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Amyloids: From molecular structure to mechanical properties
Many proteins of diverse sequence, structure and function self-assemble into morphologically similar fibrillar aggregates known as amyloids. Amyloids are remarkable polymers in several respects. First of all, amyloids can be formed from proteins with very different amino acid sequences; the common denominator is that the individual proteins constituting the amyloid fold predominantly into a β-sheet structure. Secondly, the formation of the fibril occurs through non-covalent interactions between primarily the β-sheets, causing the monomers to stack into fibrils. The fibrils are remarkably robust, considering that the monomers are bound non-covalently. Finally, a common characteristic of fibrils is their unbranched, straight, fiber-like structure arising from the intertwining of the multiple β-sheet filaments. These remarkably ordered and stable nanofibrils can be useful as building blocks for protein-based functional materials, but they are also implicated in severe neurodegenerative diseases. The overall aim of this article is to highlight recent efforts aimed at obtaining insights into amyloid proteins on different length scales. Starting from molecular information on amyloids, single fibril properties and mechanical properties of networks of fibrils are described. Specifically, we focus on the self-assembly of amyloid protein fibrils composed of peptides and denatured model proteins, as well as the influence of inhibitors of fibril formation. Additionally, we will demonstrate how the application of recently developed vibrational spectroscopic techniques has emerged as a powerful approach to gain spatially resolved information on the structure-function relation of amyloids. While spectroscopy provides information on local molecular conformations and protein secondary structure, information on the single fibril level has been developed by diverse microscopic techniques. The approaches to reveal basic mechanical properties of single fibrils like bending rigidity, shear modulus, ultimate tensile strength and fracture behavior are illustrated. Lastly, mechanics of networks of amyloid fibrils, typically forming viscoelastic gels are outlined, with a focus on (micro-) rheological properties. The resulting fundamental insights are essential for the rational design of novel edible and biodegradable protein-based polymers, but also to devise therapeutic strategies to combat amyloid assembly and accumulation during pathogenic disorders.
On the possibility of PhotoEmission Electron Microscopy for E. coli advanced studies
The novel approach was proposed for detailed high-resolution studies of morphology and physico-chemical properties concomitantly at one measurement spot of E. coli bacterial cells culture immobilized onto silicon wafer surface in UHV conditions applying PhotoEmission Electron Microscopy under Hg lamp irradiation. For the E. coli characterization scanning electron microscopy (electron beam) and X-ray photoelectron spectroscopy (X-ray tube radiation) were applied prior to PhotoEmission Electron Microscopy measurements. In spite of irradiation doses collected for the cell arrays we were successful in detection of high-resolution images even of single E. coli bacterium by PhotoEmission Electron Microscopy technique followed by detailed high-resolution morphology studies by scanning electron microscopy. These results revealed widespread stability of the E. coli membranes shape after the significant number of applied characterization techniques. © 2019 The Authors
Successful optimization of reconstruction parameters in structured illumination microscopy
The impact of the different reconstruction parameters in super-resolution structured illumination microscopy (SIM) on image artifacts is carefully analyzed. These parameters comprise the Wiener filter parameter, an apodization function, zero-frequency suppression and modifications of the optical transfer function. A detailed investigation of the reconstructed image spectrum is concluded to be suitable for identifying artifacts. For this purpose, two samples, an artificial test slide and a more realistic biological system, were used to characterize the artifact classes and their correlation with the image spectra as well as the reconstruction parameters. In addition, a guideline for efficient parameter optimization is suggested and the implementation of the parameters in selected up-to-date processing packages (proprietary and open-source) is depicted. © 2018 The Authors
Carrier Lifetime in Liquid-phase Crystallized Silicon on Glass
Liquid-phase crystallized silicon on glass (LPCSG) presents a promising material to fabricate high quality silicon thin films, e.g. for solar cells and modules. Barrier layers and a doped amorphous silicon layer are deposited on the glass substrate followed by crystallization with a line focus laser beam. In this paper we introduce injection level dependent lifetime measurements generated by the quasi steady-state photoconductance decay method (QSSPC) to characterize LPCSG absorbers. This contactless method allows a determination of the LPCSG absorber quality already at an early stage of solar cell fabrication, and provides a monitoring of the absorber quality during the solar cell fabrication steps. We found minority carrier lifetimes higher than 200ns in our layers (e.g. n-type absorber with ND=2x1015cm-3) indicating a surface recombination velocity SBL<3000cm/s at the barrier layer/Si interface.
Peculiarities of electronic structure and composition in ultrasound milled silicon nanowires
The combined X-ray absorption and emission spectroscopy approach was applied for the detailed electronic structure and composition studies of silicon nanoparticles produced by the ultrasound milling of heavily and lowly doped Si nanowires formed by metal-assisted wet chemical etching. The ultrasoft X-ray emission spectroscopy and synchrotron based X-ray absorption near edges structure spectroscopy techniques were utilize to study the valence and conduction bands electronic structure together with developed surface phase composition qualitative analysis. Our achieved results based on the implemented surface sensitive techniques strongly suggest that nanoparticles under studies show a significant presence of the silicon suboxides depending on the pre-nature of initial Si wafers. The controlled variation of the Si nanoparticles surface composition and electronic structure, including band gap engineering, can open a new prospective for a wide range Si-based nanostructures application including the integration of such structures with organic or biological systems. © 2020
Engineering an achromatic Bessel beam using a phase-only spatial light modulator and an iterative Fourier transformation algorithm
Bessel illumination is an established method in optical imaging and manipulation to achieve an extended depth of field without compromising the lateral resolution. When broadband or multicolour imaging is required, wavelength-dependent changes in the radial profile of the Bessel illumination can complicate further image processing and analysis. We present a solution for engineering a multicolour Bessel beam that is easy to implement and promises to be particularly useful for broadband imaging applications. A phase-only spatial light modulator (SLM) in the image plane and an iterative Fourier Transformation algorithm (IFTA) are used to create an annular light distribution in the back focal plane of a lens. The 2D Fourier transformation of such a light ring yields a Bessel beam with a constant radial profile for different wavelength.
Superconductivity in multi-phase Mg-B-O compounds
Structures of MgB2-based materials manufactured under pressure (up to 2 GPa) by different methods having high superconducting performance and connectivity are multiphase and contain different Mg-B-O compounds. Some oxygen can be incorporated into MgB2 and boron into MgO structures, MgBx (X=4-20) inclusions contain practically no oxygen. Regulating manufacturing temperature, pressure, introducing additions one can influence oxygen and boron distribution in the materials and thus, affect the formation, amount and sizes of Mg-B-O and MgBx inclusions and changing type of pinning, pinning force and so affect critical current density jc. The boron concentration increase in initial Mg and B mixture allows obtaining sample containing 88.5 wt% of MgB12 with Tc of 37.4 K (estimated magnetically).
Investigation of laser irradiated areas with electron backscatter diffraction
In this work, two silicon nitride (SiNx) layers with two different refraction indices, deposited on polished or damageetched silicon wafers were locally irradiated by laser pulses. The focus was set on the investigation of the ablation mechanisms. Thereby, an ultra-short laser source (pulse duration 10 ps, wavelength 532 nm, Gaussian profile) was used. The irradiated areas were investigated by electron backscatter diffraction (EBSD) in order to analyze the nearsurface crystallographic orientation and crystallinity. In this work an indirect ablation was observed for SiN x (n = 1.9). Further, a change from an indirect ablation to a partial lift-off for SiNx (n = 2.1) was determined to be fluence dependent. At low fluences, the SiNx was completely removed. However, at higher fluences, SiNx was not completely removed, due to direct ablation. The two-photonabsorption coefficient of SiNx (n = 2.1) was estimated to be 2·105 cm/TW.
Influence of nanobody binding on fluorescence emission, mobility, and organization of GFP-tagged proteins
Advanced fluorescence microscopy studies require specific and monovalent molecular labeling with bright and photostable fluorophores. This necessity led to the widespread use of fluorescently labeled nanobodies against commonly employed fluorescent proteins (FPs). However, very little is known how these nanobodies influence their target molecules. Here, we tested commercially available nanobodies and observed clear changes of the fluorescence properties, mobility and organization of green fluorescent protein (GFP) tagged proteins after labeling with the anti-GFP nanobody. Intriguingly, we did not observe any co-diffusion of fluorescently labeled nanobodies with the GFP-labeled proteins. Our results suggest significant binding of the nanobodies to a non-emissive, likely oligomerized, form of the FPs, promoting disassembly into monomeric form after binding. Our findings have significant implications on the application of nanobodies and GFP labeling for studying dynamic and quantitative protein organization in the plasma membrane of living cells using advanced imaging techniques.
Propagating and localized surface plasmon resonance sensing — A critical comparison based on measurements and theory
With its potential for ultrasensitive, label-free detection of molecular interactions, sensing methods based on the surface plasmon resonance (SPR) effect fully meet the requirements for modern analytical techniques. Already established by using propagating SPR in thin gold layers, the last years witnessed the emergence of another related technique utilizing extremely miniaturized noble metal sensor structures, based on a localized SPR. This paper provides a critical comparison of these kinds of SPR sensing, reviews the foundation of both general approaches, presents experimental data on exactly the same molecular model system using both techniques, as well as theoretical considerations in order to allow reasonable comparison. It highlights the specific features and effects, in order to provide guidance in choosing the right technique for given bioanalytical tasks. The study demonstrated the capabilities of LSPR for sensing of molecular layers even in the lower nanometer dimension. For the detection of small (bio)molecules, smaller particle diameters are favored regarding highest sensitivity. It also presents an approach to obtain refractive index and the thickness of a molecular film by analyzing the signal response of plasmonic sensors with metal nanoparticles. Moreover, an additional method for the improvement of the parameters' determination is introduced.