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Despite significant advances, the molecular identity of the cytotoxic species populated during in vivo amyloid formation crucial for the understanding of neurodegenerative disorders is yet to be revealed. In this study lysozyme prefibrillar oligomers and fibrils in both mature and sonicated states have been isolated through an optimized ultrafiltration/ultracentrifugation method and characterized with various optical spectroscopic techniques, atomic force microscopy, and transmission electron microscopy. We examined their level and mode of toxicity on rat pheochromocytoma (PC12) cells in both differentiated and undifferentiated states. We find that oligomers and fibrils display cytotoxic capabilities toward cultured cells in vitro, with oligomers producing elevated levels of cellular injury toward undifferentiated PC12 cells (PC12(undiff)). Furthermore, dual flow cytometry staining experiments demonstrate that the oligomers and mature fibrils induce divergent cellular death pathways (apoptosis and secondary necrosis, respectively) in these PC12 cells. We have also shown that oligomers but not sonicated mature fibrils inhibit hippocampal long term potentiation, a form of synaptic plasticity implicated in learning and memory, in vivo. We conclude that our in vitro and in vivo findings confer a level of resistance toward amyloid fibrils, and that the PC 12-based comparative cytotoxicity assay can provide insights into toxicity differences between differently aggregated protein species.
Assuntos
Amiloide/metabolismo , Biopolímeros/metabolismo , Morte Celular , Amiloide/química , Animais , Biopolímeros/química , Células PC12 , RatosRESUMO
Dry plasma etching for the pattern transfer of mask features is fundamental to semiconductor processing and the development of device and electrically conducting elements becomes more challenging as features reach the deep nanoscale regime. In this work, high resolution transmission electron microscopy (TEM) coupled with energy dispersive x-ray (EDX) characterization were used to analyze the pattern transfer of graphoepitaxially aligned block copolymer (BCP) features to germanium (Ge) substrates as a function of time. The BCP patterns were converted into metal oxide hardmasks in order to affect good aspect ratios of the transferred features. An unusual interface layer between metal oxide nanowires and the germanium-on-insulator substrate was observed. EDX analysis shows that the origin of this interface layer is a result of the presence of a negative tone e-beam resist material, HSQ (hydrogen silsesquioxane). HSQ was employed as a guiding material to align line-space features of poly(styrene)-block-poly(4-vinylpyridine) (PS-b-P4VP) BCP with 16 nm half-pitch topography. Additionally, the existence of a metal oxide layer (from the initial PS-b-P4VP film) is also shown through ex situ TEM and EDX characterization. Three dimensional modeling of features is also provided giving a unique insight into the arrangement and structure of BCP features prior to and after the pattern transfer process. The results presented in this article highlight the accuracy of high resolution electron microscopy and elemental mapping of BCP generated on-chip etch masks to observe and understand through-film features affecting pattern transfer.
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Connectivity in metallic nanowire networks with resistive junctions is manipulated by applying an electric field to create materials with tunable electrical conductivity. In situ electron microscope and electrical measurements visualize the activation and evolution of connectivity within these networks. Modeling nanowire networks, having a distribution of junction breakdown voltages, reveals universal scaling behavior applicable to all network materials. We demonstrate how local connectivity within these networks can be programmed and discuss material and device applications.
Assuntos
Nanopartículas Metálicas/química , Metais/química , Nanotecnologia/métodos , Nanofios/química , Condutividade Elétrica , Eletricidade , Humanos , Luz , Campos Magnéticos , Teste de Materiais , Modelos Estatísticos , Eletricidade Estática , Engenharia Tecidual/métodosRESUMO
If thermoplasmonic applications such as heat-assisted magnetic recording are to be commercially viable, it is necessary to optimize both thermal stability and plasmonic performance of the devices involved. In this work, a variety of different adhesion layers were investigated for their ability to reduce dewetting of sputtered 50 nm Au films on SiO2 substrates. Traditional adhesion layer metals Ti and Cr were compared with alternative materials of Al, Ta, and W. Film dewetting was shown to increase when the adhesion material diffuses through the Au layer. An adhesion layer thickness of 0.5 nm resulted in superior thermomechanical stability for all adhesion metals, with an enhancement factor of up to 200× over 5 nm thick analogues. The metals were ranked by their effectiveness in inhibiting dewetting, starting with the most effective, in the order Ta > Ti > W > Cr > Al. Finally, the Au surface-plasmon polariton response was compared for each adhesion layer, and it was found that 0.5 nm adhesion layers produced the best response, with W being the optimal adhesion layer material for plasmonic performance.
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In-situ transmission electron microscopy is rapidly emerging as the premier technique for characterising materials in a dynamic state on the atomic scale. The most important aspect of in-situ studies is specimen preparation. Specimens must be electron transparent and representative of the material in its operational state, amongst others. Here, a novel fabrication technique for the facile preparation of lamellae for in-situ transmission electron microscopy experimentation using focused ion beam milling is developed. This method involves the use of rotating microgrippers during the lift-out procedure, as opposed to the traditional micromanipulator needle and platinum weld. Using rotating grippers, and a unique adhesive substance, lamellae are mounted onto a MEMS device for in-situ TEM annealing experiments. We demonstrate how this technique can be used to avoid platinum deposition as well as minimising damage to the MEMS device during the thinning process. Our technique is both a cost effective and readily implementable alternative to the current generation of preparation methods for in-situ liquid, electrical, mechanical and thermal experimentation within the TEM as well as traditional cross-sectional lamella preparation.
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Nanoscale materials are frequently coated with surface stabilization layers during growth that prevent flocculation in solution and facilitate processing technologies such as ink-jet device printing. Here, we show that few-nanometer-thick stabilization layers typically used swell in the presence of certain solvents and impart significant stresses to the nanomaterial that remains even after the solvent has evaporated. Solvent swelling of the surface layer dramatically enhances nanomaterial-substrate adhesion via the collapse of the stabilization layer during solvent evaporation, preventing stress relaxation. We demonstrate the stress modulation of Ag, Au, and Si nanowires functionalised with surface polymers and surfactant layers and detect strain levels between 0.1 and 0.6% using atomic force microscopy mechanical measurement and Raman spectroscopy. Dry-transferred nanowires exhibit poor adhesion and show no evidence of incorporated stress but become stressed immediately following solvent exposure. Strain engineering is demonstrated by coating nanowires with few-nanometer-thick solvent-responsive polymer layers.
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In this article, we present a comprehensive investigation of the photothermal properties of plasmonic nanowire networks. We measure the local steady-state temperature increase, heat source density, and absorption in Ag, Au, and Ni metallic nanowire networks under optical illumination. This allows direct experimental confirmation of increased heat generation at the junction between two metallic nanowires and stacking-dependent absorption of polarized light. Due to thermal collective effects, the local temperature distribution in a network is shown to be completely delocalized on a micrometer scale, despite the nanoscale features in the heat source density. Comparison of the experimental temperature profile with numerical simulation allows an upper limit for the effective thermal conductivity of a Ag nanowire network to be established at 43 Wm(-1) K(-1) (0.1 κbulk).
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The measurement of Poisson's ratio of nanomaterials is extremely challenging. Here we report a lateral atomic force microscope experimental method to electromechanically measure the Poisson's ratio and gauge factor of individual nanowires. Under elastic loading conditions we monitor the four-point resistance of individual metallic nanowires as a function of strain and different levels of electrical stress. We determine the gauge factor of individual wires and directly measure the Poisson's ratio using a model that is independently validated for macroscopic wires. For macroscopic wires and nickel nanowires we find Poisson's ratios that closely correspond to bulk values, whereas for silver nanowires significant deviations from the bulk silver value are observed. Moreover, repeated measurements on individual silver nanowires at different levels of mechanical and electrical stress yield a small spread in Poisson ratio, with a range of mean values for different wires, all of which are distinct from the bulk value.
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Networks comprised of randomly oriented overlapping nanowires offer the possibility of simple fabrication on a variety of substrates, in contrast with the precise placement required for devices with single or aligned nanowires. Metal nanowires typically have a coating of surfactant or oxide that prevents aggregation, but also prevents electrical connection. Prohibitively high voltages can be required to electrically activate nanowire networks, and even after activation many nanowire junctions remain nonconducting. Nonelectrical activation methods can enhance conductivity but destroy the memristive behavior of the junctions that comprise the network. We show through both simulation and experiment that electrical stimulation, microstructured electrode geometry, and feature scaling can all be used to manipulate the connectivity and thus electrical conductivity of networks of silver nanowires with a nonconducting polymer coating. More generally, these results describe a strategy to integrate nanomaterials into controllable, adaptive macroscale materials.