O'Connor, Darragh (2019) Fluorescent probes for lipid droplet and lipid membrane imaging in cells and models. PhD thesis, Dublin City University.
Abstract
Cell membranes are believed to be laterally ordered into micro and nano-domains comprising of more fluid liquid-disordered (Ld) and more viscous liquid-ordered (Lo) phases. The latter subphases contain high concentrations of cholesterol and glycosphingolipids. These so call lipid rafts are experimentally distinguishable on the basis of their resistance to detergent solubilisation and are believed to play important roles in membrane function including in protein trafficking and signalling as they can drive protein-protein interactions through sequestering of proteins to these domains. Membrane domains in living cells are difficult to interrogate as they are dynamic and at sub-microscopic length scales they are outside the range of most conventional microscopies. However, they can potentially be imaged using recently developed super-resolution methods and as they are dynamic structures their diffusion can be measured using correlation methods. Therefore, new fluorescent probes are needed that can (a) partition selectively to membranous regions, (b) target the Lo and Ld phases selectively (c) that have appropriate photophysical properties compatible with the above techniques. These include large Stokes shift, high selectivity, excellent photostability, high molecular brightness, low cytotoxicity and high quantum yields. A key aim of this thesis was to design and synthesize new fluorescent probes that sequester specifically to lipid rich regions of cells or models and can distinguish Lo/Ld regions or lipid droplets, using confocal microscopy, fluorescence correlation spectroscopy (FCS), fluorescent lifetime imaging (FLIM) and the relatively new technique of super resolution microscopy, specifically, STimulated Emission Depletion (STED) microscopy.
Metadata
Item Type: | Thesis (PhD) |
---|---|
Date of Award: | November 2019 |
Refereed: | No |
Supervisor(s): | Keyes, Tia E. |
Uncontrolled Keywords: | BODIPY; Lipid Membranes; Photophysics; Liquid ordered; Phase probes; fluorescence; pyrene; charge transfer |
Subjects: | Physical Sciences > Chemistry |
DCU Faculties and Centres: | DCU Faculties and Schools > Faculty of Science and Health > School of Chemical Sciences |
Use License: | This item is licensed under a Creative Commons Attribution-NonCommercial-No Derivative Works 3.0 License. View License |
Funders: | Science Foundation Ireland under Grant No. [14/IA/2488] |
ID Code: | 23753 |
Deposited On: | 19 Nov 2019 16:00 by Tia Keyes . Last Modified 13 Dec 2019 15:34 |
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