{"id":{"repo_id":"ohiolink","oai_identifier":"oai:etd.ohiolink.edu:case1344251678"},"canonical_url":"https://search.dev.ndltd.org/etd/ohiolink/oai:etd.ohiolink.edu:case1344251678","repository":{"repo_id":"ohiolink","name":"OhioLINK","base_url":"https://etd.ohiolink.edu/acprod/odb_etd/ws/oai/oai"},"display":{"title":"Double Pulse Coulometry for Interfacial Biosensing at Platinum Microelectrodes","abstract":"This dissertation explores the use of a chronocoulometric technique to probe cholesterol efflux from the plasma membrane of cells and tissues. More specifically, a double potential pulse coulometric scheme for cholesterol sensing at platinum microelectrodes was developed and optimized to avoid amperometric measurements needed to reference to a nonzero solution baseline current and baseline shift upon repositioning the electrode in approaching and contacting with the cell plasma membrane. In this double potential pulse scheme, the first pulse (pulse 1) oxidizes hydrogen peroxide at the electrode surface and the second pulse (pulse 2) gauges the background charge at that time. The difference charge (i.e.; pulse 1 - pulse 2) signal arises from gradual accumulation of hydrogen peroxide generated at the interface between the electrode surface and cell plasma membrane. A bare electrode in contact with the plasma membrane serves as the control electrode in these experiments to remove the Faradaic background signal from enzyme-modified electrodes to provide an estimate of the hydrogen peroxide accumulated before pulse 1. The development of the coulometric scheme allows monitoring of background drift in real-time and signal averaging with electrode held in contact with the cell plasma membrane. Chapter 1 describes a general overview of the scope of this research and briefly explains previous work performed by this research group that demonstrates the use of different biosensing strategies to measure cholesterol efflux from the plasma membrane. Chapter 2 describes optimization and characterization studies of the double potential pulse waveform. Previous work with cavity enzyme-modified electrodes led to the optimization of the coulometric method with disk microelectrodes. Importantly, the characterization studies presented in this chapter will show that Faradaic background signal is influenced by platinum electrode surface modification, temperature, contact of cell plasma membrane, and solution chemistries. Chapter 3 describes the analog chronocoulometric detection of plasma membrane cholesterol of excised mouse trachea and frog muscle tissue. Chapter 4 describes the analog chronocoulometric detection of cystic fibrosis (CF) human epithelial cells. Lastly, Chapter 5 outlines future directions with cholesterol cycling from neuron stimulation.","abstract_html":"This dissertation explores the use of a chronocoulometric technique to probe cholesterol efflux from the plasma membrane of cells and tissues. More specifically, a double potential pulse coulometric scheme for cholesterol sensing at platinum microelectrodes was developed and optimized to avoid amperometric measurements needed to reference to a nonzero solution baseline current and baseline shift upon repositioning the electrode in approaching and contacting with the cell plasma membrane. In this double potential pulse scheme, the first pulse (pulse 1) oxidizes hydrogen peroxide at the electrode surface and the second pulse (pulse 2) gauges the background charge at that time. The difference charge (i.e.; pulse 1 - pulse 2) signal arises from gradual accumulation of hydrogen peroxide generated at the interface between the electrode surface and cell plasma membrane. A bare electrode in contact with the plasma membrane serves as the control electrode in these experiments to remove the Faradaic background signal from enzyme-modified electrodes to provide an estimate of the hydrogen peroxide accumulated before pulse 1. The development of the coulometric scheme allows monitoring of background drift in real-time and signal averaging with electrode held in contact with the cell plasma membrane. Chapter 1 describes a general overview of the scope of this research and briefly explains previous work performed by this research group that demonstrates the use of different biosensing strategies to measure cholesterol efflux from the plasma membrane. Chapter 2 describes optimization and characterization studies of the double potential pulse waveform. Previous work with cavity enzyme-modified electrodes led to the optimization of the coulometric method with disk microelectrodes. Importantly, the characterization studies presented in this chapter will show that Faradaic background signal is influenced by platinum electrode surface modification, temperature, contact of cell plasma membrane, and solution chemistries. Chapter 3 describes the analog chronocoulometric detection of plasma membrane cholesterol of excised mouse trachea and frog muscle tissue. Chapter 4 describes the analog chronocoulometric detection of cystic fibrosis (CF) human epithelial cells. Lastly, Chapter 5 outlines future directions with cholesterol cycling from neuron stimulation.","abstract_has_math":false,"creators":["West, Richard H."],"institution":"Case Western Reserve University School of Graduate Studies","degree_name":"Doctor of Philosophy","degree_level":"doctoral","degree_discipline":"Chemistry","degree_department":null,"school":null,"contributors":["Burgess, James","Dunbar, Robert"],"advisors":[],"committee_chairs":[],"committee_members":[],"year":2013,"date_issued":"2013-03-12","date_published":"2013-03-12","updated_at":"2026-07-24T03:35:52Z","subjects":["Physical Chemistry","Cholesterol","Coulometry","Microelectrodes"],"languages":["English"],"rights":["unrestricted","This thesis or dissertation is protected by copyright: all rights reserved. It may not be copied or redistributed beyond the terms of applicable copyright laws."],"rights_urls":[],"identifier_entries":[]},"links":{"outbound_url":"http://rave.ohiolink.edu/etdc/view?acc_num=case1344251678","outbound_label":"Repository record","outbound_source":"dc:identifier"},"metadata_groups":[{"id":"people","label":"People","entries":[{"key":"dc:contributor","label":"Contributor","values":["Burgess, James","Dunbar, Robert"]},{"key":"dc:creator","label":"Author","values":["West, Richard H."]}]},{"id":"academic_context","label":"Academic Context","entries":[{"key":"dc:date","label":"Dc Date","values":["2013-03-12"]},{"key":"dc:publisher","label":"Institution","values":["Case Western Reserve University School of Graduate Studies / OhioLINK"]},{"key":"dc:type","label":"Dc Type","values":["Electronic Thesis or Dissertation"]},{"key":"thesis:degree_discipline","label":"Discipline","values":["Chemistry"]},{"key":"thesis:degree_level","label":"Degree Level","values":["doctoral"]},{"key":"thesis:degree_name","label":"Degree Name","values":["Doctor of Philosophy"]},{"key":"thesis:institution_name","label":"Thesis Institution Name","values":["Case Western Reserve University School of Graduate Studies"]}]},{"id":"subjects_keywords","label":"Subjects and Keywords","entries":[{"key":"dc:subject","label":"Dc Subject","values":["Physical Chemistry","Cholesterol","Coulometry","Microelectrodes"]}]},{"id":"language_rights","label":"Language and Rights","entries":[{"key":"dc:language","label":"Dc Language","values":["English"]},{"key":"dc:rights","label":"Dc Rights","values":["unrestricted","This thesis or dissertation is protected by copyright: all rights reserved. It may not be copied or redistributed beyond the terms of applicable copyright laws."]}]},{"id":"identifiers","label":"Identifiers","entries":[{"key":"dc:identifier","label":"Identifier","values":["http://rave.ohiolink.edu/etdc/view?acc_num=case1344251678"]}]},{"id":"additional","label":"Additional Metadata","entries":[{"key":"dc:description","label":"Description","values":["This dissertation explores the use of a chronocoulometric technique to probe cholesterol efflux from the plasma membrane of cells and tissues. More specifically, a double potential pulse coulometric scheme for cholesterol sensing at platinum microelectrodes was developed and optimized to avoid amperometric measurements needed to reference to a nonzero solution baseline current and baseline shift upon repositioning the electrode in approaching and contacting with the cell plasma membrane. In this double potential pulse scheme, the first pulse (pulse 1) oxidizes hydrogen peroxide at the electrode surface and the second pulse (pulse 2) gauges the background charge at that time. The difference charge (i.e.; pulse 1 - pulse 2) signal arises from gradual accumulation of hydrogen peroxide generated at the interface between the electrode surface and cell plasma membrane. A bare electrode in contact with the plasma membrane serves as the control electrode in these experiments to remove the Faradaic background signal from enzyme-modified electrodes to provide an estimate of the hydrogen peroxide accumulated before pulse 1. The development of the coulometric scheme allows monitoring of background drift in real-time and signal averaging with electrode held in contact with the cell plasma membrane. Chapter 1 describes a general overview of the scope of this research and briefly explains previous work performed by this research group that demonstrates the use of different biosensing strategies to measure cholesterol efflux from the plasma membrane. Chapter 2 describes optimization and characterization studies of the double potential pulse waveform. Previous work with cavity enzyme-modified electrodes led to the optimization of the coulometric method with disk microelectrodes. Importantly, the characterization studies presented in this chapter will show that Faradaic background signal is influenced by platinum electrode surface modification, temperature, contact of cell plasma membrane, and solution chemistries. Chapter 3 describes the analog chronocoulometric detection of plasma membrane cholesterol of excised mouse trachea and frog muscle tissue. Chapter 4 describes the analog chronocoulometric detection of cystic fibrosis (CF) human epithelial cells. Lastly, Chapter 5 outlines future directions with cholesterol cycling from neuron stimulation."]},{"key":"dc:format","label":"Dc Format","values":["application/pdf","p.104","1. MB"]},{"key":"dc:title","label":"Title","values":["Double Pulse Coulometry for Interfacial Biosensing at Platinum Microelectrodes"]}]}],"canonical_facts":{"dc:contributor":["Burgess, James","Dunbar, Robert"],"dc:creator":["West, Richard H."],"dc:date":["2013-03-12"],"dc:description":["This dissertation explores the use of a chronocoulometric technique to probe cholesterol efflux from the plasma membrane of cells and tissues. More specifically, a double potential pulse coulometric scheme for cholesterol sensing at platinum microelectrodes was developed and optimized to avoid amperometric measurements needed to reference to a nonzero solution baseline current and baseline shift upon repositioning the electrode in approaching and contacting with the cell plasma membrane. In this double potential pulse scheme, the first pulse (pulse 1) oxidizes hydrogen peroxide at the electrode surface and the second pulse (pulse 2) gauges the background charge at that time. The difference charge (i.e.; pulse 1 - pulse 2) signal arises from gradual accumulation of hydrogen peroxide generated at the interface between the electrode surface and cell plasma membrane. A bare electrode in contact with the plasma membrane serves as the control electrode in these experiments to remove the Faradaic background signal from enzyme-modified electrodes to provide an estimate of the hydrogen peroxide accumulated before pulse 1. The development of the coulometric scheme allows monitoring of background drift in real-time and signal averaging with electrode held in contact with the cell plasma membrane. Chapter 1 describes a general overview of the scope of this research and briefly explains previous work performed by this research group that demonstrates the use of different biosensing strategies to measure cholesterol efflux from the plasma membrane. Chapter 2 describes optimization and characterization studies of the double potential pulse waveform. Previous work with cavity enzyme-modified electrodes led to the optimization of the coulometric method with disk microelectrodes. Importantly, the characterization studies presented in this chapter will show that Faradaic background signal is influenced by platinum electrode surface modification, temperature, contact of cell plasma membrane, and solution chemistries. Chapter 3 describes the analog chronocoulometric detection of plasma membrane cholesterol of excised mouse trachea and frog muscle tissue. Chapter 4 describes the analog chronocoulometric detection of cystic fibrosis (CF) human epithelial cells. Lastly, Chapter 5 outlines future directions with cholesterol cycling from neuron stimulation."],"dc:format":["application/pdf","p.104","1. MB"],"dc:identifier":["http://rave.ohiolink.edu/etdc/view?acc_num=case1344251678"],"dc:language":["English"],"dc:publisher":["Case Western Reserve University School of Graduate Studies / OhioLINK"],"dc:rights":["unrestricted","This thesis or dissertation is protected by copyright: all rights reserved. It may not be copied or redistributed beyond the terms of applicable copyright laws."],"dc:subject":["Physical Chemistry","Cholesterol","Coulometry","Microelectrodes"],"dc:title":["Double Pulse Coulometry for Interfacial Biosensing at Platinum Microelectrodes"],"dc:type":["Electronic Thesis or Dissertation"],"thesis:degree_discipline":["Chemistry"],"thesis:degree_level":["doctoral"],"thesis:degree_name":["Doctor of Philosophy"],"thesis:institution_name":["Case Western Reserve University School of Graduate Studies"]},"updated_at":"2026-07-24T03:35:52Z"}