prikaz prve stranice dokumenta Optimization of giant unilamellar vesicle electroformation for different lipid mixtures with a focus on high cholesterol concentrations
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doctoral thesis
Optimization of giant unilamellar vesicle electroformation for different lipid mixtures with a focus on high cholesterol concentrations
2023. urn:nbn:hr:166:553755
Boban, Zvonimir
University of Split
Faculty of Science
Department of Physics

Cite this document

Boban, Z. (2023). Optimization of giant unilamellar vesicle electroformation for different lipid mixtures with a focus on high cholesterol concentrations (Doctoral thesis). Split: University of Split, Faculty of Science. Retrieved from https://urn.nsk.hr/urn:nbn:hr:166:553755

Boban, Zvonimir. "Optimization of giant unilamellar vesicle electroformation for different lipid mixtures with a focus on high cholesterol concentrations." Doctoral thesis, University of Split, Faculty of Science, 2023. https://urn.nsk.hr/urn:nbn:hr:166:553755

Boban, Zvonimir. "Optimization of giant unilamellar vesicle electroformation for different lipid mixtures with a focus on high cholesterol concentrations." Doctoral thesis, University of Split, Faculty of Science, 2023. https://urn.nsk.hr/urn:nbn:hr:166:553755

Boban, Z. (2023). 'Optimization of giant unilamellar vesicle electroformation for different lipid mixtures with a focus on high cholesterol concentrations', Doctoral thesis, University of Split, Faculty of Science, accessed 18 April 2024, https://urn.nsk.hr/urn:nbn:hr:166:553755

Boban Z. Optimization of giant unilamellar vesicle electroformation for different lipid mixtures with a focus on high cholesterol concentrations [Doctoral thesis]. Split: University of Split, Faculty of Science; 2023 [cited 2024 April 18] Available at: https://urn.nsk.hr/urn:nbn:hr:166:553755

Z. Boban, "Optimization of giant unilamellar vesicle electroformation for different lipid mixtures with a focus on high cholesterol concentrations", Doctoral thesis, University of Split, Faculty of Science, Split, 2023. Available at: https://urn.nsk.hr/urn:nbn:hr:166:553755

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Metadata
TitleOptimization of giant unilamellar vesicle electroformation for different lipid mixtures with a focus on high cholesterol concentrations
Title (croatian)Optimizacija elektroformacije divovskih unilamelarnih vezikula za različite lipidne smjese uz naglasak na visoke koncentracije kolesterola
AuthorZvonimir Boban
MentorMarija Raguž (mentor)
Committee memberMile Dželalija (predsjednik povjerenstva)
Committee memberDamir Sapunar (član povjerenstva)
Committee memberLucija Krce (član povjerenstva)
GranterUniversity of Split
Faculty of Science
(Department of Physics)
Split
Defense date and country2023-04-27, Croatia
Scientific / art field,
discipline and subdiscipline		NATURAL SCIENCES
Physics
Universal decimal classification
(UDC)		577 - Biochemistry. Molecular biology. Biophysics
Abstract
Giant unilamellar vesicles (GUVs) are widely used as artificial cell membrane models. Electroformation is the most commonly used method for their production. The method depends on multiple parameters, so reproducible production of high quality GUVs can be challenging, especially when working with lipid mixtures containing cholesterol concentrations reaching and surpassing the bilayer saturation threshold. After analyzing existing electroformation protocols, we identified two key issues for such mixtures. The first issue is related to experiment reproducibility. Most protocols use the drop-deposition method which produces lipid films of nonuniform thickness, resulting in lower experiment reproducibility and efficiency of GUV formation. The second is cholesterol demixing in the form of anhydrous crystals during lipid film drying, resulting in an artifactual decrease of cholesterol concentration in GUVs compared to the initial lipid mixture. To deal with the first problem, we first tested electroformation from such mixtures with replacement of the drop-deposition step by spin-coating. Optimizing this process improved the GUV size and yield, but the cholesterol demixing issue remained. To address this issue, we developed a new protocol which combines the rapid solvent exchange, plasma cleaning and spin-coating techniques to produce GUVs by electroformation from damp lipid films. We believe that this new improved electroformation protocol will allow us to successfully study models of eye lens fiber cell membranes with their very high Chol content.
Abstract (croatian)
Divovske unilamelarne vezikule (DUV) naširoko se koriste kao modeli staničnih membrana. Elektroformacija je najčešće korištena metoda za njihovu proizvodnju. Metoda ovisi o više parametara, tako da ponovljiva proizvodnja visokokvalitetnih DUV-ova može biti izazovna, osobito pri radu s lipidnim smjesama koje sadrže koncentracije kolesterola koje dosežu i premašuju prag zasićenja membranskog dvosloja. Nakon analize postojećih protokola elektroformacije, identificirali smo dva ključna problema za takve smjese. Prvi se odnosi na ponovljivost eksperimenata. Većina protokola koristi metodu nakapavanja koja proizvodi lipidne filmove nejednake debljine, što rezultira smanjenom ponovljivošću eksperimenata i učinkovitošću stvaranja DUV-ova. Drugi je izlučivanje kolesterola u obliku dehidriranih kristala tijekom sušenja lipidnog filma, što dovodi do artefaktualnog smanjenja koncentracije kolesterola u DUV-ovima u usporedbi s početnom lipidnom smjesom. Kako bismo umanjili prvi problem, testirali smo elektroformaciju iz takvih smjesa uz zamjenu koraka nakapavanja tehnikom brzog razmazivanja. Optimizacija ovog procesa poboljšala je veličinu i prinos DUV-ova, ali i dalje je preostalo pitanje izlučivanja kolesterola. Potaknuti time, razvili smo novi protokol koji kombinira tehnike brze izmjene otapala, čišćenja plazmom i brzog razmazivanja za proizvodnju DUV-ova elektroformacijom iz vlažnih lipidnih filmova. Smatramo da će nam ovaj novi protokol pomoći u daljnjem istraživanju svojstava membrana s vrlo visokim udjelom kolesterola, poput vlaknastih stanica leće oka.
Keywords
Keywords (croatian)
Languageenglish
URN:NBNurn:nbn:hr:166:553755
Study programmeTitle: Postgraduate University Study of Biophysics
Study programme type: university
Study level: postgraduate
Academic / professional title: doktor/doktorica znanosti, područje prirodnih znanosti, polje fizika (doktor/doktorica znanosti, područje prirodnih znanosti, polje fizika)
Type of resourceText
Extent84 str. ; 30 cm
File originBorn digital
Access conditionsOpen access
Terms of useLicence Attribution-NoDerivatives 4.0 International (CC BY-ND 4.0) icon
Created on2023-05-04 12:04:35