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Title: Phase separation in binary phospholipid vesicles studied using fluorescence microscopy techniques
Author: Beales, Paul
Awarding Body: University of Edinburgh
Current Institution: University of Edinburgh
Date of Award: 2005
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In this thesis I study Giant Unilamellar Vesicles (GUVs) consisting of a mixture of two phospholipids with identical hydrophobic tails but different hydrophilic head groups. GUVs are between 5 and 200 microns in diameter so can be studied using optical observation techniques. This size range and membrane geometry is also comparable to that of the cell membranes of living organisms, hence the study of simple lipid model membrane systems in the form of GUVs may give insight into any heterogeneous structure of living membranes such as the possible existence of ‘lipid rafts’. Improved understanding of GUVs with only a small number of lipid components could also be of use for in vivo drug delivery systems where drugs are transported in encapsulated form to a target site where they are released. The binary lipid GUVs were studied by fluorescence multiphoton and confocal microscopy, techniques which allow the collection of digital images in the form of thin, in-focus sections through the sample. This results in clear, high resolution images of the GUVs. Amphiphilic fluorophores, which preferentially partition between lipid phases, were added to the lipids in trace amounts to enable the detection of domains of different phases in the membrane of the vesicles. GUVs were obtained by electroformation in formation chambers which were designed and constructed especially for this project. Domain formation was observed as the vesicles were slowly cooled from the high temperature fluid (Lα) phase through a region of gel – fluid phase coexistence.
Supervisor: Not available Sponsor: Not available
Qualification Name: Thesis (Ph.D.) Qualification Level: Doctoral
EThOS ID:  DOI: Not available