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- Akhtar, Sultan
(författare)
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Transmission Electron Microscopy of Graphene and Hydrated Biomaterial Nanostructures : Novel Techniques and Analysis
- 2012
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Doktorsavhandling (övrigt vetenskapligt/konstnärligt)abstract
- Transmission Electron Microscopy (TEM) on light element materials and soft matters is problematic due to electron irradiation damage and low contrast. In this doctoral thesis techniques were developed to address some of those issues and successfully characterize these materials at high resolution. These techniques were demonstrated on graphene flakes, DNA/magnetic beads and a number of water containing biomaterials. The details of these studies are given below.A TEM based method was presented for thickness characterization of graphene flakes. For the thickness characterization, the dynamical theory of electron diffraction is used to obtain an analytical expression for the intensity of the transmitted electron beam as a function of thickness. From JEMS simulations (experiments) the absorption constant λ in a low symmetry orientation was found to be ~ 208 nm (225 ± 9 nm). When compared to standard techniques for thickness determination of graphene/graphite, the method has the advantage of being relatively simple, fast and requiring only the acquisition of bright-field (BF) images. Using the proposed method, it is possible to measure the thickness change due to one monolayer of graphene if the flake has uniform thickness over a larger area.A real-space TEM study on magnetic bead-DNA coil interaction was conducted and a statistical analysis of the number of beads attached to the DNA-coils was performed. The average number of beads per DNA coil was calculated around 6 and slightly above 2 for samples with 40 nm and 130 nm beads, respectively. These results are in good agreement with magnetic measurements. In addition, the TEM analysis supported an earlier hypothesis that 40 nm beads are preferably attached interior of the DNA-coils while 130 nm beads closer to the exterior of the coils.A focused ion-beam in-situ lift-out technique for hydrated biological specimens was developed for cryo-TEM. The technique was demonstrated on frozen Aspergillus niger spores which were frozen with liquid nitrogen to preserve their cellular structures. A thin lamella was prepared, lifted out and welded to a TEM grid. Once the lamella was thinned to electron transparency, the grid was cryogenically transferred to the TEM using a cryo-transfer bath. The structure of the cells was revealed by BF imaging. Also, a series of energy filtered images was acquired and C, N and Mn elemental maps were produced. Furthermore, 3 Å lattice fringes of the underlying Al support were successfully resolved by high resolution imaging, confirming that the technique has the potential to extract structural information down to the atomic scale. The experimental protocol is ready now to be employed on a large variety of samples e.g. soft/hard matter interfaces.
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- Akhtar, Sultan, et al.
(författare)
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Visualization of functionalization of nano-particles and graphene in the TEM
- 2010
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Ingår i: Advanced Materials Workshop 2010.
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Konferensbidrag (refereegranskat)abstract
- Recently, the activity on functionalized nano-objects has strongly increased. Yet, there are, to our knowledge no techniques available that visualize the attachment of molecules to nano-entities such as nanoparticles and graphene. In this work, we show a methodology to analyse the attachment of molecules to nanoparticles and graphene. The difficulty of such transmission electron microscopy (TEM) characterization consists in the high beam sensitivity of these nanoobjects. We employed a high resolution- as well as diffraction contrast-imaging methods to characterize graphene. First, we have developed a method to measure the thickness of free-standing graphene-like layers. The refinement of these imaging techniques enabled the imaging of functionalized C60 (fullerene) on top of a few-layer graphene flake by TEM. We also developed a methodology to visualize the attachment of functionalized gold and magnetic nanoparticles (different sizes) to nonstained and unlabeled single strand DNA-coils. This technique can be used to understand the interaction of a large variety of functionalized nanoparticles with their solution environment and/or macromolecular structures for their large applications.
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| 8. |
- Ferrari, Raffaele, et al.
(författare)
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Frontotemporal dementia and its subtypes: a genome-wide association study.
- 2014
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Ingår i: Lancet Neurology. - 1474-4465. ; 13:7, s. 686-699
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Tidskriftsartikel (refereegranskat)abstract
- Frontotemporal dementia (FTD) is a complex disorder characterised by a broad range of clinical manifestations, differential pathological signatures, and genetic variability. Mutations in three genes-MAPT, GRN, and C9orf72-have been associated with FTD. We sought to identify novel genetic risk loci associated with the disorder.
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| 16. |
- Li, Ling-Guang, 1982-, et al.
(författare)
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Dynamics of SiO2 Buried Layer Removal from Si-SiO2-Si and Si-SiO2-SiC Bonded Substrates by Annealing in Ar
- 2014
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Ingår i: Journal of Electronic Materials. - : Springer Berlin/Heidelberg. - 0361-5235 .- 1543-186X. ; 43:2, s. 541-547
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Tidskriftsartikel (refereegranskat)abstract
- Silicon-on-silicon-carbide substrates could be ideal for high-power and radiofrequency silicon devices. Such hybrid wafers, when made by wafer bonding, contain an intermediate silicon dioxide layer with poor thermal characteristics, which can be removed by high-temperature annealing in an inert atmosphere. To understand the dynamics of this process, removal of 2.4-nm-thick SiO2 layers from Si-SiO2-Si and Si-SiO2-SiC substrates has been studied at temperatures ranging from 1100A degrees C to 1200A degrees C. The substrates were analyzed by transmission electron microscopy, electron energy-loss spectroscopy, secondary-ion mass spectroscopy, and ellipsometry, before and after annealing. For oxide thickness less than 2.4 nm, the activation energy for oxide removal was estimated to be 6.4 eV, being larger than the activation energy reported for removal of thicker oxides (4.1 eV). Under the same conditions, the SiO2 layer became discontinuous. In the time domain, three steps could be distinguished: bulk diffusion, bulk diffusion with void formation, and bulk diffusion with disintegration. The void formation, predominant here, has an energetic cost that could explain the larger activation energy. The oxide remaining after prolonged annealing corresponds to one layer of oxygen atoms.
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| 18. |
- Lidbaum, Hans, et al.
(författare)
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Reciprocal and real space maps for EMCD experiments
- 2010
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Ingår i: Ultramicroscopy. - : Elsevier BV. - 0304-3991 .- 1879-2723. ; 110:11, s. 1380-1389
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Tidskriftsartikel (refereegranskat)abstract
- Electron magnetic chiral dichroism (EMCD) is an emerging tool for quantitative measurements of magnetic properties using the transmission electron microscope (TEM), with the possibility of nanometer resolution. The geometrical conditions, data treatment and electron gun settings are found to influence the EMCD signal. In this article, particular care is taken to obtain a reliable quantitative measurement of the ratio of orbital to spin magnetic moment using energy filtered diffraction patterns. For this purpose, we describe a method for data treatment, normalization and selection of mirror axis. The experimental results are supported by theoretical simulations based on dynamical diffraction and density functional theory. Special settings of the electron gun, so called telefocus mode, enable a higher intensity of the electron beam, as well as a reduction of the influence from artifacts on the signal. Using these settings, we demonstrate the principle of acquiring real space maps of the EMCD signal. This enables advanced characterization of magnetic materials with superior spatial resolution.
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| 20. |
- Pujari-Palmer, Shiuli, et al.
(författare)
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In vivo and in vitro evaluation of hydroxyapatite nanoparticle morphology on the acute inflammatory response
- 2016
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Ingår i: Biomaterials. - : Elsevier BV. - 0142-9612 .- 1878-5905. ; 90, s. 1-11
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Tidskriftsartikel (refereegranskat)abstract
- Biomedical implants have been widely used in bone repair applications. However, nanosized degradation products from these implants could elicit an inflammatory reaction, which may lead to implant failure. It is well known that the size, chemistry, and charge of these nanoparticles can modulate this response, but little is known regarding the role that the particle's morphology plays in inducing inflammation. The present study aims to investigate the effect of hydroxyapatite nanoparticle (HANPs) morphology on inflammation, in-vitro and in-vivo. Four distinct HANP morphologies were fabricated and characterized: long rods, dots, sheets, and fibers. Primary human polymorphonuclear cells (PMNCs), mononuclear cells (MNCs), and human dermal fibroblasts (hDFs) were exposed to HANPs and alterations in cell viability, morphology, apoptotic activity, and reactive oxygen species (ROS) production were evaluated, in vitro. PMNCs and hDFs experienced a 2-fold decrease in viability following exposure to fibers, while MNC viability decreased 5-fold after treatment with the dots. Additionally, the fibers stimulated an elevated ROS response in both PMNCs and MNCs, and the largest apoptotic behavior for all cell types. Furthermore, exposure to fibers and dots resulted in greater capsule thickness when implanted subcutaneously in mice. Collectively, these results suggest that nanoparticle morphology can significantly impact the inflammatory response.
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| 21. |
- Rubino, Stefano, et al.
(författare)
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A Simple Transmission Electron Microscopy Method for Fast Thickness Characterization of Suspended Graphene and Graphite Flakes
- 2016
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Ingår i: Microscopy and Microanalysis. - 1431-9276 .- 1435-8115. ; 22:1, s. 250-256
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Tidskriftsartikel (refereegranskat)abstract
- We present a simple, fast method for thickness characterization of suspended graphene/graphite flakes that is based on transmission electron microscopy (TEM). We derive an analytical expression for the intensity of the transmitted electron beam I-0(t), as a function of the specimen thickness t (t < < lambda; where lambda is the absorption constant for graphite). We show that in thin graphite crystals the transmitted intensity is a linear function of t. Furthermore, high-resolution (HR) TEM simulations are performed to obtain lambda for a 001 zone axis orientation, in a two-beam case and in a low symmetry orientation. Subsequently, HR (used to determine t) and bright-field (to measure I-0(0) and I-0(t)) images were acquired to experimentally determine lambda. The experimental value measured in low symmetry orientation matches the calculated value (i.e., lambda = 225 +/- 9 nm). The simulations also show that the linear approximation is valid up to a sample thickness of 3-4 nm regardless of the orientation and up to several ten nanometers for a low symmetry orientation. When compared with standard techniques for thickness determination of graphene/graphite, the method we propose has the advantage of being simple and fast, requiring only the acquisition of bright-field images.
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| 23. |
- Rubino, Stefano, et al.
(författare)
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A site-specific focused-ion-beam lift-out method for cryo Transmission Electron Microscopy
- 2012
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Ingår i: Journal of Structural Biology. - : Elsevier BV. - 1047-8477 .- 1095-8657. ; 180:3, s. 572-576
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Tidskriftsartikel (refereegranskat)abstract
- The focused-ion-beam (FIB) is the method of choice for site-specific sample preparation for Transmission Electron Microscopy (TEM) in material sciences. A lamella can be physically lifted out from a specific region of a bulk specimen with submicrometer precision and thinned to electron transparency for high-resolution imaging in the TEM. The possibility to use this tool in life sciences applications has been limited by the lack of lift-out capabilities at the cryogenic temperatures often needed for biological samples. Conventional cryo-TEM sample preparation is mostly based on ultramicrotomy, a procedure that is not site-specific and known to produce artifacts. Here we demonstrate how a cooled nanomanipulator and a custom-built transfer station can be used to achieve cryo-preparation of TEM samples with the FIB, enabling high-resolution investigation of frozen-hydrated specimens in the TEM.
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| 24. |
- Rubino, Stefano, et al.
(författare)
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Cryo-electron Microscopy Specimen Preparation By Means Of a Focused Ion Beam
- 2014
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Ingår i: Journal of Visualized Experiments. - : MyJove Corporation. - 1940-087X. ; :89
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Tidskriftsartikel (refereegranskat)abstract
- Here we present a protocol used to prepare cryo-TEM samples of Aspergillus niger spores, but which can easily be adapted for any number of microorganisms or solutions. We make use of a custom built cryo-transfer station and a modified cryo-SEM preparation chamber2. The spores are taken from a culture, plunge-frozen in a liquid nitrogen slush and observed in the cryo-SEM to select a region of interest. A thin lamella is then extracted using the FIB, attached to a TEM grid and subsequently thinned to electron transparency. The grid is transferred to a cryo-TEM holder and into a TEM for high resolution studies. Thanks to the introduction of a cooled nanomanipulator tip and a cryo-transfer station, this protocol is a straightforward adaptation to cryogenic temperature of the routinely used FIB preparation of TEM samples. As such it has the advantages of requiring a small amount of modifications to existing instruments, setups and procedures; it is easy to implement; it has a broad range of applications, in principle the same as for cryo-TEM sample preparation. One limitation is that it requires skillful handling of the specimens at critical steps to avoid or minimize contaminations.
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