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Small-angle scattering (SAS) is a technique commonly used in the soft matter sciences to characterize the structure of molecules self-assembling in solution. As such, it is suitable to study preparations of homogeneous liposomes to extract simple structural parameters such as radius and lipid bilayer thickness in solution. Here, we describe the preparation of liposomes for small-angle neutron scattering via extrusion and present the simplest way the resulting data can be modeled.

Decagram scale syntheses of deuterated mineral oil (d-MO) and poly(1-decene) (d-PD) were achieved by direct catalytic hydrothermal H/D exchange reaction in D2O. Nuclear magnetic resonance (NMR), mass density and small angle neutron scattering (SANS) analyses collectively revealed deuteration ratios of 54 ± 12% and 88 ± 4% for d-MO and d-PD, respectively. Further SANS analyses of polyethylene (PE) dissolved in d-MO revealed that the oil behaved as a theta solvent at temperatures of 100 and 120 °C, based on assessments of the radii of gyration and mass-fractal dimensions in the PE coils. These results suggest that PE chains in d-MO likely adopt the same coiled molecular conformations as those reported for PE in the molten state. This approach of deuterating the oil solvent, in lieu of the polymer analyte, may have far reaching applications for the SANS assessments of oil additives in polymer materials. Additionally, such deuterated oils are expected to exhibit improved stability and oxidative resistance, which will benefit many applications given the ubiquitous use of mineral oil in industry.

AlphaB crystallin (αB-crystallin) is a key protein for maintaining the long-term transparency of the eye lens. In the eye lens, αB-crystallin is a “dynamical” oligomer regulated by subunit exchange between the oligomers. To elucidate the unsettled mechanism of subunit exchange in αB-crystallin oligomers, the study was carried out at two different protein concentrations, 28.5 mg/mL (dense sample) and 0.45 mg/mL (dilute sample), through inverse contrast matching small-angle neutron scattering. Interestingly, the exchange rate of the dense sample was the same as that of the dilute sample. From analytical ultracentrifuge measurements, the coexistence of small molecular weight components and oligomers was detected, regardless of the protein concentration. The model proposed that subunit exchange could proceed through the assistance of monomers and other small oligomers; the key mechanism is attaching/detaching monomers and other small oligomers to/from oligomers. Moreover, this model successfully reproduced the experimental results for both dense and dilute solutions. It is concluded that the monomer and other small oligomers attaching/detaching mainly regulates the subunit exchange in αB-crystallin oligomer.

QUOKKA is a 40 m pinhole small-angle neutron scattering instrument in routine user operation at the OPAL research reactor at the Australian Nuclear Science and Technology Organisation. Operating with a neutron velocity selector enabling variable wavelength, QUOKKA has an adjustable collimation system providing source–sample distances of up to 20 m. Following the large-area sample position, a two-dimensional 1 m2 position-sensitive detector measures neutrons scattered from the sample over a secondary flight path of up to 20 m. Also offering incident beam polarization and analysis capability as well as lens focusing optics, QUOKKA has been designed as a general purpose SANS instrument to conduct research across a broad range of scientific disciplines, from structural biology to magnetism. As it has recently generated its first 100 publications through serving the needs of the domestic and international user communities, it is timely to detail a description of its as-built design, performance and operation as well as its scientific highlights. Scientific examples presented here reflect the Australian context, as do the industrial applications, many combined with innovative and unique sample environments.

The isothermal evolution of nanometre-sized precipitates formed in a Ti-Mo microalloyed steel through interphase precipitation has been investigated using atom probe tomography and small-angle neutron scattering. The coiling time and applied strain have been varied to observe the precipitate evolution at a constant coiling temperature of 650 °C, where various evolution parameters such as particle radius, number density, volume fraction and chemical composition have been evaluated and compared. The possibility of early stage solute clustering and its effect on precipitate formation have also been investigated. Clustering of Ti, Mo and C atoms as Ti-C and Mo-C has been observed at the shortest coiling time of 5 min. These clusters are assumed to be precursors to the carbide precipitates observed in the system, which exhibit a metastable composition, containing a carbon fraction (C/Ti+Mo ratio) in the range of 0.2–1. In particles having a Guinier radius > 3 nm, however, the average chemical composition approached the stable MC carbide stoichiometry with Ti/Mo ratio ~2.5 and C/(Ti+Mo) ratio ~0.55. This study reveals that the precipitate coarsening kinetics are very slow, with average particle diameter 10 h) in both the undeformed and deformed conditions. This is believed to be due to the reduction in equilibrium Ti content in the matrix as a result of partial replacement of Ti by Mo (Ti/Mo ratio > 2) in the precipitate lattice, in the presence of excess C in the system.

The formation of bilayer-based lyotropic liquid crystals and vesicle dispersions by phospholipids in a range of protic ionic liquids has been investigated by polarizing optical microscopy using isothermal penetration scans, differential scanning calorimetry, and small angle X-ray and neutron scattering. The stability and structure of both lamellar phases and vesicle dispersions is found to depend primarily on the underlying amphiphilic nanostructure of the ionic liquid itself. This finding has significant implica-tions for the use of ionic liquids in soft and biological materials and for biopreservation, and demon-strates how vesicle structure and properties can be controlled through selection of cation and anion. For a given ionic liquid, systematic trends in bilayer thickness, chain-melting temperature and enthalpy in-crease with phospholipid acyl chain length, paralleling behaviour in aqueous systems. Australian Research Council, ANSTO, AINSE

Measurements of a well-characterised standard sample can verify the performance of an instrument. Typically, small-angle neutron scattering instruments are used to investigate a wide range of samples and may often be used in a number of configurations. Appropriate standard samples are useful to test different aspects of the performance of hardware as well as that of the data reduction and analysis software. Measurements on a number of instruments with different intrinsic characteristics and designs in a round robin can not only better characterise the performance for a wider range of conditions but also, perhaps more importantly, reveal the limits of the current state of the art of small-angle scattering. The exercise, followed by detailed analysis, tests the limits of current understanding as well as uncovers often forgotten assumptions, simplifications and approximations that underpin the current practice of the technique. This paper describes measurements of polystyrene latex, radius 72 nm with a number of instruments. Scattering from monodisperse, uniform spherical particles is simple to calculate and displays sharp minima. Such data test the calibrations of intensity, wavelength and resolution as well as the detector response. Smoothing due to resolution, multiple scattering and polydispersity has been determined. Sources of uncertainty are often related to systematic deviations and calibrations rather than random counting errors. The study has prompted development of software to treat modest multiple scattering and to better model the instrument resolution. These measurements also allow checks of data reduction algorithms and have identified how they can be improved. The reproducibility and the reliability of instruments and the accuracy of parameters derived from the data are described., Comment: Journal of Applied Crystallography - accepted for publication

Mesoporous metals that combine catalytic activity and high surface area can provide more opportunities for electrochemical applications. Various synthetic methods, including hard and soft templating, have been developed to prepare mesoporous/nanoporous metals. Micelle assembly, typically involved in soft-templates, is flexible and convenient for such purposes. It is, however, difficult to control, and the ordering is significantly destroyed during the metal deposition process, which is detrimental when it comes to designing precisely mesostructured materials. In the present work, mesoporous Pd films were uniformly electrodeposited using a nonionic surfactant, triblock copolymer poly(ethylene oxide)-b-poly(propylene oxide)-b-poly(ethylene oxide), as a pore-directing agent. The interaction between micelles and metal precursors greatly influences the metal growth and determines the final structure. The water-coordinated species interact with the ethylene oxide moiety of the micelles to effectively drive the Pd(II) species toward the working electrode surface. From small-angle neutron scattering data, it is found that spherical P123 micelles, with an average diameter of ∼14 nm, are formed in the electrolyte, and the addition of Pd ions does not significantly modify their structure, which is the essence of the micelle assembly approach. The uniformly sized mesopores are formed over the entire mesoporous Pd film and have an average pore diameter of 10.9 nm. Cross-sectional observation of the film also shows mesopores spanning continuously from the bottom to the top of the film. The crystallinity, crystal phase, and electronic coordination state of the Pd film are also confirmed. Through this study, it is found that the optimized surfactant concentration and applied deposition potential are the key factors to govern the formation of homogeneous and well-distributed pores over the entire film. Interestingly, the as-prepared mesoporous Pd films exhibit superior electrocatalytic activity toward the ethanol oxidation reaction by fully utilizing the accessible active surface area. Our approach combines electrochemistry with colloidal and coordination chemistry and is widely applicable to other promising metals and alloy electrocatalysts.

The stable dispersion of nanoparticles in organic solvents, essential for their practical application, is often achieved by the grafting of organic dispersants. Good dispersion is usually assumed when a transparent colloidal sol is formed. However, even in such a case, the dispersion may not be in the form of distinct nanoparticles. In this study, zirconia nanoparticles have been modified with a series of carboxylic acids so that they can be dispersed in polar protic/aprotic solvents or nonpolar organic solvents as transparent colloidal sols. The dispersed state in these solvents was characterized by SAXS and SANS. In most cases, the transparent sol consists of “soft” secondary clusters generated from the ZrO2 nanoparticles. The size and fractal structure of the dispersed clusters varied with the acid modifier. Only in the linoleic acid-modified case and with chloroform as the solvent was the ZrO2 dispersed as primary particles. However, the same modification leads to secondary clusters in other solvents,...

We used neutron scattering and specific hydrogen-deuterium labeling to investigate the thermal dynamics of isotope-labeled amino acids and retinal, predominantly in the active core and extracellular moiety of bacteriorhodopsin (BR) in the purple membrane and the dynamical response to hydration. Measurements on two neutron spectrometers allowed two populations of motions to be characterized. The lower amplitude motions were found to be the same for both the labeled amino acids and retinal of BR and the global membrane. The larger amplitude dynamics of the labeled part, however, were found to be more resilient than the average membrane, suggesting their functional importance. The response to hydration was characterized, showing that the labeled part of BR is not shielded from hydration effects. The results suggest that the inhibition of high-amplitude motions by lowering hydration may play a key role in the slowing down of the photocycle and the proton pumping activity of BR.

The function and dynamics of proteins depend on their direct environment, and much evidence has pointed to a strong coupling between water and protein motions. Recently however, neutron scattering measurements on deuterated and natural-abundance purple membrane (PM), hydrated in H2O and D2O, respectively, revealed that membrane and water motions on the ns–ps time scale are not directly coupled below 260 K (Wood et al. in Proc Natl Acad Sci USA 104:18049–18054, 2007). In the initial study, samples with a high level of hydration were measured. Here, we have measured the dynamics of PM and water separately, at a low-hydration level corresponding to the first layer of hydration water only. As in the case of the higher hydration samples previously studied, the dynamics of PM and water display different temperature dependencies, with a transition in the hydration water at 200 K not triggering a transition in the membrane at the same temperature. Furthermore, neutron diffraction experiments were carried out to monitor the lamellar spacing of a flash-cooled deuterated PM stack hydrated in H2O as a function of temperature. At 200 K, a sudden decrease in lamellar spacing indicated the onset of long-range translational water diffusion in the second hydration layer as has already been observed on flash-cooled natural-abundance PM stacks hydrated in D2O (Weik et al. in J Mol Biol 275:632–634, 2005), excluding thus a notable isotope effect. Our results reinforce the notion that membrane-protein dynamics may be less strongly coupled to hydration water motions than the dynamics of soluble proteins.

The dynamical coupling between proteins and their hydration water is important for the understanding of macromolecular function in a cellular context. In the case of membrane proteins, the environment is heterogeneous, composed of lipids and hydration water, and the dynamical coupling might be more complex than in the case of the extensively studied soluble proteins. Here, we examine the dynamical coupling between a biological membrane, the purple membrane (PM), and its hydration water by a combination of elastic incoherent neutron scattering, specific deuteration, and molecular dynamics simulations. Examining completely deuterated PM, hydrated in H 2 O, allowed the direct experimental exploration of water dynamics. The study of natural abundance PM in D 2 O focused on membrane dynamics. The temperature-dependence of atomic mean-square displacements shows inflections at 120 K and 260 K for the membrane and at 200 K and 260 K for the hydration water. Because transition temperatures are different for PM and hydration water, we conclude that ps–ns hydration water dynamics are not directly coupled to membrane motions on the same time scale at temperatures

Hydration water is the natural matrix of biological macromolecules and is essential for their activity in cells. The coupling between water and protein dynamics has been intensively studied, yet it remains controversial. Here we combine protein perdeuteration, neutron scattering and molecular dynamics simulations to explore the nature of hydration water motions at temperatures between 200 and 300 K, across the so-called protein dynamical transition, in the intrinsically disordered human protein tau and the globular maltose binding protein. Quasi-elastic broadening is fitted with a model of translating, rotating and immobile water molecules. In both experiment and simulation, the translational component markedly increases at the protein dynamical transition (around 240 K), regardless of whether the protein is intrinsically disordered or folded. Thus, we generalize the notion that the translational diffusion of water molecules on a protein surface promotes the large-amplitude motions of proteins that are required for their biological activity., Hydration water plasticizes protein structures and is essential for their biological functions, such as enzymatic catalysis. Here, the authors use neutron scattering and molecular dynamics simulations to study hydration water at the dynamical transition of folded and disordered proteins.

There is increasing interest in the contribution of methyl groups to the overall dynamics measured by neutron scattering experiments of proteins. In particular an inflection observed in atomic mean square displacements measured as a function of temperature on high resolution spectrometers (similar to 1 mu eV) was explained by the onset of methyl group rotations. By specifically labeling a non-methyl-containing side-chain in a native protein system, the purple membrane, and performing neutron scattering measurements, we here provide direct experimental evidence that the observed inflection is indeed due to methyl group rotations. Molecular dynamics simulations reproduce the experimental data, and their analysis suggests that the apparent transition is due to methyl group rotation entering the finite instrumental resolution of the spectrometer. Methyl group correlation times measured by solid state NMR in the purple membrane, taken from previous work, support the interpretation.

An integrated picture of hydration shell dynamics and of its coupling to functional macromolecular motions is proposed from studies on a soluble protein, on a membrane protein in its natural lipid environment, and on the intracellular environment in bacteria and red blood cells. Water dynamics in multimolar salt solutions was also examined, in the context of the very slow water component previously discovered in the cytoplasm of extreme halophilic archaea. The data were obtained from neutron scattering by using deuterium labelling to focus on the dynamics of different parts of the complex systems examined.

Intracellular water dynamics in Haloarcula marismortui , an extremely halophilic organism originally isolated from the Dead Sea, was studied by neutron scattering. The water in centrifuged cell pellets was examined by means of two spectrometers, IN6 and IN16, sensitive to motions with time scales of 10 ps and 1 ns, respectively. From IN6 data, a translational diffusion constant of 1.3 × 10 −5 cm 2 s −1 was determined at 285 K. This value is close to that found previously for other cells and close to that for bulk water, as well as that of the water in the 3.5 M NaCl solution bathing the cells. A very slow water component was discovered from the IN16 data. At 285 K the water-protons of this component displays a residence time of 411 ps (compared with a few ps in bulk water). At 300 K, the residence time dropped to 243 ps and was associated with a translational diffusion of 9.3 × 10 −8 cm 2 s −1 , or 250 times lower than that of bulk water. This slow water accounts for ≈76% of cell water in H. marismortui . No such water was found in Escherichia coli measured on BSS, a neutron spectrometer with properties similar to those of IN16. It is hypothesized that the slow mobility of a large part of H. marismortui cell water indicates a specific water structure responsible for the large amounts of K + bound within these extremophile cells.