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Title page for ETD etd-03232011-210812

Type of Document Dissertation
Author McKenzie, Jennifer Robin
Author's Email Address jennifer.mckenzie@vanderbilt.edu
URN etd-03232011-210812
Title Novel Electrochemical Techniques for Metabolic Profiling of Cellular Stress
Degree PhD
Department Chemistry
Advisory Committee
Advisor Name Title
David Cliffel Committee Chair
David Wright Committee Member
John McLean Committee Member
John Wikswo Committee Member
  • microphysiometry
  • toxicology
  • electroanalytical chemistry
Date of Defense 2011-03-02
Availability unrestricted
Real-time electrochemical monitoring of cellular metabolism has revolutionized traditional toxicology by providing immediate insights into the effects of cellular stress on metabolism. The multi-analyte microphysiometer (MAMP) allows for simultaneous measurement of extracellular glucose, lactate, oxygen, and acidification in a microfluidic environment. In this study, microphysiometry techniques provided new insights into the metabolic state of ischemic neurons, revealing acid as the analyte predictive of neuronal survival. This technique also furthered understanding of the metabolic action of Cholera toxin, demonstrating that toxin mediated activation of cAMP production occurrs in shorter time-frames than previously observed. Additionally, the effects of SEB exposure on several intestinal and immune cell lines were explored to create a profile of the metabolic effects of this toxin as it transcytoses through the intestine to the immune system. Finally, a screen-printed electrode (SPE) with five modifiable platinum electrodes was designed and found to be a highly adaptable tool for the simultaneous, real-time detection of a range of analytes in bulk and microfluidic environments. Microfluidic cell housing was created using polydimethylsiloxane (PDMS) to increase portability and sensitivity over the current MAMP system for monitoring metabolic flux. With the culmination of these works, the wealth of techniques possible with the MAMP has been expanded through the design and implementation of new sensor platforms, sensing elements, and instrumentation for multi-analyte data collection from current and future sensors. Based on these advancements, the future of the MAMP technology will continue to expand and provide new and exciting insights into the role of metabolism in cellular disease and stress.
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