Electroformation of giant liposomes from spin-coated films of lipids

This paper describes spin-coating of solutions of lipids and using the resulting thin films for electroformation of giant liposomes. Spin-coating made it possible to generate uniform films of lipids with controllable thickness over large surfaces (>25 cm(2)) of indium tin oxide. Establishing a range of thicknesses optimal for electroformation (25-50 nm), we demonstrate formation of giant liposomes from lipids (such as asolectin, phosphatidylserine, and phosphatidylglycerol) that do not readily form giant liposomes from traditional, droplet-derived films. We compared liposomes from a spin-coated film of lipids to liposomes formed from traditional droplet-derived films and found that spin-coated films produced larger (by factor of 2-5) and more abundant liposomes than droplet-derived films of lipids. Electroformation from spin-coated, homogenous lipid films of optimal thickness provided a reproducible way to obtain liposomes with diameters that are predominantly larger than 30 microm over the entire surface of formation.     

Phospholipid bilayer formation on hydroxyapatite sol-gel synthesized films

Lipid bilayers supported by porous biomaterials are being explored as models for cell membranes. Hydroxyapatite is a relevant material currently being used extensively for biomedical applications. In this study, hydroxyapatite films produced via a sol-gel chemistry route have been characterized and explored as a scaffolding material for lipid membranes. The hydroxyapatite has been characterized using XRD, SEM, and AFM, followed by vesicle-fusion of lipids characterized by fluorescence microscopy and fluorescence recovery after photobleaching (FRAP) to determine the diffusion coefficient of this system. The HA films produced in this work were found to produce slow lateral diffusion and, in the two-phase lipid systems, some domains were observed. The low lateral diffusion coefficients were believed to be a result of the large undulations present on the hydroxyapatite film surface.     

Structure of organized organic mono- and multi-layer films

A review is presented of some interesting structural properties of monomolecular layers at the air/water interface (Langmuir films) and of organized multilayer structures of organic molecules (Langmuir-Blodgett films). In particular multilayer films of bipolar lipids from archaebacteria and of proteins are considered, together with single-electron junction quantum effects in parallel, special emphasis is given to some recently developed experimental techniques based on X-ray scattering. supramolecular assemblies consisting of multilayers containing nanoparticles.     

Voltammetric study of levomepromazine induced ionic permeability in a model lipid membrane system

The interaction of solid supported liquid films of lipids with levomepromazine is investigated in the present work by voltammetry. It is shown that the levomepromazine penetrates into the films, which results in a readily detectable change of the film permeability to electroactive species. In this respect, possibilities for the employment of supported lipid films as model systems in the research of biomembrane active compounds are briefly discussed.     

Self‐Assembled Aggregates and Molecular Bilayer Films of a Double‐Chain Fullerene Lipid: Structure and Electrochemistry

The self‐aggregation behavior of C₆₀ fullerenes that bear two octadecyl chains (lipid 1 ) as well as the structures and electrochemical properties of cast films of 1 are described. We also examined the self‐aggregation behavior in organic solvents of three previously reported compounds: C₆₀ with three each of hexadecyl (lipid 2 ), tetradecyl (lipid 3 ), or dodecyl (lipid 4 ) chains. The fullerene lipids in alcohols spontaneously formed spherical aggregates, whose diameters are related to the alkyl‐chain lengths, concentrations of the fullerene lipids, and the solvent polarity. The morphologies of the aggregates showed temperature dependence. Cast films of 1 formed multimolecular bilayer structures that undergo a phase transition typical of lipid bilayer membranes. The electrochemistry of cast films of 1 on an electrode in aqueous medium exhibits temperature dependence.     

Synchrotron X-ray techniques for the investigation of structures and dynamics in interfacial systems

This review focuses on recent results obtained by synchrotron X-ray techniques applied to the characterization of interfacial systems, with main emphasis on flat interfaces and on colloidal systems. The techniques covered are, for structural determinations: X-ray reflectivity (XRR), grazing incidence X-ray diffraction (GIXRD) and grazing incidence X-ray excited fluorescence (GIXF), while dynamics are investigated by X-ray photon correlation spectroscopy (XPCS) mainly in the grazing-incidence geometry (GIXPCS).The systems reviewed are, in order of growing complexity, floating Langmuir monolayers, supported films of lipids and proteins, polymeric films, buried interfaces, colloidal systems and gels formed by colloids either in 3D or in the form of 2D interfacial layers. Recent results are critically discussed, and some interesting directions of development are outlined, having also in mind new technical developments such as X-ray free electron laser sources and micro-focused synchrotron beamlines.     

MINOR SURFACE-ACTIVE LIPIDS OF OLIVE OIL AND VISCOELASTICITY OF PROTEIN FILMS AT THE OLIVE OIL-WATER INTERFACE

The influence of the minor surface-active lipids of olive oil on the viscoelastic parameters of bovine serum albumin, sodium caseinate and egg yolk films, formed following adsorption at the olive oil-protein solution interface, was studied. All the parameters substantialy decreased in the case of bovine serum albumin when surface-active lipids were removed from the olive oil while the absence of these components resulted in purely viscous films in the cases of sodium caseinate and egg yolk. Protein film viscoelasticity is probably influenced by olive oil surface-active lipids as a result of their interactions with adsorbed and unfolded protein molecules at the oil-protein solution interface.     

533—Structure and transport properties of artificial bipolar lipid membranes

The membranes of thermophilic archaebacteria are characterized by the presence of unusual bipolar saturated isoprenoid lipids. In order to investigate their molecular arrangement in the membrane and a possible influence on transport properties, we studied black films made from lipids of Caldariella acidophila, one of the most thermophilic archaebacteria. Details on the kinetics of formation at various temperatures are presented.Capacitance, compressibility and valinomycin-induced conductance values are compared with the corresponding data for a glycerol-monooleate (GMO) bilayer. A very peculiar behavior is presented by the bipolar lipid films studied. In fact, the values of conductance are more than two orders of magnitude lower than those of a GMO bilayer, while the values of capacitance and compressibility do not depend appreciably on the solvent in which the lipid is dispersed (in contrast with a GMO bilayer, where there is a 100% change).The results are discussed in terms of a proposed model of a monolayer organization of bipolar lipids and of the unusual composition of the hydrophobic core of the membrane.     

Thin film formation of silica nanoparticle/lipid composite films at the fluid-fluid interface

We report a new and simple method for the formation of thin films at the interface between aqueous silica Ludox dispersions and lipid solutions in decane. The lipids used are stearic acid, stearyl amine, and stearyl alcohol alongside silica Ludox nanoparticle dispersions of varying pH. At basic pH thin films consisting of a mixture of stearic acid and silica nanoparticles precipitate at the interface. At acidic and neutral pH we were able to produce thin films consisting of stearyl amine and silica particles. The film growth was studied in situ with interfacial shear rheology. In addition to that, surface pressure isotherm and dynamic light scattering experiments were performed. The films all exhibit strong dynamic rheological moduli, rendering them an interesting material for applications such as capsule formation, surface coating, or as functional membranes.     

An atomic force microscope study of thermal behavior of phospholipid monolayers on mica

We observed by using atomic force microscope (AFM) phospholipid (1,2-dipalmitoyl-sn-glycero-3-phosphocholine) monolayers on mica being annealed and cooled to a selection of temperatures through steps of 2-4 degrees C/min. The annealed phospholipid monolayers started to disappear at 45-50 degrees C and disappeared completely above 60-63 degrees C under AFM observation. The phospholipid monolayers reformed when the samples were cooled below 60 degrees C and developed from fractal into compact monolayer films with decreasing temperatures. Simultaneously the height of the reformed phospholipid films also increased with decreasing temperatures from 0.4 nm to the value before annealing. The observed thermal features are attributed to a phase-transition process that upon heating to above 45-50 degrees C, the lipids condensed in the monolayers transform into a low-density expanded phase in which the lipids are invisible to AFM, and the transformation continues and completes at 60-63 degrees C. The lipid densities of the expanded phase inferred from the dissociated area of the condensed phase are observed to be a function of the temperature. The behavior contrasts with a conventional first-order phase transition commonly seen in the Langmuir films. The temperature-dependent height and shape of the reformed phospholipid films during cooling are argued to arise from the adjustment of the packing and molecular tilting (with respect to the mica surface) of the phospholipids in order to accommodate more condensed phospholipids.     

Nanometer-thick surficial films in oxides as a case of prewetting

Stable, nanometer-thick films are observed to form at the {1120} facets of Bi(2)O(3)-doped ZnO in several bulk-phase stability fields. Electron microscopy shows these surficial films to exhibit some degree of partial order in quenched samples. The equilibrium film thickness, corresponding to the Gibbs excess solute, decreases monotonically with decreasing temperature until vanishing at a dewetting temperature, well below the eutectic. Assuming that perfect wetting occurs at some higher temperature above the eutectic, as is observed on polycrystal surfaces and at grain boundaries in the same system, the adsorption and wetting events in this system illustrate temperature- and composition-dependent prewetting. The observation of a second class of thicker films coexisting with nanodroplets and a numerical evaluation of thickness versus temperature elucidate the critical role of volumetric thermodynamic terms in determining film stability and thickness. Analogous temperature-dependent surface films involving adsorbed MoO(3) on Al(2)O(3) were also observed.     

Effects of divalent cations on the formation and structure of solid supported lipid films

The interaction of glassy carbon-supported thin wetting films of lecithin with some divalent cations is investigated by impedimetry and voltammetry. The influence of Ca2+, Mg2+, and Mn2+ on the film structure is explored in two different cases--the divalent cations are added to the electrolyte either before or after the formation of the film. When the film has been previously formed, the addition of divalent cations in millimolar concentrations leads to changes in the passive electrical parameters and the blocking properties of the films. On the one hand the dielectric properties of the film measured in 0.1 M KCl seem to improve after the interaction with divalent cations--the film capacitance decreases, the resistance and resistivity of the film increase. On the other hand the increase of the redox current in the presence of 1 mM Fe(CN)6(3-/4-) in the electrolyte suggests the formation of some defects in the lipid structure of the film after the action of divalent cations. It is shown that the amount of these defects could be significantly decreased when the divalent cations are present in the electrolyte solution before the film formation. The effect of divalent cations on the film stability is tested by applying negative potential to the film. In 0.1 M KCl the films are not stable at potential of - 0.8 V (vs. Ag/AgCl) and are destroyed. The addition of divalent cations stabilizes the films and at certain millimolar concentrations the films remain intact after the action of the negative potential. The effect of Mn2+ is more pronounced, the Ca2+ and Mg2+ have smaller commensurate effect. It is proposed that the changes in the films' properties could be related with more tight packing of the lipid molecules with the divalent cations inserted in the film and that some defects could be opened during the rearrangement of the lipids when the film has been previously formed.     

Wetting properties of model biological membranes

Various aspects of native and model biological membrane wettability are discussed. Among others hydration of mono-, bi-, and multi-layers of lipids as well as wettability of macroscopic surfaces of solid supported lipid films was investigated via apparent contact angle measurements and calculation of the apparent surface free energy of the films. The effects of relative humidity on the layer hydration and contact angle changes are also discussed. Finally, the effect of liposomes and enzymes (due to the hydrolysis reactions) on the hydrophobic/hydrophilic character of the film surfaces is overviewed.     

Mixed monolayers of Bauhinia monandra and concanavalin A lectins with phospholipids, part II

Isotherms of surface pressure and surface potential versus mean molecular area for dibehenoylphosphatidylcholine (DBPC), dipalmitoylphosphatidylcholine (DPPC), dipalmitoylphosphatidylethanolamine (DPPE), and dioleoylphosphatidylcholine (DOPC) monolayers were shown to be greatly modified when these lipids were cospread with either Bauhinia monandra (BmoLL) or Concanavalin A (Con A) lectins. For the binary films of DBPC, DPPC, and DPPE cospread with each of these two lectins, there was both a displacement of the Pi-A and DeltaV-A isotherms toward higher molecular areas relative to pure lipids and an increase in the maximum surface potential values relative to the DeltaV-A relationships observed for the corresponding single-lectin systems. Both effects can be understood in terms of the occurrence of an explicit interaction between the lipids and the lectins. The plots of the corresponding compressibilities versus molecular areas reveal that, for all lipids but DOPC, the extent of this interaction was always larger for BmoLL than for Con A. The DPPC and DPPE mixed films with BmoLL differed in compressibility. Owing to the small DPPE polar headgroup, the DPPE-BmoLL film was much more incompressible than the DPPC-BmoLL mixed monolayer. However, for the DOPC-BmoLL and DOPC-Con A mixed films there was no evidence that an interaction between the lectins and the lipid took place, a fact attributed to the unsaturated character in the DOPC aliphatic chains, which leads to an expanded Pi-A isotherm.     

Colorimetric detection and fingerprinting of bacteria by glass-supported lipid/polydiacetylene films

Glass-supported films of lipids and polydiacetylene were applied for visual detection and colorimetric fingerprinting of bacteria. The sensor films comprise polydiacetylene domains serving as the chromatic reporter interspersed within lipid monolayers that function as a biomimetic membrane platform. The detection schemes are based on either visible blue-red transitions or fluorescence transformations of polydiacetylene, induced by amphiphilic molecules secreted by proliferating bacteria. An important feature of the new film platform is the feasibility of either naked-eye detection of bacteria or color analysis using conventional scanners. Furthermore, we find that the degrees of bacterially induced color transformations depend both on the bacterial strains examined and the lipid compositions of the films. Accordingly, bacterial fingerprinting can be achieved through pattern recognition obtained by recording the chromatic transformations in an array of lipid/PDA films having different lipid components.     

Thin films of discotic liquid crystals and their applications

ABSTRACT

Discotic liquid crystals (DLCs) are considered as fascinating systems due to their unique property of self-assembly to yield different columnar structures. DLCs are organic semiconductors and create pathways for the development of numerous optical and electrical devices. The thin films of DLCs can be considered as low dimensional system which can exhibit remarkable optical and physical properties. In this article, we present a review on ultrathin films of some interesting DLC molecules at air–water and air–solid interfaces. The Langmuir monolayer and Langmuir–Blodgett films of DLC molecules are extensively studied. The ultrathin films of DLC molecules can yield highly anisotropic layer wherein the molecular orientation and aggregation can have large impact on the physicochemical properties of the film. Different surface phases with different molecular orientations as function of surface density and temperature can be obtained by forming the Langmuir monolayer of the DLC molecules at the air–water interface. The Langmuir monolayer in a particular phase can be deposited onto the active area of a device layer-by-layer by employing a highly controlled Langmuir–Blodgett technique. Here, we report some interesting results related on molecular orientation of the DLC molecules at different interfaces. Such aggregation of DLC molecules in ultrathin films may find applications in thin film-based electro-optical devices.     

Sphingomyelin/phosphatidylcholine and cholesterol interactions studied by imaging mass spectrometry

Label-free imaging mass spectrometry is utilized the first time to study lipid-lipid interactions in a model membrane system. Ternary lipid mixtures of cholesterol (CH), sphingomyelin (SM), and phosphatidylcholine (PC) on supported Langmuir-Blodgett films are investigated as a mimic of the cellular membrane. The unique chemical specificity and imaging capability allow identification and localization of each lipid molecule in the membranes. The SM and PC in each ternary mixture vary in their acyl chain saturation with both, either, or neither one double bonded at the same position of their acyl chain. For the ternary mixtures with SM and PC both saturated or unsaturated, all the lipids are evenly distributed in the molecule-specific images. However, domain structures were observed for the two mixtures with either SM or PC unsaturated. In both films, the saturated lipid, whether it is SM or PC, colocalized with CH while the unsaturated lipid was excluded from the CH domains. These results strongly suggest that acyl chain saturation, rather than the specific interactions between SM and CH, is the dominating factor for SM colocalization with CH in the raft areas of the cellular membranes.     

Determination of symmetry and molecular arrangements of free-standing liquid crystal films using null-transmission ellipsometry

A high resolution null-transmission ellipsometry system has been developed for measurement of the optical properties of liquid crystal free-standing films. The ellipsometric parameters are measured to within a resolution of 0.002° from samples held in a sealed, temperature-controlled environment. By measuring the ellipsometric parameters as a function of applied electric field orientation, the optical symmetry and molecular orientations of free-standing liquid crystal films can be determined in some cases. Three carefully selected applications of this technique are presented.     

Thermochromism of bacteriorhodopsin and its pH dependence

Purple membranes (PMs), which consist of the photochromic membrane protein bacteriorhodopsin (BR) and lipids only, show complex thermochromic properties. Three different types of reversible temperature-dependent spectral transitions were found, involving spectral states absorbing at 460, 519, and 630 nm. These thermochromic absorption changes were analyzed in the range from 10 to 80 degrees C. In dependence on the bulk pH value, hypsochromic or bathochromic shifts in the BR absorption spectra are observed in BR gels as well as in BR films. The thermochromic changes between both purple and blue or purple and red were quantified in the CIE color system. The molecular changes causing these effects are discussed, and a model is presented in terms of intramolecular protonation equilibriums. The thermochromic properties of BR may be of interest in applications like security tags, as this feature may complement the well-known photochromic properties of BR.     

Structure of spin-coated lipid films and domain formation in supported membranes formed by hydration

An atomic force and fluorescence microscopy investigation of the structure of spin-coated lipid films is presented. In the surface of the dry film, lipids are found to orient in a conformation where acyl chains are pointing outward while laterally the individual layers of the multilamellar film exhibit a dewetting pattern similar to what is found in polymer thin films. Hydration of the film in liquid water promotes detachment of bilayers from the surface while a single membrane remains on the mica substrate. This supported membrane is highly uniform and defect-free as compared to supported membranes prepared by conventional methods. It is further demonstrated that supported membranes of binary lipid mixtures prepared by this method exhibit gel-fluid domain coexistence in accordance with expectations from the phase diagrams.