Permeability of phospholipid membrane for small polar molecules determined from osmotic swelling of giant phospholipid vesicles

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 mounted on a phase-contrast microscope. Using a least-squares procedure for circle fitting, vesicle radius R is computed from the recorded images of vesicle cross-section. Two methods for determining membrane permeability from the obtained R(t) dependence are described: the first one uses the slope of R(t) for a spherical GUV, and the second one the R(t) dependence around the transition point at which a flaccid vesicle transforms into a spherical one. We demonstrate that both methods give consistent estimates for membrane permeability.Comment: 40 pages, 8 figures, to appear in Advances in Planar Lipid Membranes and Liposomes vol. 1

Osmotically induced optical changes in unilamellar vesicles: Experiment and theory

Previous light scattering studies have shown increases in scattering from vesicles prepared in isotonically increasing concentrations of NaCl. It is believed that changes in NaCl concentration induce physical changes in the membrane, seen either as changes to membrane refractive index or thickness. However, the specific nature of these changes is not known. The purpose of the research described in this thesis was the use of neutron scattering to obtain information complementary to previous light scattering studies. Small angle and contrast variation neutron scattering were used to obtain measurements of membrane thickness and scattering length density as a function of isotonic NaCl concentration. Results showed changes in scattering as a function of isotonic NaCl concentration. Results from modelling and analysis have suggested that the most significant change in scattering is due to changes occurring in the headgroup region of the bilayer and that these changes are ionic in nature

Scattering from laterally heterogeneous vesicles. II. The form factor

Despite growing interest in the formation of domains or `rafts' in cell and model membranes, there have been relatively few attempts to characterize such systems via scattering techniques. Previously [Pencer et al. (2006). J. Appl. Cryst. 39, 293-303], it was demonstrated that the Porod invariant, Q, could be used to detect lateral segregation. Here, the general theory for scattering from laterally heterogeneous vesicles is outlined and form factors are derived for vesicles containing either single circular or annular domains. These form factors are then applied to the analysis of neutron scattering data from heterogeneous vesicles. Potential advantages and limitations of this technique are also discussed.NRC publication: Ye

A Sucrose Solution Application to the Study of Model Biological Membranes

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 region of sucrose concentrations 30%-40% created the best experimental conditions for X-ray small-angle experiments with phospholipid vesicles.Comment: PDF: 10 pages, 6 figures. MS Word sours

Adolescent substance use in first episode psychosis: a test of three models

Bibliography: p. 89-11