Formation of giant unilamellar vesicles from spin-coated lipid films by localized IR heating

Billerit, Céline; Jeffries, Gavin D. M.; Orwar, Owe; Jesorka, Aldo

doi:10.1039/c2sm26394g

Cited by 25 publications

(17 citation statements)

References 27 publications

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“…1b). The formed compartments are strictly unilamellar 3 which is in contrast to the vesicles reported by Billerit et al 10,11 where a lamellarity distribution typical for swelling of stacked bilayers was observed. The majority of techniques for artificial vesicle generation feature this distribution 12,13 .…”

Section: Enhanced Protocell Formation and Growthcontrasting

confidence: 61%

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Rapid growth and fusion of protocells in surface-adhered membrane networks

Köksal

Liese

Xue

et al. 2020

Preprint

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Elevated temperatures might have promoted the nucleation, growth and replication of protocells on the early Earth. Recent reports have shown evidence that moderately high temperatures not only permit protocell assembly at the origin of life, but could have actively supported it. Here we show the fast nucleation and growth of vesicular compartments from autonomously formed lipid networks on solid surfaces, induced by a moderate increase in temperature. Branches of the networks, initially consisting of self-assembled interconnected nanotubes, rapidly swell into microcompartments which can spontaneously encapsulate RNA fragments. The increase in temperature further causes fusion of adjacent networkconnected compartments, resulting in the redistribution of the RNA. The experimental observations and the mathematical model indicate that the presence of nanotubular interconnections between protocells facilitates the fusion process.The important role of solid surface support for the autonomous formation of primitive protocells has been suggested earlier in the context of the origin of life 1,2 . Hanczyc, Szostak et al. showed that vesicle formation from fatty acids was significantly enhanced in the presence of solid particle surfaces consisting of natural minerals or synthetic materials 1,2 . Particularly the silicate-based minerals accelerated the vesicle generation.In a recent report, we showed the autonomous formation and growth of surface adhered protocell populations as a result of a sequence of topological transformations on a solid substrate 3 . Briefly, upon contact with a mineral-like solid substrate, a lipid reservoir spreads as a double bilayer membrane. The distal membrane (upper -with respect to the surface-) ruptures and forms a carpet of lipid nanotubes. Over the course of a few hours, fragments of these nanotubes swell into giant, strictly unilamellar vesicular compartments. This relatively slow process is entirely self-driven and only requires a lipid reservoir as source, a solid surface, and surrounding aqueous media. The resulting structure consists of thousands of lipid compartments, which are physically connected to each other via a

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Section: Enhanced Protocell Formation and Growthcontrasting

confidence: 61%

“…S1). The formation of giant unilamellar vesicles from disordered membrane layers induced by localized heating (>25 °C) has been reported before 10,11 . In our experiments, the thermal gradient leads to the instant formation and growth of vesicular compartments exclusively from the lipid nanotubes ( Fig.…”

Section: Enhanced Protocell Formation and Growthsupporting

confidence: 53%

Rapid growth and fusion of protocells in surface-adhered membrane networks

Köksal

Liese

Xue

et al. 2020

Preprint

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“…Electroformation, polymer assisted swelling and microfluidic approaches are the most common methods used to form GUVs for studying basic properties of cellular membranes. Several variations for each method exist together with other approaches such as rapid evaporation (Moscho et al, 1996 ), local heating (Billerit et al, 2012 ), reverse emulsion (Pautot et al, 2003a , b ) or cDICE (continuous droplet interface crossing encapsulation) (Abkarian et al, 2011 ; Blosser et al, 2016 ). Each method has different advantages and disadvantages, which can influence the experiments in various ways.…”

Section: Discussionmentioning

confidence: 99%

Production of Isolated Giant Unilamellar Vesicles under High Salt Concentrations

et al. 2017

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The cell membrane forms a dynamic and complex barrier between the living cell and its environment. However, its in vivo studies are difficult because it consists of a high variety of lipids and proteins and is continuously reorganized by the cell. Therefore, membrane model systems with precisely controlled composition are used to investigate fundamental interactions of membrane components under well-defined conditions. Giant unilamellar vesicles (GUVs) offer a powerful model system for the cell membrane, but many previous studies have been performed in unphysiologically low ionic strength solutions which might lead to altered membrane properties, protein stability and lipid-protein interaction. In the present work, we give an overview of the existing methods for GUV production and present our efforts on forming single, free floating vesicles up to several tens of μm in diameter and at high yield in various buffer solutions with physiological ionic strength and pH.

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“…There are several methods for lipid membrane film preparations, such as casting [ 13 ], the spin coating [ 10 , 14 ], Langmuir-Blodgett membrane formation [ 15 – 17 ], vesicle rupture [ 18 – 20 ], and water-in-oil droplet template methods [ 21 ]. The casting and spin coating methods are conventionally adopted in requesting membrane films composed of any kinds of lipids or mixtures.…”

Section: Discussionmentioning

confidence: 99%

Autonomous buckling of micrometer-sized lipid-protein membrane patches constructed by Dictyostelium discoideum

Takahashi

Toyota

2015

J Biol Eng

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BackgroundThe cytosol of amoeba cells controls the membrane deformation during their motion in vivo. To investigate such ability of the cytosol of amoeba cell, Dictyostelium discoideum (Dictyostelium), in vitro, we used lipids extracted from Dictyostelium and commercially available phospholipids, and prepared substrate-supported lipid membrane patches on the micrometer scale by spin coating.ResultsWe found that the spin coater holder, which has pores (pore size = 3.1 mm) of negative pressure to hold the cover glass induced the concave surface of the cover glass. The membrane lipid patches were formed at each position in the vicinity of the holder pores and their sizes were in the range of 2.7 to 3.2 × 104 μm2. After addition of the cytosol extracted from Dictyostelium to the lipid membrane patches, through time-lapse observation with a confocal laser scanning fluorescence microscope, we observed an autonomous buckling of the Dictyostelium lipid patches and localized behaviours of proteins found within.ConclusionThe current method serves as the novel technique for the preparation of film patches in which the positions of patches are controlled by the holder pores without fabricating, modifying, and arranging the chemical properties of the solution components of lipids. The findings imply that lipid-binding proteins in the cytosol were adsorbed and accumulated within the Dictyostelium lipid patches, inducing the transformation of the cell-sized patch.Electronic supplementary materialThe online version of this article (doi:10.1186/1754-1611-9-3) contains supplementary material, which is available to authorized users.

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Formation of giant unilamellar vesicles from spin-coated lipid films by localized IR heating

Cited by 25 publications

References 27 publications

Rapid growth and fusion of protocells in surface-adhered membrane networks

Rapid growth and fusion of protocells in surface-adhered membrane networks

Production of Isolated Giant Unilamellar Vesicles under High Salt Concentrations

Autonomous buckling of micrometer-sized lipid-protein membrane patches constructed by Dictyostelium discoideum

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