The details of a newly constructed small-angle X-ray scattering instrument are presented. The geometry of the instrument is highly customizable, enabling it to address vastly different experimental situations from academic research to industrial problems. The high degree of motorization and automation compared to conventional laboratory-scale SAXS instruments facilitates the alignment and daily use. Data reduction routines are incorporated in the instrument control software, yielding fully corrected and calibrated results promptly after the end of measurements. Optimization of the fluxversus resolution balance can be done routinely for each measurement task. A wide, continuous range ofq= 4πsinθ/λ can be reached, from below 0.02 nm⁻¹up to 30 nm⁻¹, corresponding to periodic distances ofca 350 nm down to 0.2 nm. A few representative results obtained from samples of different research fields demonstrate the versatility of the instrument. Scattering curves are routinely calibrated into absolute units using a glassy carbon secondary standard. More information and recent developments can be found on the web page of the instrument at http://credo.ttk.mta.hu.

Silica nanoparticles (SNPs) belong to the most widely produced nanomaterials nowadays. Particle size distribution (PSD) is a key property of SNPs that needs to be accurately determined for a successful application. Many single particle and ensemble characterization methods are available for the determination of the PSD of SNPs, each having different advantages and limitations. Since most preparation protocols for SNPs can yield bimodal or heterogeneous PSDs, the capability of a given method to resolve bimodal PSD is of great importance. In this work, four different methods, namely transmission electron microscopy (TEM), dynamic light scattering (DLS), microfluidic resistive pulse sensing (MRPS) and small-angle X-ray scattering (SAXS) were used to characterize three different, inherently bimodal SNP samples. We found that DLS is unsuitable to resolve bimodal PSDs, while MRPS has proven to be an accurate single-particle size and concentration characterization method, although it is limited to sizes above 50 nm. SAXS was found to be the only method which provided statistically significant description of the bimodal PSDs. However, the analysis of SAXS curves becomes an ill-posed inverse mathematical problem for broad size distributions, therefore the use of orthogonal techniques is required for the reliable description of the PSD of SNPs.

Colloidal stabile nanoerythrosomes with 200 nm average diameter were formed from hemoglobin-free erythrocyte ghost membrane via sonication and membrane extrusion. The incorporation of extra lipid (1,2-dipalmitoyl-sn-glycero-3-phosphocholine, DPPC), added to the sonicated ghosts, caused significant changes in the thermotropic character of the original membranes. As a result of the increased DPPC ratio the chain melting of the hydrated DPPC system and the characteristic small angle X-ray scattering (SAXS) of the lipid bilayers appeared. Significant morphological changes were followed by transmission electron microscopy combined with freeze fracture method (FF-TEM). After the ultrasonic treatment the large entities of erythrocyte ghosts transformed into nearly spherical nanoerythrosomes with diameters between 100 and 300 nm and at the same time a great number of 10-30 nm large membrane proteins or protein clusters were dispersed in the aqueous medium. The infrared spectroscopy (FT-IR) pointed out, that the sonication did not cause changes in the secondary structures of the membrane proteins under our preparation conditions. About fivefold of extra lipid--compared to the lipid content of the original membrane--caused homogeneous dispersion of nanoerythrosomes however the shape of the vesicles was not uniform. After the addition of about tenfold of DPPC, monoform and monodisperse nanoerythrosomes became typical. The outer surfaces of these roughly spherical objects were frequently polygonal, consisting of a net of pentagons and hexagons.

Fibrin, the main scaffold of thrombi, is susceptible to citrullination by PAD (peptidyl arginine deiminase) 4, secreted from neutrophils during the formation of neutrophil extracellular traps. Citrullinated fibrinogen (citFg) has been detected in human plasma as well as in murine venous thrombi, and it decreases the lysability and mechanical resistance of fibrin clots. To investigate the effect of fibrinogen citrullination on the structure of fibrin clots. Fibrinogen was citrullinated with PAD4 and clotted with thrombin. Scanning electron microscopy (SEM) and atomic force microscopy (AFM) were used to measure fiber thickness, fiber height/width ratio, and fiber persistence length in clots containing citFg. Fiber density was measured with laser scanning microscopy (LSM) and permeability measurements were carried out to estimate the porosity of the clots. The intra‐fiber structure of fibrin was analyzed with small‐angle X‐ray scattering (SAXS). SEM images revealed a decrease in the median fiber diameter that correlated with the fraction of citFg in the clot, while the fiber width/length ratio remained unchanged according to AFM. With SAXS we observed that citrullination resulted in the formation of denser clots in line with increased fiber density shown by LSM. The permeability constant of citrullinated fibrin decreased more than 3‐fold indicating significantly decreased porosity. SAXS also showed largely preserved periodicity in the longitudinal assembly of fibrin monomers. The current observations of thin fibers combined with dense packing and low porosity in the presence of citFg can provide a structural framework for the mechanical fragility and lytic resistance of citrullinated fibrin.

Cochleates, calcium-stabilized membrane rolls of nanoscale diameter, promise a unique and efficient way of delivering lipid-soluble drugs, proteins or nucleic acids into biological systems because they protect the encapsulated material against enzymatic or chemical degradation. Self-aggregation, which typically arises during production and storage is a major obstacle that has so far precluded the development of an efficient cochleate-based drug-delivery system. Here we show that citric acid, added transiently in a narrow concentration range, effectively disperses cochleate aggregates, stabilizes the disperse state for long-term storage and preserves the canonical ultrastructure and topological characteristics of cochleate nanoparticles.

Novel pathways of the synthesis of photoluminescent gold quantum clusters (AuQCs) using biomolecules as reactants provide biocompatible products for biological imaging techniques. In order to rationalize the rules for the preparation of red-emitting AuQCs in aqueous phase using proteins or peptides, the role of different organic structural units was investigated. Three systems were studied: proteins, peptides, and amino acid mixtures, respectively. We have found that cysteine and tyrosine are indispensable residues. The SH/S-S ratio in a single molecule is not a critical factor in the synthesis, but on the other hand, the stoichiometry of cysteine residues and the gold precursor is crucial. These observations indicate the importance of proper chemical behavior of all species in a wide size range extending from the atomic distances (in the Au^I-S semi ring) to nanometer distances covering the larger sizes of proteins assuring the hierarchical structure of the whole self-assembled system.