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Quantitative analysis of the lamellarity of giant liposomes prepared by the inverted emulsion method.
Chiba M
,
Miyazaki M
,
Ishiwata S
.
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The inverted emulsion method is used to prepare giant liposomes by pushing water-in-oil droplets through the oil/water interface into an aqueous medium. Due to the high encapsulation efficiency of proteins under physiological conditions and the simplicity of the protocol, it has been widely used to prepare various cell models. However, the lamellarity of liposomes prepared by this method has not been evaluated quantitatively. Here, we prepared liposomes that were partially stained with a fluorescent dye, and analyzed their fluorescence intensity under an epifluorescence microscope. The fluorescence intensities of the membranes of individual liposomes were plotted against their diameter. The plots showed discrete distributions, which were classified into several groups. The group with the lowest fluorescence intensity was determined to be unilamellar by monitoring the exchangeability of the inner and the outer solutions of the liposomes in the presence of the pore-forming toxin α-hemolysin. Increasing the lipid concentration dissolved in oil increased the number of liposomes ∼100 times. However, almost all the liposomes were unilamellar even at saturating lipid concentrations. We also investigated the effects of lipid composition and liposome content, such as highly concentrated actin filaments and Xenopus egg extracts, on the lamellarity of the liposomes. Remarkably, over 90% of the liposomes were unilamellar under all conditions examined. We conclude that the inverted emulsion method can be used to efficiently prepare giant unilamellar liposomes and is useful for designing cell models.
Akashi,
Preparation of giant liposomes in physiological conditions and their characterization under an optical microscope.
1996, Pubmed
Akashi,
Preparation of giant liposomes in physiological conditions and their characterization under an optical microscope.
1996,
Pubmed
Campillo,
Unexpected membrane dynamics unveiled by membrane nanotube extrusion.
2013,
Pubmed
Carvalho,
Cell-sized liposomes reveal how actomyosin cortical tension drives shape change.
2013,
Pubmed
Desai,
The use of Xenopus egg extracts to study mitotic spindle assembly and function in vitro.
1999,
Pubmed
,
Xenbase
Emoto,
Redistribution of phosphatidylethanolamine at the cleavage furrow of dividing cells during cytokinesis.
1996,
Pubmed
Funakoshi,
Formation of giant lipid vesiclelike compartments from a planar lipid membrane by a pulsed jet flow.
2007,
Pubmed
Hamada,
Construction of asymmetric cell-sized lipid vesicles from lipid-coated water-in-oil microdroplets.
2008,
Pubmed
Honda,
Morphogenesis of liposomes encapsulating actin depends on the type of actin-crosslinking.
1999,
Pubmed
Hotani,
Dynamic features of microtubules as visualized by dark-field microscopy.
1990,
Pubmed
Hu,
Microfluidic fabrication of asymmetric giant lipid vesicles.
2011,
Pubmed
Ito,
Dynamical formation of lipid bilayer vesicles from lipid-coated droplets across a planar monolayer at an oil/water interface.
2013,
Pubmed
Jimenez,
Towards high throughput production of artificial egg oocytes using microfluidics.
2011,
Pubmed
,
Xenbase
Limozin,
On the organization of self-assembled actin networks in giant vesicles.
2003,
Pubmed
Limozin,
Polymorphism of cross-linked actin networks in giant vesicles.
2002,
Pubmed
Liu,
Membrane-induced bundling of actin filaments.
2008,
Pubmed
Liu,
Biology under construction: in vitro reconstitution of cellular function.
2009,
Pubmed
Macdonald,
Calcium binding to mixed phosphatidylglycerol-phosphatidylcholine bilayers as studied by deuterium nuclear magnetic resonance.
1987,
Pubmed
Maeda,
Assembly of MreB filaments on liposome membranes: a synthetic biology approach.
2012,
Pubmed
Matosevic,
Stepwise synthesis of giant unilamellar vesicles on a microfluidic assembly line.
2011,
Pubmed
McPhee,
Measuring the lamellarity of giant lipid vesicles with differential interference contrast microscopy.
2013,
Pubmed
Miyata,
Protrusive growth from giant liposomes driven by actin polymerization.
1999,
Pubmed
Miyata,
Morphological changes in liposomes caused by polymerization of encapsulated actin and spontaneous formation of actin bundles.
1992,
Pubmed
Montes,
Giant unilamellar vesicles electroformed from native membranes and organic lipid mixtures under physiological conditions.
2007,
Pubmed
Murrell,
Spreading dynamics of biomimetic actin cortices.
2011,
Pubmed
Noireaux,
A vesicle bioreactor as a step toward an artificial cell assembly.
2004,
Pubmed
Ohvo-Rekilä,
Cholesterol interactions with phospholipids in membranes.
2002,
Pubmed
Osawa,
Liposome division by a simple bacterial division machinery.
2013,
Pubmed
Ota,
Microfluidic formation of monodisperse, cell-sized, and unilamellar vesicles.
2009,
Pubmed
Pautot,
Engineering asymmetric vesicles.
2003,
Pubmed
Pinot,
Effects of confinement on the self-organization of microtubules and motors.
2009,
Pubmed
Pontani,
Reconstitution of an actin cortex inside a liposome.
2009,
Pubmed
Pott,
Giant unilamellar vesicle formation under physiologically relevant conditions.
2008,
Pubmed
Richmond,
Forming giant vesicles with controlled membrane composition, asymmetry, and contents.
2011,
Pubmed
Robertson,
Membrane structure.
1981,
Pubmed
Song,
Structure of staphylococcal alpha-hemolysin, a heptameric transmembrane pore.
1996,
Pubmed
Stachowiak,
Inkjet formation of unilamellar lipid vesicles for cell-like encapsulation.
2009,
Pubmed
Stachowiak,
Unilamellar vesicle formation and encapsulation by microfluidic jetting.
2008,
Pubmed
Stachowiak,
Membrane bending by protein-protein crowding.
2012,
Pubmed
Suzuki,
Preparation of bead-tailed actin filaments: estimation of the torque produced by the sliding force in an in vitro motility assay.
1996,
Pubmed
Takiguchi,
Entrapping desired amounts of actin filaments and molecular motor proteins in giant liposomes.
2008,
Pubmed
Takiguchi,
Transformation of actoHMM assembly confined in cell-sized liposome.
2011,
Pubmed
Tanaka-Takiguchi,
Septin-mediated uniform bracing of phospholipid membranes.
2009,
Pubmed
Terasawa,
Coupling of the fusion and budding of giant phospholipid vesicles containing macromolecules.
2012,
Pubmed
Tsai,
Encapsulation of active cytoskeletal protein networks in cell-sized liposomes.
2011,
Pubmed
van Meer,
Membrane lipids: where they are and how they behave.
2008,
Pubmed
van Swaay,
Microfluidic methods for forming liposomes.
2013,
Pubmed
Weinberger,
Gel-assisted formation of giant unilamellar vesicles.
2013,
Pubmed
Yamada,
Spontaneous transfer of phospholipid-coated oil-in-oil and water-in-oil micro-droplets through an oil/water interface.
2006,
Pubmed
Yamashita,
A new method for the preparation of giant liposomes in high salt concentrations and growth of protein microcrystals in them.
2002,
Pubmed
Yanagisawa,
Oriented reconstitution of a membrane protein in a giant unilamellar vesicle: experimental verification with the potassium channel KcsA.
2011,
Pubmed
