Contrast Match Point Research Articles

Deuteration of non-labile protium in starch: Biosynthesis and characterisation from yeast-derived starch granules

Deuterium labelling of the non-labile protium atoms in starch granules has been achieved for the first time, by growing genetically modified yeast on deuterated media. Mass spectrometry of the glucose monomers from digested starch showed 44 % average deuteration of the non-labile protium when grown on partially deuterated raffinose (with average deuteration 48 %); yielding starch with 26 % average overall deuteration. Non-labile deuteration was also demonstrated using D2O solvent in the culture medium. Solid-state NMR revealed that deuteration was not evenly distributed across the monomer, being highest at the C6 carbon and lowest at the C1 carbon. SANS revealed two structural features at q = 0.05 Å−1 and 0.4 Å−1, the first corresponding to a lamellar repeat of approximately 12–13 nm while the latter is consistent with B-type crystalline polymer packing. Furthermore, solvent contrast variation SANS analysis yielded a contrast match point of 66 mol% D2O indicative of approximately 30–35 % average deuteration of the bulk granules, consistent with mass spectroscopy. When coupled with the more traditional process of exchange of labile protium in the hydroxyl groups by D2O solvent exchange, the biosynthesis of highly deuterated starch opens new opportunities for neutron scattering experiments involving multicomponent starch-based systems.

Small angle neutron scattering and lipidomic analysis of a native, trimeric PSI-SMALP from a thermophilic cyanobacteria

The use of styrene-maleic acid copolymers (SMAs) to produce membrane protein-containing nanodiscs without the initial detergent isolation has gained significant interest over the last decade. We have previously shown that a Photosystem I SMALP from the thermophilic cyanobacterium, Thermosynechococcus elongatus (PSI-SMALP), has much more rapid energy transfer and charge separation in vitro than detergent isolated PSI complexes. In this study, we have utilized small-angle neutron scattering (SANS) to better understand the geometry of these SMALPs. These techniques allow us to investigate the size and shape of these particles in their fully solvated state. Further, the particle's proteolipid core and detergent shell or copolymer belt can be interrogated separately using contrast variation, a capability unique to SANS. Here we report the dimensions of the Thermosynechococcus elongatus PSI-SMALP containing a PSI trimer. At ~1.5 MDa, PSI-SMALP is the largest SMALP to be isolated; our lipidomic analysis indicates it contains ~1300 lipids/per trimeric particle, >40-fold more than the PSI-DDM particle and > 100 fold more than identified in the 1JB0 crystal structure. Interestingly, the lipid composition to the PSI trimer in the PSI-SMALP differs significantly from bulk thylakoid composition, being enriched ~50 % in the anionic sulfolipid, SQDG. Finally, utilizing the contrast match point for the SMA 1440 copolymer, we also can observe the ~1 nm SMA copolymer belt surrounding this SMALP for the first time, consistent with most models of SMA organization.

Structure of Diisobutylene Maleic Acid Copolymer (DIBMA) and Its Lipid Particle as a "Stealth" Membrane-Mimetic for Membrane Protein Research.

The study of membrane proteins remains challenging, especially in a native membrane environment. Recently, major progress has been made using maleic acid copolymers, such as styrene maleic acid, to purify membrane proteins and study them directly with native lipids associated with the membrane. Additional maleic acid copolymers, such as diisobutylene maleic acid (DIBMA) membrane-mimetic systems, are being developed and found to have improved spectroscopic properties and pH stability. We studied DIBMA and its lipid particles in solution to better understand its assembly, without and with the lipids, to provide an insight regarding how to use it in solution for better membrane extraction. Using small-angle neutron and X-ray scattering (SANS/SAXS), we show that DIBMA organizes into structures of different size scales at various concentrations and ionic strengths. The polymer performed reasonably well under most solvent conditions except in very low concentrations and high-salt conditions that could result in limited interaction with lipids. To explore DIBMA lipid particles as a suitable membrane-mimetic system for neutron scattering studies of membrane proteins, we measured and determined the contrast-matching point of DIBMA to be ∼12% (v/v) D2O - similar to that of most protiated lipid molecules but distinct from that of regular protiated proteins - providing a natural contrast for separating their neutron scattering signals. Using SANS contrast variation, we demonstrated that the scattering from the whole lipid particle can be annihilated. Further, we determined that a well-defined lipid nanodisc structure with DIBMA was contrast-matched. These results demonstrate that the DIBMA lipid particle is an outstanding "stealth" membrane-mimetic for membrane proteins. The results provide a structural framework for understanding the organization and assembly process of the polymer itself and the lipid molecules. Such an understanding is imperative for structural techniques such as cryo-electron microscopy, nuclear magnetic resonance, small-angle scattering, and other biophysical techniques.

Effects of soil particles and convective transport on dispersion and aggregation of nanoplastics via small-angle neutron scattering (SANS) and ultra SANS (USANS).

Terrestrial nanoplastics (NPs) pose a serious threat to agricultural food production systems due to the potential harm of soil-born micro- and macroorganisms that promote soil fertility and ability of NPs to adsorb onto and penetrate into vegetables and other crops. Very little is known about the dispersion, fate and transport of NPs in soils. This is because of the challenges of analyzing terrestrial NPs by conventional microscopic techniques due to the low concentrations of NPs and absence of optical transparency in these systems. Herein, we investigate the potential utility of small-angle neutron scattering (SANS) and Ultra SANS (USANS) to probe the agglomeration behavior of NPs prepared from polybutyrate adipate terephthalate, a prominent biodegradable plastic used in agricultural mulching, in the presence of vermiculite, an artificial soil. SANS with the contrast matching technique was used to study the aggregation of NPs co-dispersed with vermiculite in aqueous media. We determined the contrast match point for vermiculite was 66% D2O / 33% H2O. At this condition, the signal for vermiculite was ~50-100%-fold lower that obtained using neat H2O or D2O as solvent. According to SANS and USANS, smaller-sized NPs (50 nm) remained dispersed in water and did not undergo size reduction or self-agglomeration, nor formed agglomerates with vermiculite. Larger-sized NPs (300-1000 nm) formed self-agglomerates and agglomerates with vermiculite, demonstrating their significant adhesion with soil. However, employment of convective transport (simulated by ex situ stirring of the slurries prior to SANS and USANS analyses) reduced the self-agglomeration, demonstrating weak NP-NP interactions. Convective transport also led to size reduction of the larger-sized NPs. Therefore, this study demonstrates the potential utility of SANS and USANS with contrast matching technique for investigating behavior of terrestrial NPs in complex soil systems.

Structural Properties of Janus Particles with Nano- and Mesoscale Anisotropy.

Synthesis of anisotropic Janus particles (AnJPs) is crucial for understanding the fundamental principles behind non-equilibrium self-organization of cells, bacteria, or enzymes, and for the design of novel multicomponent carriers for guided self-assembly, drug delivery or molecular imaging. Their catalytic activity, as well as many other chemical and physical properties are intimately related to the nano- and mesoscale structure. An efficient and fast in situ monitoring of the structural changes involves non-destructive techniques which can probe macroscopic volumes of multicomponent systems, such as small-angle scattering (SAS). However, the interpretation of scattering data is often a difficult task since the existing models deal only with symmetric AnJPs, thus greatly restricting their applicability. Here, a general theoretical framework is developed, which describes scattering from a system containing randomly oriented and placed two-phase AnJPs with arbitrarily tunable geometric and chemical asymmetries embedded in a solution/matrix of different chemical composition. This approach allows an analytic description of the contrast matching point, and it is shown that the interplay between the scattering curves of the two phases gives rise to a rich scaling behavior which allows extracting structural information about each individual phase. To illustrate the above findings, analytic expression for the scattering curves of asymmetric AnJPs are derived, and the results are validated by Monte-Carlo simulations. The broad general features of the scattering curves are explained by using a simple scaling approach which allows gaining more physical insight into the scattering processes as well as for the interpretation of SAS intensity.

Designing Mixed Detergent Micelles for Uniform Neutron Contrast.

Micelle-forming detergents provide an amphipathic environment that mimics lipid bilayers and are important tools used to solubilize and stabilize membrane proteins in solution for in vitro structural investigations. Small-angle neutron scattering (SANS) at the neutron contrast match point of detergent molecules allows observing the signal from membrane proteins unobstructed by contributions from the detergent. However, we show that even for a perfectly average-contrast matched detergent there arises significant core-shell scattering from the contrast difference between aliphatic detergent tails and hydrophilic head groups. This residual signal interferes with interpreting structural data of membrane proteins. This complication is often made worse by the presence of excess empty (protein-free) micelles. We present an approach for the rational design of mixed micelles containing a deuterated detergent analog, which eliminates neutron contrast between core and shell and allows the micelle scattering to be fully contrast-matched to unambiguously resolve membrane protein structure using solution SANS.

Single-Chain Conformation for Interacting Poly(N-isopropylacrylamide) in Aqueous Solution

The demixing phase behavior of Poly(N-isopropylacrylamide) (PNIPAM) aqueous solution is investigated using small-angle neutron scattering. This polymer phase separates upon heating and demixes around 32 ∘C. The pre-transition temperature range is characterized by two scattering modes; a low-Q (large-scale) signal and a high-Q dissolved chains signal. In order to get insight into this pre-transition region, especially the origin of the low-Q (large-scale) structure, the zero average contrast method is used in order to isolate single-chain conformations even in the demixing polymers transition region. This method consists of measuring deuterated and non-deuterated polymers dissolved in mixtures of deuterated and non-deuterated water for which the polymer scattering length density matches the solvent scattering length density. A fixed 4% polymer mass fraction is used in a contrast variation series where the d-water/h-water fraction is varied in order to determine the match point. The zero average contrast (match point) sample displays pure single-chain scattering with no interchain contributions. Our measurements prove that the large scale structure in this polymer solution is due to a transient polymer network formed through hydrophobic segment-segment interactions. Scattering intensity increases when the temperature gets close to the phase boundary. While the apparent radius of gyration increases substantially at the Lower Critical Solution Temperature (LCST) transition due to strong interchain correlation, the single-chain true radius of gyration has been found to decrease slightly with temperature when approaching the transition.

Synthesis, Characterization and Applications of a Perdeuterated Amphipol

Amphipols are short amphipathic polymers that can substitute for detergents at the hydrophobic surface of membrane proteins (MPs), keeping them soluble in the absence of detergents while stabilizing them. The most widely used amphipol, known as A8-35, is comprised of a polyacrylic acid (PAA) main chain grafted with octylamine and isopropylamine. Among its many applications, A8-35 has proven particularly useful for solution-state NMR studies of MPs, for which it can be desirable to eliminate signals originating from the protons of the surfactant. In the present work, we describe the synthesis and properties of perdeuterated A8-35 (perDAPol). Perdeuterated PAA was obtained by radical polymerization of deuterated acrylic acid. It was subsequently grafted with deuterated amines, yielding perDAPol. The number-average molar mass of hydrogenated and perDAPol, ~4 and ~5 kDa, respectively, was deduced from that of their PAA precursors, determined by size exclusion chromatography in tetrahydrofuran following permethylation. Electrospray ionization-ion mobility spectrometry-mass spectrometry measurements show the molar mass and distribution of the two APols to be very similar. Upon neutron scattering, the contrast match point of perDAPol is found to be ~120% D2O. In (1)H-(1)H nuclear overhauser effect NMR spectra, its contribution is reduced to ~6% of that of hydrogenated A8-35, making it suitable for extended uses in NMR spectroscopy. PerDAPol ought to also be of use for inelastic neutron scattering studies of the dynamics of APol-trapped MPs, as well as small-angle neutron scattering and analytical ultracentrifugation.

In-situ vapor small angle neutron scattering of sulfonated polyarylenethioethersulfone random copolymer (SPTES 70) fuel cells membrane

Highly sulfonated polyarylenethioethersulfone copolymer (SPTES 70) with excellent thermal stability, high proton conductivity and good mechanical properties is an excellent candidate as high temperature fuel cells membrane. The morphology of the SPTES 70 was examined with in situ vapor small angle neutron scattering (iVSANS), and transmission electron microscopy. An experimental neutron contrast match point was obtained with swelling SPTES 70 in a series of D2O/H2O ratios and compared with the calculated scattering contrast. The calculated scattering length density of SPTES 70 was in excellent agreement with the experiment scattering length density. Small angle neutron scattering (SANS) data of the magnesium, calcium, and silver exchanged hydrated SPTES 70 membranes exhibited nanostructured morphology which was compared with the reported nanostructure of D2O hydrated SPTES 70. SANS data showed that the average ionic domain size changes with the size and valence of the counter ion.

Analysis of the solution structure of Thermosynechococcus elongatus photosystem I in n-dodecyl-β-d-maltoside using small-angle neutron scattering and molecular dynamics simulation

Small-angle neutron scattering (SANS) and molecular dynamics (MD) simulation were used to investigate the structure of trimeric photosystem I (PSI) from Thermosynechococcus elongatus (T. elongatus) stabilized in n-dodecyl-β-d-maltoside (DDM) detergent solution. Scattering curves of detergent and protein–detergent complexes were measured at 18% D2O, the contrast match point for the detergent, and 100% D2O, allowing observation of the structures of protein/detergent complexes. It was determined that the maximum dimension of the PSI–DDM complex was consistent with the presence of a monolayer belt of detergent around the periphery of PSI. A dummy-atom reconstruction of the shape of the complex from the SANS data indicates that the detergent envelope has an irregular shape around the hydrophobic periphery of the PSI trimer rather than a uniform, toroidal belt around the complex. A 50ns MD simulation model (a DDM ring surrounding the PSI complex with extra interstitial DDM) of the PSI–DDM complex was developed for comparison with the SANS data. The results suggest that DDM undergoes additional structuring around the membrane-spanning surface of the complex instead of a simple, relatively uniform belt, as is generally assumed for studies that use detergents to solubilize membrane proteins.

The pore wall structure of porous semi-crystalline anatase TiO2

The structure of porous TiO2prepared by electrochemical anodization in a fluoride-containing ethylene glycol electrolyte solution was quantitatively studied using small-angle neutron scattering (SANS) and ultra-small-angle neutron scattering (USANS). The cylindrical pores along the coaxial direction were somewhat irregular in shape, were widely distributed in diameter, and seemed to have a broadly pseudo-hexagonal arrangement. The scattering from the pore wall showed a negative deviation from Porod scattering, indicating that the interface between TiO2and the pore was not sharp. A density gradient of around 40–60 Å at the pore wall (i.e.the interface between the pore and the TiO2matrix) was estimated using both constant and semi-sigmoidal interface models. This gradient may be due to the presence of fluorine and carbon partially absorbed by the pore wall from the fluoride-containing electrolyte or to sorbed water molecules on the wall. The neutron contrast-matching point between the TiO2matrix and the pores filled with liquid H2O/D2O mixtures was 51/49%(v/v) H2O/D2O, yielding an estimated mass density of 3.32 g cm−3. The specific surface area of the sample derived from the (U)SANS data was around 939–1003 m2 cm−3(283–302 m2 g−1).

Three-dimensional Model of Human Platelet Integrin αIIbβ3 in Solution Obtained by Small Angle Neutron Scattering

Integrin alphaIIbbeta3 is the major membrane protein and adhesion receptor at the surface of blood platelets, which after activation plays a key role in platelet plug formation in hemostasis and thrombosis. Small angle neutron scattering (SANS) and shape reconstruction algorithms allowed formation of a low resolution three-dimensional model of whole alphaIIb beta3 in Ca(2+)/detergent solutions. Model projections after 90 degrees rotation along its long axis show an elongated and "arched" form (135 degrees) not observed before and a "handgun" form. This 20-nm-long structure is well defined, despite alphaIIb beta3 multidomain nature and expected segmental flexibility, with the largest region at the top, followed by two narrower and smaller regions at the bottom. Docking of this SANS envelope into the high resolution structure of alphaIIb beta3, reconstructed from crystallographic and NMR data, shows that the solution structure is less constrained, allows tentative assignment of the disposition of the alphaIIb and beta3 subunits and their domains within the model, and points out the structural analogies and differences of the SANS model with the crystallographic models of the recombinant ectodomains of alphaIIb beta3 and alphaV beta3 and with the cryo-electron microscopy model of whole alphaIIb beta3. The ectodomain is in the bent configuration at the top of the model, where alphaIIb and beta3 occupy the concave and convex sides, respectively, at the arched projection, with their bent knees at its apex. It follows the narrower transmembrane region and the cytoplasmic domains at the bottom end. AlphaIIb beta3 aggregated in Mn(2+)/detergent solutions, which impeded to get its SANS model.

Insight into the Structure of Light-Harvesting Complex II and Its Stabilization in Detergent Solution

The structure of spinach light-harvesting complex II (LHC II), stabilized in a solution of the detergent n-octyl-beta-D-glucoside (BOG), was investigated by small-angle neutron scattering (SANS). Physicochemical characterization of the isolated complex indicated that it was pure (>95%) and also in its native trimeric state. SANS with contrast variation was used to investigate the properties of the protein-detergent complex at three different H(2)O/D(2)O contrast match points, enabling the scattering properties of the protein and detergent to be investigated independently. The topological shape of LHC II, determined using ab initio shape restoration methods from the SANS data at the contrast match point of BOG, was consistent with the X-ray crystallographic structure of LHC II (Liu et al. Nature 2004 428, 287-292). The interactions of the protein and detergent were investigated at the contrast match point for the protein and also in 100% D(2)O. The data suggested that BOG micelle structure was altered by its interaction with LHC II, but large aggregate structures were not formed. Indirect Fourier transform analysis of the LHC II/BOG scattering curves showed that the increase in the maximum dimension of the protein-detergent complex was consistent with the presence of a monolayer of detergent surrounding the protein. A model of the LHC II/BOG complex was generated to interpret the measurements made in 100% D(2)O. This model adequately reproduced the overall size of the LHC II/BOG complex, but demonstrated that the detergent does not have a highly regular shape that surrounds the hydrophobic periphery of LHC II. In addition to demonstrating that natively structured LHC II can be produced for functional characterization and for use in artificial solar energy applications, the analysis and modeling approaches described here can be used for characterizing detergent-associated alpha-helical transmembrane proteins.

Characterization of Sol−Gel-Encapsulated Proteins Using Small-Angle Neutron Scattering

Entrapment of biomolecules in silica-derived sol-gels has grown into a vibrant area of research since it was originally demonstrated. However, accessing the consequences of entrapment on biomolecules and the gel structure remains a major challenge in characterizing these biohybrid materials. We present the first demonstration that it is possible with small-angle neutron scattering (SANS) to study the conformation of dilute proteins that are entrapped in transparent and dense sol-gels. Using deuterium-labeled green fluorescent protein (GFP) and SANS with contrast variation, we demonstrate that the scattering signatures of the sol-gel and the protein can be separated. Analysis of the scattering curves of the sol-gels using a mass-fractal model shows that the size of the colloidal silica particles and the fractal dimensions of the gels were similar in the absence and presence of protein, demonstrating that GFP did not influence the reaction pathway for the formation of the gel. The major structural difference in the gels was that the pore size was increased 2-fold in the presence of the protein. At the contrast match point for silica, the scattering signal from GFP inside the gel became distinguishable over a wide q range. Simulated scattering curves representing a monomer, end-to-end dimer, and parallel dimer of the protein were calculated and compared to the experimental data. Our results show that the most likely structure of GFP is that of an end-to-end dimer. This approach can be readily applied and holds great potential for the structural characterization of complex biohybrid and other materials.

Structural evolution and stability of sol–gel biocatalysts

Immobilisation strategies for catalytic enzymes are important as they allow recovery and reuse of the biocatalysts. In this work, sol–gel matrices have been used to immobilise Candida antarctica lipase B (CALB), a commonly used industrial enzyme. The sol–gel bioencapsulate is produced through fluoride-catalysed hydrolysis of mixtures of tetramethylorthosilicate (TMOS) and methyltrimethoxysilane (MTMS) in the presence of CALB, yielding materials with controlled pore sizes and surface chemistries. Sol–gel matrices prolong the catalytic life and enhance the activity of CALB, although the molecular basis for this effect has yet to be elucidated due to the limitations of analytical techniques applied to date. Small angle neutron scattering (SANS) allows such multi-component systems to be characterised through contrast matching. In the sol–gel bioencapsulate system at the contrast match point for silica, residual scattering intensity is due to the CALB and density fluctuations in the matrix. A SANS contrast variation series found the match point for the silica matrix, both with and without enzyme present, to be around 35%. The model presented here proposes a mechanism for the interaction between CALB and the surrounding sol–gel matrix, and the observed improvement in enzyme activity and matrix strength. Essentially, the inclusion of CALB modulates silicate speciation during evolution of the inorganic network, leading to associated variations in SANS contrast. The SANS protocol developed here may be applied more generally to other encapsulated enzyme systems.

Preliminary neutron scattering studies of the Type I restriction-modification enzyme M.Ahdl

Type I restriction-modification (R-M) systems encode multisubunit/multidomain enzymes. Two genes (M and S) are required to form the 160 kDa methyltransferase that methylates a specific base within the recognition sequence and protects DNA from cleavage by the endonuclease. Small angle neutron scattering (SANS) revealed an unusually large structural change in the EcoR124I methyltransferase following DNA binding; this involves a major repositioning of the subunits of the enzyme, resulting in a 60 Å reduction in the dimensions of the enzyme on forming a complex with DNA. The related methyltransferase M.Ahdl, with stoichiometry M 2S 2 has been prepared in two protonated/deuterated states (S and M subunits protonated, S deuterated and M protonated) for which SANS data have been collected in a number of H:D solvent contrasts. The contrast match point of the selectively deuterated enzyme confirms the successful reconstitution of the enzyme with deuterated S subunits. Ab initio shape determination using this contrast matched data is in progress to determine the subunit organization of M.Ahdl and the large change in structure that occurs on DNA binding.

In situ vapor sorption apparatus for small-angle neutron scattering and its application

An in situ vapor sorption apparatus has been constructed for use in small-angle neutron scattering (SANS) measurements. The apparatus adapts two independent operating mechanisms, with and without a carrier gas, to control relative or absolute pressure, respectively, in the SANS sorption cell. By controlling the absolute pressure, a target relative vapor pressure between 0% and 90% can be reached within 1–2 min. The short response time makes it possible to correlate diffusion kinetics and/or sorption/desorption isotherms with structure evolution during wetting/drying, which is not possible in gravimetric methods. Also, one can extract diffusion coefficients and interaction parameters. Other uses include the enhancement of scattering contrast in the study of semicrystalline polymers by the preferential vapor sorption of deuterated vapor into the amorphous regions. Thus, one can obtain the same structural information as small-angle x-ray scattering measurements on dry samples. Also, the apparatus has the capability to inject a pore-masking liquid into the sample cell while under vacuum to ensure the filling of all open porosities in a sample. The capability to mix two vapors in various ratios facilitates the determination of a contrast-matching point using a single sample, which also eliminates a major source of systematic error. The performance of the apparatus is demonstrated using a polyelectrolyte membrane and semicrystalline polyethylene. Additionally, technical points relating to controlling the relative vapor pressure, relative humidity, and regarding the vapor distribution in the cell are discussed.

Partial specific volume and solvent interactions of amphipol A8-35

Amphipols are small amphiphilic polymers that can stabilize and keep soluble membrane proteins in aqueous solutions in the absence of detergent. A prerequisite to solution studies of membrane protein/amphipol complexes is the determination of the partial specific volume v ¯ 2 and effective charge z of the polymer. The ratio ( R) of the buoyant molar masses of particles in D 2O and H 2O solutions, obtained from sedimentation velocity ( s H/ s D method) and sedimentation equilibrium experiments, and their contrast match point (CMP), determined in small-angle neutron scattering experiments, depend on v ¯ 2 and z. When z is known, v ¯ 2 can be estimated from R with a good accuracy as long as v ¯ 2 is close to 1. The effects of labile H/D exchange and of polyelectrolyte counter-ion dissociation in general cannot be neglected. The accuracy, advantages, and limits of the s H/ s D method have been studied in details using model macromolecules (DNA, protein, and polysaccharide). The s H/ s D method appears particularly advantageous for the study of heterogeneous samples. Measurements of density, s D/s H buoyant molar masses in H 2O, D 2O, and D 2 18O, and CMP of hydrogenated and partially deuterated A8-35, a polyacrylate-based amphipol containing 35 underivatized carboxylates per 100 monomers, led to a consistent description of its buoyancy and charge properties.

Characterization of Nanoporous Low-k Thin Films by Small-Angle Neutron Scattering Contrast Variation

Small-angle neutron scattering contrast variation is applied to characterization of nanoporous low-dielectric constant (low-k) thin films. Films are exposed to saturated solvent vapor in air, whereby the pores fill with liquid by capillary condensation. The pores are filled with mixtures of hydrogen- and deuterium-containing solvents to vary the neutron contrast with the matrix (wall). The composition of the solvent mixture is systematically varied to identify a composition that minimizes the scattered intensity (contrast match point). From the contrast match point composition, film characteristics including matrix density and homogeneity are assessed. Four spin-on low-k materials including a methylsilsesquioxane, an organic polymer, a xerogel, and a hydrogensilsesquioxane are characterized by the new technique. Calculated matrix mass densities are compared to independent density measurements obtained by an established specular X-ray reflectivity technique. We find no evidence of "closed pores", defined here as pores inaccessible to the probe solvent, in any of the materials studied.

The structure of colloidal alloy crystals revealed by ultra-small-angle neutron scattering

We investigated the structure of colloidal crystals in binary colloidal dispersion by the ultra-small-angle neutron scattering (USANS) technique. The mixed dispersion of deutrated polystyrene latex ( d-PS) and hydrogenated polystyrene latex ( h-PS) dispersions (the size ratio α=0.54) was prepared for the USANS experiment to apply the contrast matching technique. The scattering from the each component could be detected separately. We also extracted the partial scattering intensity I dPS-hPS( q) at three mixing ratios. The USANS scattering profiles at the contrast matching point for each component were changed clearly with the change of the mixing ratio of d-PS and h-PS lattices. The I dPS-hPS( q) profile obtained for the d-PS number fraction x dPS03=0.39 indicated the correlation between d-PS and h-PS particles. These USANS observation suggested strongly the formation of the liquid-like colloidal alloy structure in the binary colloidal dispersion.