Pregled bibliografske jedinice broj: 1266021
Electroformation of Giant Unilamellar Vesicles from Damp Lipid Films Formed by Vesicle Fusion
Electroformation of Giant Unilamellar Vesicles from Damp Lipid Films Formed by Vesicle Fusion // Membranes, 13 (2023), 3; 352, 12 doi:10.3390/membranes13030352 (međunarodna recenzija, članak, znanstveni)
CROSBI ID: 1266021 Za ispravke kontaktirajte CROSBI podršku putem web obrasca
Naslov
Electroformation of Giant Unilamellar
Vesicles from Damp Lipid Films Formed by
Vesicle Fusion
Autori
Boban, Zvonimir ; Mardešić, Ivan ; Jozić, Sanja Perinović ; Šumanovac, Josipa ; Subczynski, Witold Karol ; Raguž, Marija
Izvornik
Membranes (2077-0375) 13
(2023), 3;
352, 12
Vrsta, podvrsta i kategorija rada
Radovi u časopisima, članak, znanstveni
Ključne riječi
GUV ; electroformation ; cholesterol ; damp lipid film ; rapid solvent exchange ; plasma cleaning ; cholesterol crystals ; vesicle fusion
Sažetak
Giant unilamellar vesicles (GUVs) are artificial membrane models which are of special interest to researchers because of their similarity in size to eukaryotic cells. The most commonly used method for GUVs production is electroformation. However, the traditional electroformation protocol involves a step in which the organic solvent is completely evaporated, leaving behind a dry lipid film. This leads to artifactual demixing of cholesterol (Chol) in the form of anhydrous crystals. These crystals do not participate in the formation of the lipid bilayer, resulting in a decrease of Chol concentration in the bilayer compared to the initial lipid solution. We propose a novel electroformation protocol which addresses this issue by combining the rapid solvent exchange, plasma cleaning and spin-coating techniques to produce GUVs from damp lipid films in a fast and reproducible manner. We have tested the protocol efficiency using 1/1 phosphatidylcholine/Chol and 1/1/1 phosphatidylcholine/sphingomyelin/Chol lipid mixtures and managed to produce a GUV population of an average diameter around 40 µm, with many GUVs being larger than 100 µm. Additionally, compared to protocols that include the dry film step, the sizes and quality of vesicles determined from fluorescence microscopy images were similar or better, confirming the benefits of our protocol in that regard as well.
Izvorni jezik
Engleski
POVEZANOST RADA
Projekti:
IP-2019-04-1958 - Otkrivanje organizacije i funkcije kolesterolove domene u plasma membrani fibroznih stanica leće oka uz primjenu fluorescentnih metoda (BioPhyCBD) (Raguž, Marija, HRZZ - 2019-04) ( CroRIS)
Ustanove:
Kemijsko-tehnološki fakultet, Split,
Prirodoslovno-matematički fakultet, Split,
Medicinski fakultet, Split
Citiraj ovu publikaciju:
Časopis indeksira:
- Current Contents Connect (CCC)
- Web of Science Core Collection (WoSCC)
- Science Citation Index Expanded (SCI-EXP)
- SCI-EXP, SSCI i/ili A&HCI
- Scopus