Ortega, Eduardo | de Jonge, Niels
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DOI:Ortega, Eduardo | Nicholls, Daniel | Browning, Nigel D. | de Jonge, Niels
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Scanning transmission electron microscopy (STEM) provides structural analysis with sub-angstrom resolution. But the pixel-by-pixel scanning process is a limiting factor in acquiring high-speed data. Different strategies have been implemented to increase scanning speeds while at the same time minimizing beam damage via optimizing the scanning strategy. Here, we achieve the highest possible scanning speed by eliminating the image acquisition dead time induced by the beam flyback time combined with reducing the amount of scanning pixels via sparse imaging. A calibration procedure was developed to compensate for the hysteresis of the magnetic scan coils. A combination of sparse and serpentine scanning routines was tested for a crystalline thin film, gold nanoparticles, and in an in-situ liquid phase STEM experiment. Frame rates of 92, 23 and 5.8 s-1 were achieved for images of a width of 128, 256, and 512 pixels, respectively. The methods described here can be applied to single-particle tracking and analysis of radiation sensitive materials.
Peckys, Diana B. | Gaa, Daniel | de Jonge, Niels
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Currently, breast cancer patients are classified uniquely according to the expression level of hormone receptors, and human epidermal growth factor receptor 2 (HER2). This coarse classification is insufficient to capture the phenotypic complexity and heterogeneity of the disease. A methodology was developed for absolute quantification of receptor surface density ρR, and molecular interaction (dimerization), as well as the associated heterogeneities, of HER2 and its family member, the epidermal growth factor receptor (EGFR) in the plasma membrane of HER2 overexpressing breast cancer cells. Quantitative, correlative light microscopy (LM) and liquid-phase electron microscopy (LPEM) were combined with quantum dot (QD) labeling. Single-molecule position data of receptors were obtained from scanning transmission electron microscopy (STEM) images of intact cancer cells. Over 280,000 receptor positions were detected and statistically analyzed. An important finding was the subcellular heterogeneity in heterodimer shares with respect to plasma membrane regions with different dynamic properties. Deriving quantitative information about EGFR and HER2 ρR, as well as their dimer percentages, and the heterogeneities thereof, in single cancer cells, is potentially relevant for early identification of patients with HER2 overexpressing tumors comprising an enhanced share of EGFR dimers, likely increasing the risk for drug resistance, and thus requiring additional targeted therapeutic strategies
Peckys, Diana B. | Gaa, Daniel | Weinberg, Florian | de Jonge, Niels
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DOI:Ma, Haoran | Bennewitz, Roland
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Metallic glasses are excellent materials for micromechanical systems, where miniature components involving mechanical contact require control of friction at the microscopic scale. We report on an in-situ study of the structure of oxide films formed upon electrochemical polarization and their role in nanoscale friction on a metallic glass in aqueous environment using atomic force microscopy. The oxide film has a bilayer structure, as revealed by repeated scanning with the tip of an atomic force microscope. The dependence of friction on electrochemical potential reveals the growth mechanism and highlights the role of the oxide films for the frictional response of metallic glasses. The chemical sensitivity of nanotribology studies under electrochemical control contributes to the understanding of corrosion mechanisms on metallic glasses.
Penth, Michael | Schnellnhuber, Kordula | Bennewitz, Roland | Blass, Johanna
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DNA has become a powerful platform to design functional nanodevices. DNA nanodevices are often composed of self-assembled DNA building blocks that differ significantly from the structure of native DNA. In this study, we present Flow Force Microscopy as a massively parallel approach to study the nanomechanics of DNA self-assemblies on the single-molecular level. The high-throughput experiments performed in a simple microfluidic channel enable statistically meaningful studies with nanometer scale precision in a time frame of several minutes. A surprisingly high flexibility was observed for a typical construct used in DNA origami, reflected in a persistence length of 10.2 nm, a factor of five smaller than for native DNA. The enhanced flexibility is attributed to the discontinuous backbone of DNA self-assemblies that facilitate base pair opening by thermal fluctuations at the end of hybridized oligomers. We believe that the results will contribute to the fundamental understanding of DNA nanomechanics and help to improve the design of DNA nanodevices with applications in biological analysis and clinical research.
Elahi, Mahdi Amne | Koch, Marcus | Bardon, Julien | Addiego, Frédéric | Plapper, Peter
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The development of strong metal to polymer assemblies is currently an important research subject thanks to its prominence to develop lightweight structures. Furthermore, laser welding is known to be a fast, reliable, and versatile joining process, and it was demonstrated recently that it can be applied to such metal to polymer systems. To enhance the mechanical properties of the laser-joined aluminum-polyamide (Al-PA) specimens, laser polishing and laser ablation processes have been implemented on the aluminum surface before joining. The polyamide surface was also treated with the laser beam, separately. The surfaces were tested by several characterization techniques before and after each surface treatment. Then aluminum and polyamide samples with different surface treatments have been joined with an identical laser joining process. The mechanical properties of the joints in single lap shear configuration are reported and the failure mechanisms are discussed based on micro-computed x-ray tomography imaging of joined specimens and microscopic analysis before failure. Results show that both surface treatments of aluminum significantly improve the shear load of the joint; however, with different failure mechanisms. Polyamide surface treatment and increasing degree of crystallinity are effective when combined with the laser polishing of the Al surface. This combination is responsible for further enhancement of the shear load of the joint to the limit of base metal strength which is approximately 60 % improvement compared to the untreated samples. Finally, energy dispersive X-ray mapping shows the physicochemical bonding between aluminum oxide and polyamide at the interface.
Elahi, Mahdi Amne | Koch, Marcus | Plapper, Peter
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Laser welding is one of the most promising joining techniques to realize hybrid joints between metals and polymers in order to achieve weight reduction and functionalization of the parts. The surface treatment of the metal has a decisive effect on the joint quality and thus on the mechanical properties. In the present study, different mechanical and laser-based surface treatments have been investigated to develop diverse surface conditions on aluminum. Abrasive blasting and laser ablation were used to increase the surface roughness, while abrasive polishing and laser polishing were applied to minimize the surface roughness. In contrast to abrasive surface treatments, laser-based ones were implemented to create artificial oxide layers on the aluminum surface. The surface structures of pretreated samples have been studied with scanning electron microscopy and roughness test. The laser welding of pretreated aluminum with polyamide was achieved with the heat conduction joining technique. To enlarge the welding area and control the heat input, spatial and temporal modulations of the laser beam were implemented. Finally, a single lap tensile-shear test, microscopic analysis of fractured surfaces, and welding cross sections were employed to evaluate the joints. Results show that the presence of an artificial aluminum oxide layer and low roughness are essential to achieve a superior joint between aluminum and polyamide (improvement of approximately 58% in the shear load of the joint compared to as-received welded samples). The cross section of the superior joint which is the laser-polished aluminum welded to polyamide is studied with transmission electron microscopy.
Flormann, Daniel A. | Schu, Moritz | Terriac, Emmanuel | Thalla, Divyendu G. | Kainka, Lucina | Koch, Marcus | Gad, Annica K. B. | Lautenschläger, Franziska
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Abstract The rapid development of advanced microscopy techniques over recent decades has significantly increased the quality of imaging and our understanding of subcellular structures, such as the organization of the filaments of the cytoskeleton using fluorescence and electron microscopy. However, these recent improvements in imaging techniques have not been matched by similar development of techniques for computational analysis of the images of filament networks that can now be obtained. Hence, for a wide range of applications, reliable computational analysis of such two-dimensional methods remains challenging. Here, we present a new algorithm for tracing of filament networks. This software can extract many important parameters from grayscale images of filament networks, including the mesh hole size, and filament length and connectivity (also known as Coordination Number). In addition, the method allows sub-networks to be distinguished in two-dimensional images using intensity thresholding. We show that the algorithm can be used to analyze images of cytoskeleton networks obtained using different advanced microscopy methods. We have thus developed a new improved method for computational analysis of two-dimensional images of filamentous networks that has wide applications for existing imaging techniques. The algorithm is available as open-source software.
Konakov, Artem O. | Dremova, Nadejda N. | Khodos, Igor I. | Koch, Marcus | Zolotukhina, Ekaterina V. | Silina, Yuliya E.
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A novel one-pot chemical synthesis of functional copper iodide-polypyrrole composites, CuI-PPy, has been proposed. The fabrication process allows the formation of nanodimensional metal salt/polymer hybrid structures in a fully controlled time- and concentration-dependent manner. The impact of certain experimental conditions, viz., duration of synthesis, sequence of component addition and concentrations of the intact reagents on the structure, dimensionality and yield of the end-product was evaluated in detail. More specifically, the amount of marshite CuI within the hybrid composite can be ranged from 60 to 90 wt.%, depending on synthetic conditions (type and concentration of components, process duration). In addition, the conditions allowing the synthesis of nano-sized CuI distributed inside the polypyrrole matrix were found. A high morphological stability and reproducibility of the synthesized nanodimensional metal-polymer hybrid materials were approved. Finally, the electrochemical activity of the formed composites was verified by cyclic voltammetry studies. The stability of CuI-PPy composite deposited on the electrodes was strongly affected by the applied anodic limit. The proposed one-pot synthesis of the hybrid nanodimensional copper iodide-polypyrrole composites is highly innovative, meets the requirements of Green Chemistry and is potentially useful for future biosensor development. In addition, this study is expected to generally contribute to the knowledge on the hybrid nano-based composites with tailored properties.
