scholarly journals Studying the effects of asymmetry on the bending rigidity of lipid membranes formed by microfluidics

2016 ◽  
Vol 52 (30) ◽  
pp. 5277-5280 ◽  
Author(s):  
K. Karamdad ◽  
R. V. Law ◽  
J. M. Seddon ◽  
N. J. Brooks ◽  
O. Ces
Keyword(s):  
Mechanical Properties ◽  
High Throughput ◽  
Lipid Membranes ◽  
Bending Rigidity ◽  
Giant Unilamellar Vesicles ◽  
Unilamellar Vesicles ◽  
Microfluidic Platform

In this article we detail a robust high-throughput microfluidic platform capable of fabricating either symmetric or asymmetric giant unilamellar vesicles (GUVs) and characterise the mechanical properties of their membranes.

scholarly journals Surfactant-free production of biomimetic giant unilamellar vesicles using PDMS-based microfluidics

2021 ◽  
Vol 4 (1) ◽  
Author(s):  
Naresh Yandrapalli ◽  
Julien Petit ◽  
Oliver Bäumchen ◽  
Tom Robinson
Keyword(s):  
High Throughput ◽  
Paradigm Shift ◽  
Mammalian Cells ◽  
Lipid Vesicles ◽  
Giant Unilamellar Vesicles ◽  
Unilamellar Vesicles ◽  
Artificial Cells ◽  
Membrane Function ◽  
Lipid Diffusion ◽  
Protein Incorporation

AbstractMicrofluidic production of giant lipid vesicles presents a paradigm-shift in the development of artificial cells. While production is high-throughput and the lipid vesicles are mono-disperse compared to bulk methods, current technologies rely heavily on the addition of additives such as surfactants, glycerol and even ethanol. Here we present a microfluidic method for producing biomimetic surfactant-free and additive-free giant unilamellar vesicles. The versatile design allows for the production of vesicle sizes ranging anywhere from ~10 to 130 µm with either neutral or charged lipids, and in physiological buffer conditions. Purity, functionality, and stability of the membranes are validated by lipid diffusion, protein incorporation, and leakage assays. Usability as artificial cells is demonstrated by increasing their complexity, i.e., by encapsulating plasmids, smaller liposomes, mammalian cells, and microspheres. This robust method capable of creating truly biomimetic artificial cells in high-throughput will prove valuable for bottom-up synthetic biology and the understanding of membrane function.

Magainin-H2 effects on the permeabilization and mechanical properties of giant unilamellar vesicles

2019 ◽  
Vol 553 ◽  
pp. 247-258 ◽  
Author(s):  
Andrea Mescola ◽  
Nathaly Marín-Medina ◽  
Gregorio Ragazzini ◽  
Maurizio Accolla ◽  
Andrea Alessandrini
Keyword(s):  
Mechanical Properties ◽  
Giant Unilamellar Vesicles ◽  
Unilamellar Vesicles

Dynamic behaviour of giant unilamellar vesicles induced by the uptake of [70]fullerene

2014 ◽  
Vol 50 (11) ◽  
pp. 1288-1291 ◽  
Author(s):  
Atsushi Ikeda ◽  
Tomohiro Hida ◽  
Tatsuya Iizuka ◽  
Manami Tsukamoto ◽  
Jun-ichi Kikuchi ◽  
...  
Keyword(s):  
Exchange Reaction ◽  
Lipid Membranes ◽  
Phase Contrast ◽  
Dynamic Behaviour ◽  
Phase Contrast Microscopy ◽  
Giant Unilamellar Vesicles ◽  
Unilamellar Vesicles ◽  
Contrast Microscopy

We have directly observed the processes involved in a C70-exchange reaction from γ-cyclodextrin cavities to lipid membranes using GUVs by phase contrast microscopy.

scholarly journals Mechanical properties of plasma membrane vesicles correlate with lipid order and viscosity and depend on cell density

2019 ◽  
Author(s):  
Jan Steinkühler ◽  
Erdinc Sezgin ◽  
Iztok Urbančič ◽  
Christian Eggeling ◽  
Rumiana Dimova
Keyword(s):  
Mechanical Properties ◽  
Plasma Membrane ◽  
Lipid Membranes ◽  
Plasma Membranes ◽  
Fluorescence Emission ◽  
Membrane Vesicles ◽  
Seeding Density ◽  
Bending Rigidity ◽  
Plasma Membrane Vesicles ◽  
Lipid Order

AbstractPlasma membranes dynamically respond to external cues and changing environment. Quantitative measurements of these adaptations can elucidate the mechanism that cells exploit to survive, adapt and function. However, cell-based assays are affected by active processes while measurements on synthetic models suffer from compositional limitations. Here, as a model system we employ giant plasma membrane vesicles (GPMVs), which largely preserve the plasma membrane lipidome and proteome. From analysis of fluorescence emission and lifetime of environment-sensitive dyes, and membrane shape fluctuations, we investigate how plasma membrane order, viscosity and bending rigidity are affected by different stimuli such as cell seeding density in three different cell models. Our studies reveal that bending rigidity of plasma membranes vary with lipid order and microviscosity in a highly correlated fashion. Thus, readouts from polarity- and viscosity-sensitive probes represent a promising indicator of membrane mechanical properties. Quantitative analysis of the data allows for comparison to synthetic lipid membranes as plasma membrane mimetics.

scholarly journals Giant unilamellar vesicles - a perfect tool to visualize phase separation and lipid rafts in model systems.

2009 ◽  
Vol 56 (1) ◽  
Author(s):  
Olga Wesołowska ◽  
Krystyna Michalak ◽  
Jadwiga Maniewska ◽  
Andrzej B Hendrich
Keyword(s):  
Phase Separation ◽  
Lipid Membranes ◽  
Average Diameter ◽  
Model Systems ◽  
Membrane Properties ◽  
Lipid Phase ◽  
Giant Unilamellar Vesicles ◽  
Unilamellar Vesicles ◽  
Simple Method ◽  
Wide Range

Model systems such as black lipid membranes or conventional uni- or multilamellar liposomes are commonly used to study membrane properties and structure. However, the construction and dimensions of these models excluded their direct optical microscopic observation. Since the introduction of the simple method of liposome electroformation in alternating electric field giant unilamellar vesicles (GUVs) have become an important model imitating biological membranes. Due to the average diameter of GUVs reaching up to 100 microm, they can be easily observed under a fluorescent or confocal microscope provided that the appropriate fluorescent probe was incorporated into the lipid phase during vesicle formation. GUVs can be formed from different lipid mixtures and they are stable in a wide range of physical conditions such as pH, pressure or temperature. This mini-review presents information about the methods of GUV production and their usage. Particularly, the use of GUVs in studying lipid phase separation and the appearance and behavior of lipid domains (rafts) in membranes is discussed but also other examples of GUVs use in membrane research are given. The experience of the authors in setting up the GUV-forming equipment and production of GUVs is also presented.

scholarly journals Preparation and mechanical characterisation of giant unilamellar vesicles by a microfluidic method

Lab on a Chip ◽  
2015 ◽  
Vol 15 (2) ◽  
pp. 557-562 ◽  
Author(s):  
K. Karamdad ◽  
R. V. Law ◽  
J. M. Seddon ◽  
N. J. Brooks ◽  
O. Ces
Keyword(s):  
Mechanical Properties ◽  
Fluctuation Analysis ◽  
Giant Unilamellar Vesicles ◽  
Unilamellar Vesicles ◽  
Mechanical Characterisation ◽  
Novel Method

We present a novel method for the generation of giant unilamellar vesicles and the use of fluctuation analysis to characterise their mechanical properties.

scholarly journals Charge-controlled microfluidic formation of lipid-based single- and multicompartment systems

Lab on a Chip ◽  
2018 ◽  
Vol 18 (17) ◽  
pp. 2665-2674 ◽  
Author(s):  
Barbara Haller ◽  
Kerstin Göpfrich ◽  
Martin Schröter ◽  
Jan-Willi Janiesch ◽  
Ilia Platzman ◽  
...  
Keyword(s):  
High Throughput ◽  
Cell Model ◽  
Model Systems ◽  
Giant Unilamellar Vesicles ◽  
Unilamellar Vesicles ◽  
On Demand ◽  
Demand Creation ◽  
Synthetic Cell ◽  
Multicompartment Systems

We introduce a high-throughput microfluidic off-the-shelf approach for the on-demand creation of giant unilamellar vesicles (GUVs) or multicompartment synthetic cell model systems.

scholarly journals Interaction of a Polyarginine Peptide with Membranes of Different Mechanical Properties

Biomolecules ◽  
2019 ◽  
Vol 9 (10) ◽  
pp. 625 ◽  
Author(s):  
Matías A. Crosio ◽  
Matías A. Via ◽  
Candelaria I. Cámara ◽  
Agustin Mangiarotti ◽  
Mario G. Del Pópolo ◽  
...  
Keyword(s):  
Mechanical Properties ◽  
Lipid Membranes ◽  
Langmuir Monolayers ◽  
Phase Segregation ◽  
Cell Penetrating Peptides ◽  
Unilamellar Vesicles ◽  
Black Lipid Membranes ◽  
Large Unilamellar Vesicles ◽  
Host Membrane ◽  
Subsequent Effect

The membrane translocation efficiency of cell penetrating peptides (CPPs) has been largely studied, and poly-arginines have been highlighted as particularly active CPPs, especially upon negatively charged membranes. Here we inquire about the influence of membrane mechanical properties in poly-arginine adsorption, penetration and translocation, as well as the subsequent effect on the host membrane. For this, we selected anionic membranes exhibiting different rigidity and fluidity, and exposed them to the nona-arginine KR9C. Three different membrane compositions were investigated, all of them having 50% of the anionic lipid 1,2-dioleoyl-sn-glycero-3-phospho-(1’-rac-glycerol) (DOPG), thus, ensuring a high affinity of the peptide for membrane surfaces. The remaining 50% was a saturated PC (1,2-dipalmitoyl-sn-glycero-3-phosphocholine, DPPC), an unsaturated PC (1,2-dioleoyl-sn-glycero-3-phosphocholine, DOPC) or a mixture of DOPC with cholesterol. Peptide-membrane interactions were studied using four complementary models for membranes: Langmuir monolayers, Large Unilamellar Vesicles, Black Lipid Membranes and Giant Unilamellar Vesicles. The patterns of interaction of KR9C varied within the different membrane compositions. The peptide strongly adsorbed on membranes with cholesterol, but did not incorporate or translocate them. KR9C stabilized phase segregation in DPPC/DOPG films and promoted vesicle rupture. DOPC/DOPG appeared like the better host for peptide translocation: KR9C adsorbed, inserted and translocated these membranes without breaking them, despite softening was observed.

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