Small-angle Neutron Scattering Profiles Research Articles

Alcohol-Induced Denaturation of Hen Egg White Lysozyme Studied by Infrared, Circular Dichroism, and Small-Angle Neutron Scattering.

In aqueous binary solvents with fluorinated alcohols, 2,2,2-trifluoroethanol (TFE) and 1,1,1,3,3,3-hexafluoroisopropanol (HFIP), and aliphatic alcohols, ethanol (EtOH) and 2-propanol (2-PrOH), the denaturation of hen egg white lysozyme (HEWL) with increasing alcohol mole fraction xA has been investigated in a wide view from the molecular vibration to the secondary and ternary structures. Circular dichroism (CD) measurement showed that the secondary structure of α-helix content of HEWL increases on adding a small amount of the fluorinated alcohol to the aqueous solution, while the β-sheet content decreases. On the contrary, the secondary structure does not significantly change by the addition of the aliphatic alcohols. Correspondingly, the infrared (IR) spectroscopic measurements revealed that the amide I band red-shifts on the addition of the fluorinated alcohol. However, the band remains unchanged in the aliphatic alcohol systems with increasing alcohol content. To observe the ternary structure of HEWL, small-angle neutron scattering (SANS) experiments with H/D substitution technique have been applied to the HEWL solutions. The SANS experiments were successful in revealing the details of how the geometry of the HEWL changes as a function of xA. The SANS profiles indicated the spherical structure of HEWL in all of the alcohol systems in the xA range examined. The mean radius of HEWL in the two fluorinated alcohol systems increases from ∼16 to ∼18 Å during the change in the secondary structure against the increase in the fluorinated alcohol content. On contrast, the radius does not significantly change in both aliphatic alcohol systems below xA = 0.3 but expands to ∼19 Å as the alcohol content is close to the limitation of the HEWL solubility. According to the present results, together with our knowledge of the alcohol cluster formation and the interaction of the trifluoromethyl (CF3) groups with the hydrophobic moieties of biomolecules, the effects of alcohols on the denaturation of the protein have been discussed on a molecular scale.

Contrast dependence of scattering profiles for poly(ethylene glycol) in water: Investigation by small-angle neutron scattering with 3He spin filter and small-angle x-ray scattering.

The small-angle neutron scattering (SANS) and small-angle x-ray scattering (SAXS) measurements were performed for deuterated and non-deuterated poly(ethylene glycol) (d-PEG and h-PEG, respectively) in D2O and a D2O/H2O mixed solvent (Mix) to compare the scattering profiles. To determine the coherent scattering intensity of SANS, a 3He spin filter was utilized. The scattering profiles determined by the SANS measurements were analyzed in terms of the wormlike chain model with touched beads along the contour of the chain. However, the SAXS profiles were not explained by the same model with uniform beads but with beads each consisting of a core and a shell having different electron densities. To explore the chain thickness determined from the SANS profile, the scattering intensities for different combinations of d-PEG/D2O, d-PEG/Mix, h-PEG/D2O, and h-PEG/Mix were also examined.

Structure of Single-Chain Nanoparticles under Crowding Conditions: A Random Phase Approximation Approach.

The conformation of poly(methyl methacrylate) (PMMA)-based single-chain nanoparticles (SCNPs) and their corresponding linear precursors in the presence of deuterated linear PMMA in deuterated dimethylformamide (DMF) solutions has been studied by small-angle neutron scattering (SANS). The SANS profiles were analyzed in terms of a three-component random phase approximation (RPA) model. The RPA approach described well the scattering profiles in dilute and crowded solutions. Considering all the contributions of the RPA leads to an accurate estimation of the single chain form factor parameters and the Flory-Huggins interaction parameter between PMMA and DMF. The value of the latter in the dilute regime indicates that the precursors and the SCNPs are in good solvent conditions, while in crowding conditions, the polymer becomes less soluble.

Water distribution in human hair microstructure elucidated by spin contrast variation small-angle neutron scattering

Dynamic nuclear polarization (DNP) is effective for controlling the neutron scattering length of protons and can be utilized for contrast variation in small-angle neutron scattering (SANS). Using the TEMPOL solution soaking method as electron spin doping, the DNP–SANS technique was applied to human hair fiber for the first time. For dry and D2O-swollen hair samples, a drastic change in the SANS profile was observed at high polarization conditions (|P H P N| ∼ 60%, where P H and P N are the proton and neutron spin polarization, respectively). The SANS profile as a function of the magnitude of the scattering vector, q, was composed of a low-q upturn, a middle-q oscillation and a high-q flat region. The low-q upturn was assumed to be a combination of two power-law functions, q −4 due to a large structure interface (Porod's law) and q −2 due to random coil. The middle-q oscillation was well reproduced by numerical calculation based on the structure model of intermediate filaments (IFs) as proposed by Er Rafik et al. [Biophys. J. (2004), 86, 3893–3904]: one pair of keratin coiled-coils is located at the center and surrounded by seven pairs of keratin coiled-coils located in a circle (called the `7 + 1' model), and a collection of IFs is arranged in a quasi-hexagonal manner. For the observed SANS profiles for different P H P N, the IF term contribution maintained a constant q-dependent profile, despite significant changes in intensity. This indicates that the macrofibril is composed of two domains (keratin coiled-coils and matrix). In addition, D2O swelling enhanced the IF term intensity and shifted the polarization-dependent local minimum to higher P H P N. This behavior was reproduced by contrast factor calculation based on the two-domain model. Scattering length densities of keratin coiled-coil and surrounding matrix domains were calculated by use of the known amino acid composition, considering the hydrogen–deuterium exchange reaction during soaking with D2O solution of TEMPOL. As a result, it was found that for keratin coiled-coil domains, about 40% of the peptide backbone amide NH protons were replaced with deuterons. This means that 68% of the α-helix domain is rigid, but the rest is flexible to allow dynamic dissociation of the hydrogen bond. Furthermore, the local mass density of each domain was precisely evaluated. The obtained data are expected to be a guide for further detailed investigation of keratin and keratin-associated protein distribution. This approach is expected to be applied to a wide variety of bio-derived materials, which are water absorbing in general.

Biophysical characterization of calcium-binding and modulatory-domain dynamics in a pentameric ligand-gated ion channel

Pentameric ligand-gated ion channels (pLGICs) perform electrochemical signal transduction in organisms ranging from bacteria to humans. Among the prokaryotic pLGICs, there is architectural diversity involving N-terminal domains (NTDs) not found in eukaryotic relatives, exemplified by the calcium-sensitive channel (DeCLIC) from a Desulfofustis deltaproteobacterium, which has an NTD in addition to the canonical pLGIC structure. Here, we have characterized the structure and dynamics of DeCLIC through cryoelectron microscopy (cryo-EM), small-angle neutron scattering (SANS), and molecular dynamics (MD) simulations. In the presence and absence of calcium, cryo-EM yielded structures with alternative conformations of the calcium-binding site. SANS profiles further revealed conformational diversity at room temperature beyond that observed in static structures, shown through MD to be largely attributable to rigid-body motions of the NTD relative to the protein core, with expanded and asymmetric conformations improving the fit of the SANS data. This work reveals the range of motion available to the DeCLIC NTD and calcium-binding site, expanding the conformational landscape of the pLGIC family. Further, these findings demonstrate the power of combining low-resolution scattering, high-resolution structural, and MD simulation data to elucidate interfacial interactions that are highly conserved in the pLGIC family.

Contrast Variation Small-Angle Neutron Scattering Study of Solubilization of Perfumes in Cationic Surfactant Micelles.

Perfume solubilization is an important process in the production of commercial products such as beverages, foods, and cosmetics. In the present study, small-angle neutron scattering (SANS) experiments were performed to investigate the solubilization behavior of perfumes in cetyltrimethylammonium bromide (CTAB) micelles. The solubilization of linalool (LL) and l-menthol (MT), which are relatively hydrophilic perfumes, did not change the size of the CTAB micelles although the perfumes were incorporated in the micelles, as indicated by a decrease in scattering length density. On the other hand, the solubilization of d-limonene (LN), a hydrophobic perfume, led to the swelling of CTAB micelles. An internal contrast variation SANS study was performed by the deuteration of CTAB molecules to directly observe the perfumes in the micelles. The radius of d-CTAB micelles solubilizing LL or MT corresponds to that of h-CTAB, which indicates that these perfumes are accommodated in the palisade layers of the micelles and are homogeneously distributed in the micelles. On the other hand, LN formed small droplets, as indicated by the SANS profile, which implies the solubilization of LN molecules in the core of the CTAB micelles. We found that the relatively hydrophilic perfumes (LL and MT) show less impact on the sizes of the cationic micelles in comparison to nonionic micelles. Thus, the internal contrast variation method of SANS allowed the direct observation of the solubilization sites of perfumes with different hydrophilicity-hydrophobicity balances. This method is a powerful tool to determine the solubilization states that affect the solubilization capacity, volatilization, or release speed of perfumes.

A new simultaneous measurement system of wide Q-range small angle neutron scattering combined with polarized Fourier transform infrared spectroscopy.

Small angle neutron scattering (SANS) is a versatile and convenient method to investigate the higher order structure of molecular assembly systems. However, the more complicated a system of interest, the more difficult the interpretation in the SANS profile. In order to increase the reliability of structural analysis on a complicated system, it is desirable to obtain different kinds of structural information from the same sample simultaneously. Polarized infrared spectroscopy is able to provide information about the molecular structure, concentration, and orientation of each chemical species in a system. In order to utilize these advantages of polarized infrared spectroscopy, a simultaneous measurement system was built by incorporating a Fourier transform infrared (FTIR) spectrometer into a time-of-flight small angle neutron scattering instrument covering a wide Q range. Using this system, simultaneous measurements of wide- and small-angle neutron scattering and polarized FTIR spectroscopy was realized for the first time.

Imidazolium-Based Grafted Anion Exchange Membranes: Interplay between the Morphology and Anion Transport Behavior

Anion-exchange membranes (AEMs) have been regarded as an alternative to proton-exchange membranes (PEMs) in energy conversion devices, due to the advantage of saving expensive platinum catalysts. However, neither the molecular design nor the property understanding is sufficient for developing AEMs capable of practical fuel cell applications. It is crucial to thoroughly study the current AEMs in terms of microphase structures and conducting properties. Recently, we developed a new graft type of AEMs by radiation-induced grafting method, where imidazolium and styrene monomers were grafted into poly(ethylene-co-tetrafluoroethylene) (ETFE) base films under a dose of 80 kGy from the 60Co γ-ray source (QST, Takasaki), followed by alkylation and ion exchange reactions. These AEMs exhibit well-balanced properties of high ion conductivity (> 100 mS/cm at 80oC) and good stability, which are believed to be controlled by the microphase structure of the membrane. In this report, we investigate the memrbane structure and reveal the relationship between structure and properties, using small-angle neutron scattering (SANS) method. Fig. 1 shows typical SANS intensity profiles, I(q), of pristine ETFE films (profile 1), grafted-ETFE membranes (profile 2), dry AEM (profile 3) and AEM equilibrated in D2O (profile 4) as a function of scattering vector q. The different morphologies which can be deduced from the SANS profiles are also shown in the figure. The experiments confirm the semi-crystalline feature of AEM conserved from the original ETFE base film, which is the key factor for the high membrane mechanical stability, and show the dependence of the structure on hydration. Contrast variation experiments performed on AEM equilibrated in water mixtures of H2O and D2O, further show that graft polymers and water consist the hydrophilic ion channels [1]. Since graft polymers distribute randomly in AEM, well-connected hydrophilic ion channels are feasibly formed, which is the key factor for the high membrane conductivity. Above all, the interplay between the morphology and properties can be characterized as follows: 1) semi-crystalline base film offers good mechanical properties; 2) graft polymers and water form ion channels to promote the conductivity. Figure 1

Structural investigation of hectorite aqueous suspensions by dielectric microscopy and small-angle neutron scattering coupling with rheological measurement

Abstract The microstructure of synthetic hectorite aqueous suspensions was investigated by scanning electron-assisted dielectric microscopy (SE-ADM) and small angle neutron scattering (SANS) coupled with rheological measurements (Rheo-SANS). The direct observations of the aqueous suspensions by SE-ADM confirmed the array structure of the hectorite discs with ~100–300 nm scale periodic length. The suspensions demonstrated a typical thixotropic flow curve with hysteresis. No change in the SANS profiles at high shear rate (1000 s−1) was observed by the Rheo-SANS. These results suggested that the array was destroyed by the shear rate but the microstructure in the arrays was preserved in flow condition.

Energy-resolved small-angle neutron scattering from steel

Recent progress of pulsed neutron sources has enabled energy-resolved analysis of neutron attenuation spectra, which include information on neutron scattering. In this study, a new analysis technique for small-angle neutron scattering (SANS) is demonstrated. A clear difference is observed in the neutron attenuation spectra between steels with different nanostructures; this difference can be understood as arising from attenuation due to SANS. The neutron attenuation spectra calculated from the corresponding SANS profiles agree well with the experimentally observed attenuation spectra. This result indicates that measurement of neutron attenuation spectra may enable the development of a novel experimental technique,i.e.energy-resolved SANS.

Morphological Study of Poly(vinylbenzyl chloride)-Grafted Poly(ethylene-co-tetrafluoroethylene) [ETFE-g-PVBC] Films Using Small-Angle Neutron Scattering Analysis.

In this study, the effect of degree of the grafting and crosslinking on the morphology of the crystalline domain in poly(vinylbenzyl chloride)-grafted poly(ethylene-co-tetrafluoroethylene) [ETFE-g-PVBC] films was investigated using a SANS (small-angle neutron scattering) analysis. The grafted films can be used as a precursor for ion-exchange membrane. ETFE-g-PVBC films with various degrees of cross-linking were prepared by a simultaneous irradiation grafting of vinylbenzyl chloride (VBC) and divinylbenzene (DVB) onto an ETFE film. The SEM-EDX (scanning electron microscopy-energy dispersive X-ray spectroscopy) results of a cross-sectional distribution of ETFE-g-PVBC films showed that the chlorine atoms were well-distributed throughout the films. SANS profiles of the PVBC-grafted films in the absence of a DVB crosslinker showed that the crystalline domain peaks were observed and the peak maximum position shifted significantly from 0.032 Å-(-1) to 0.02 Å(-1) with an increase in the degree of grafting. However, peak maximum positions of the PVBC-grafted films in the presence of a DVB crosslinker shifted slightly from 0.02 Å(-1) to 0.024 Å(-1) with an increase in the amount of DVB monomer at same degree of grafting. These results indicate that the degree of grafting and crosslinking affect the morphology of the crystalline domain in the ETFE-g-PVBC films.

Small-angle neutron scattering study of specific interaction and coordination structure formed by mono-acetyl-substituted dibenzo-20-crown-6-ether and cesium ions

This study uses small-angle neutron scattering (SANS) to elucidate the coordination structure of the complex of mono-acetyl-substituted dibenzo-20-crown-6-ether (ace-DB20C6) with cesium ions (Cs+). SANS profiles obtained for the complex of ace-DB20C6 and Cs+ (ace-DB20C6/Cs) in deuterated dimethyl sulfoxide indicated that Cs+ coordination resulted in a more compact structure than the free ace-DB20C6. The data were fitted well with SANS profiles calculated using Debye function for scattering on an absolute scattering intensity scale. For this theoretical calculation of the scattering profiles, the coordination structure proposed based on density functional theory calculation was used. Consequently, we conclude that the SANS analysis experimentally supports the proposed coordination structure of ace-DB20C6/Cs and suggests the following: (1) the complex of ace-DB20C6 and Cs+ is formed with an ace-DB20C6/Cs molar ratio of 1/1 and (2) the two benzene rings of ace-DB20C6 fold around Cs+ above the center of the crown ether ring of ace-DB20C6.

Quantitative Analysis of Cementite Spheroidization in Pearlite by Small-Angle Neutron Scattering

Small-angle neutron scattering (SANS) was used to investigate the spheroidization of cementite in pearlite upon annealing. Globally averaged information on the shape and size of cementite particles was collected by using the SANS profile analysis of 0.8 mass pct C pearlitic steel (0.8C) samples annealed at 973 K (700 °C) for 3.6 to 86.4 ks. The change in the total area of the ferrite–cementite interface or aspect ratio determined by ex situ SANS exhibits excellent correspondence with scanning electron microscopy observations. Then, in situ SANS data were collected during isothermal annealing of the 0.8C steel and the commercially available 0.45C carbon steel (JIS-S45C). The shape change of the cementite plate was monitored, and the spheroidization rate of the 0.8C steel was found to be faster than that of the 0.45C steel.

Small-angle neutron scattering observation of the surface structure of Ti0.31Cr0.33V0.36 alloy in hydrogenation

Abstract The surface structure of Ti 0.31 Cr 0.33 V 0.36 , which is an excellent hydrogen absorbing alloy, was directly observed in the first absorption–desorption cycle of hydrogen by small-angle neutron scattering (SANS). In addition, characterizations by pressure–composition isotherm ( P – C – T ) measurements and X-ray diffraction (XRD) were also carried out. The results of these experiments revealed the relationship between the slope of a SANS profile ( α ) and the hydrogen content (H/M). We suggest that the variation of α with H/M can be understood by considering the volume expansion of the surface during hydrogenation, which may exercise a significant role in the change from rough to smooth surface.

Spin Contrast Variation Study of Fuel-efficient Tire Rubber

The scattering length of a proton against a polarized neutron depends strongly on the polarization of proton spins (PH). This dependence can be utilized for contrast variation in small angle neutron scattering (SANS). We applied this spin contrast variation technique to a silica-filled SBR rubber specimen, which is widely used for tread rubber of fuel-efficient tires. For realizing high PH, we used dynamic nuclear polarization (DNP) technique, in which large polarization of electron spin at low temperature and high magnetic field is transferred to proton spin by microwave irradiation with a tuned frequency. As this electron spin source, we doped stable radical TEMPO (2,2,6,6-tetramethyl piperidine 1-oxyl) into the rubber sample by use of a vapor sorption technique. For the TEMPO-doped rubber sample, SANS measurements were conducted at PH = −20%, 0%, and +13%, with almost fully polarized neutron beam %) with its wavelength of 6.5±0.6 Å. The SANS profile clearly changed as a function of PH, which can be explained by the PH dependence of the neutron scattering length densities of the main three components (SBR, silica and zinc oxide). By a linear transformation of the profiles obtained at the three different PH values, we successfully determined the partial scattering function of silica, which reflects the aggregation of silica particles.

Structural Transitions of CTAB Micelles in a Protic Ionic Liquid

Micellar solutions of hexadecyltrimethylammonium bromide (CTAB) in a protic ionic liquid, ethylammonium nitrate (EAN), are studied by shear rheology, polarizing optical microscopy (POM), conductivity measurements, and small angle neutron scattering (SANS). Three concentration regimes are examined: A dilute regime (with concentrations [CTAB] < 5 wt %) consisting of noninteracting spherical micelles, a semidilute regime (5 wt % ≤ [CTAB] ≤ 45 wt %) where micelles interact via electrostatic repulsions, and a concentrated regime (45 wt % < [CTAB] ≤ 62 wt %) where a reversible, temperature-dependent isotropic (L(1)) to hexatic (Hex) phase transition is observed. The L(1)-Hex transition, which has been predicted but not previously observed, is characterized by (1) a sharp increase in the shear viscosity, (2) the formation of focal conical birefringence textures (observed by POM), and (3) enhancement of the crystalline order, evidenced by the appearance of Bragg reflections in the SANS profiles. Ionic conductivity is not sensitive to the L(1)-Hex transition, which corroborates the absence of topological transitions.

A Structural Study of an Organogel Investigated by Small-Angle Neutron Scattering and Synchrotron Small-Angle X-ray Scattering

We investigated structures of 12-hydroxystearic acid (12-HSA) gels in a wide temperature range from the gel to the sol states and the gelation process by means of small-angle neutron scattering (SANS) and time-resolved synchrotron small-angle X-ray scattering (SAXS), respectively. The SANS result shows that the size of the cross section of the crystalline fibers in the gel was not affected much by temperature change, although the density of the fibers decreased with the increase of temperature even in the gel state. The SANS profiles at various temperatures and compositions were well reduced by the product of the volume fraction of the fiber and the square of the scattering contrast. At temperatures above the melting point of the gels obtained from differential scanning calorimeter, the gel turned into a homogeneous solution. The time-resolved small-angle X-ray scattering experiments revealed the gelation process as mentioned below. At the early stage, the scattering behavior did not show any change (induction period). The induction period was longer at higher temperatures. In the second stage, an amorphous precursor appeared as an initial structure of crystalline nucleus. In the next stage, the fiber was developed via crystalline nucleation and growth. In this stage, the fibers grew with keeping the size of the cross section (the radius of gyration R ≅ 82 -84 Å) constant, and finally, the fiber growth was arrested because of gelation.

Anomaly in SANS χ for polydisperse polystyrene-b-poly(isooctyl acrylate)

Small-angle neutron scattering (SANS) measurements have been performed on a disordered block copolymer from deuterated polystyrene (dPS) and self-adhesive poly(isooctyl acrylate) (POA) in order to elicit the effective Flory–Huggins χ, which carries the essence of the copolymer phase behavior. The copolymer sample for the measurement was prepared by blending two polydisperse dPS-b-POAs of different molecular weights, where the overall average size of the blend was low enough to ensure to be in the mean-field region but high enough to have discernible scattering intensities. The SANS profiles for the copolymer were fitted to Leibler's scattering function for a polydisperse copolymer system described by Schulz-Zimm distribution. The resultant χ as a function of inverse temperature was shown to have a strong entropic contribution and a weak enthalpic contribution. By adopting Sanchez-Balasz or ten Brinke-Karasz-type simple analysis for specific interactions, it was found that the entropically dominated χ for dPS-b-POA arises from the steric hindrance of long alkyl side groups of POA.

Super-Efficient Surfactant for Stabilizing Water-in-Carbon Dioxide Microemulsions

The fluorinated double-tailed glutarate anionic surfactant, sodium 1,5-bis[(1H,1H,2H,2H-perfluorodecyl)oxy]-1,5-dioxopentane-2-sulfonate (8FG(EO)(2)), was found to stabilize water-in-supercritical CO(2) microemulsions with high water-to-surfactant molar ratios (W(0)). Studies were carried out here to obtain detailed information on the phase stability and nanostructure of the microemulsions by using a high-pressure UV-vis dye probe and small-angle neutron scattering (SANS) measurements. The UV-vis spectra, with methyl orange as a reporter dye, indicated a maximum attainable W(0) of 60 at 45 and 75 °C, and SANS profiles indicated regular droplet swelling with a linear relationship between the water core nanodroplet radius and W(0). This represents the highest water solubilization reported to date for any water-in-CO(2) microemulsion. Further analysis of the SANS data indicated critical packing parameters for 8FG(EO)(2) at the microemulsion interface >1.34, representing approximately 1.1 times the value for common aerosol-OT in water-in-heptane microemulsions under equivalent conditions.

2D-Ising-like critical behavior in mixtures of water and 3-methylpyridine including antagonistic salt or ionic surfactant

The effect of an antagonistic salt on the phase behavior and nanoscale structure of a mixture of D2O and 3-methylpyridine was investigated by visual inspection and small-angle neutron scattering (SANS). The addition of the antagonistic salt, namely sodium tetraphenylborate (NaBPh4), induces the shrinking of the two-phase region in contrast to the case in which a normal (hydrophilic) salt is added. Below the phase separation point, the SANS profiles cannot be described by the Ornstein–Zernike function owing to the existence of a long-range periodic structure. With increasing salt concentration, the critical exponents change from the values of 3D-Ising and approach those of 2D-Ising. These results suggest that the concentration fluctuation of the mixture of solvents is limited to a quasi two-dimensional space by the periodic structure induced by the adding the salt. The same behaviors were also observed in mixtures composed of water, 3-methylpyridine, and ionic surfactant.