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Li Li thesis whole 4-5.pdf (3.55 MB)

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Optimization of Micro-manufactured Human Sensing Platform

Li, Li
http://rave.ohiolink.edu/etdc/view?acc_num=case152294755902939

Abstract Details

2018, Doctor of Philosophy, Case Western Reserve University, Chemistry.
This dissertation explores the fabrication of nano-platinum modified micro-electrodes and array platinum chips to probe cholesterol diffusion from the cell plasma membrane (PM) of human mucosa. Cholesterol oxidase was covalently linked to the platinum sites of the micro-electrodes or chips. With the sensor touching the cell PM, active “free cholesterol” diffuses and then is oxidized producing hydrogen peroxide. The hydrogen peroxide (HOOH) generated was catalytically electro-oxidized at platinum as the electrochemical signal transduction mechanism in a double potential pulse method. In brief, HOOH was allowed to accumulate and was then detected by the first potential pulse. The second potential pulse gauged background charge for each consecutive measurement of HOOH. This difference charge (¿Q) was used to calculate the rate of cholesterol diffusion from the cell PM which is a measure of cholesterol thermodynamic activity in the PM. The double potential pulse method corrects for baseline drift by subtracting the background in real time. To advance this technology and its application to cystic fibrosis (CF), my work involved optimizing the double potential pulse method by improving micro-electrode based sensor fabrication and performance, carrying out in vivo studies on CF patients for comparison to healthy subjects, measuring PM cholesterol in CF and wild type (WT) human bronchial epithelial (HBE) cells including the consequences of treatment with drugs used to treat CF, and fabricating and characterizing micro-array sensor chips for human cholesterol measurements. Chapter 2 describes improving the micro-electrode fabrication, including trying different methods to deposit platinum nanoparticles on disk micro-electrodes. Here, the work for characterizing these micro-electrode-based sensors for in vivo PM cholesterol measurements is discussed. Chapter 3 describes fabricating carbon disk micro-electrodes containing platinum particles of nanometer dimensions and the application of these sensors to measure human mucosa cholesterol for CF patients vs. healthy subjects as controls. Chapter 4 describes measuring PM cholesterol in live CF and WT HBE cells, and studying the effects of drugs used for the treatment of CF on those cells with respect to PM cholesterol. The relationship between CF and elevated cholesterol diffusion observed in this cell study is similar to what was observed in the human studies presented in Chapter 3 and provides cell and tissue level evidence concerning PM cholesterol diffusion in CF models relative to the human disease state. Chapter 5 describes carrying out human cholesterol measurements with wafer-batch fabricated platinum micro-array chips through collaboration of the Burgess and Kelley groups with the Shin laboratory at Georgia Southern University. Finally, Chapter 6 gives a summary of possible future directions for this science. Real time analysis of cholesterol in human subjects is described and emphasized. Single cell cholesterol methodology and some other biomarker measurements were included. Some initial work demonstrating the feasibility of implementing this general sensing platform as a non-blood glucose test at human mucosa is presented in the Appendix.
Thomas Gray (Committee Chair)
Robert Salomon (Committee Member)
Michael Zagorski (Committee Member)
James Burgess (Committee Member)
Thomas Kelley (Advisor)
130 p.
Analytical Chemistry; Physical Chemistry

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Citations

  • Li, L. (2018). Optimization of Micro-manufactured Human Sensing Platform [Doctoral dissertation, Case Western Reserve University]. OhioLINK Electronic Theses and Dissertations Center. http://rave.ohiolink.edu/etdc/view?acc_num=case152294755902939

    APA Style (7th edition)

  • Li, Li. Optimization of Micro-manufactured Human Sensing Platform. 2018. Case Western Reserve University, Doctoral dissertation. OhioLINK Electronic Theses and Dissertations Center, http://rave.ohiolink.edu/etdc/view?acc_num=case152294755902939.

    MLA Style (8th edition)

  • Li, Li. "Optimization of Micro-manufactured Human Sensing Platform." Doctoral dissertation, Case Western Reserve University, 2018. http://rave.ohiolink.edu/etdc/view?acc_num=case152294755902939

    Chicago Manual of Style (17th edition)

case152294755902939
99
© 2018, some rights reserved.Creative Commons License Optimization of Micro-manufactured Human Sensing Platform by Li Li is licensed under a Creative Commons Attribution-NonCommercial-NoDerivs 3.0 Unported License. Based on a work at etd.ohiolink.edu.
This open access ETD is published by Case Western Reserve University School of Graduate Studies and OhioLINK.