Naslov
Optimizacija elektroformacije divovskih unilamelarnih vezikula za različite lipidne smjese uz naglasak na visoke koncentracije kolesterola
(Optimization of giant unilamellar vesicle electroformation for different lipid mixtures with a focus on high cholesterol concentrations)

Autori
Zvonimir Boban

Vrsta, podvrsta i kategorija rada
Ocjenski radovi, doktorska disertacija

Mjesto
Split

Datum
27.04

Godina
2023

Stranica
83

Mentor
Raguž Marija

Ključne riječi
DUV, elektroformacija, kolesterol, fosfolipid, debljina filma, brza izmjena otapala, tehnika brzog razmazivanja, čišćenje plazmom, vlažni lipidni film, fluorescencijska mikroskopija
(GUV, electroformation, cholesterol, phospholipid, film thickness, rapid solvent exchange, spin-coating, plasma cleaning, damp lipid film, fluorescence microscopy)

Sažetak
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.

Izvorni jezik
Engleski

Znanstvena područja
Fizika, Interdisciplinarne prirodne znanosti, Temeljne medicinske znanosti

POVEZANOST RADA