Neutron Reflectivity Studies Research Articles

Spectro-Electro-Gravimetry of the Compactation Effect of Poly-(ortho-toluidine)

An example of an electroactive polymer is poly(ortho-toluidine) (POT), whose monomer has been widely used as aniline derivative for dyestuff, pesticides, pharmaceutical drugs and organic intermediates. POT’s properties can be looked into by spectroelectrochemical means, in that they are directly related to the variations of colour associated to the oxidation states of its active sites [1-4]. POT displays three main oxidation forms: fully reduced state (leucoemeraldine, LE), partially oxidized (emeraldine, EM), and fully oxidised (pernigraniline, PN). Among these, only the EM form is conductive.The 100 CV cycles obtained during electropolymerisation of o-toluidine are shown in Figure 1. At about 0.85 V, the sharp peaks observed correspond to the oxidation of o-toluidine and the fact that their maximum current density increases as cycling increases suggests that the polymer is forming. In this work, the behavior of POT during compaction and subsequent swelling has been reported on. Basically, electropolymerisation of a monomer, ortho-toluidine, was executed on a flat electrode. Then, Electrochemical Quartz Crystal Microbalance (EQCM) coupled with in situ Vis-NIR spectroscopy were employed to characterize the as-formed polymer. The compaction-decompaction process of poly(o-toluidine) (POT) films synthesized from H2SO4 solution by cyclic voltammetry has been looked into. We present a detailed electrochemical mechanism for the insulating-to-conductor change of POT films. Oxidation of POT films involves their transformation into the conducting (i.e. emeraldine) form, with simultaneous anion insertion into the polymeric matrix. Coupling of electrochemical quartz crystal microbalance (EQCM) with vis-NIR spectroscopy provided singular information about the global response of current during the doping processes. Isolated polarons (P*) could form because of trapped anions when the film is reducing and shrinking (compaction). During the decompaction (i.e. relaxation), protons are expulsed when the isolated polarons detected at 420 nm are oxidized. Anions insert during the formation of conducting polarons (PC), which consist of coil structures due to the strong interaction of anions with the C-NH+ -C group of polymer detected at 840 nm. This mechanism can be used to understand the relaxation process in other polyaniline-derived films.During the LE → EM change, POT can show two different intermediate states, known as polaron and bipolaron forms, which are considered as well-defined species with their own characteristics. The transition mechanism between such intermediate states of POT, with corresponding intake or loss of ions A- from the medium. It can be observed that POT presents two successive redox processes, the first of which implies an anion transfer, whilst the second one is brought about by proton transfer [5]. the wavelength centered at 840 nm is suitable for studying the so-called isolated polaron. This species differs from others in that, during polymer oxidation, it forms somewhere in the polymer chain along with a trapped anion. As this polaron-anion pair does not conduct, its presence diminishes the conductivity of the polymer and, actually, if it gets to interact with other charges, therefore, prevents electron flow across polymer chains. The electrochemical behavior of POT is reproducible and the polymer can be stabilized through a compaction process during a given time. As demonstrated before, the electrochemical activity depends upon the compaction time. From analysis of the F(dm/dQ) function, it can be inferred that POT can be reduced and shrink with either concomitant expulsion of anions or intake of H3O+ cations, or both processes simultaneously. When POT underwent relaxation, the values of current, peak potential charge, as well as the variations of mass and absorbance (at 840 nm) with respect to time showed linear response. [1] J. Agrisuelas, C. Gabrielli, J.J. Garcia-Jareno, H. Perrot, F. Vicente, Kinetic and Mechanistic Aspects of a Poly(o-toluidine)-Modified Gold Electrode. 1. Simultaneous Cyclic Spectroelectrochemistry and Electrogravimetry Studies in H2SO4 Solutions, J. Phys. Chem. C. 116 (2012) 15620–15629.[2] C. Hu, Y. Li, Y. Kong, Y. Ding, Preparation of poly(o-toluidine)/nano ZnO/epoxy composite coating and evaluation of its corrosion resistance properties, Synth. Met. 214 (2016) 62–70. [3] S. Bilal, A.-H.A. Shah, R. Holze, A correlation of electrochemical and spectroelectrochemical properties of poly(o-toluidine), Electrochim. Acta 54 (2009) 4851–4856. [4] L. Bailey, M.J. Henderson, A.R. Hillman, N. Gadegaard, A. Glidle, In situ neutron reflectivity studies of poly-o-toluidine films, J. Phys. B-At. Mol. Opt. 276 (2000) 373–374. [5] P. Herrasti, P. Ocon, Study of the electropolymerization of o-toluidine and the effect of the temperature on the resistance of the polymer, Port. Electrochim. Acta 9 (1991) 233-266.Part of this work was supported by MINECO-FEDER CTQ2015-71794-R.

Influence of the Surfactant Structure on Photoluminescent π-Conjugated Polymer Nanoparticles: Interfacial Properties and Protein Binding

π-Conjugated polymer nanoparticles (CPNs) are under investigation as photoluminescent agents for diagnostics and bioimaging. To determine whether the choice of surfactant can improve CPN properties and prevent protein adsorption, five nonionic polyethylene glycol alkyl ether surfactants were used to produce CPNs from three representative π-conjugated polymers. The surfactant structure did not influence size or yield, which was dependent on the nature of the conjugated polymer. Hydrophobic interaction chromatography, contact angle, quartz crystal microbalance, and neutron reflectivity studies were used to assess the affinity of the surfactant to the conjugated polymer surface and indicated that all surfactants were displaced by the addition of a model serum protein. In summary, CPN preparation methods which rely on surface coating of a conjugated polymer core with amphiphilic surfactants may produce systems with good yields and colloidal stability in vitro, but may be susceptible to significant surface alterations in physiological fluids.

Understanding Silicon Electrode Surface Reactivity through Model Silicate Thin Film Layers

Development of higher capacity anodes in lithium ion batteries for use in electric vehicles is necessary in order to further enhance their energy density. Silicon anodes are being considered for these next generation lithium ion batteries due to its exceptionally high specific capacity (3579 mAh/g). One main drawback to silicon anodes is the formation of an unstable solid electrolyte interface (SEI), a surface film formed by decomposition of the electrolyte components during cycling of the battery. This unstable SEI continuously consumes Li ions and solvents from the electrolyte and stunts battery life. One major cause of this SEI instability is due to silicon anode volume expansion and contraction of up to 300% during cycling[1]. However, there is still much to learn about the chemical reactions occurring at the silicon surface before, during and after the first cycle of the battery. A useful way to narrow observations to just silicon surface reactions with no convolution from binders or conductive additives is by studying thin film silicon anodes. A neutron reflectivity study done by Veith et al utilized thin film silicon to note that the complexity of the reactions at the surface of silicon is increased by the chemical reactivity of the with the electrolyte at open circuit voltage before the anode was even cycled[2]. This implies that the length of time and composition of the surface of the anode before cycling will have an impact on how the SEI forms during the first cycle. To better understand the nature and evolution of this SEI layer formed prior to any cycling of the silicon anode and how it impacts the performance of the silicon anode, model SEI layers were deposited on clean 50 nm thick silicon thin films using RF magnetron co-sputtering. Thin film chemistries from SiO2 to Li4SiO4 were synthesized in order to model the proposed lithiation of the oxide layer during the first cycle. Model thin films were soaked in 1.2M LiPF6 in EC:EMC 3:7 wt% electrolyte from 30 minutes to 3 days in an inert atmosphere glovebox, removed and rinsed with DMC and studied using ATR IR, XPS, XRR and FIB CS. Once the pre-lithiatied film was studied using these techniques, half cells with these same silicate model films were cycled in order to observe any differences in SEI formation or cell performance. Impedance measurements were taken on model films before and after lithiation in order to better understand lithium transport of these model lithium silicates during cycling. Initial results on these model films after soaking in the electrolyte indicate a dependence on stoichiometry for the time resolved surface reactivity. While electrodes with unlithiated oxide (SiO2) soaked in electrolyte for up to 24 hours before discernable surface reaction peaks presented in their ATR IR spectra, the electrodes with lithium silicates showed significantly faster appearance of reaction peaks. This could indicate that lithium silicates passivate more quickly against the electrolyte. Preliminary FIB CS data also indicates that the SiO2 layer may be consumed with time, perhaps producing gaseous products instead of a passivating surface film. These model systems have started to tease apart the complexity of the surface reactivity and lithiation kinetics that manifest during storage and cycling of silicon anodes and could provide key insight into the low cycling stability of silicon anodes observed in practice. Ko, M., S. Chae, and J. Cho, Challenges in Accommodating Volume Change of Si Anodes for Li-Ion Batteries. Chemelectrochem, 2015. 2(11): p. 1645-1651.Veith, G.M., et al., Direct measurement of the chemical reactivity of silicon electrodes with LiPF6-based battery electrolytes. Chemical Communications, 2014. 50(23): p. 3081-3084. Figure 1

X-ray and Neutron Reflectivity Study Shows That CLIC1 Undergoes Cholesterol-Dependent Structural Reorganization in Lipid Monolayers.

CLIC1 belongs to the ubiquitous family of chloride intracellular ion channel proteins that are evolutionarily conserved across species. The CLICs are unusual in that they exist mainly as soluble proteins but possess the intriguing property of spontaneous conversion from the soluble to an integral membrane-bound form. This conversion is regulated by the membrane lipid composition, especially by cholesterol, together with external factors such as oxidation and pH. However, the precise physiological mechanism regulating CLIC1 membrane insertion is currently unknown. In this study, X-ray and neutron reflectivity experiments were performed to study the interaction of CLIC1 with different phospholipid monolayers prepared using POPC, POPE, or POPS with and without cholesterol in order to better understand the regulatory role of cholesterol in CLIC1 membrane insertion. Our findings demonstrate for the first time two different structural orientations of CLIC1 within phospholipid monolayers, dependent upon the absence or presence of cholesterol. In phospholipid monolayers devoid of cholesterol, CLIC1 was unable to insert into the lipid acyl chain region. However, in the presence of cholesterol, CLIC1 showed significant insertion within the phospholipid acyl chains occupying an area per protein molecule of 6-7 nm2 with a total CLIC1 thickness ranging from ∼50 to 56 Å across the entire monolayer. Our data strongly suggests that cholesterol not only facilitates the initial docking or binding of CLIC1 to the membrane but also promotes deeper penetration of CLIC1 into the hydrophobic tails of the lipid monolayer.

Neutron Reflection Study of Surface Adsorption of Fc, Fab, and the Whole mAb.

Characterizing the influence of fragment crystallization (Fc) and antigen-binding fragment (Fab) on monoclonal antibody (mAb) adsorption at the air/water interface is an important step to understanding liquid mAb drug product stability during manufacture, shipping, and storage. Here, neutron reflection is used to study the air/water adsorption of a mAb and its Fc and Fab fragments. By varying the isotopic contrast, the adsorbed amount, thickness, orientation, and immersion of the adsorbed layers could be determined unambiguously. While Fc adsorption reached saturation within the hour, its surface adsorbed amount showed little variation with bulk concentration. In contrast, Fab adsorption was slower and the adsorbed amount was concentration dependent. The much higher Fc adsorption, as compared to Fab, was linked to its lower surface charge. Time and concentration dependence of mAb adsorption was dominated by Fab behavior, although both Fab and Fc behaviors contributed to the amount of mAb adsorbed. Changing the pH from 5.5 to 8.8 did not much perturb the adsorbed amount of Fc, Fab, or mAb. However, a small decrease in adsorption was observed for the Fc over pH 8-8.8 and vice versa for the Fab and mAb, consistent with a dominant Fab behavior. As bulk concentration increased from 5 to 50 ppm, the thicknesses of the Fc layers were almost constant at 40 Å, while Fab and mAb layers increased from 45 to 50 Å. These results imply that the adsorbed mAb, Fc, and Fab all retained their globular structures and were oriented with their short axial lengths perpendicular to the interface.

The internal structure of PMETAC brush/gold nanoparticle composites: a neutron and X-ray reflectivity study.

This work addresses the pH-triggered distribution and relocation of charge-stabilized gold nanoparticles (AuNPs) incorporated into strong polyelectrolyte brushes. Brush/particle composite materials were investigated under aqueous conditions and at different humidities using neutron and X-ray reflectivity, respectively. X-ray reflectivity measurements complement neutron reflectivity measurements and reveal results that could not be observed by neutron reflectivity measurements. Both methods allow scanning the particle density profile, but due to different contrasts, they are sensitive to different regions within the brush. More specifically, 3-mercaptopropionic acid (MPA)-coated AuNPs were incorporated into poly-[2-(methacryloyloxy)ethyl]trimethylammonium chloride (PMETAC) polyelectrolyte brushes at different pH values. The pH value triggers a change in the AuNP surface charge caused by the pH-sensitivity of the MPA ligands, while the charge of the PMETAC brush is not affected by pH variations. The particle number density as well as the particle distribution depend strongly on the pH value of the incubation medium: a rather non-homogeneous assembly (2D assembly) is found when the PMETAC brush is incubated in AuNP suspension at pH 4, while a more homogeneous assembly (3D assembly) is found when the PMETAC brush is incubated in AuNP suspension at pH 8. The main factor dominating the formation of 2D or 3D assembly is assigned to the particle-particle interaction and not to the particle-polymer interaction. No significant relocation of AuNPs within the brush can be found by changing the environmental conditions. The control of particle amount and distribution within the polymer brush has a strong impact on the optical properties of those composite materials, which is crucial for the fabrication of colorimetric sensors.

Interaction of the Antimicrobial Peptides Rhesus θ-Defensin and Porcine Protegrin-1 with Anionic Phospholipid Monolayers.

A combination of Langmuir isotherm, Brewster angle microscopy (BAM), and neutron reflectivity studies have been performed to gain insight into the effects on model bacterial cell membranes of the antimicrobial peptides, Rhesus θ-defensin 1 (RTD-1), and porcine protegrin 1 (PG-1). The peptides were interacted with monolayers spread at the air-water interface and prepared from a 3:1 molar mixture of phosphatidylethanolamine and phosphatidylglycerol used to approximate the cell membranes of Gram positive bacteria. The Langmuir film balance measurements show that both peptides perturb the lipid monolayers causing an increase in surface pressure, and the BAM studies show that each results in the formation of small domains within the lipid films, around 5 μm diameter. The overall change in monolayer surface pressure caused by PG-1, however, is a little more pronounced than that due to RTD-1 (+8.5 mN·m(-1) vs +5.5 mN·m(-1)), and the rate of its initial interaction with the monolayer is a little more rapid than that for RTD-1. The neutron reflectivity studies also show differences for PG-1 and RTD-1, with the model fits to these data showing that the more amphiphilic PG-1 becomes fully embedded within the lipid film-causing an extension of the lipid acyl chains but leaving the thickness of the lipid headgroup layer unaffected-while RTD-1 is seen to insert less deeply-causing the same extension of the lipid acyl chains as PG-1 but also causing a significant increase in thickness of the lipid headgroup layer. The various differing effects of the two peptides on anionic lipid monolayers are discussed in the context of their differing hemolytic activities, and their proposed differing propensities to form transmembrane pores.

VERITAS: a high-flux neutron reflectometer with vertical sample geometry for a long pulse spallation source

An instrument concept of a reflectometer with a vertical sample geometry fitted to the long pulse structure of a spallation source, called “VERITAS” at the ESS, is presented. It focuses on designing a reflectometer with high intensity at the lowest possible background following the users' demand to investigate thin layers or interfacial areas in the sub-nanometer length scale. The high intensity approach of the vertical reflectometer fits very well to the long pulse structure of the ESS. Its main goal is to deliver as much usable intensity as possible at the sample position and be able to access a reflectivity range of 8 orders of magnitude and more. The concept assures that the reflectivity measurements can be performed in its best way to maximize the flux delivered to the sample. The reflectometer is optimized for studies of (magnetic) layers having thicknesses down to 5Å and a surface area of 1x1cm2. With reflectivity measurements the depth-resolved, laterally averaged chemical and magnetic profile can be investigated. By using polarised neutrons, additional vector information on the in-plane magnetic correlations (off-specular scattering at the pm length scale, GISANS at the nm length scale) can be studied. The full polarisation analysis could be used for soft matter samples to correct for incoherent scattering which is presently limiting neutron reflectivity studies to a reflectivity range on the order of 10-6.

Neutron reflection study of the interaction of the eukaryotic pore-forming actinoporin equinatoxin II with lipid membranes reveals intermediate states in pore formation

Equinatoxin II (EqtII), a eukaryotic pore-forming toxin, lyses cell membranes through a mechanism involving the insertion of its N-terminal α-helix into the membrane. EqtII pore formation is dependent on sphingomyelin (SM), although cholesterol (Chol) and membrane microdomains have also been suggested to enhance its activity. We have investigated the mechanism of EqtII binding and insertion by using neutron reflection to determine the structures of EqtII–membrane assemblies in situ. EqtII has several different modes of binding to membranes depending on the lipid composition. In pure dimyristoyl-phosphatidylcholine (DMPC) membranes, EqtII interacts weakly and reversibly with the lipid head groups in an orientation approximately parallel to the membrane surface. The presence of sphingomyelin (SM) gives rise to a more upright orientation of EqtII, but Chol is required for insertion into the core of the membrane. Cooling the EqtII–lipid assembly below the lipid phase transition temperature leads to deep water penetration and a significant reduction in the extension of the protein outside the membrane, indicating that phase-separation plays a role in EqtII pore-formation. An inactive double-cysteine mutant of EqtII in which the α-helix is covalently tethered to the rest of the protein, interacts only reversibly with all the membranes. Releasing the α-helix in situ by reduction of the disulphide bridge, however, causes the mutant protein to penetrate in DMPC–SM–Chol membranes in a manner identical to that of the wild-type protein. Our results help clarify the early steps in pore formation by EqtII and highlight the valuable information on protein–membrane interactions available from neutron reflection measurements.

Complexation of phospholipids and cholesterol by triterpenic saponins in bulk and in monolayers

The interactions between three triterpene saponins: α-hederin, hederacoside C and ammonium glycyrrhizate with model lipids: cholesterol and dipalmitoylphosphatidylcholine (DPPC) are described. The oleanolic acid-type saponins (α-hederin and hederacoside C) were shown to form 1:1 complexes with lipids in bulk, characterized by stability constants in the range (4.0±0.2)·103–(5.0±0.4)·104M−1. The complexes with cholesterol are generally stronger than those with DPPC. On the contrary, ammonium glycyrrhizate does not form complexes with any of the lipids in solution. The saponin–lipid interactions were also studied in a confined environment of Langmuir monolayers of DPPC and DPPC/cholesterol with the saponins present in the subphase. A combined monolayer relaxation, surface dilational rheology, fluorescence microscopy and neutron reflectivity (NR) study showed that all three saponins are able to penetrate pure DPPC and mixed DPPC/cholesterol monolayers. Overall, the effect of the saponins on the model lipid monolayers does not fully correlate with the lipid–saponin complex formation in the homogeneous solution. The best correlation was found for α-hederin, for which even the preference for cholesterol over DPPC observed in bulk is well reflected in the monolayer studies and the literature data on its membranolytic activity. Similarly, the lack of interaction of ammonium glycyrrhizate with both lipids is evident equally in bulk and monolayer experiments, as well as in its weak membranolytic activity. The combined bulk and monolayer results are discussed in view of the role of confinement in modulating the saponin–lipid interactions and possible mechanism of membranolytic activity of saponins.

Neutron Scattering Studies of the Effects of Formulating Amphotericin B with Cholesteryl Sulfate on the Drug’s Interactions with Phospholipid and Phospholipid–Sterol Membranes

Langmuir surface pressure, small-angle neutron scattering (SANS), and neutron reflectivity (NR) studies have been performed to determine how formulation of the antifungal drug amphotericin B (AmB), with sodium cholesteryl sulfate (SCS)-as in Amphotec-affects its interactions with ergosterol-containing (model fungal cell) and cholesterol-containing (model mammalian cell) membranes. The effects of mixing AmB in 1:1 molar ratio with cholesteryl sulfate (yielding AmB-SCS micelles) are compared against those of free AmB, using monolayers and bilayers formed from palmitoyloleoylphosphatidylcholine (POPC) in the absence and presence of 30 mol % ergosterol or cholesterol, in all cases employing a 1:0.05 molar ratio of lipid:AmB. Analyses of the (bilayer) SANS and (monolayer) NR data indicate that the equilibrium changes in membrane structure induced in sterol-free and sterol-containing membranes are the same for free AmB and AmB-SCS. Stopped-flow SANS experiments, however, reveal that the structural changes to vesicle membranes occur far more rapidly following exposure to AmB-SCS vs free drug, with the kinetics of these changes varying with membrane composition. With POPC vesicles, the structural changes induced by AmB-SCS become apparent only after several minutes, and equilibrium is reached after ∼30 min. The corresponding onset of changes in POPC-ergosterol and POPC-cholesterol vesicles, however, occurs within ∼5 s, with equilibrium reached after 10 and 120 s, respectively. The rate of insertion of AmB into POPC-sterol membranes is thus increased through formulation as AmB-SCS. Moreover, the differences in monolayer surface pressure and SANS structure-change equilibration times suggest significant rearrangement of AmB within these membranes following insertion. The reduced times to equilibrium for the POPC-ergosterol vs POPC-cholesterol systems are consistent with the known differences in affinity of AmB for these two sterols, and the reduced time to equilibrium for AmB-SCS interaction with POPC-ergosterol membranes vs that for free AmB is consistent with the reduced host toxicity of Amphotec.

Probing the position of resveratrol in lipid bilayers: A neutron reflectivity study

The effect of the natural antioxidant resveratrol on the structure of solid supported di-palmitoyl-phosphatidyl-choline (DPPC) bilayers in their fluid state was investigated by neutron reflectometry. Results reveal an accumulation of resveratrol (up to 25%, mol/mol) inside the headgroups and they exclude its presence in the hydrophobic core. The presence of resveratrol induces an increase of the average thickness and of the interfacial roughness of the headgroup layer. This may be due to a change of the tilt angle of the phosphocholine headgroups residing next to the resveratrol to a more upright orientation and leading to a reduction of the projected area per headgroup. This effect is propagated into the hydrophobic core, where the chain packing is modified despite the absence of resveratrol. When interacting with a DPPC/cholesterol membrane, resveratrol has a similar effect on the neighboring PC headgroups as in the cholesterol free membrane. The almost precise 1:1 insertion ratio (resveratrol:cholesterol) suggests that resveratrol is most probably inserted on top of the hydroxyl group of the cholesterol in between the PC headgroups. The ordering effect of cholesterol on the hydrophobic core is absent when both cholesterol and resveratrol are present. Most probably, the interaction of resveratrol with lipid membranes is non-specific.

From Homogeneous to Segregated Structure of Poly(dimethylsiloxane)/Cellulose Derivative Mixed Langmuir Films Depending on Composition: An in Situ Neutron Reflectivity Study.

The mixing behavior of deuterated polydimethylsiloxane (PDMSd) and cellulose acetate butyrate (CAB) spread as Langmuir films at the air-water interface was studied by means of surface pressure-area isotherms, Brewster angle microscopy (BAM) observations, and in situ neutron reflectivity. The contrast variation method was used with different D2O/H2O mixtures as subphase, allowing contrast matching to either CAB, PDMSd, or PDMSd/CAB mixed film if homogeneous. At PDMSd volume fractions Φ lower than 0.6, the mixed film is a homogeneous monolayer throughout the film compression, in agreement with the monophasic film observed by BAM and the attractive interactions between PDMSd and CAB evidenced from the isotherm measurements. In contrast, at PDMSd volume fractions Φ higher than 0.6, a vertically segregated structure of the mixed film is highlighted. Indeed, whatever the surface pressure, a bilayer structure is observed with a PDMSd layer in contact with the air over a thin CAB layer in contact with the subphase. These results show that the structure of the film is mainly driven by the PDMSd volume fraction which allows obtaining either a homogeneous membrane which composition can be tuned or a vertically segregated system. In contrast, only the thickness of the layers varies with the surface pressure while the structure of the film is not affected.

Antimicrobial peptide dendrimer interacts with phosphocholine membranes in a fluidity dependent manner: A neutron reflection study combined with molecular dynamics simulations

The interaction mechanism of a novel amphiphilic antimicrobial peptide dendrimer, BALY, with model lipid bilayers was explored through a combination of neutron reflection and molecular dynamics simulations. 1-Palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC) and 1,2-dipalmitoyl-sn-glycero-3-phos-phocholine (DPPC) lipid bilayers were examined at room temperature to extract information on the interaction of BALY with fluid and gel phases, respectively. Furthermore, a 1:4 mixture of POPC and DPPC was used as a model of a phase-separated membrane. Upon interaction with fluid membranes, BALY inserted in the distal leaflet and caused thinning and disordering of the headgroups. Membrane thinning and expansion of the lipid cross-sectional area were observed for gel phase membranes, also with limited insertion to the distal leaflet. However, dendrimer insertion through the entire lipid tail region was observed upon crossing the lipid phase transition temperature of DPPC and in phase separated membranes. The results show clear differences in the interaction mechanism of the dendrimer depending on the lipid membrane fluidity, and suggest a role for lipid phase separation in promoting its antimicrobial activity.

The adsorption of fluorinated dopants at the surface of 5CB: a neutron reflection study

The adsorption of dopants at the surface of 5CB has been studied using neutron reflection. The dopants were versions of 11OCB with partly fluorinated chains, and the 5CB was interfaced with air or with silica treated with fluorocarbon or hydrocarbon coatings. At the air interface, the F17-11OCB adsorbed homeotropically, and the amount increased on cooling into the nematic phase of 5CB. At low temperatures, the adsorption from the nematic phase appeared to saturate at a bilayer. At the solid interfaces, there was some evidence for a thin planar layer at the surface, but most of the adsorbed dopant was in the form of diffuse layers. In both cases, an increase in dopant concentration caused higher adsorbed amounts. For F17-11OCB, the amount absorbed was much greater for a fluorinated coating as opposed to a hydrocarbon-coated surface. Also no adsorption of the F3-11OCB could be detected, suggesting that the adsorption process is driven by the oleophobic effect. The results demonstrate that it is possible to manipulate surface properties of liquid crystals using surfactant-like dopants.

Neutron reflection study of the adsorption of the phosphate surfactant NaDEHP onto alumina from water.

The adsorption of a phosphorus analogue of the surfactant AOT, sodium bis(2-ethylhexyl) phosphate (NaDEHP), at the water/alumina interface is described. The material is found to adsorb as an essentially water-free bilayer from neutron reflection measurements. This is similar to the behavior of AOT under comparable conditions, although AOT forms a thicker, more hydrated layer. The NaDEHP shows rather little variation with added salt, but a small thickening of the layer on increasing the pH, in contrast to the behavior of AOT.

Neutron reflectivity study of the swollen structure of polyzwitterion and polyeletrolyte brushes in aqueous solution

The swollen brush structures of polycation and zwitterionic polymer brushes, such as poly(2-methacryloyloxyethyltrimethylammonium chloride) (PMTAC), poly(2-methacryloyloxyethyl phosphorylcholine) (PMPC), and poly[3-(N-2-methacryloyloxyethyl-N,N-dimethyl)ammonatopropanesulfonate] (PMAPS), in aqueous solutions of various ionic strengths were characterized by neutron reflectivity (NR) measurements. A series of the polyelectrolyte brushes were prepared by surface-initiated controlled radical polymerization on silicon substrates. A high-graft-density PMTAC brush in salt-free water (D2O) adopted a two-region step-like structure consisting of a shrunk region near the Si substrate surface and a diffuse brush region with a relatively stretched chain structure at the solution interface. The diffuse region of PMTAC was reduced with increase in salt (NaCl) concentration. The PMAPS brush in D2O formed a collapsed structure due to the strong molecular interaction between betaine groups, while significant increase in the swollen thickness was observed in salt aqueous solution. In contrast, no change was observed in the depth profile of the swollen PMPC brush in D2O with various salt concentrations. The unique solution behaviors of zwitterionic polymer brushes were described.

Neutron reflectivity of electrodeposited thin magnetic films

We present a polarised neutron reflectivity (PNR) study of magnetic/non-magnetic (CoNiCu/Cu) thin films grown by single bath electrodeposition. We find that the composition is neither homogeneous with time, nor consistent with bulk values. Instead an initial, non-magnetic copper rich layer is formed, around 2nm thick. This layer is formed by the deposition of the dilute, but rapidly diffusing, Cu2+ ions near the electrode surface at the start of growth, before the region is depleted and the deposition becomes mass transport limited. After the region has been depleted, by growth etc., this layer does not form and thus may be prevented by growing a copper buffer layer immediately preceding the magnetic layer growth. As has been previously found, cobalt deposits anomalously compared to nickel, and even inhibits Ni deposition in thin films. The layer magnetisation and average neutron scattering length are fitted independently but both depend upon the alloy composition. Thus these parameters can be used to check for model self-consistency, increasing confidence in the derived composition.

Interfacial structure of immobilized antibodies and perdeuterated HSA in model pregnancy tests measured with neutron reflectivity.

Experimental studies of antibody adsorption and antigen binding that mimicked pregnancy test immunoassays have been performed using neutron reflectivity studies of a model antibody/antigen system immobilized on the silica/water interface. The study revealed the nature of the antibody/antigen interaction and also the importance of a blocking protein, in this case human serum albumin (HSA), that enhances the immunoassay's specificity and efficiency. Of central importance to this study has been the use of a perdeuterated human serum albumin (d-HSA), providing contrast that highlights the orientation and position of the blocking agent within the adsorbed layer. It was found that the adsorbed HSA filled the gaps between the preadsorbed antibodies on the substrate, with decreased adsorption occurring as a function of increased antibody surface coverage. In addition, the antigen binding capacity of the adsorbed antibodies was investigated as a function of antibody surface coverage. The amount of specifically bound antigen was found to saturate at approximately 0.17 mg/m(2) and became independent of the antibody surface coverage. The ratio of bound antigen to immobilized antibody decreased with increased antibody surface coverage. These results are of importance for a full understanding of immunoassay systems that are widely used in clinical tests and in the detection of environmental contaminants.

Stereoselective synthesis of perdeuterated phytanic acid, its phospholipid derivatives and their formation into lipid model membranes for neutron reflectivity studies

We describe a straightforward method, for synthesis of large scale (gram quantities) of highly deuterated phytanic acid from commercially available phytol while preserving the stereochemistry around the chiral centres. The subsequent synthesis of tail-deuterated analogues of the archeabacterial membrane lipids 1,2-di(3RS,7R,11R-phytanyl)-sn-glycero-3-phosphocholine (DPEPC) and 1,2-di(3RS,7R,11R-phytanoyl)-sn-glycero-3-phosphocholine (DPhyPC) from perdeuterated phytanic acid is also described. Both lipids were employed in construction of two different model membranes, namely Langmuir monolayers and a tethered bilayer membrane (TBM) on a solid substrate, characterised by pressure area isotherm and neutron reflectometry techniques. At 10mN/m pressure the head-group thickness of both monolayers was similar while the thickness of the tail region was significantly larger for tail-deuterated DPhyPC, which was evident from a smaller area per molecule. At 20mN/m the thickness of the head and tail regions in both lipids was comparable, yet the area per molecule of tail-deuterated DPhyPC was 10% smaller than tail-deuterated DPEPC. In the TBM bilayer model membrane, the thickness of the lipid tails in both inner and outer leaflets was 8.2Å, giving a total of 16.4Å. Deuteration enabled unambiguous determination of the relative proportion of the hydrogenous tether, phospholipid and subphase.