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Measurement and manipulation of mitochondrial membrane potential in cancer cells
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Skates, Emily (2022) Measurement and manipulation of mitochondrial membrane potential in cancer cells. PhD thesis, University of Warwick.
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Official URL: http://webcat.warwick.ac.uk/record=b3956954
Abstract
This thesis focuses on the measurement and the manipulation of the mitochondrial membrane potential. Major findings in this work include the presentation of a novel fluorescent dye for the measurement of the mitochondrial membrane potential. We demonstrate this dyes ability to report qualitative dynamics of mitochondrial membrane potential with the use of real-time fluorescence confocal microscopy. In our investigations, we also identified experimental conditions where the photosensitisation of this dye caused it to become an actuator of the mitochondrial membrane potential. Therefore, we created a simple mathematical model that described this behaviour. The work in this thesis highlights the importance of microscopy imaging and the impact that this has on biological samples and fluorescent dye usability.
Furthermore, we follow up investigations into this dye and determined that its photoactivation can reduce viability in a cancer cell line, by loss of the mitochondrial membrane potential, possibly by a mechanism that involves reactive oxygen species. In summary, we identified a dye that has the dual ability for not only the measurement of the mitochondrial membrane potential in mammalian cells but also its manipulation to incur cancer cell death. Additionally, we further developed methods of membrane potential manipulation by designing a microfluidic device with integrated electrodes for the electrical stimulation of single-cells via capactive coupling. This worked aimed to create a device that enabled easy cell loading and electrode creation for the user in order to improve reliability and repeatability of this method. This setup is also in combination with a simple FEM model for determining the electric field strength that each cell is exposed to.
Item Type: | Thesis (PhD) | ||||
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Subjects: | Q Science > QH Natural history Q Science > QP Physiology R Medicine > RC Internal medicine > RC0254 Neoplasms. Tumors. Oncology (including Cancer) |
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Library of Congress Subject Headings (LCSH): | Mitochondrial membranes, Membrane potentials (Electrophysiology), Cell membranes -- Electric properties, Fluorescent labeling, Cancer cells | ||||
Official Date: | December 2022 | ||||
Dates: |
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Institution: | University of Warwick | ||||
Theses Department: | School of Life Sciences | ||||
Thesis Type: | PhD | ||||
Publication Status: | Unpublished | ||||
Supervisor(s)/Advisor: | Soyer, Orkun S. ; Asally, Munehiro | ||||
Format of File: | |||||
Extent: | xv, 175 pages : illustrations | ||||
Language: | eng |
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