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Now showing 1 - 6 of 6
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    Combining super-resolution microcopy with neuronal network recording using magnesium fluoride thin films as cover layer for multi-electrode array technology
    (Berlin : Nature Publishing, 2019) Schmidl, Lars; Schmidl, Gabriele; Gawlik, Annett; Dellith, Jan; Hübner, Uwe; Tympel, Volker; Schmidl, Frank; Plentz, Jonathan; Geis, Christian; Haselmann, Holger
    We present an approach for fabrication of reproducible, chemically and mechanically robust functionalized layers based on MgF2 thin films on thin glass substrates. These show great advantages for use in super-resolution microscopy as well as for multi-electrode-array fabrication and are especially suited for combination of these techniques. The transparency of the coated substrates with the low refractive index material is adjustable by the layer thickness and can be increased above 92%. Due to the hydrophobic and lipophilic properties of the thin crystalline MgF2 layers, the temporal stable adhesion needed for fixation of thin tissue, e.g. cryogenic brain slices is given. This has been tested using localization-based super-resolution microscopy with currently highest spatial resolution in light microscopy. We demonstrated that direct stochastic optical reconstruction microscopy revealed in reliable imaging of structures of central synapses by use of double immunostaining of post- (homer1 and GluA2) and presynaptic (bassoon) marker structure in a 10 µm brain slice without additional fixing of the slices. Due to the proven additional electrical insulating effect of MgF2 layers, surfaces of multi-electrode-arrays were coated with this material and tested by voltage-current-measurements. MgF2 coated multi-electrode-arrays can be used as a functionalized microscope cover slip for combination with live-cell super-resolution microscopy.
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    New perspectives for viability studies with high-content analysis Raman spectroscopy (HCA-RS)
    (Berlin : Nature Publishing, 2019) Mondol, Abdullah S.; Töpfer, Natalie; Rüger, Jan; Neugebauer, Ute; Popp, Jürgen; Schie, Iwan W.
    Raman spectroscopy has been widely used in clinical and molecular biological studies, providing high chemical specificity without the necessity of labels and with little-to-no sample preparation. However, currently performed Raman-based studies of eukaryotic cells are still very laborious and time-consuming, resulting in a low number of sampled cells and questionable statistical validations. Furthermore, the approach requires a trained specialist to perform and analyze the experiments, rendering the method less attractive for most laboratories. In this work, we present a new high-content analysis Raman spectroscopy (HCA-RS) platform that overcomes the current challenges of conventional Raman spectroscopy implementations. HCA-RS allows sampling of a large number of cells under different physiological conditions without any user interaction. The performance of the approach is successfully demonstrated by the development of a Raman-based cell viability assay, i.e., the effect of doxorubicin concentration on monocytic THP-1 cells. A statistical model, principal component analysis combined with support vector machine (PCA-SVM), was found to successfully predict the percentage of viable cells in a mixed population and is in good agreement to results obtained by a standard cell viability assay. This study demonstrates the potential of Raman spectroscopy as a standard high-throughput tool for clinical and biological applications.
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    Surface deep profile synchrotron studies of mechanically modified top-down silicon nanowires array using ultrasoft X-ray absorption near edge structure spectroscopy
    (Berlin : Nature Publishing, 2019) Turishchev, S.Yu.; Parinova, V.E.; Pisliaruka, Aleksandra; Koyuda, D.A.; Yermukhamed, Dana; Ming, Tingsen; Ovsyannikov, Ruslan; Smirnov, Dmitriy; Makarova, Anna; Sivakov, Vladimir
    Atomic, electronic structure and composition of top-down metal-assisted wet-chemically etched silicon nanowires were studied by synchrotron radiation based X-ray absorption near edge structure technique. Local surrounding of the silicon and oxygen atoms in silicon nanowires array was studied on as-prepared nanostructured surfaces (atop part of nanowires) and their bulk part after, first time applied, in-situ mechanical removal atop part of the formed silicon nanowires. Silicon suboxides together with disturbed silicon dioxide were found in the composition of the formed arrays that affects the electronic structure of silicon nanowires. The results obtained by us convincingly testify to the homogeneity of the phase composition of the side walls of silicon nanowires and the electronic structure in the entire length of the nanowire. The controlled formation of the silicon nanowires array may lead to smart engineering of its atomic and electronic structure that influences the exploiting strategy of metal-assisted wet-chemically etched silicon nanowires as universal matrices for different applications.
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    Structural insights into heme binding to IL-36α proinflammatory cytokine
    (Berlin : Nature Publishing, 2019) Wißbrock, Amelie; Goradia, Nishit; Kumar, Amit; Paul George, Ajay Abisheck; Kühl, Toni; Bellstedt, Peter; Ramachandran, Ramadurai; Hoffmann, Patrick; Galler, Kerstin; Popp, Jürgen; Neugebauer, Ute; Hampel, Kornelia; Zimmermann, Bastian; Adam, Susanne; Wiendl, Maximilian; Krönke, Gerhard; Hamza, Iqbal; Heinemann, Stefan H.; Frey, Silke; Hueber, Axel J.; Ohlenschläger, Oliver; Imhof, Diana
    Cytokines of the interleukin (IL)-1 family regulate immune and inflammatory responses. The recently discovered IL-36 family members are involved in psoriasis, rheumatoid arthritis, and pulmonary diseases. Here, we show that IL-36α interacts with heme thereby contributing to its regulation. Based on in-depth spectroscopic analyses, we describe two heme-binding sites in IL-36α that associate with heme in a pentacoordinated fashion. Solution NMR analysis reveals structural features of IL-36α and its complex with heme. Structural investigation of a truncated IL-36α supports the notion that the N-terminus is necessary for association with its cognate receptor. Consistent with our structural studies, IL-36-mediated signal transduction was negatively regulated by heme in synovial fibroblast-like synoviocytes from rheumatoid arthritis patients. Taken together, our results provide a structural framework for heme-binding proteins and add IL-1 cytokines to the group of potentially heme-regulated proteins.
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    Symmetry breaking-induced magnetic Fano resonances in densely packed arrays of symmetric nanotrimers
    (Berlin : Nature Publishing, 2019) Wang, Ning; Zeisberger, Matthias; Hübner, Uwe; Giannini, Vincenzo; Schmidt, Markus A.
    Due to unique properties and great design flexibilities, Fano resonances represent one of the most promising optical features mediated by metallic nanostructures, while the excitation of some Fano modes is impossible due to symmetry reasons. The aim of this work is to show that dense lattice arrangements can have a profound impact on the optical properties of nanostructures and, in particular, can enable the excitation of otherwise dark modes. Here, we demonstrate this concept using the example of rectangular arrays of symmetric trimers packed so densely that the coupling between neighbouring unit cells imposes a symmetry break, enabling the excitation of magnetic Fano resonances. We found that in experiments as well as in simulations, electric and magnetic Fano resonances can be simultaneously formed in cases where the inter-trimer distances are sufficiently small. By analysing the transition from an isolated trimer mode into a regime of strong near-field coupling, we show that by modifying the rectangular unit cell lengths due to the symmetry mismatch between lattice and trimer, two types of Fano resonances can be found, especially magnetic Fano resonances with loop-type magnetic field distributions within the centre of each trimer, which can be either enhanced or suppressed. In addition, the influence of the refractive index environment was measured, showing sensitivity values of approximately 300 nm/RIU. Our work provides fundamental insights into the interaction of the lattice and nanostructure response and paves the way towards the observation of novel optical excitations.
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    Autofluorescence lifetime augmented reality as a means for real-time robotic surgery guidance in human patients
    (Berlin : Nature Publishing, 2019) Gorpas, Dimitris; Phipps, Jennifer E.; Bec, Julien; Ma, Dinglong; Dochow, Sebastian; Yankelevich, Diego R.; Sorger, Jonathan; Popp, Jürgen; Bewley, Arnaud Fassett; Gandour-Edwards, Regina F.; Marcu, Laura; Farwell, D. Gregory
    Due to loss of tactile feedback the assessment of tumor margins during robotic surgery is based only on visual inspection, which is neither significantly sensitive nor specific. Here we demonstrate time-resolved fluorescence spectroscopy (TRFS) as a novel technique to complement the visual inspection of oral cancers during transoral robotic surgery (TORS) in real-time and without the need for exogenous contrast agents. TRFS enables identification of cancerous tissue by its distinct autofluorescence signature that is associated with the alteration of tissue structure and biochemical profile. A prototype TRFS instrument was integrated synergistically with the da Vinci Surgical robot and the combined system was validated in swine and human patients. Label-free and real-time assessment and visualization of tissue biochemical features during robotic surgery procedure, as demonstrated here, not only has the potential to improve the intraoperative decision making during TORS but also other robotic procedures without modification of conventional clinical protocols.