M. A. Kiselev, P. Lesieur, A. M. Kisselev, D. Lombardo, V. L. Aksenov
New model of separated form factors is proposed for the evaluation of small-angle neutron scattering curves from large unilamellar vesicles. The validity of the model was checked by comparison to the model of hollow sphere. The model of separated form factors and hollow sphere model give reasonable agreement in the evaluation of vesicle parameters.
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A widely used method to measure the bending rigidity of bilayer membranes is fluctuation spectroscopy, which analyses the thermally-driven membrane undulations of giant unilamellar vesicles recorded with either phase-contrast or confocal microscopy. Here, we analyze the fluctuations of the same vesicle using both techniques and obtain consistent values for the bending modulus. We discuss the factors that may lead to discrepancies.
The parameters of the polydispersed unilamellar vesicle population are analyzed on the basis of the separated form-factors model for three different type of membranes. The neutron scattering length density across the membrane is simulated on the base of the HH-model with the linear water distribution.
We investigated the thermodynamic relaxation of giant unilamellar vesicles (GUVs) which contained small vesicles within their interior. Quenching these vesicles from their fluid phase (T>Tm) through the phase transition in the gel state (T<Tm) drives the inner vesicles to be expelled from the larger mother vesicle via the accompanying decrease in vesicle area by ~25% which forces a pore to open in the mother vesicle. We demonstrate that the proceeding time evolution of the resulting efflux follows the relaxation of the membrane area and describe the entire relaxation process using an Onsager-l…
On the basis of the separated form-factors model, parameters of the polydispersed unilamellar DMPC vesicle population are analyzed. The neutron scattering length density across the membrane is simulated on the basis of fluctuated model of lipid bilayer. The hydration of vesicle is described by sigmoid distribution function of the water molecules. The results of fitting of the experimental data obtained at the small angle spectrometer SANS-I, PSI (Switzerland) are: average vesicle radius 272 angstrom, polydispersity of the radius 27%, membrane thickness 50.6 angstrom, thickness of hydrocarbon c…
On the basis of the separated form-factor model, a code for fitting the small-angle neutron scattering spectra of the polydispersed vesicle population has been developed. Vesicle and membrane bilayer parameters are analyzed for various hierarchical models of the neutron scattering length density across the membrane. It is shown that hydration of vesicle can be described by a linear distribution function of water molecules. For the first time, the average radius and polydispersity of the vesicle population, thickness of the membrane bilayer, thickness of hydrophobic and hydrophilic parts of bil…
Wrapping pathways of anisotropic dumbbell particles by giant unilamellar vesicles
该研究结合GUV实验、理论与分子动力学模拟，揭示膜张力及哑铃颈部曲率变化如何决定各向异性颗粒的包裹路径、速度与最终状态。
High-Throughput Mechanical Characterization of Giant Unilamellar Vesicles by Real-Time Deformability Cytometry
本文将实时可变形细胞术（RT-DC）应用于巨型单层囊泡（GUVs），提出模拟模型与两种互补拟合策略提取面积膨胀模量，实现高通量无接触力学表征，通量较传统技术高数量级，还支持群体分选。
In a previous work, we have shown that a spatially localized transmembrane pH gradient, produced by acid micro-injection near the external side of cardiolipin-containing giant unilamellar vesicles, leads to the formation of tubules that retract after the dissipation of this gradient. These tubules have morphologies similar to mitochondrial cristae. The tubulation effect is due to direct phospholipid packing modification in the outer leaflet that is promoted by protonation of cardiolipin headgroups. Here we compare the case of cardiolipin-containing giant unilamellar vesicles with that of phosp…
The small-angle X-ray and neutron scattering, time resolved X-ray small-angle and wide-angle diffraction coupled with differential scanning calorimetry have been applied to the investigation of unilamellar and multilamellar dimyristoylphosphatidylcholine (DMPC) vesicles in sucrose buffers with sucrose concentrations from 0 to 60%. Sucrose buffer decreased vesicle size and polydispersity and increased an X-ray contrast between phospholipid membrane and bulk solvent sufficiently. No influence of sucrose on the membrane thickness or mutual packing of hydrocarbon chains has been detected. The regi…
Small angle neutron scattering (SANS) on the unilamellar vesicle populations (diameter 500 and 1000 angstrom) was used to characterize lipid vesicles from dimyristoylphosphatidylcholine (DMPC) at three phases (gel, ripple, and liquid). Parameters of vesicle populations and internal structure of the DMPC bilayer were characterized on the basis of the Separated Form Factor (SFF) model. Parameters of the internal bilayer structure (thickness of the membrane and the hydrophobic core, hydration, and surface area of lipid molecule) were determined on the basis of the Hydrophobic-Hydrophilic (HH) app…
A method for determining permeability of phospholipid bilayer based on the osmotic swelling of micrometer-sized giant unilamellar vesicles (GUVs) is presented as an alternative to the two established techniques, dynamic light scattering on liposome suspension, and electrical measurements on planar lipid bilayers. In the described technique, an individual GUV is transferred using a micropipette from a sucrose/glucose solution into an isomolar solution containing the solute under investigation. Throughout the experiment, vesicle cross-section is monitored and recorded using a digital camera moun…
We use computer simulations and a simple free energy model to study the response of a bilayer membrane to the application of a negative (compressive) mechanical tension. Such a tension destabilizes the long wavelength undulation modes of giant vesicles, but it can be sustained when small membranes and vesicles are considered. Our negative tension simulation results reveal two regimes - (i) a weak negative tension regime characterized by stretching-dominated elasticity, and (ii) a strong negative tension regime featuring bending-dominated elastic behavior. This resembles the findings of the cla…
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