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PubMed2001

Phase separation in AlxGa1-xAs nanowhiskers grown by the solution-liquid-solid mechanism.

Low-temperature growth of AlxGa1-xAs by the solution-liquid-solid mechanism affords nanowhiskers having two specific overall compositions: x = 0.1 and 0.8. Intermediate compositions (0.1 < x < 0.8) are inaccessible by the method employed. Examination of the nanowhiskers by transmission electron microscopy, photoluminescence spectroscopy, electron energy loss spectroscopy, and annealing studies indicates that they possess compositionally modulated nanostructures. The observed compositional modulation consists of alternating comparatively Al-rich and Ga-rich AlxGa1-xAs domains with dimensions of 3-20 nm. The spontaneous phase separation, which occurs during growth, appears to be kinetically driven, and the resulting phase-separated nanostructures appear to be, therefore, metastable rather than stable. Spontaneous phase separation in AlxGa1-xAs, which strongly influences its photoemission behavior, is not well understood theoretically.

Journal Article↗

PubMed2005

Phase separation patterns for diblock copolymers on spherical surfaces: a finite volume method.

We explore phase separation on spherical surfaces by solving the Cahn-Hilliard equation modified for diblock copolymers using a finite volume method. The spherical surface is discretized into almost uniform triangles by employing successive dyadic refinements of the spherical icosahedron, a methodology that avoids potential mathematical and numerical problems related to the poles in spherical coordinates. The finite volume method is based on averaging Voronoi cells built from triangular meshes to calculate the Laplace-Beltrami operator on the curved surface, which greatly improves both the accuracy and speed of calculation as compared to the conventional finite difference method. By using this method we simulate the phase separation of diblock copolymers on a spherical surface. It is found that stable and intrinsic defects, which would not occur in a flat space after sufficient annealing, appear in the periodic arrangement of the domains on the curved surface due to the distinct Euler characteristic of the surface.

Journal Article↗

PubMed1999

Conformational stability of lyophilized PEGylated proteins in a phase-separating system.

PEGylation of proteins is of great interest to the pharmaceutical industry as covalent attachment of poly(ethylene glycol) (PEG) molecules can increase protein sera half-lives and reduce antigenicity. Not surprisingly, PEGylation significantly alters the surface characteristics of a protein, and consequently, its conformational stability during freezing and drying. Freeze concentration-induced phase separation between excipients has been previously shown to cause degradation of the secondary structure in lyophilized hemoglobin. In this report we show how PEGylation of two proteins, hemoglobin- and brain-derived neurotrophic factor (BDNF), influences partitioning and protein secondary structure as determined by FTIR spectroscopy in a system prone to freezing-induced phase separation. PEGylation of hemoglobin reduces the loss of structure induced by lyophilization in a PEG/dextran system that phase separates during freezing, perhaps due to altered partitioning. The partition coefficient for native hemoglobin favors the dextran-rich phase (PEG/dextran partition coefficient = 0.3), while PEGylated hemoglobin favors the PEG phase (partition coefficient = 3.1). In addition, we demonstrate that PEGylation alters hemoglobin's stability during lyophilization in the absence of other excipients. In contrast, because native BDNF already partitions into the PEG-rich phase, PEGylation of BDNF has a less dramatic effect on both partition coefficients and conformational stability during lyophilization. This is the first report on the effects of PEGylation on protein structural stability during lyophilization and points out the need to consider modification of formulations in response to changing protein surface characteristics.

Brain-Derived Neurotrophic Factor↗

PubMedDate not supplied

Lipid phase separation correlates with activation in platelets during chilling.

When human platelets are chilled below 22 degrees C, they spontaneously activate, a phenomenon that severely limits their storage life. It has previously been proposed that there is a correlation between cold-induced platelet activation and passage of the membranes through a liquid-crystalline to gel phase transition. Because animal models are essential for developing methods for cold storage of platelets, it is necessary to investigate such a correlation in animal platelets. In this work, horse platelets were used as a model, and it was found that cold-induced morphological activation is related to the lipid phase transition. Using fluorescence microscopy with the lipophilic fluorescent dye 1,1'-dioctadecyl-3,3,3',3'-tetramethyl-indocarbocyanine perchlorate (Dil-C18), and Fourier transform infrared spectroscopy (FTIR), it was found that lipid phase separation occurs during cooling and low temperature storage. Furthermore, removal of cholesterol from the plasma membrane also induced a phase separation, possibly between specific phospholipid classes. Steady-state fluorescence anisotropy of 1,6-diphenyl-1,3,5-hexatriene (DPH) and trimethylammonium-DPH (TMA-DPH) were compared in cells and multilamellar vesicles (MLV) composed of platelet lipids. Cholesterol depletion led to a decrease in the fluorescence anisotropy of the two probes, which can be explained by changes in the order of the phospholipid molecules. In addition, the lipid composition and fatty acid profile of the cellular phospholipids were determined. Based of the similarities between horse and human platelets, it is suggested that horse platelets may be used as a model for studying cold-stored platelets. The results are discussed in relation to the possible role of phase separation during cell signalling.

Animals↗

PubMed1996

Lateral phase separation in interfacial films of pulmonary surfactant.

To determine if lateral phase separation occurs in films of pulmonary surfactant, we used epifluorescence microscopy and Brewster angle microscopy (BAM) to study spread films of calf lung surfactant extract (CLSE). Both microscopic methods demonstrated that compression produced domains of liquid-condensed lipids surrounded by a liquid-expanded film. The temperature dependence of the pressure at which domains first emerged for CLSE paralleled the behavior of its most prevalent component, dipalmitoyl phosphatidylcholine (DPPC), although the domains appeared at pressures 8-10 mN/m higher than for DPPC over the range of 20-37 degrees C. The total area occupied by the domains at room temperature increased to a maximum value at 35 mN/m during compression. The area of domains reached 25 +/- 5% of the interface, which corresponds to the predicted area of DPPC in the monolayer. At pressures above 35 mN/m, however, both epifluorescence and BAM showed that the area of the domains decreased dramatically. These studies therefore demonstrate a pressure-dependent gap in the miscibility of surfactant constituents. The monolayers separate into two phases during compression but remain largely miscible at higher and lower surface pressures.

Animals↗

PubMed1985

Adenylate cyclase and a fatty acid spin probe detect changes in plasma membrane lipid phase separations induced by dietary manipulation of the cholesterol:phospholipid ratio.

Rats fed with a cholesterol supplement to their diet exhibited an increase in their plasma membrane cholesterol phospholipid (C/P)-lipid molar ratio from 0.72 to 0.98, whereas those fed the hypocholesterolaemic drug clofibrate in their diet exhibited a decrease in this ratio to 0.62. The properties of these membranes were analysed with regard to ligand-stimulated adenylate cyclase activity and the mobility of a fatty acid spin probe which allowed lipid phase separations to be identified. Membranes with elevated C/P ratios exhibited two distinct lipid phase separations, one at around 36 degrees C that was attributed to the external half of the bilayer and one at around 22 degrees C which was attributed to the inner half of the bilayer. Membranes with lowered C/P ratios exhibited a single lipid phase separation occurring at around 21 degrees C which was attributed to the lipids of the inner half of the bilayer. These results were compared with those obtained by manipulation of C/P ratios in vitro using liposome-cholesterol exchange techniques. Dietary manipulation of the C/P ratio of plasma membranes in vivo led to alterations in the fold stimulation of adenylate cyclase by various stimulatory ligands.

Adenylyl Cyclases↗

PubMed2006

Thermodynamic stability of fluid-fluid phase separation in binary athermal mixtures: the role of nonadditivity.

We studied the thermodynamic stability of fluid-fluid phase separation in binary nonadditive mixtures of hard-spheres for moderate size ratios. We are interested in elucidating the role played by small amounts of nonadditivity in determining the stability of fluid-fluid phase separation with respect to the fluid-solid phase transition. The demixing curves are built in the framework of the modified-hypernetted chain and of the Rogers-Young integral equation theories through the calculation of the Gibbs free energy. We also evaluated fluid-fluid phase equilibria within a first-order thermodynamic perturbation theory applied to an effective one-component potential obtained by integrating out the degrees of freedom of the small spheres. A qualitative agreement emerges between the two different approaches. We also addressed the determination of the freezing line by applying the first-order thermodynamic perturbation theory to the effective interaction between large spheres. Our results suggest that for intermediate size ratios a modest amount of nonadditivity, smaller than earlier thought, can be sufficient to drive the fluid-fluid critical point into the thermodinamically stable region of the phase diagram. These findings could be significant for rare-gas mixtures in extreme pressure and temperature conditions, where nonadditivity is expected to be rather small.

Journal Article↗

PubMed2006

Transmission electron microscopy analysis of phase separation in GaInAsSb films grown on GaSb substrate.

The GaSb-based quaternary alloys are a good choice for thermophotovoltaic applications. The thermophotovoltaic cell converts infrared radiation to electricity, using the same principles as photovoltaic devices. The aim of the present work was the microstructural study of such an alloy, namely Ga(0.84)In(0.16)As(0.12)Sb(0.88). A thin film of the material was grown by metal organic vapour phase epitaxy on a (100)alpha-->[111]B (alpha = 2 degrees, 4 degrees, 6 degrees) GaSb substrate. The GaInAsSb alloy has an appropriate band gap, but suffers from a phase separation consisting of GaAs-rich and InSb-rich regions that is disadvantageous for cell efficiency. In this work, we employed a morphological approach to phase separation, with the use of conventional transmission electron microscopy and atomic force microscopy. The phase separation occurs in two different orientations: parallel to the growth direction (vertical) and inclined (lateral). After application of fast Fourier transformation filtering, the vertical periodicity was found to be lambda = 5 nm for the pair (black and white) of layers independently of the cut-off angle, whereas the lateral periodicity was related to it.

Journal Article↗

PubMed1998

Lalpha-phase separation in phosphatidylcholine-water systems induced by alkali chlorides.

The effects of alkali chlorides on phosphatidylcholine-water bilayer systems in the Lalpha-phase were investigated by using small- and wide-angle X-ray scattering. The ternary system LiCl-POPC-H2O under isothermal conditions has shown that above Li+/POPC molar ratios of 0.1 and a lipid concentration above 5% (w/w), a splitting of the lamellar Bragg diffraction peaks into discrete components indicates a phase separation into different lamellar liquid crystalline (smectic A) phases. It is also shown that in saturated distearoyl phosphatidylcholine and in egg phosphatidylcholine, alkali chlorides induce Lalpha-phase separation. The number and repeat distance of the coexisting lamellar phases depend on the nature and concentration of the alkali chloride, the concentration of the phosphatidylcholine, and the degree of the acyl chain unsaturation.

Chemical Phenomena↗

PubMed2002

Interfacial tension in phase-separated gelatin/dextran aqueous mixtures.

The effect of solute concentrations on interfacial tension was investigated in phase-separated mixtures of dextran and gelatin over a range of concentrations that covered different tie-lines and different positions on one tie-line. The investigations were carried out using equilibrated gelatin-rich and dextran-rich phases in a computer-controlled Couette device at 40 degrees C (above the gelation point of gelatin) and interfacial tensions were measured using the retracting drop method. The results show that the interfacial tension can be related to the length of the tie-line or to the difference in the concentration of dextran (or gelatin) in the separated phases. Interfacial tension increases as either of these parameters increases. For concentrations lying on any single tie-line, the interfacial tension is constant and independent of the concentration of biopolymers. Also, the addition of small amounts of low molecular weight dextran to a dextran-rich phase does not significantly affect the interfacial tension between the gelatine-rich and dextran-rich phases. Experimental results were also compared with theoretical predictions of the interfacial tension using a Flory-Huggins based analysis of the measured tie-line data. Reasonable agreement was found between predicted and measured values, indicating that this approach captures the basic physics of the system.

Dextrans↗

PubMed2002

Self-assembling morphology induced by nanoscale rods in a phase-separating mixture.

A symmetric binary mixture containing mobile nanoscale rods is quenched from a one-phase state into an unstable phase-separating region. Incorporating the motion of rods, the system undergoes spinodal decomposition to form a dropletlike structure. The rod-rod interaction and the preferential adsorption of one of the two immiscible phases onto the mobile rods play an important role in driving the system to self-assemble into this special structure. Within each cluster of the wetting phase, the rods align parallel to each other as in a nematic liquid crystal, while the rod orientations between these clusters are randomly distributed. However, an interconnected structure is recovered in the presence of hydrodynamic interaction. In the present phase-separating mixture with rods, the growth dynamics is examined in detail, and our simulations reveal a crossover from a rod-mobility-dependent to an independent regime. The system always exhibits slowing-down growth behavior where the well-known diffusive Lifshitz-Slyozov mechanism and hydrodynamic effects are both suppressed.

Journal Article↗

PubMed1985

Small-angle neutron scattering study of lateral phase separation in dimyristoylphosphatidylcholine-cholesterol mixed membranes.

The small-angle neutron scattering (SANS) technique developed previously is used to study the lateral phase separation in dimyristoylphosphatidylcholine (DMPC)-cholesterol mixed vesicles in the L alpha (35 degrees C) and L beta' (7 degrees C) phase of DMPC. To increase the sensitivity of the previous method, we apply the so-called inverse contrast variation technique where contrast matching is performed at a constant H2O/D2O ratio by varying the ratio of DMPC with deuterated and protonated hydrocarbon chains. Phase boundaries can be determined to an accuracy of +/- 0.5 mol %. In parallel experiments phase separation in the L beta' phase was also studied by freeze-fracture electron microscopy. For DMPC in the L alpha phase complete miscibility is clearly established up to cholesterol molar fractions of xc = 0.14. Strong evidence is provided that this is also the case up to xc approximately equal to 0.45. Cholesterol is no longer soluble above this limit and precipitates as small crystallites. For the L beta' phase (7 degrees C) phase boundaries are clearly established at xc1 = 0.08 and xc2 = 0.24, and very strong evidence is provided for two additional boundaries at xc3 = 0.435 and xc4 approximately equal to 1.0. At 0 less than or equal to xc less than or equal to xc1 the mixture forms a tilted solid solution in both the L beta' and P beta' phase while at xc1 less than or equal to xc less than or equal to xc2 this phase coexists with a nontilted mixture containing 24 mol % cholesterol.(ABSTRACT TRUNCATED AT 250 WORDS)

Cholesterol↗

PubMed1986

Fusion and phase separation monitored by lifetime changes of a fluorescent phospholipid probe.

The sensitivity of the fluorescence lifetime of 1-palmitoyl-2-[[2-[4- (6-phenyl-trans-1,3,5-hexatrienyl)phenyl]ethyl]carbonyl]- 3-sn-phosphatidylcholine (DPHpPC) to its local concentration in lipid bilayers was used to monitor both lipid mixing and phase separation occurring during membrane vesicle fusion. Vesicles containing 2 mol % DPHpPC were mixed with a 10-fold excess of vesicles devoid of probe. Upon addition of a fusogen, mixing of bilayer lipids associated with fusion was followed as an increase in the fluorescence lifetime of DPHpPC. Ca2+-induced fusion of phosphatidylserine vesicles served to test the method and was shown to have an exponential half-time of 7 s. Phase separation (between the phosphatidylserine head groups of bulk lipid and the phosphatidylcholine head groups of the probe) was monitored by DPHpPC under the same conditions used to follow lipid mixing due to fusion. Phase separation was not significant until 10 min after Ca2+ addition and was completely reversible by disodium ethylenediaminetetraacetate addition. Vesicle aggregation induced by Ca2+ addition to mixed phosphatidylserine/phosphatidylcholine vesicles did not alter the DPHpPC lifetime, indicating that close association of vesicles did not promote intervesicular exchange of the probe. In addition, we have investigated the effects of CA2+ on the fluorescence properties of this probe and of the head-group-labeled fluorescent probes N-(4-nitro-2,1,3-benzoxadiazolyl)phosphatidylethanolamine and N-(lissamine Rhodamine B sulfonyl)dioleoyl-phosphatidylethanolamine, which are used in the fluorescence energy transfer assay of Struck et al.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

PubMed2002

Lateral organization of GM1 in phase-separated monolayers visualized by scanning force microscopy.

Phase separation of glycolipids in lipid mono- and bilayers is of great interest for the understanding of membrane function. The distribution of the ganglioside GM1 in sphingomyelin (SM)/1-palmitoyl-2-oleoyl- sn-glycero-3-phosphocholine (POPC), SM/1,2-dipalmitoyl- sn-glycero-3-phosphocholine (DOPC) and SM/cholesterol/POPC Langmuir-Blodgett (LB) monolayers transferred at 36 mN/m has been studied by scanning force microscopy. Besides lateral organization of the glycolipid in LB monolayers as deduced from topography, material properties have been investigated by phase imaging, pulsed force mode and force modulation microscopy. It was shown that GM1 preferentially clusters in an ordered lipid matrix, i.e. the SM phase in the case of the SM/POPC and SM/DOPC mixture or in the ordered phase of POPC/SM/cholesterol monolayers. At higher local concentrations, three-dimensional protrusions enriched in GM1 occur, which may represent a precursor for the formation of micelles budding into the aqueous subphase. Electronic supplementary material to this paper can be obtained by using the Springer Link server located at http://dx.doi.org/10.1007/s00249-002-0232-4.

Brain↗

PubMed2004

Template-directed patterning using phase-separated Langmuir-Blodgett films.

The structures of the mixed Langmuir-Blodgett (LB) films of conventional amphiphiles (CAs) and amphiphilic silane-coupling agents (SCAs) were investigated using IR spectroscopy, atomic force microscopy, and friction force microscopy. By using CAs having hydrogenated alkyl chains and SCAs having perfluorinated alkyl chains, phase-separated structures were formed with domains consisting of CAs surrounded by SCAs. The size and shape of the domains depended strongly on the mixed components, the mixing ratios, and the subphase temperature. In particular, usage of a CA having hydrogenated and perfluorinated portions in the hydrophobic group as one of the components led to the formation of nanothreads. When the phase-separated mixed LB films were heated, SCAs formed covalent bonds with the substrates having silanol groups whereas CAs did not have such ability. Rinsing the heat-treated LB films with ethanol selectively removed CAs with the SCA regions intact, resulting in the fabrication of templates. The structures of the templates reflected those of the original phase-separated LB films. LB transfer of amphiphiles on the templates led to the confinement of the amphiphiles in regions with the size and shape delineated by the templates. These results demonstrate that a variety of amphiphiles can be confined two-dimensionally in a designed manner.

Journal Article↗

PubMed2004

Phase separation in mixtures of Yukawa and charged Yukawa particles from Gibbs ensemble Monte Carlo simulations and the mean spherical approximation.

The phase equilibrium of mixtures of Yukawa and charged Yukawa particles is studied by means of Gibbs ensemble Monte Carlo (GEMC) simulation method and the mean spherical approximation (MSA). The strength of the Coulomb energy compared to that of the Yukawa attraction is characterized by a coupling constant. For low coupling constants a classical vapor--liquid phase separation appears with a good agreement between GEMC and the MSA. For high coupling constant, a phase separation between a salt poor and a salt rich phase occurs that resembles the phase equilibrium behavior of the solvent primitive model.

Journal Article↗

PubMedDate not supplied

The atomic force microscope as a tool for studying phase separation in lipid membranes.

Atomic force microscopy has developed into a powerful tool in the study of phase separation in lipid bilayers. Its ability to image a semi-fluid surface under buffer at nanometre lateral resolution and Angstrom resolution vertically allows us to distinguish phase separated lipid domains, models of the elusive rafts postulated to exist as functional platforms in the cellular membrane, which may only rise 0.3 nm above the surrounding membrane. This review charts the history of this development, and includes a description of sample preparation techniques, factors affecting image contrast mechanisms, its use in the investigation of the pre-transition ripple phase, and in the localization of cell surface proteins.

Animals↗

PubMed1994

Is lateral phase separation required for fatty acid to stimulate lipases in a phosphatidylcholine interface?

Lipase-catalyzed oxygen exchange between 13,16-cis,cis-docosadienoic acid and water in liquid-expanded monolayers with 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine exhibits abrupt, lipid composition-dependent changes in extent and mechanism [e.g., Muderhwa, J. M. and Brockman, H. L. (1992) J. Biol. Chem. 267, 24184-24192]. The critical nature of this transition suggests possible lateral phase separation of the lipids. This has been addressed by substituting for either lipid species one which can exist in more condensed monolayer states. Analysis of phase transition surface pressures as a function of lipid composition shows that each set of fatty acid-phosphatidylcholine mixtures exhibits a finite range of miscibility in liquid-expanded monolayers. These results strongly suggest that 13,16-cis,cis-docosadienoic acid and 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine are miscible under the conditions of the oxygen-exchange experiments. Furthermore, to address more directly the relation of lateral lipid phase separation to lipase regulation, oxygen exchange catalyzed by pancreatic carboxylester and triglyceride lipases was studied using mixed monolayers of [18O]2-docosadienoic acid and 1-myristoyl-2-palmitoyl-sn-glycero-3-phosphocholine. These lipids are miscible in the liquid-expanded state at all compositions. The lipid composition dependencies of both the extent and mechanism of lipase-catalyzed oxygen exchange were essentially identical to those obtained earlier. Thus, lateral lipid phase separation is not required for the critical transition in substrate accessibility to lipases. This finding supports a percolation-based model of lipase regulation within a single surface phase and suggests the "topo-temporal" regulation of lipid-mediated signaling in cells.

Chemical Phenomena↗