From atoms to layers: in situ gold cluster growth kinetics during sputter deposition.

The adjustment of size-dependent catalytic, electrical and optical properties of gold cluster assemblies is a very significant issue in modern applied nanotechnology. We present a real-time investigation of the growth kinetics of gold nanostructures from small nuclei to a complete gold layer during magnetron sputter deposition with high time resolution by means of in situ microbeam grazing incidence small-angle X-ray scattering (µGISAXS). We specify the four-stage growth including their thresholds with sub-monolayer resolution and identify phase transitions monitored in Yoneda intensity as a material-specific characteristic. An innovative and flexible geometrical model enables the extraction of morphological real space parameters, such as cluster size and shape, correlation distance, layer porosity and surface coverage, directly from reciprocal space scattering data. This approach enables a large variety of future investigations of the influence of different process parameters on the thin metal film morphology. Furthermore, our study allows for deducing the wetting behavior of gold cluster films on solid substrates and provides a better understanding of the growth kinetics in general, which is essential for optimization of manufacturing parameters, saving energy and resources.

P03, the microfocus and nanofocus X-ray scattering (MiNaXS) beamline of the PETRA III storage ring: the microfocus endstation.

The P03 beamline, also called the microfocus and nanofocus X-ray scattering (MiNaXS) beamline, exploits the excellent photon beam properties of the low-emittance source PETRA III to provide a microfocused/nanofocused beam with ultra-high intensity for time-resolved X-ray scattering experiments. The beamline has been designed to perform X-ray scattering in both transmission and reflection geometries. The microfocus endstation started user operation in May 2011. An overview of the beamline status and of some representative results highlighting the performance of the microfocus endstation at MiNaXS are given.

Influence of nanoparticle surface functionalization on the thermal stability of colloidal polystyrene films.

The installation of large scale colloidal nanoparticle thin films is of great interest in sensor technology or data storage. Often, such devices are operated at elevated temperatures. In the present study, we investigate the effect of heat treatment on the structure of colloidal thin films of polystyrene (PS) nanoparticles in situ by using the combination of grazing incidence small-angle X-ray scattering (GISAXS) and optical ellipsometry. In addition, the samples are investigated with optical microscopy, atomic force microscopy (AFM), and field emission scanning electron microscopy (FESEM). To install large scale coatings on silicon wafers, spin-coating of colloidal pure PS nanoparticles and carboxylated PS nanoparticles is used. Our results indicate that thermal annealing in the vicinity of the glass transition temperature T(g) of pure PS leads to a rapid loss in the ordering of the nanoparticles in spin-coated films. For carboxylated particles, this loss of order is shifted to a higher temperature, which can be useful for applications at elevated temperatures. Our model assumes a softening of the boundaries between the individual colloidal spheres, leading to strong changes in the nanostructure morphology. While the nanostructure changes drastically, the macroscopic morphology remains unaffected by annealing near T(g).

Effect of solvent annealing on the tensile deformation mechanism of a colloidal crystalline polymeric latex film.

The influence of solvent annealing on microscopic deformational behavior of a styrene/n-butyl acrylate copolymer latex film subjected to uniaxial tensile deformation was studied by small-angle X-ray scattering. It was demonstrated that the microscopic deformation mechanism of the latex films transformed from a nonaffine deformation behavior to an affine deformation behavior after solvent annealing. This was attributed to the interdiffusion of polymeric chains between adjacent swollen latex particles in the film. It turns out that solvent annealing is much more efficient than thermal annealing due to a much slow evaporation process after solvent annealing.

Time-resolved ultrathin cobalt film growth on a colloidal polymer template.

Cobalt (Co) sputter deposition onto a colloidal polymer template is investigated using grazing incidence small-angle X-ray scattering (GISAXS), scanning electron microscopy (SEM), and atomic force microscopy (AFM). SEM and AFM data picture the sample topography, GISAXS the surface and near-surface film structure. A two-phase model is proposed to describe the time evolution of the Co growth. The presence of the colloidal template results in the correlated deposition of an ultrathin Co film on the sample surface and thus in the creation of Co capped polystyrene (PS) colloids. Well below the percolation threshold, the radial growth is restricted and only height growth is observed.

Structural evolution of a colloidal crystal fiber during heating and annealing studied by in situ synchrotron small angle X-ray scattering.

The structural evolution of a colloidal crystal fiber during heating and annealing was followed by in situ synchrotron small-angle X-ray scattering. The polymer dispersion (with a particle size of 118 nm) from which the fibers were formed by directed drying contained emulsifier and salt. A cellular structure formed upon drying in which the percolating phase (the "membrane phase") is composed from these components; this membrane phase gives rise to the scattering contrast on which the present observations build. Changes of the lattice constant of the colloidal crystallites and the intensity evolution of the scattering from the crystalline and the amorphous phases during heating and annealing indicate characteristic temperatures where the system exhibits pronounced structural changes. The first characteristic temperature was identified as 125 degrees C above which residue water in the membrane material was evaporated leading to shrinkage of the colloidal crystalline lattice. At a temperature above about 140 degrees C the membrane material was expelled out of the crystalline domains. This effect is accompanied by the progressive interdiffusion of polymer chains between adjacent latex particles and leads to further thermal shrinkage of the colloidal crystals. The second characteristic temperature is defined by a rapid increase in isotropic scattering. This effect is attributed to the formation of increasingly large domains of the membrane material and the concomitant disappearance of the membrane phase from the former crystal domains.

Polymorphic transformation of isotactic poly(1-butene) in form III upon heating: in situ synchrotron small- and wide-angle X-ray scattering studies.

The phase transformation of form III isotactic poly(1-butene) was investigated as a function of temperature. The polymer was isothermally precipitated from a dilute solution in iso-amyl acetate and observed with real-time synchrotron small- and wide-angle X-ray scattering techniques. The results confirmed that the polymorphic transition of form III was strongly dependent on temperature. The phase transformation from form III to form I' proceeded at a temperature of ca. 80 degrees C. This was accompanied by the presence of two distinctly different lamellar periodicities arising from form III and I' crystals, respectively. The coexistence of form III and I' crystals can persist up to 103 degrees C, followed by melting and recrystallizing into form II crystals. Finally, the reflections resulting from form II crystals disappeared at 118 degrees C.

Spatially resolved investigation of solution cast nanoparticle films by X-ray scattering and multidimensional data set classification.

A combinatorial high-throughput approach is used to investigate a solution cast gradient consisting of colloidal gold nanoparticles on top of a silicon substrate by means of a X-ray nanobeam. Classification algorithms are used to reveal and visualize structural transitions from a frozen colloidal solution to a well-defined nanostructure. Prominent length scales on the order of 100 nm are observed. A periodic change in the nanostructure along the gradient is explained by a simplified stick-slip model.

Combining imaging ellipsometry and grazing incidence small angle X-ray scattering for in situ characterization of polymer nanostructures.

A combination of microbeam grazing incidence small angle X-ray scattering (muGISAXS) and imaging ellipsometry is introduced as a new versatile tool for the characterization of nanostructures. muGISAXS provides a local lateral and depth-sensitive structural characterization, and imaging ellipsometry adds the position-sensitive determination of the three-dimensional morphology in terms of thickness, roughness, refractive index, and extinction coefficient. Together muGISAXS and imaging ellipsometry enable a complete characterization of structure and morphology. On the basis of an example of buildup of nanostructures from monodisperse colloidal polystyrene nanospheres on a rough solid support, the scope of this new combination is demonstrated. Roughness is introduced by a dewetting structure of a diblock copolymer film with one block being compatible with the colloidal nanoparticles and one block being incompatible. To demonstrate the potential for kinetic investigations, muGISAXS and imaging ellipsometry are applied to probe the drying process of an aqueous dispersion of nanospheres on such a type of rough substrate.

Grazing incidence small-angle X-ray scattering microtomography demonstrated on a self-ordered dried drop of nanoparticles.

We combine grazing-incidence small-angle X-ray scattering (GISAXS) with scanning X-ray microtomography to investigate the nanostructure in a dried gold/polystyrene nanocomposite drop. Local GISAXS structure factors are reconstructed at each position on the surface of this two-dimensionally heterogeneous sample with 30 microm pixel size. Evidence for four types of self-assembled colloidal crystalline structures is provided by the reconstructed data of the drop demonstrating the feasibility of the method.

Small angle x-ray scattering with a beryllium compound refractive lens as focusing optic.

At BW4 at HASYLAB a beryllium compound refractive lens (Be-CRL) is used for the focusing in small-angle x-ray scattering experiments. Using it provides the advantages of higher long-term stability and a much easier alignment compared to a setup with focusing mirrors. In our investigations presented here, we show the advantages of using a Be-CRL in small-angle and also ultra small-angle x-ray scattering. We investigated the beam characteristics at the sample position with respect to spot size and photon flux. The spot size is comparable to that of a setup with focusing mirrors but with a gain in flux and better long-term stability. It is also shown that plane mirrors are still necessary to suppress higher order energies passing the monochromator.

GIUSAXS and AFM studies on surface reconstruction of latex thin films during thermal treatment.

The structural evolution of a single-layer latex film during annealing was studied via grazing incidence ultrasmall-angle X-ray scattering (GIUSAXS) and atomic force microscopy (AFM). The latex particles were composed of a low-Tg (-54 degrees C) core (n-butylacrylate, 30 wt %) and a high-Tg (41 degrees C) shell (t-butylacrylate, 70 wt %) and had an overall diameter of about 500 nm. GIUSAXS data indicate that the q(y) scan at q(z) = 0.27 nm(-1) (out-of-plane scan) contains information about both the structure factor and the form factor. The GIUSAXS data on latex films annealed at various temperatures ranging from room temperature to 140 degrees C indicate that the structure of the latex thin film beneath the surface changed significantly. The evolution of the out-of-plane scan plot reveals the surface reconstruction of the film. Furthermore, we also followed the time-dependent behavior of structural evolution when the latex film was annealed at a relatively low temperature (60 degrees C) where restructuring within the film can be followed that cannot be detected by AFM, which detects only surface morphology. Moreover, compared to AFM studies GIUSAXS provides averaged information covering larger areas.

A detailed analysis of biodegradable nanospheres by different techniques--a combined approach to detect particle sizes and size distributions.

Poly(d,l-lactide-co-glycolide) nanosupensions as intravenous nanosphere systems were produced by solvent deposition in aqueous Poloxamer 188 solutions. Light scattering techniques were applied to these colloidal systems to characterize particle sizes. Regularly shaped spherical particles were received as proved by freeze fracture replica and small-angle X-ray scattering (SAXS). SAXS was performed using intensive synchrotron radiation. Particle sizes were calculated from the small-angle part of scattering curve that were in good agreement with z-average values received from photon correlation spectroscopy (PCS). The flow field-flow fractionation (FlFFF) fractograms in combination with multi-angle light scattering (MALS) allowed an easy detection of maximum particle sizes what is most important for parenteral systems. Furthermore, high quality size distributions were received due to the separation step prior to size characterization. The calculated average size values exhibited a good correlation with z-averages determined by PCS. Only for suspensions of broader size distributions, higher deviations were observed. Comparing particle sizes with and without Poloxamer, differences in diameters resulted that were quantified. The additional Poloxamer shell was not able to be removed by an intensive washing during FlFFF focusing and separation. Especially FlFFF/MALS proved to be a valuable tool to characterize the pharmaceutical nanosuspensions in detail what is of great importance especially for controlled drug delivery systems.

Facile preparation of macroscopic soft colloidal crystals with fiber symmetry.

A facile, efficient way to fabricate macroscopic soft colloidal crystals with fiber symmetry by drying a latex dispersion in a tube is presented. A transparent, stable colloidal crystal was obtained from a 25 wt % latex dispersion by complete water evaporation for 4 days. The centimeter-long sample was investigated by means of synchrotron small-angle X-ray diffraction (SAXD). Analysis of a large number of distinct Bragg peaks reveals that uniaxially oriented colloidal crystals with face-centered cubic lattice structure were formed. The measurement of evaporation rates under different conditions indicates that the water evaporates primarily through the optically clear regions (i.e., via the solid material) even when the region is more than 2 mm thick.

Non-affine structural evolution of soft colloidal crystalline latex films under stretching as observed via synchrotron X-ray scattering.

A polymer dispersion consisting of soft latex spheres with a diameter of 135 nm was used to produce a crystalline film with face-centered cubic (fcc) packing of the spheres. Different from conventional small-molecule and hard-sphere colloidal crystals, the crystalline latex film in the present case is soft (i.e., easily deformable). The structural evolution of this soft colloidal latex film under stretching was investigated by in-situ synchrotron ultra-small-angle X-ray scattering. The film exhibits polycrystalline scattering behavior corresponding to fcc structure. Stretching results not only in a large deformation of the crystallographic structure but also in considerable nonaffine deformation at high draw ratios. The unexpected nonaffine deformation was attributed to slippage between rows of particles and crystalline grain boundaries. The crystalline structure remains intact even at high deformation, suggesting that directional anisotropic colloidal crystallites can be easily produced.

Heterologous gene expression by infectious and replicon vectors derived from tick-borne encephalitis virus and direct comparison of this flavivirus system with an alphavirus replicon.

The flavivirus tick-borne encephaltis virus (TBEV) was established as a vector system for heterologous gene expression. The variable region of the genomic 3' non-coding region was replaced by an expression cassette consisting of the reporter gene enhanced green fluorescent protein (EGFP) under the translational control of an internal ribosomal entry site element, both in the context of an infectious virus genome and of a replicon lacking the genes of the surface proteins prM/M and E. The expression level and the stability of expression were measured by fluorescence-activated cell-sorting analysis and compared to an established alphavirus replicon vector derived from Venezuelan equine encephaltis virus (VEEV), expressing EGFP under the control of its natural subgenomic promoter. On the first day, the alphavirus replicon exhibited an approximately 180-fold higher expression level than the flavivirus replicon, but this difference decreased to about 20- and 10-fold on days 2 and 3, respectively. Four to six days post-transfection, foreign gene expression by the VEEV replicon vanished almost completely, due to extensive cell killing. In contrast, in the case of the TBEV replicon, the percentage of positive cells and the amount of EGFP expression exhibited only a moderate decline over a time period of almost 4 weeks. The infectious TBEV vector expressed less EGFP than the TBEV replicon at all times. Significant expression from the infectious vector was maintained for four cell-culture passages. The results indicate that the VEEV vector is superior with respect to achieving high expression levels, but the TBEV system may be advantageous for applications that require a moderate, but more enduring, gene expression.

Incorporation of tick-borne encephalitis virus replicons into virus-like particles by a packaging cell line.

RNA replicons derived from flavivirus genomes show considerable potential as gene transfer and immunization vectors. A convenient and efficient encapsidation system is an important prerequisite for the practical application of such vectors. In this work, tick-borne encephalitis (TBE) virus replicons and an appropriate packaging cell line were constructed and characterized. A stable CHO cell line constitutively expressing the two surface proteins prM/M and E (named CHO-ME cells) was generated and shown to efficiently export mature recombinant subviral particles (RSPs). When replicon NdDeltaME lacking the prM/M and E genes was introduced into CHO-ME cells, virus-like particles (VLPs) capable of initiating a single round of infection were released, yielding titers of up to 5 x 10(7)/ml in the supernatant of these cells. Another replicon (NdDeltaCME) lacking the region encoding most of the capsid protein C in addition to proteins prM/M and E was not packaged by CHO-ME cells. As observed with other flavivirus replicons, both TBE virus replicons appeared to exert no cytopathic effect on their host cells. Sedimentation analysis revealed that the NdDeltaME-containing VLPs were physically distinct from RSPs and similar to infectious virions. VLPs could be repeatedly passaged in CHO-ME cells but maintained the property of being able to initiate only a single round of infection in other cells during these passages. CHO-ME cells can thus be used both as a source for mature TBE virus RSPs and as a safe and convenient replicon packaging cell line, providing the TBE virus surface proteins prM/M and E in trans.

Online SAXS investigations of polymeric hollow fibre membranes.

Polymeric membranes are used in industrial and analytical separation techniques. In this study small-angle X-ray scattering (SAXS) with synchrotron radiation has been applied for in-situ characterisation during formation of polymeric membranes. The spinning of a polyetherimide (PEI) hollow fibre membrane was chosen for investigation of dynamic aggregation processes during membrane formation, because it allows the measurement of the dynamic equilibrium at different distances from the spinning nozzle. With this system it is possible to resolve structural changes in the nm-size range which occur during membrane formation on the time-scale of milliseconds. Integral structural parameters, like radius of gyration and pair-distance distribution, were determined. Depending on the chosen spinning parameters, e.g. the flow ratio between polymer solution and coagulant water, significant changes in the scattering curves have been observed. The data are correlated with the distance from the spinning nozzle in order to get information about the kinetics of membrane formation which has fundamental influence on structure and properties of the membrane.

Characterisation of structure and aggregation processes of aquatic humic substances using small-angle scattering and X-ray microscopy.

Aquatic humic substances (HS), an important part of the dissolved organic carbon in freshwater systems, are polyfunctional natural compounds with polydisperse structure showing strong aggregation/coagulation behaviour at high HS concentrations and in the presence of metal ions. In this study, small-angle neutron scattering (SANS), small-angle X-ray scattering (SAXS) and X-ray microscopy (XRM) were applied to characterise the structure and aggregation processes of HS in solution. In SAXS and XRM the high brilliant synchrotron radiation was used as X-ray source. Applying small-angle scattering, information about the size distribution and shape of aquatic HS was obtained. Spherical HS units were found which were stable in a wide concentration range in a kind of "monomeric" state almost independent of pH and ionic strength. At higher concentrations they formed chain-like agglomerates or disordered HS structures. In studies on the coagulation behaviour of HS after addition of copper ions, a linear relationship between Cu(2+) concentration and the formation of large disordered HS-Cu(2+) agglomerates was obtained. By using X-ray microscopy, single "huge" particles were found in older solutions and in solutions with high HS concentrations. Over a threshold Cu(2+) concentration of approx. 300 mg/L, the formation of an extensive HS-Cu(2+) network structure was observed within a few minutes. The presented structures show the ability of the methods used to characterise processes between diluted phase and suspended matter, which play an important role particularly in the region of phase interfaces.