2nd 2008, 2:00 pm
refreshments served at 1:45 pm
Location: McCullough building, room 122 (map attached)
Abstract:
Microfluidic devices have been particularly attractive for separation based chemical and biological analysis since the small length scales bring fundamental improvements in reagent volume, analysis time, resolution and separation efficiency. However, smaller length scales and volumes are also associated with lower detection sensitivity and therefore, microchip electrophoresis analysis is often less sensitive and is more commonly used for fluorescent analytes since fluorescence detection platform offers higher sensitivity. This presentation will focus on leveraging an electrophoresis technique termed isotachophoresis (ITP) for improving the detection sensitivity of on-chip electrophoresis assays and extending its scope to non-fluorescent analytes.
ITP is a robust sample preconcentration technique focuses analytes into zones that are ~10 µm wide. Such extreme compression of analytes results in drastic improvement in the detection sensitivity and resolution of electrophoretic separation system. We present a theoretical and experimental study of dynamics of ITP preconcentration that helps identify and optimize experiment parameters to achieve high sample preconcentration We have also demonstrated an indirect detection technique based on ITP to detect non fluorescent analytes on a standard fluorescence detection platforms. We leverage ITP to preconcentrate and separate analytes into distinct analyte zones and use a set of fluorescent species with different electrophoretic mobilities to demarcate the boundaries of these analyte zones and thereby, indirectly detect the non-fluorescent analytes present. We obtain ~1 µM detection sensitivity with this assay with high repeatability and have demonstrated indirect detection of a variety of analytes such as amino acids, organic acids and environmental toxins such as phenols and cresol.
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