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Sample records for high-resolution cryo-scanning electron

  1. Correlated cryo-fluorescence and cryo-electron microscopy with high spatial precision and improved sensitivity

    International Nuclear Information System (INIS)

    Schorb, Martin; Briggs, John A.G.

    2014-01-01

    Performing fluorescence microscopy and electron microscopy on the same sample allows fluorescent signals to be used to identify and locate features of interest for subsequent imaging by electron microscopy. To carry out such correlative microscopy on vitrified samples appropriate for structural cryo-electron microscopy it is necessary to perform fluorescence microscopy at liquid-nitrogen temperatures. Here we describe an adaptation of a cryo-light microscopy stage to permit use of high-numerical aperture objectives. This allows high-sensitivity and high-resolution fluorescence microscopy of vitrified samples. We describe and apply a correlative cryo-fluorescence and cryo-electron microscopy workflow together with a fiducial bead-based image correlation procedure. This procedure allows us to locate fluorescent bacteriophages in cryo-electron microscopy images with an accuracy on the order of 50 nm, based on their fluorescent signal. It will allow the user to precisely and unambiguously identify and locate objects and events for subsequent high-resolution structural study, based on fluorescent signals. - Highlights: • Workflow for correlated cryo-fluorescence and cryo-electron microscopy. • Cryo-fluorescence microscopy setup incorporating a high numerical aperture objective. • Fluorescent signals located in cryo-electron micrographs with 50 nm spatial precision

  2. Correlated cryo-fluorescence and cryo-electron microscopy with high spatial precision and improved sensitivity

    Energy Technology Data Exchange (ETDEWEB)

    Schorb, Martin [Structural and Computational Biology Unit, European Molecular Biology Laboratory, D-69117 Heidelberg (Germany); Briggs, John A.G., E-mail: john.briggs@embl.de [Structural and Computational Biology Unit, European Molecular Biology Laboratory, D-69117 Heidelberg (Germany); Cell Biology and Biophysics Unit, European Molecular Biology Laboratory, D-69117 Heidelberg (Germany)

    2014-08-01

    Performing fluorescence microscopy and electron microscopy on the same sample allows fluorescent signals to be used to identify and locate features of interest for subsequent imaging by electron microscopy. To carry out such correlative microscopy on vitrified samples appropriate for structural cryo-electron microscopy it is necessary to perform fluorescence microscopy at liquid-nitrogen temperatures. Here we describe an adaptation of a cryo-light microscopy stage to permit use of high-numerical aperture objectives. This allows high-sensitivity and high-resolution fluorescence microscopy of vitrified samples. We describe and apply a correlative cryo-fluorescence and cryo-electron microscopy workflow together with a fiducial bead-based image correlation procedure. This procedure allows us to locate fluorescent bacteriophages in cryo-electron microscopy images with an accuracy on the order of 50 nm, based on their fluorescent signal. It will allow the user to precisely and unambiguously identify and locate objects and events for subsequent high-resolution structural study, based on fluorescent signals. - Highlights: • Workflow for correlated cryo-fluorescence and cryo-electron microscopy. • Cryo-fluorescence microscopy setup incorporating a high numerical aperture objective. • Fluorescent signals located in cryo-electron micrographs with 50 nm spatial precision.

  3. Preservation of high resolution protein structure by cryo-electron microscopy of vitreous sections

    International Nuclear Information System (INIS)

    Sader, Kasim; Studer, Daniel; Zuber, Benoit; Gnaegi, Helmut; Trinick, John

    2009-01-01

    We have quantitated the degree of structural preservation in cryo-sections of a vitrified biological specimen. Previous studies have used sections of periodic specimens to assess the resolution present, but preservation before sectioning was not assessed and so the damage due particularly to cutting was not clear. In this study large single crystals of lysozyme were vitrified and from these X-ray diffraction patterns extending to better than 2.1 A were obtained. The crystals were high pressure frozen in 30% dextran, and cryo-sectioned using a diamond knife. In the best case, preservation to a resolution of 7.9 A was shown by electron diffraction, the first observation of sub-nanometre structural preservation in a vitreous section.

  4. Cryo-electron microscopy of membrane proteins.

    Science.gov (United States)

    Goldie, Kenneth N; Abeyrathne, Priyanka; Kebbel, Fabian; Chami, Mohamed; Ringler, Philippe; Stahlberg, Henning

    2014-01-01

    Electron crystallography is used to study membrane proteins in the form of planar, two-dimensional (2D) crystals, or other crystalline arrays such as tubular crystals. This method has been used to determine the atomic resolution structures of bacteriorhodopsin, tubulin, aquaporins, and several other membrane proteins. In addition, a large number of membrane protein structures were studied at a slightly lower resolution, whereby at least secondary structure motifs could be identified.In order to conserve the structural details of delicate crystalline arrays, cryo-electron microscopy (cryo-EM) allows imaging and/or electron diffraction of membrane proteins in their close-to-native state within a lipid bilayer membrane.To achieve ultimate high-resolution structural information of 2D crystals, meticulous sample preparation for electron crystallography is of outmost importance. Beam-induced specimen drift and lack of specimen flatness can severely affect the attainable resolution of images for tilted samples. Sample preparations that sandwich the 2D crystals between symmetrical carbon films reduce the beam-induced specimen drift, and the flatness of the preparations can be optimized by the choice of the grid material and the preparation protocol.Data collection in the cryo-electron microscope using either the imaging or the electron diffraction mode has to be performed applying low-dose procedures. Spot-scanning further reduces the effects of beam-induced drift. Data collection using automated acquisition schemes, along with improved and user-friendlier data processing software, is increasingly being used and is likely to bring the technique to a wider user base.

  5. Rapid increase of near atomic resolution virus capsid structures determined by cryo-electron microscopy.

    Science.gov (United States)

    Ho, Phuong T; Reddy, Vijay S

    2018-01-01

    The recent technological advances in electron microscopes, detectors, as well as image processing and reconstruction software have brought single particle cryo-electron microscopy (cryo-EM) into prominence for determining structures of bio-molecules at near atomic resolution. This has been particularly true for virus capsids, ribosomes, and other large assemblies, which have been the ideal specimens for structural studies by cryo-EM approaches. An analysis of time series metadata of virus structures on the methods of structure determination, resolution of the structures, and size of the virus particles revealed a rapid increase in the virus structures determined by cryo-EM at near atomic resolution since 2010. In addition, the data highlight the median resolution (∼3.0 Å) and size (∼310.0 Å in diameter) of the virus particles determined by X-ray crystallography while no such limits exist for cryo-EM structures, which have a median diameter of 508 Å. Notably, cryo-EM virus structures in the last four years have a median resolution of 3.9 Å. Taken together with minimal sample requirements, not needing diffraction quality crystals, and being able to achieve similar resolutions of the crystal structures makes cryo-EM the method of choice for current and future virus capsid structure determinations. Copyright © 2017 Elsevier Inc. All rights reserved.

  6. Coordinate transformation based cryo-correlative methods for electron tomography and focused ion beam milling

    International Nuclear Information System (INIS)

    Fukuda, Yoshiyuki; Schrod, Nikolas; Schaffer, Miroslava; Feng, Li Rebekah; Baumeister, Wolfgang; Lucic, Vladan

    2014-01-01

    Correlative microscopy allows imaging of the same feature over multiple length scales, combining light microscopy with high resolution information provided by electron microscopy. We demonstrate two procedures for coordinate transformation based correlative microscopy of vitrified biological samples applicable to different imaging modes. The first procedure aims at navigating cryo-electron tomography to cellular regions identified by fluorescent labels. The second procedure, allowing navigation of focused ion beam milling to fluorescently labeled molecules, is based on the introduction of an intermediate scanning electron microscopy imaging step to overcome the large difference between cryo-light microscopy and focused ion beam imaging modes. These methods make it possible to image fluorescently labeled macromolecular complexes in their natural environments by cryo-electron tomography, while minimizing exposure to the electron beam during the search for features of interest. - Highlights: • Correlative light microscopy and focused ion beam milling of vitrified samples. • Coordinate transformation based cryo-correlative method. • Improved correlative light microscopy and cryo-electron tomography

  7. High-precision correlative fluorescence and electron cryo microscopy using two independent alignment markers

    International Nuclear Information System (INIS)

    Schellenberger, Pascale; Kaufmann, Rainer; Siebert, C. Alistair; Hagen, Christoph; Wodrich, Harald; Grünewald, Kay

    2014-01-01

    Correlative light and electron microscopy (CLEM) is an emerging technique which combines functional information provided by fluorescence microscopy (FM) with the high-resolution structural information of electron microscopy (EM). So far, correlative cryo microscopy of frozen-hydrated samples has not reached better than micrometre range accuracy. Here, a method is presented that enables the correlation between fluorescently tagged proteins and electron cryo tomography (cryoET) data with nanometre range precision. Specifically, thin areas of vitrified whole cells are examined by correlative fluorescence cryo microscopy (cryoFM) and cryoET. Novel aspects of the presented cryoCLEM workflow not only include the implementation of two independent electron dense fluorescent markers to improve the precision of the alignment, but also the ability of obtaining an estimate of the correlation accuracy for each individual object of interest. The correlative workflow from plunge-freezing to cryoET is detailed step-by-step for the example of locating fluorescence-labelled adenovirus particles trafficking inside a cell. - Highlights: • Vitrified mammalian cell were imaged by fluorescence and electron cryo microscopy. • TetraSpeck fluorescence markers were added to correct shifts between cryo fluorescence channels. • FluoSpheres fiducials were used as reference points to assign new coordinates to cryoEM images. • Adenovirus particles were localised with an average correlation precision of 63 nm

  8. High-precision correlative fluorescence and electron cryo microscopy using two independent alignment markers

    Energy Technology Data Exchange (ETDEWEB)

    Schellenberger, Pascale [Oxford Particle Imaging Centre, Division of Structural Biology, Wellcome Trust Centre for Human Genetics, University of Oxford, Roosevelt Drive, Oxford OX3 7BN (United Kingdom); Kaufmann, Rainer [Oxford Particle Imaging Centre, Division of Structural Biology, Wellcome Trust Centre for Human Genetics, University of Oxford, Roosevelt Drive, Oxford OX3 7BN (United Kingdom); Department of Biochemistry, University of Oxford, South Parks Road, Oxford OX1 3QU (United Kingdom); Siebert, C. Alistair; Hagen, Christoph [Oxford Particle Imaging Centre, Division of Structural Biology, Wellcome Trust Centre for Human Genetics, University of Oxford, Roosevelt Drive, Oxford OX3 7BN (United Kingdom); Wodrich, Harald [Microbiologie Fondamentale et Pathogénicité, MFP CNRS UMR 5234, University of Bordeaux SEGALEN, 146 rue Leo Seignat, 33076 Bordeaux (France); Grünewald, Kay, E-mail: kay@strubi.ox.ac.uk [Oxford Particle Imaging Centre, Division of Structural Biology, Wellcome Trust Centre for Human Genetics, University of Oxford, Roosevelt Drive, Oxford OX3 7BN (United Kingdom)

    2014-08-01

    Correlative light and electron microscopy (CLEM) is an emerging technique which combines functional information provided by fluorescence microscopy (FM) with the high-resolution structural information of electron microscopy (EM). So far, correlative cryo microscopy of frozen-hydrated samples has not reached better than micrometre range accuracy. Here, a method is presented that enables the correlation between fluorescently tagged proteins and electron cryo tomography (cryoET) data with nanometre range precision. Specifically, thin areas of vitrified whole cells are examined by correlative fluorescence cryo microscopy (cryoFM) and cryoET. Novel aspects of the presented cryoCLEM workflow not only include the implementation of two independent electron dense fluorescent markers to improve the precision of the alignment, but also the ability of obtaining an estimate of the correlation accuracy for each individual object of interest. The correlative workflow from plunge-freezing to cryoET is detailed step-by-step for the example of locating fluorescence-labelled adenovirus particles trafficking inside a cell. - Highlights: • Vitrified mammalian cell were imaged by fluorescence and electron cryo microscopy. • TetraSpeck fluorescence markers were added to correct shifts between cryo fluorescence channels. • FluoSpheres fiducials were used as reference points to assign new coordinates to cryoEM images. • Adenovirus particles were localised with an average correlation precision of 63 nm.

  9. Hydration Effects on Skin Microstructure as Probed by High-Resolution Cryo-Scanning Electron Microscopy and Mechanistic Implications to Enhanced Transcutaneous Delivery of Biomacromolecules

    Science.gov (United States)

    Tan, Grace; Xu, Peng; Lawson, Louise B.; He, Jibao; Freytag, Lucia C.; Clements, John D.; John, Vijay T.

    2010-01-01

    Although hydration is long known to improve the permeability of skin, penetration of macromolecules such as proteins is limited and the understanding of enhanced transport is based on empirical observations. This study uses high-resolution cryo-scanning electron microscopy to visualize microstructural changes in the stratum corneum (SC) and enable a mechanistic interpretation of biomacromolecule penetration through highly hydrated porcine skin. Swollen corneocytes, separation of lipid bilayers in the SC intercellular space to form cisternae, and networks of spherical particulates are observed in porcine skin tissue hydrated for a period of 4–10 h. This is explained through compaction of skin lipids when hydrated, a reversal in the conformational transition from unilamellar liposomes in lamellar granules to lamellae between keratinocytes when the SC skin barrier is initially established. Confocal microscopy studies show distinct enhancement in penetration of fluorescein isothiocyanate-bovine serum albumin (FITC-BSA) through skin hydrated for 4–10 h, and limited penetration of FITC-BSA once skin is restored to its natively hydrated structure when exposed to the environment for 2–3 h. These results demonstrate the effectiveness of a 4–10 h hydration period to enhance transcutaneous penetration of large biomacromolecules without permanently damaging the skin. PMID:19582754

  10. High-resolution structure of the Shigella type-III secretion needle by solid-state NMR and cryo-electron microscopy

    Science.gov (United States)

    Demers, Jean-Philippe; Habenstein, Birgit; Loquet, Antoine; Kumar Vasa, Suresh; Giller, Karin; Becker, Stefan; Baker, David; Lange, Adam; Sgourakis, Nikolaos G.

    2014-09-01

    We introduce a general hybrid approach for determining the structures of supramolecular assemblies. Cryo-electron microscopy (cryo-EM) data define the overall envelope of the assembly and rigid-body orientation of the subunits while solid-state nuclear magnetic resonance (ssNMR) chemical shifts and distance constraints define the local secondary structure, protein fold and inter-subunit interactions. Finally, Rosetta structure calculations provide a general framework to integrate the different sources of structural information. Combining a 7.7-Å cryo-EM density map and 996 ssNMR distance constraints, the structure of the type-III secretion system needle of Shigella flexneri is determined to a precision of 0.4 Å. The calculated structures are cross-validated using an independent data set of 691 ssNMR constraints and scanning transmission electron microscopy measurements. The hybrid model resolves the conformation of the non-conserved N terminus, which occupies a protrusion in the cryo-EM density, and reveals conserved pore residues forming a continuous pattern of electrostatic interactions, thereby suggesting a mechanism for effector protein translocation.

  11. A simple cryo-holder facilitates specimen observation under a conventional scanning electron microscope.

    Science.gov (United States)

    Tang, Chih-Yuan; Huang, Rong-Nan; Kuo-Huang, Ling-Long; Kuo, Tai-Chih; Yang, Ya-Yun; Lin, Ching-Yeh; Jane, Wann-Neng; Chen, Shiang-Jiuun

    2012-02-01

    A pre-cryogenic holder (cryo-holder) facilitating cryo-specimen observation under a conventional scanning electron microscope (SEM) is described. This cryo-holder includes a specimen-holding unit (the stub) and a cryogenic energy-storing unit (a composite of three cylinders assembled with a screw). After cooling, the cryo-holder can continue supplying cryogenic energy to extend the observation time for the specimen in a conventional SEM. Moreover, the cryogenic energy-storing unit could retain appropriate liquid nitrogen that can evaporate to prevent frost deposition on the surface of the specimen. This device is proved feasible for various tissues and cells, and can be applied to the fields of both biology and material science. We have employed this novel cryo-holder for observation of yeast cells, trichome, and epidermal cells in the leaf of Arabidopsis thaliana, compound eyes of insects, red blood cells, filiform papillae on the surface of rat tongue, agar medium, water molecules, penicillium, etc. All results suggested that the newly designed cryo-holder is applicable for cryo-specimen observation under a conventional SEM without cooling system. Most importantly, the design of this cryo-holder is simple and easy to operate and could adapt a conventional SEM to a plain type cryo-SEM affordable for most laboratories. Copyright © 2011 Wiley Periodicals, Inc.

  12. Correlative cryo-fluorescence light microscopy and cryo-electron tomography of Streptomyces.

    Science.gov (United States)

    Koning, Roman I; Celler, Katherine; Willemse, Joost; Bos, Erik; van Wezel, Gilles P; Koster, Abraham J

    2014-01-01

    Light microscopy and electron microscopy are complementary techniques that in a correlative approach enable identification and targeting of fluorescently labeled structures in situ for three-dimensional imaging at nanometer resolution. Correlative imaging allows electron microscopic images to be positioned in a broader temporal and spatial context. We employed cryo-correlative light and electron microscopy (cryo-CLEM), combining cryo-fluorescence light microscopy and cryo-electron tomography, on vitrified Streptomyces bacteria to study cell division. Streptomycetes are mycelial bacteria that grow as long hyphae and reproduce via sporulation. On solid media, Streptomyces subsequently form distinct aerial mycelia where cell division leads to the formation of unigenomic spores which separate and disperse to form new colonies. In liquid media, only vegetative hyphae are present divided by noncell separating crosswalls. Their multicellular life style makes them exciting model systems for the study of bacterial development and cell division. Complex intracellular structures have been visualized with transmission electron microscopy. Here, we describe the methods for cryo-CLEM that we applied for studying Streptomyces. These methods include cell growth, fluorescent labeling, cryo-fixation by vitrification, cryo-light microscopy using a Linkam cryo-stage, image overlay and relocation, cryo-electron tomography using a Titan Krios, and tomographic reconstruction. Additionally, methods for segmentation, volume rendering, and visualization of the correlative data are described. © 2014 Elsevier Inc. All rights reserved.

  13. Imaging of Norway spruce early somatic embryos with the ESEM, Cryo-SEM and laser scanning microscope.

    Science.gov (United States)

    Neděla, Vilém; Hřib, Jiří; Havel, Ladislav; Hudec, Jiří; Runštuk, Jiří

    2016-05-01

    This article describes the surface structure of Norway spruce early somatic embryos (ESEs) as a typical culture with asynchronous development. The microstructure of extracellular matrix covering ESEs were observed using the environmental scanning electron microscope as a primary tool and using the scanning electron microscope with cryo attachment and laser electron microscope as a complementary tool allowing our results to be proven independently. The fresh samples were observed in conditions of the air environment of the environmental scanning electron microscope (ESEM) with the pressure from 550Pa to 690Pa and the low temperature of the sample from -18°C to -22°C. The samples were studied using two different types of detector to allow studying either the thin surface structure or material composition. The scanning electron microscope with cryo attachment was used for imaging frozen extracellular matrix microstructure with higher resolution. The combination of both electron microscopy methods was suitable for observation of "native" plant samples, allowing correct evaluation of our results, free of error and artifacts. Copyright © 2016 Elsevier Ltd. All rights reserved.

  14. Advances in cryo-electron tomography for biology and medicine.

    Science.gov (United States)

    Koning, Roman I; Koster, Abraham J; Sharp, Thomas H

    2018-05-01

    Cryo-electron tomography (CET) utilizes a combination of specimen cryo-fixation and multi-angle electron microscopy imaging to produce three-dimensional (3D) volume reconstructions of native-state macromolecular and subcellular biological structures with nanometer-scale resolution. In recent years, cryo-electron microscopy (cryoEM) has experienced a dramatic increase in the attainable resolution of 3D reconstructions, resulting from technical improvements of electron microscopes, improved detector sensitivity, the implementation of phase plates, automated data acquisition schemes, and improved image reconstruction software and hardware. These developments also greatly increased the usability and applicability of CET as a diagnostic and research tool, which is now enabling structural biologists to determine the structure of proteins in their native cellular environment to sub-nanometer resolution. These recent technical developments have stimulated us to update on our previous review (Koning, R.I., Koster, A.J., 2009. Cryo-electron tomography in biology and medicine. Ann Anat 191, 427-445) in which we described the fundamentals of CET. In this follow-up, we extend this basic description in order to explain the aforementioned recent advances, and describe related 3D techniques that can be applied to the anatomy of biological systems that are relevant for medicine. Copyright © 2018 Elsevier GmbH. All rights reserved.

  15. Secondary electron spectroscopy and Auger microscopy at high spatial resolution. Application to scanning electron microscopy

    International Nuclear Information System (INIS)

    Le Gressus, Claude; Massignon, Daniel; Sopizet, Rene

    1979-01-01

    Secondary electron spectroscopy (SES), Auger electron spectroscopy (AES) and electron energy loss spectroscopy (ELS) are combined with ultra high vacuum scanning microscopy (SEM) for surface analysis at high spatial resolution. Reliability tests for the optical column for the vacuum and for the spectrometer are discussed. Furthermore the sensitivity threshold in AES which is compatible with a non destructive surface analysis at high spatial resolution is evaluated. This combination of all spectroscopies is used in the study of the beam damage correlated with the well known secondary electron image (SEI) darkening still observed in ultra high vacuum. The darkening is explained as a bulk decontamination of the sample rather than as a surface contamination from the residual vacuum gas [fr

  16. National Cryo-Electron Microscopy Facility

    Science.gov (United States)

    Information about the National Cryo-EM Facility at NCI, created to provide researchers access to the latest cryo-EM technology for high resolution imaging. Includes timeline for installation and how to access the facility.

  17. Pleomorphism and Viability of the Lyme Disease Pathogen Borrelia burgdorferi Exposed to Physiological Stress Conditions: A Correlative Cryo-Fluorescence and Cryo-Scanning Electron Microscopy Study

    Czech Academy of Sciences Publication Activity Database

    Vancová, Marie; Rudenko, Natalia; Vaněček, Jiří; Golovchenko, Maryna; Strnad, Martin; Rego, Ryan O. M.; Tichá, Lucie; Grubhoffer, Libor; Nebesářová, Jana

    2017-01-01

    Roč. 8, 11 April (2017), č. článku 596. ISSN 1664-302X R&D Projects: GA TA ČR(CZ) TE01020118; GA MŠk(CZ) LM2015062 EU Projects: European Commission(XE) 278976 - ANTIGONE Institutional support: RVO:60077344 Keywords : cryo-fluorescence * cryo-scanning electron microscopy * Borrelia burgdorferi * Lyme disease * round body * pleomorphism * viability staining Subject RIV: EB - Genetics ; Molecular Biology OBOR OECD: 2.11 Other engineering and technologies Impact factor: 4.076, year: 2016

  18. Zernike phase contrast cryo-electron tomography of whole bacterial cells.

    Science.gov (United States)

    Guerrero-Ferreira, Ricardo C; Wright, Elizabeth R

    2014-01-01

    Cryo-electron tomography (cryo-ET) provides three-dimensional (3D) structural information of bacteria preserved in a native, frozen-hydrated state. The typical low contrast of tilt-series images, a result of both the need for a low electron dose and the use of conventional defocus phase-contrast imaging, is a challenge for high-quality tomograms. We show that Zernike phase-contrast imaging allows the electron dose to be reduced. This limits movement of gold fiducials during the tilt series, which leads to better alignment and a higher-resolution reconstruction. Contrast is also enhanced, improving visibility of weak features. The reduced electron dose also means that more images at more tilt angles could be recorded, further increasing resolution. Copyright © 2013 Elsevier Inc. All rights reserved.

  19. Cryo-electron tomography of bacterial viruses

    Energy Technology Data Exchange (ETDEWEB)

    Guerrero-Ferreira, Ricardo C. [Division of Pediatric Infectious Diseases, Emory University School of Medicine, Children' s Healthcare of Atlanta, Atlanta, GA 30322 (United States); Wright, Elizabeth R., E-mail: erwrigh@emory.edu [Division of Pediatric Infectious Diseases, Emory University School of Medicine, Children' s Healthcare of Atlanta, Atlanta, GA 30322 (United States)

    2013-01-05

    Bacteriophage particles contain both simple and complex macromolecular assemblages and machines that enable them to regulate the infection process under diverse environmental conditions with a broad range of bacterial hosts. Recent developments in cryo-electron tomography (cryo-ET) make it possible to observe the interactions of bacteriophages with their host cells under native-state conditions at unprecedented resolution and in three-dimensions. This review describes the application of cryo-ET to studies of bacteriophage attachment, genome ejection, assembly and egress. Current topics of investigation and future directions in the field are also discussed.

  20. Cryo-Scanning Electron Microscopy (SEM) and Scanning Transmission Electron Microscopy (STEM)-in-SEM for Bio- and Organo-Mineral Interface Characterization in the Environment.

    Science.gov (United States)

    Wille, Guillaume; Hellal, Jennifer; Ollivier, Patrick; Richard, Annie; Burel, Agnes; Jolly, Louis; Crampon, Marc; Michel, Caroline

    2017-12-01

    Understanding biofilm interactions with surrounding substratum and pollutants/particles can benefit from the application of existing microscopy tools. Using the example of biofilm interactions with zero-valent iron nanoparticles (nZVI), this study aims to apply various approaches in biofilm preparation and labeling for fluorescent or electron microscopy and energy dispersive X-ray spectrometry (EDS) microanalysis for accurate observations. According to the targeted microscopy method, biofilms were sampled as flocs or attached biofilm, submitted to labeling using 4',6-diamidino-2-phenylindol, lectins PNA and ConA coupled to fluorescent dye or gold nanoparticles, and prepared for observation (fixation, cross-section, freezing, ultramicrotomy). Fluorescent microscopy revealed that nZVI were embedded in the biofilm structure as aggregates but the resolution was insufficient to observe individual nZVI. Cryo-scanning electron microscopy (SEM) observations showed nZVI aggregates close to bacteria, but it was not possible to confirm direct interactions between nZVI and cell membranes. Scanning transmission electron microscopy in the SEM (STEM-in-SEM) showed that nZVI aggregates could enter the biofilm to a depth of 7-11 µm. Bacteria were surrounded by a ring of extracellular polymeric substances (EPS) preventing direct nZVI/membrane interactions. STEM/EDS mapping revealed a co-localization of nZVI aggregates with lectins suggesting a potential role of EPS in nZVI embedding. Thus, the combination of divergent microscopy approaches is a good approach to better understand and characterize biofilm/metal interactions.

  1. Collaborative Computational Project for Electron cryo-Microscopy

    Energy Technology Data Exchange (ETDEWEB)

    Wood, Chris; Burnley, Tom [Science and Technology Facilities Council, Research Complex at Harwell, Didcot OX11 0FA (United Kingdom); Patwardhan, Ardan [European Molecular Biology Laboratory, Wellcome Trust Genome Campus, Hinxton, Cambridge CB10 1SD (United Kingdom); Scheres, Sjors [MRC Laboratory of Molecular Biology, Francis Crick Avenue, Cambridge Biomedical Campus, Cambridge CB2 0QH (United Kingdom); Topf, Maya [University of London, Malet Street, London WC1E 7HX (United Kingdom); Roseman, Alan [University of Manchester, Oxford Road, Manchester M13 9PT (United Kingdom); Winn, Martyn, E-mail: martyn.winn@stfc.ac.uk [Science and Technology Facilities Council, Daresbury Laboratory, Warrington WA4 4AD (United Kingdom); Science and Technology Facilities Council, Research Complex at Harwell, Didcot OX11 0FA (United Kingdom)

    2015-01-01

    The Collaborative Computational Project for Electron cryo-Microscopy (CCP-EM) is a new initiative for the structural biology community, following the success of CCP4 for macromolecular crystallography. Progress in supporting the users and developers of cryoEM software is reported. The Collaborative Computational Project for Electron cryo-Microscopy (CCP-EM) has recently been established. The aims of the project are threefold: to build a coherent cryoEM community which will provide support for individual scientists and will act as a focal point for liaising with other communities, to support practising scientists in their use of cryoEM software and finally to support software developers in producing and disseminating robust and user-friendly programs. The project is closely modelled on CCP4 for macromolecular crystallography, and areas of common interest such as model fitting, underlying software libraries and tools for building program packages are being exploited. Nevertheless, cryoEM includes a number of techniques covering a large range of resolutions and a distinct project is required. In this article, progress so far is reported and future plans are discussed.

  2. Collaborative Computational Project for Electron cryo-Microscopy

    International Nuclear Information System (INIS)

    Wood, Chris; Burnley, Tom; Patwardhan, Ardan; Scheres, Sjors; Topf, Maya; Roseman, Alan; Winn, Martyn

    2015-01-01

    The Collaborative Computational Project for Electron cryo-Microscopy (CCP-EM) is a new initiative for the structural biology community, following the success of CCP4 for macromolecular crystallography. Progress in supporting the users and developers of cryoEM software is reported. The Collaborative Computational Project for Electron cryo-Microscopy (CCP-EM) has recently been established. The aims of the project are threefold: to build a coherent cryoEM community which will provide support for individual scientists and will act as a focal point for liaising with other communities, to support practising scientists in their use of cryoEM software and finally to support software developers in producing and disseminating robust and user-friendly programs. The project is closely modelled on CCP4 for macromolecular crystallography, and areas of common interest such as model fitting, underlying software libraries and tools for building program packages are being exploited. Nevertheless, cryoEM includes a number of techniques covering a large range of resolutions and a distinct project is required. In this article, progress so far is reported and future plans are discussed

  3. High-resolution electron microscopy

    CERN Document Server

    Spence, John C H

    2013-01-01

    This new fourth edition of the standard text on atomic-resolution transmission electron microscopy (TEM) retains previous material on the fundamentals of electron optics and aberration correction, linear imaging theory (including wave aberrations to fifth order) with partial coherence, and multiple-scattering theory. Also preserved are updated earlier sections on practical methods, with detailed step-by-step accounts of the procedures needed to obtain the highest quality images of atoms and molecules using a modern TEM or STEM electron microscope. Applications sections have been updated - these include the semiconductor industry, superconductor research, solid state chemistry and nanoscience, and metallurgy, mineralogy, condensed matter physics, materials science and material on cryo-electron microscopy for structural biology. New or expanded sections have been added on electron holography, aberration correction, field-emission guns, imaging filters, super-resolution methods, Ptychography, Ronchigrams, tomogr...

  4. Challenges and opportunities in the high-resolution cryo-EM visualization of microtubules and their binding partners.

    Science.gov (United States)

    Nogales, Eva; Kellogg, Elizabeth H

    2017-10-01

    As non-crystallizable polymers, microtubules have been the target of cryo-electron microscopy (cryo-EM) studies since the technique was first established. Over the years, image processing strategies have been developed that take care of the unique, pseudo-helical symmetry of the microtubule. With recent progress in data quality and data processing, cryo-EM reconstructions are now reaching resolutions that allow the generation of atomic models of microtubules and the factors that bind them. These include cellular partners that contribute to microtubule cellular functions, or small ligands that interfere with those functions in the treatment of cancer. The stage is set to generate a family portrait for all identified microtubule interacting proteins and to use cryo-EM as a drug development tool in the targeting of tubulin. Copyright © 2017 The Authors. Published by Elsevier Ltd.. All rights reserved.

  5. The innovation of cryo-SEM freeze-fracturing methodology demonstrated on high pressure frozen biofilm.

    Science.gov (United States)

    Hrubanova, Kamila; Nebesarova, Jana; Ruzicka, Filip; Krzyzanek, Vladislav

    2018-04-22

    In this study we present an innovative method for the preparation of fully hydrated samples of microbial biofilms of cultures Staphylococcus epidermidis, Candida parapsilosis and Candida albicans. Cryo-scanning electron microscopy (cryo-SEM) and high-pressure freezing (HPF) rank among cutting edge techniques in the electron microscopy of hydrated samples such as biofilms. However, the combination of these techniques is not always easily applicable. Therefore, we present a method of combining high-pressure freezing using EM PACT2 (Leica Microsystems), which fixes hydrated samples on small sapphire discs, with a high resolution SEM equipped with the widely used cryo-preparation system ALTO 2500 (Gatan). Using a holder developed in house, a freeze-fracturing technique was applied to image and investigate microbial cultures cultivated on the sapphire discs. In our experiments, we focused on the ultrastructure of the extracellular matrix produced during cultivation and the relationships among microbial cells in the biofilm. The main goal of our investigations was the detailed visualization of areas of the biofilm where the microbial cells adhere to the substrate/surface. We show the feasibility of this technique, which is clearly demonstrated in experiments with various freeze-etching times. Copyright © 2018 Elsevier Ltd. All rights reserved.

  6. High-resolution imaging in the scanning transmission electron microscope

    International Nuclear Information System (INIS)

    Pennycook, S.J.; Jesson, D.E.

    1992-03-01

    The high-resolution imaging of crystalline materials in the scanning transmission electron microscopy (STEM) is reviewed with particular emphasis on the conditions under which an incoherent image can be obtained. It is shown that a high-angle annular detector can be used to break the coherence of the imaging process, in the transverse plane through the geometry of the detector, or in three dimensions if multiphonon diffuse scattering is detected. In the latter case, each atom can be treated as a highly independent source of high-angle scattering. The most effective fast electron states are therefore tightly bound s-type Bloch states. Furthermore, they add constructively for each incident angle in the coherent STEM probe, so that s states are responsible for practically the entire image contrast. Dynamical effects are largely removed, and almost perfect incoherent imaging is achieved. s states are relatively insensitive to neighboring strings, so that incoherent imaging is maintained for superlattice and interfaces, and supercell calculations are unnecessary. With an optimum probe profile, the incoherent image represents a direct image of the crystal projection, with compositional sensitivity built in through the strong dependence of the scattering cross sections on atomic number Z

  7. Spatial and temporal resolution in cryo-electron microscopy : a scope for nano-chemistry

    NARCIS (Netherlands)

    Frederik, P.M.; Sommerdijk, N.A.J.M.

    2005-01-01

    Cryo-electron microscopy has evolved in an established approach to study the structure of bio-colloids. Recent developments in instrumentation and automation, often demanded by life sciences, made cryo-EM a general tool in colloid chemistry. Recently improved instrumentation for vitrification has

  8. High resolution surface scanning of Thick-GEM for single photo-electron detection

    International Nuclear Information System (INIS)

    Hamar, G.; Varga, D.

    2012-01-01

    An optical system for high resolution scanning of TGEM UV photon detection systems is introduced. The structure exploits the combination of a single Au-coated TGEM under study, and an asymmetric MWPC (Close Cathode Chamber) as post-amplification stage. A pulsed UV LED source with emission down to 240 nm has been focused to a spot of 0.07 mm on the TGEM surface, and single photo-electron charge spectra has been recorded over selected two dimensional regions. This way, the TGEM gain (order of 10–100) and TGEM photo-electron detection efficiency is clearly separated, unlike in case of continuous illumination. The surface structure connected to the TGEM photon detection is well observable, including inefficiencies in the holes and at the symmetry points between holes. The detection efficiency as well as the gas gain are fluctuating from hole to hole. The gain is constant in the hexagon around any hole, pointing to the fact that the gain depends on hole geometry, and less on the position where the electron enters. The detection probability map strongly changes with the field strength above the TGEM surface, in relation to the change of the actual surface field configuration. The results can be confronted with position-dependent simulations of TGEM electron transfer and gas multiplication. -- Highlights: ► First demonstration of Thick GEM surface scanning with single photo-electrons. ► Resolution of 0.1 mm is sufficient to identify structures connected to TGEM surface field structure. ► Gain and detection efficiency and separately measurable. ► Detection efficiency is high in a ring around the holes, and gain is constant in the hexagonal collection regions.

  9. A deep convolutional neural network approach to single-particle recognition in cryo-electron microscopy.

    Science.gov (United States)

    Zhu, Yanan; Ouyang, Qi; Mao, Youdong

    2017-07-21

    Single-particle cryo-electron microscopy (cryo-EM) has become a mainstream tool for the structural determination of biological macromolecular complexes. However, high-resolution cryo-EM reconstruction often requires hundreds of thousands of single-particle images. Particle extraction from experimental micrographs thus can be laborious and presents a major practical bottleneck in cryo-EM structural determination. Existing computational methods for particle picking often use low-resolution templates for particle matching, making them susceptible to reference-dependent bias. It is critical to develop a highly efficient template-free method for the automatic recognition of particle images from cryo-EM micrographs. We developed a deep learning-based algorithmic framework, DeepEM, for single-particle recognition from noisy cryo-EM micrographs, enabling automated particle picking, selection and verification in an integrated fashion. The kernel of DeepEM is built upon a convolutional neural network (CNN) composed of eight layers, which can be recursively trained to be highly "knowledgeable". Our approach exhibits an improved performance and accuracy when tested on the standard KLH dataset. Application of DeepEM to several challenging experimental cryo-EM datasets demonstrated its ability to avoid the selection of un-wanted particles and non-particles even when true particles contain fewer features. The DeepEM methodology, derived from a deep CNN, allows automated particle extraction from raw cryo-EM micrographs in the absence of a template. It demonstrates an improved performance, objectivity and accuracy. Application of this novel method is expected to free the labor involved in single-particle verification, significantly improving the efficiency of cryo-EM data processing.

  10. High resolution surface scanning of Thick-GEM for single photo-electron detection

    Energy Technology Data Exchange (ETDEWEB)

    Hamar, G., E-mail: hamar.gergo@wigner.mta.hu [Wigner Research Centre for Physics, Budapest (Hungary); Varga, D., E-mail: vdezso@mail.cern.ch [Eoetvoes Lorand University, Budapest (Hungary)

    2012-12-01

    An optical system for high resolution scanning of TGEM UV photon detection systems is introduced. The structure exploits the combination of a single Au-coated TGEM under study, and an asymmetric MWPC (Close Cathode Chamber) as post-amplification stage. A pulsed UV LED source with emission down to 240 nm has been focused to a spot of 0.07 mm on the TGEM surface, and single photo-electron charge spectra has been recorded over selected two dimensional regions. This way, the TGEM gain (order of 10-100) and TGEM photo-electron detection efficiency is clearly separated, unlike in case of continuous illumination. The surface structure connected to the TGEM photon detection is well observable, including inefficiencies in the holes and at the symmetry points between holes. The detection efficiency as well as the gas gain are fluctuating from hole to hole. The gain is constant in the hexagon around any hole, pointing to the fact that the gain depends on hole geometry, and less on the position where the electron enters. The detection probability map strongly changes with the field strength above the TGEM surface, in relation to the change of the actual surface field configuration. The results can be confronted with position-dependent simulations of TGEM electron transfer and gas multiplication. -- Highlights: Black-Right-Pointing-Pointer First demonstration of Thick GEM surface scanning with single photo-electrons. Black-Right-Pointing-Pointer Resolution of 0.1 mm is sufficient to identify structures connected to TGEM surface field structure. Black-Right-Pointing-Pointer Gain and detection efficiency and separately measurable. Black-Right-Pointing-Pointer Detection efficiency is high in a ring around the holes, and gain is constant in the hexagonal collection regions.

  11. Development of superconducting cryo-electron microscope and its applications

    International Nuclear Information System (INIS)

    Iwatsuki, Masashi

    1988-01-01

    Recently, a superconducting cryo-electron microscope in which specimens are cooled to the liquid helium temperature (4.2 K) has been developed. The main components and functional features of this new microscope are reported together with application data on polyethylene, poly (4-methyl-1-pentene), valonia cellulose, rock salt, ice crystallites and ceramic superconductor. The resistance to electron radiation damage, of beam-sensitive specimens including polymers has been increased more than ten times. Thus, the microscope has made it possible to take high resolution images and to analyze the crystal-structure of micro-areas. (orig.) [de

  12. High-resolution noise substitution to measure overfitting and validate resolution in 3D structure determination by single particle electron cryomicroscopy

    International Nuclear Information System (INIS)

    Chen, Shaoxia; McMullan, Greg; Faruqi, Abdul R.; Murshudov, Garib N.; Short, Judith M.; Scheres, Sjors H.W.; Henderson, Richard

    2013-01-01

    Three-dimensional (3D) structure determination by single particle electron cryomicroscopy (cryoEM) involves the calculation of an initial 3D model, followed by extensive iterative improvement of the orientation determination of the individual particle images and the resulting 3D map. Because there is much more noise than signal at high resolution in the images, this creates the possibility of noise reinforcement in the 3D map, which can give a false impression of the resolution attained. The balance between signal and noise in the final map at its limiting resolution depends on the image processing procedure and is not easily predicted. There is a growing awareness in the cryoEM community of how to avoid such over-fitting and over-estimation of resolution. Equally, there has been a reluctance to use the two principal methods of avoidance because they give lower resolution estimates, which some people believe are too pessimistic. Here we describe a simple test that is compatible with any image processing protocol. The test allows measurement of the amount of signal and the amount of noise from overfitting that is present in the final 3D map. We have applied the method to two different sets of cryoEM images of the enzyme beta-galactosidase using several image processing packages. Our procedure involves substituting the Fourier components of the initial particle image stack beyond a chosen resolution by either the Fourier components from an adjacent area of background, or by simple randomisation of the phases of the particle structure factors. This substituted noise thus has the same spectral power distribution as the original data. Comparison of the Fourier Shell Correlation (FSC) plots from the 3D map obtained using the experimental data with that from the same data with high-resolution noise (HR-noise) substituted allows an unambiguous measurement of the amount of overfitting and an accompanying resolution assessment. A simple formula can be used to calculate an

  13. High-resolution noise substitution to measure overfitting and validate resolution in 3D structure determination by single particle electron cryomicroscopy

    Energy Technology Data Exchange (ETDEWEB)

    Chen, Shaoxia; McMullan, Greg; Faruqi, Abdul R.; Murshudov, Garib N.; Short, Judith M.; Scheres, Sjors H.W.; Henderson, Richard, E-mail: rh15@mrc-lmb.cam.ac.uk

    2013-12-15

    Three-dimensional (3D) structure determination by single particle electron cryomicroscopy (cryoEM) involves the calculation of an initial 3D model, followed by extensive iterative improvement of the orientation determination of the individual particle images and the resulting 3D map. Because there is much more noise than signal at high resolution in the images, this creates the possibility of noise reinforcement in the 3D map, which can give a false impression of the resolution attained. The balance between signal and noise in the final map at its limiting resolution depends on the image processing procedure and is not easily predicted. There is a growing awareness in the cryoEM community of how to avoid such over-fitting and over-estimation of resolution. Equally, there has been a reluctance to use the two principal methods of avoidance because they give lower resolution estimates, which some people believe are too pessimistic. Here we describe a simple test that is compatible with any image processing protocol. The test allows measurement of the amount of signal and the amount of noise from overfitting that is present in the final 3D map. We have applied the method to two different sets of cryoEM images of the enzyme beta-galactosidase using several image processing packages. Our procedure involves substituting the Fourier components of the initial particle image stack beyond a chosen resolution by either the Fourier components from an adjacent area of background, or by simple randomisation of the phases of the particle structure factors. This substituted noise thus has the same spectral power distribution as the original data. Comparison of the Fourier Shell Correlation (FSC) plots from the 3D map obtained using the experimental data with that from the same data with high-resolution noise (HR-noise) substituted allows an unambiguous measurement of the amount of overfitting and an accompanying resolution assessment. A simple formula can be used to calculate an

  14. Nano, Queensland and cryo-electron microscopy

    International Nuclear Information System (INIS)

    McDowall, A.W.

    2002-01-01

    Full text: In a recent review the authors, Wolfgang Baumeister and Alasdair Steven wrote, '....there is immense opportunity for Cryo-EM, especially as boosted by merging crystallographic structures of individual subunits into moderate resolution Cryo-EM density maps of whole complexes. Electron tomography has now advanced to the point where it is a realistic goal to glimpse molecular machines operating inside cells....' This statement gives testament to the advances made over the past 25 years by many labs around the world to the area of microscopy referred to as Cryo-EM and related 3-D computing technologies. Australian scientific societies have been eager followers of this progress and heard first hand of the new developments in the field at the 1984 ACEM-8 (2). Since those early days the ACEM and other Australian/NZ societies have sponsored numerous researchers and workshops in the field of Cryo-EM to their conferences, Helin Sabil, Wah Chiu, Ron Milligan, Richard Henderson and Werner Kuhlbrandt to name only a few. These visits have stimulated a desire from Australian/NZ researchers to establish collaborations and access to prominent labs in the USA and Europe, where the means and knowledge to provide Cryo EM and 3D reconstruction technology for studying macromolecular complexes is well established. However, Australia has not been backward in seeking to provide its home research community with access to a base in biological molecular microscopy and electron crystallography technology. Since the last ACEM we have seen the emergence of a number of crucial factors, which will make the establishment of a national research facility in this field an operational reality in early 2003. Well publicized is the development of Australia's newest and perhaps most unique research institute, the institute for Molecular Bioscience (IMB) to open at the University of Queensland (UQ) in 2002. The IMB will be the platform for a new research group in advanced computational 3D

  15. An extended model of electrons: experimental evidence from high-resolution scanning tunneling microscopy

    International Nuclear Information System (INIS)

    Hofer, Werner A

    2012-01-01

    In a recent paper we introduced a model of extended electrons, which is fully compatible with quantum mechanics in the formulation of Schrödinger. However, it contradicts the current interpretation of electrons as point-particles. Here, we show by a statistical analysis of high-resolution scanning tunneling microscopy (STM) experiments, that the interpretation of electrons as point particles and, consequently, the interpretation of the density of electron charge as a statistical quantity will lead to a conflict with the Heisenberg uncertainty principle. Given the precision in these experiments we find that the uncertainty principle would be violated by close to two orders of magnitude, if this interpretation were correct. We are thus forced to conclude that the density of electron charge is a physically real, i.e. in principle precisely measurable quantity, as derived in a recent paper. Experimental evidence to the contrary, in particular high-energy scattering experiments, is briefly discussed. The finding is expected to have wide implications in condensed matter physics, chemistry, and biology, scientific disciplines which are based on the properties and interactions of electrons.

  16. High-resolution electron microscopy of advanced materials

    Energy Technology Data Exchange (ETDEWEB)

    Mitchell, T.E.; Kung, H.H.; Sickafus, K.E.; Gray, G.T. III; Field, R.D.; Smith, J.F. [Los Alamos National Lab., NM (United States). Materials Science and Technology Div.

    1997-11-01

    This final report chronicles a three-year, Laboratory Directed Research and Development (LDRD) project at Los Alamos National Laboratory (LANL). The High-Resolution Electron Microscopy Facility has doubled in size and tripled in quality since the beginning of the three-year period. The facility now includes a field-emission scanning electron microscope, a 100 kV field-emission scanning transmission electron microscope (FE-STEM), a 300 kV field-emission high-resolution transmission electron microscope (FE-HRTEM), and a 300 kV analytical transmission electron microscope. A new orientation imaging microscope is being installed. X-ray energy dispersive spectrometers for chemical analysis are available on all four microscopes; parallel electron energy loss spectrometers are operational on the FE-STEM and FE-HRTEM. These systems enable evaluation of local atomic bonding, as well as chemical composition in nanometer-scale regions. The FE-HRTEM has a point-to-point resolution of 1.6 {angstrom}, but the resolution can be pushed to its information limit of 1 {angstrom} by computer reconstruction of a focal series of images. HRTEM has been used to image the atomic structure of defects such as dislocations, grain boundaries, and interfaces in a variety of materials from superconductors and ferroelectrics to structural ceramics and intermetallics.

  17. Precision controlled atomic resolution scanning transmission electron microscopy using spiral scan pathways

    Science.gov (United States)

    Sang, Xiahan; Lupini, Andrew R.; Ding, Jilai; Kalinin, Sergei V.; Jesse, Stephen; Unocic, Raymond R.

    2017-03-01

    Atomic-resolution imaging in an aberration-corrected scanning transmission electron microscope (STEM) can enable direct correlation between atomic structure and materials functionality. The fast and precise control of the STEM probe is, however, challenging because the true beam location deviates from the assigned location depending on the properties of the deflectors. To reduce these deviations, i.e. image distortions, we use spiral scanning paths, allowing precise control of a sub-Å sized electron probe within an aberration-corrected STEM. Although spiral scanning avoids the sudden changes in the beam location (fly-back distortion) present in conventional raster scans, it is not distortion-free. “Archimedean” spirals, with a constant angular frequency within each scan, are used to determine the characteristic response at different frequencies. We then show that such characteristic functions can be used to correct image distortions present in more complicated constant linear velocity spirals, where the frequency varies within each scan. Through the combined application of constant linear velocity scanning and beam path corrections, spiral scan images are shown to exhibit less scan distortion than conventional raster scan images. The methodology presented here will be useful for in situ STEM imaging at higher temporal resolution and for imaging beam sensitive materials.

  18. Ultrastable gold substrates: Properties of a support for high-resolution electron cryomicroscopy of biological specimens

    Science.gov (United States)

    Russo, Christopher J.; Passmore, Lori A.

    2016-01-01

    Electron cryomicroscopy (cryo-EM) allows structure determination of a wide range of biological molecules and specimens. All-gold supports improve cryo-EM images by reducing radiation-induced motion and image blurring. Here we compare the mechanical and electrical properties of all-gold supports to amorphous carbon foils. Gold supports are more conductive, and have suspended foils that are not compressed by differential contraction when cooled to liquid nitrogen temperatures. These measurements show how the choice of support material and geometry can reduce specimen movement by more than an order of magnitude during low-dose imaging. We provide methods for fabrication of all-gold supports and preparation of vitrified specimens. We also analyse illumination geometry for optimal collection of high resolution, low-dose data. Together, the support structures and methods herein can improve the resolution and quality of images from any electron cryomicroscope. PMID:26592474

  19. New insights into the structural organization of eukaryotic and prokaryotic cytoskeletons using cryo-electron tomography

    International Nuclear Information System (INIS)

    Kuerner, Julia; Medalia, Ohad; Linaroudis, Alexandros A.; Baumeister, Wolfgang

    2004-01-01

    Cryo-electron tomography (cryo-ET) is an emerging imaging technology that combines the potential of three-dimensional (3-D) imaging at molecular resolution (<5 nm) with a close-to-life preservation of the specimen. In conjunction with pattern recognition techniques, it enables us to map the molecular landscape inside cells. The application of cryo-ET to intact cells provides novel insights into the structure and the spatial organization of the cytoskeleton in prokaryotic and eukaryotic cells

  20. Structure of a headful DNA-packaging bacterial virus at 2.9 Å resolution by electron cryo-microscopy.

    Science.gov (United States)

    Zhao, Haiyan; Li, Kunpeng; Lynn, Anna Y; Aron, Keith E; Yu, Guimei; Jiang, Wen; Tang, Liang

    2017-04-04

    The enormous prevalence of tailed DNA bacteriophages on this planet is enabled by highly efficient self-assembly of hundreds of protein subunits into highly stable capsids. These capsids can stand with an internal pressure as high as ∼50 atmospheres as a result of the phage DNA-packaging process. Here we report the complete atomic model of the headful DNA-packaging bacteriophage Sf6 at 2.9 Å resolution determined by electron cryo-microscopy. The structure reveals the DNA-inflated, tensed state of a robust protein shell assembled via noncovalent interactions. Remarkable global conformational polymorphism of capsid proteins, a network formed by extended N arms, mortise-and-tenon-like intercapsomer joints, and abundant β-sheet-like mainchain:mainchain intermolecular interactions, confers significant strength yet also flexibility required for capsid assembly and DNA packaging. Differential formations of the hexon and penton are mediated by a drastic α-helix-to-β-strand structural transition. The assembly scheme revealed here may be common among tailed DNA phages and herpesviruses.

  1. Three-dimensional structural analysis of eukaryotic flagella/cilia by electron cryo-tomography

    International Nuclear Information System (INIS)

    Bui, Khanh Huy; Pigino, Gaia; Ishikawa, Takashi

    2011-01-01

    Based on the molecular architecture revealed by electron cryo-tomography, the mechanism of the bending motion of eukaryotic flagella/cilia is discussed. Electron cryo-tomography is a potential approach to analyzing the three-dimensional conformation of frozen hydrated biological macromolecules using electron microscopy. Since projections of each individual object illuminated from different orientations are merged, electron tomography is capable of structural analysis of such heterogeneous environments as in vivo or with polymorphism, although radiation damage and the missing wedge are severe problems. Here, recent results on the structure of eukaryotic flagella, which is an ATP-driven bending organelle, from green algae Chlamydomonas are presented. Tomographic analysis reveals asymmetric molecular arrangements, especially that of the dynein motor proteins, in flagella, giving insight into the mechanism of planar asymmetric bending motion. Methodological challenges to obtaining higher-resolution structures from this technique are also discussed

  2. Beam-induced motion correction for sub-megadalton cryo-EM particles.

    Science.gov (United States)

    Scheres, Sjors Hw

    2014-08-13

    In electron cryo-microscopy (cryo-EM), the electron beam that is used for imaging also causes the sample to move. This motion blurs the images and limits the resolution attainable by single-particle analysis. In a previous Research article (Bai et al., 2013) we showed that correcting for this motion by processing movies from fast direct-electron detectors allowed structure determination to near-atomic resolution from 35,000 ribosome particles. In this Research advance article, we show that an improved movie processing algorithm is applicable to a much wider range of specimens. The new algorithm estimates straight movement tracks by considering multiple particles that are close to each other in the field of view, and models the fall-off of high-resolution information content by radiation damage in a dose-dependent manner. Application of the new algorithm to four data sets illustrates its potential for significantly improving cryo-EM structures, even for particles that are smaller than 200 kDa. Copyright © 2014, Scheres.

  3. Resolution and Probabilistic Models of Components in CryoEM Maps of Mature P22 Bacteriophage

    Science.gov (United States)

    Pintilie, Grigore; Chen, Dong-Hua; Haase-Pettingell, Cameron A.; King, Jonathan A.; Chiu, Wah

    2016-01-01

    CryoEM continues to produce density maps of larger and more complex assemblies with multiple protein components of mixed symmetries. Resolution is not always uniform throughout a cryoEM map, and it can be useful to estimate the resolution in specific molecular components of a large assembly. In this study, we present procedures to 1) estimate the resolution in subcomponents by gold-standard Fourier shell correlation (FSC); 2) validate modeling procedures, particularly at medium resolutions, which can include loop modeling and flexible fitting; and 3) build probabilistic models that combine high-accuracy priors (such as crystallographic structures) with medium-resolution cryoEM densities. As an example, we apply these methods to new cryoEM maps of the mature bacteriophage P22, reconstructed without imposing icosahedral symmetry. Resolution estimates based on gold-standard FSC show the highest resolution in the coat region (7.6 Å), whereas other components are at slightly lower resolutions: portal (9.2 Å), hub (8.5 Å), tailspike (10.9 Å), and needle (10.5 Å). These differences are indicative of inherent structural heterogeneity and/or reconstruction accuracy in different subcomponents of the map. Probabilistic models for these subcomponents provide new insights, to our knowledge, and structural information when taking into account uncertainty given the limitations of the observed density. PMID:26743049

  4. The 2017 Nobel Prize in Chemistry: cryo-EM comes of age.

    Science.gov (United States)

    Shen, Peter S

    2018-03-01

    The 2017 Nobel Prize in Chemistry was awarded to Jacques Dubochet, Joachim Frank, and Richard Henderson for "developing cryo-electron microscopy (cryo-EM) for the high-resolution structure determination of biomolecules in solution." This feature article summarizes some of the major achievements leading to the development of cryo-EM and recent technological breakthroughs that have transformed the method into a mainstream tool for structure determination.

  5. A cylindrical specimen holder for electron cryo-tomography

    International Nuclear Information System (INIS)

    Palmer, Colin M.; Löwe, Jan

    2014-01-01

    The use of slab-like flat specimens for electron cryo-tomography restricts the range of viewing angles that can be used. This leads to the “missing wedge” problem, which causes artefacts and anisotropic resolution in reconstructed tomograms. Cylindrical specimens provide a way to eliminate the problem, since they allow imaging from a full range of viewing angles around the tilt axis. Such specimens have been used before for tomography of radiation-insensitive samples at room temperature, but never for frozen-hydrated specimens. Here, we demonstrate the use of thin-walled carbon tubes as specimen holders, allowing the preparation of cylindrical frozen-hydrated samples of ribosomes, liposomes and whole bacterial cells. Images acquired from these cylinders have equal quality at all viewing angles, and the accessible tilt range is restricted only by the physical limits of the microscope. Tomographic reconstructions of these specimens demonstrate that the effects of the missing wedge are substantially reduced, and could be completely eliminated if a full tilt range was used. The overall quality of these tomograms is still lower than that obtained by existing methods, but improvements are likely in future. - Highlights: • The missing wedge is a serious problem for electron cryo-tomography. • Cylindrical specimens allow the missing wedge to be eliminated. • Carbon nanopipettes can be used as cylindrical holders for tomography of frozen-hydrated specimens. • Cryo-tomography of cylindrical biological samples demonstrates a reduction of deleterious effects associated with the missing wedge

  6. Phasing of the Triatoma virus diffraction data using a cryo-electron microscopy reconstruction

    International Nuclear Information System (INIS)

    Estrozi, L.F.; Neumann, E.; Squires, G.; Rozas-Dennis, G.; Costabel, M.; Rey, F.A.; Guerin, D.M.A.; Navaza, J.

    2008-01-01

    The blood-sucking reduviid bug Triatoma infestans, one of the most important vector of American human trypanosomiasis (Chagas disease) is infected by the Triatoma virus (TrV). TrV has been classified as a member of the Cripavirus genus (type cricket paralysis virus) in the Dicistroviridae family. This work presents the three-dimensional cryo-electron microscopy (cryo-EM) reconstruction of the TrV capsid at about 25 A resolution and its use as a template for phasing the available crystallographic data by the molecular replacement method. The main structural differences between the cryo-EM reconstruction of TrV and other two viruses, one from the same family, the cricket paralysis virus (CrPV) and the human rhinovirus 16 from the Picornaviridae family are presented and discussed

  7. High-resolution noise substitution to measure overfitting and validate resolution in 3D structure determination by single particle electron cryomicroscopy.

    Science.gov (United States)

    Chen, Shaoxia; McMullan, Greg; Faruqi, Abdul R; Murshudov, Garib N; Short, Judith M; Scheres, Sjors H W; Henderson, Richard

    2013-12-01

    Three-dimensional (3D) structure determination by single particle electron cryomicroscopy (cryoEM) involves the calculation of an initial 3D model, followed by extensive iterative improvement of the orientation determination of the individual particle images and the resulting 3D map. Because there is much more noise than signal at high resolution in the images, this creates the possibility of noise reinforcement in the 3D map, which can give a false impression of the resolution attained. The balance between signal and noise in the final map at its limiting resolution depends on the image processing procedure and is not easily predicted. There is a growing awareness in the cryoEM community of how to avoid such over-fitting and over-estimation of resolution. Equally, there has been a reluctance to use the two principal methods of avoidance because they give lower resolution estimates, which some people believe are too pessimistic. Here we describe a simple test that is compatible with any image processing protocol. The test allows measurement of the amount of signal and the amount of noise from overfitting that is present in the final 3D map. We have applied the method to two different sets of cryoEM images of the enzyme beta-galactosidase using several image processing packages. Our procedure involves substituting the Fourier components of the initial particle image stack beyond a chosen resolution by either the Fourier components from an adjacent area of background, or by simple randomisation of the phases of the particle structure factors. This substituted noise thus has the same spectral power distribution as the original data. Comparison of the Fourier Shell Correlation (FSC) plots from the 3D map obtained using the experimental data with that from the same data with high-resolution noise (HR-noise) substituted allows an unambiguous measurement of the amount of overfitting and an accompanying resolution assessment. A simple formula can be used to calculate an

  8. Sub-nanometre resolution imaging of polymer-fullerene photovoltaic blends using energy-filtered scanning electron microscopy.

    Science.gov (United States)

    Masters, Robert C; Pearson, Andrew J; Glen, Tom S; Sasam, Fabian-Cyril; Li, Letian; Dapor, Maurizio; Donald, Athene M; Lidzey, David G; Rodenburg, Cornelia

    2015-04-24

    The resolution capability of the scanning electron microscope has increased immensely in recent years, and is now within the sub-nanometre range, at least for inorganic materials. An equivalent advance has not yet been achieved for imaging the morphologies of nanostructured organic materials, such as organic photovoltaic blends. Here we show that energy-selective secondary electron detection can be used to obtain high-contrast, material-specific images of an organic photovoltaic blend. We also find that we can differentiate mixed phases from pure material phases in our data. The lateral resolution demonstrated is twice that previously reported from secondary electron imaging. Our results suggest that our energy-filtered scanning electron microscopy approach will be able to make major inroads into the understanding of complex, nano-structured organic materials.

  9. Sub-nanometre resolution imaging of polymer–fullerene photovoltaic blends using energy-filtered scanning electron microscopy

    Science.gov (United States)

    Masters, Robert C.; Pearson, Andrew J.; Glen, Tom S.; Sasam, Fabian-Cyril; Li, Letian; Dapor, Maurizio; Donald, Athene M.; Lidzey, David G.; Rodenburg, Cornelia

    2015-01-01

    The resolution capability of the scanning electron microscope has increased immensely in recent years, and is now within the sub-nanometre range, at least for inorganic materials. An equivalent advance has not yet been achieved for imaging the morphologies of nanostructured organic materials, such as organic photovoltaic blends. Here we show that energy-selective secondary electron detection can be used to obtain high-contrast, material-specific images of an organic photovoltaic blend. We also find that we can differentiate mixed phases from pure material phases in our data. The lateral resolution demonstrated is twice that previously reported from secondary electron imaging. Our results suggest that our energy-filtered scanning electron microscopy approach will be able to make major inroads into the understanding of complex, nano-structured organic materials. PMID:25906738

  10. High-resolution, high-throughput imaging with a multibeam scanning electron microscope.

    Science.gov (United States)

    Eberle, A L; Mikula, S; Schalek, R; Lichtman, J; Knothe Tate, M L; Zeidler, D

    2015-08-01

    Electron-electron interactions and detector bandwidth limit the maximal imaging speed of single-beam scanning electron microscopes. We use multiple electron beams in a single column and detect secondary electrons in parallel to increase the imaging speed by close to two orders of magnitude and demonstrate imaging for a variety of samples ranging from biological brain tissue to semiconductor wafers. © 2015 The Authors Journal of Microscopy © 2015 Royal Microscopical Society.

  11. Investigations in space-related molecular biology. [cryo-electron microscopic and diffraction studies on terrestrial and extraterrestrial specimens

    Science.gov (United States)

    Fernandez-Moran, H.; Pritzker, A. N.

    1974-01-01

    Improved instrumentation and preparation techniques for high resolution, high voltage cryo-electron microscopic and diffraction studies on terrestrial and extraterrestrial specimens are reported. Computer correlated ultrastructural and biochemical work on hydrated and dried cell membranes and related biological systems provided information on membrane organization, ice crystal formation and ordered water, RNA virus linked to cancer, lunar rock samples, and organometallic superconducting compounds. Apollo 11, 12, 14, and 15 specimens were analyzed

  12. Combining scanning tunneling microscopy and synchrotron radiation for high-resolution imaging and spectroscopy with chemical, electronic, and magnetic contrast

    International Nuclear Information System (INIS)

    Cummings, M.L.; Chien, T.Y.; Preissner, C.; Madhavan, V.; Diesing, D.; Bode, M.; Freeland, J.W.; Rose, V.

    2012-01-01

    The combination of high-brilliance synchrotron radiation with scanning tunneling microscopy opens the path to high-resolution imaging with chemical, electronic, and magnetic contrast. Here, the design and experimental results of an in-situ synchrotron enhanced x-ray scanning tunneling microscope (SXSTM) system are presented. The system is designed to allow monochromatic synchrotron radiation to enter the chamber, illuminating the sample with x-ray radiation, while an insulator-coated tip (metallic tip apex open for tunneling, electron collection) is scanned over the surface. A unique feature of the SXSTM is the STM mount assembly, designed with a two free-flex pivot, providing an angular degree of freedom for the alignment of the tip and sample with respect to the incoming x-ray beam. The system designed successfully demonstrates the ability to resolve atomic-scale corrugations. In addition, experiments with synchrotron x-ray radiation validate the SXSTM system as an accurate analysis technique for the study of local magnetic and chemical properties on sample surfaces. The SXSTM system's capabilities have the potential to broaden and deepen the general understanding of surface phenomena by adding elemental contrast to the high-resolution of STM. -- Highlights: ► Synchrotron enhanced x-ray scanning tunneling microscope (SXSTM) system designed. ► Unique STM mount design allows angular DOF for tip alignment with x-ray beam. ► System demonstrates ability to resolve atomic corrugations on HOPG. ► Studies show chemical sensitivity with STM tip from photocurrent and tunneling. ► Results show system's ability to study local magnetic (XMCD) properties on Fe films.

  13. Limiting factors in single particle cryo electron tomography

    Directory of Open Access Journals (Sweden)

    Mikhail Kudryashev

    2012-07-01

    Full Text Available Modern methods of cryo electron microscopy and tomography allow visualization of protein nanomachines in their native state at the nanometer scale. Image processing methods including sub-volume averaging applied to repeating macromolecular elements within tomograms allow exploring their structures within the native context of the cell, avoiding the need for protein isolation and purification. Today, many different data acquisition protocols and software solutions are available to researchers to determine average structures of macromolecular complexes and potentially to classify structural intermediates. Here, we list the density maps reported in the literature, and analyze each structure for the chosen instrumental settings, sample conditions, main processing steps, and obtained resolution. We present conclusions that identify factors currently limiting the resolution gained by this approach.

  14. Modelling high-resolution electron microscopy based on core-loss spectroscopy

    International Nuclear Information System (INIS)

    Allen, L.J.; Findlay, S.D.; Oxley, M.P.; Witte, C.; Zaluzec, N.J.

    2006-01-01

    There are a number of factors affecting the formation of images based on core-loss spectroscopy in high-resolution electron microscopy. We demonstrate unambiguously the need to use a full nonlocal description of the effective core-loss interaction for experimental results obtained from high angular resolution electron channelling electron spectroscopy. The implications of this model are investigated for atomic resolution scanning transmission electron microscopy. Simulations are used to demonstrate that core-loss spectroscopy images formed using fine probes proposed for future microscopes can result in images that do not correspond visually with the structure that has led to their formation. In this context, we also examine the effect of varying detector geometries. The importance of the contribution to core-loss spectroscopy images by dechannelled or diffusely scattered electrons is reiterated here

  15. Modeling of Image Formation in Cryo-Electron Microscopy

    NARCIS (Netherlands)

    Vulovic, M.

    2013-01-01

    Knowledge of the structure of biological specimens is crucial for understanding life. Cryo-electron microscopy (cryo-EM) permits structural studies of biological specimen at their near-native state. The research performed in this thesis represents one of two subprojects of the FOM industrial

  16. Tools for macromolecular model building and refinement into electron cryo-microscopy reconstructions

    Energy Technology Data Exchange (ETDEWEB)

    Brown, Alan; Long, Fei; Nicholls, Robert A.; Toots, Jaan; Emsley, Paul; Murshudov, Garib, E-mail: garib@mrc-lmb.cam.ac.uk [MRC Laboratory of Molecular Biology, Francis Crick Avenue, Cambridge CB2 0QH (United Kingdom)

    2015-01-01

    A description is given of new tools to facilitate model building and refinement into electron cryo-microscopy reconstructions. The recent rapid development of single-particle electron cryo-microscopy (cryo-EM) now allows structures to be solved by this method at resolutions close to 3 Å. Here, a number of tools to facilitate the interpretation of EM reconstructions with stereochemically reasonable all-atom models are described. The BALBES database has been repurposed as a tool for identifying protein folds from density maps. Modifications to Coot, including new Jiggle Fit and morphing tools and improved handling of nucleic acids, enhance its functionality for interpreting EM maps. REFMAC has been modified for optimal fitting of atomic models into EM maps. As external structural information can enhance the reliability of the derived atomic models, stabilize refinement and reduce overfitting, ProSMART has been extended to generate interatomic distance restraints from nucleic acid reference structures, and a new tool, LIBG, has been developed to generate nucleic acid base-pair and parallel-plane restraints. Furthermore, restraint generation has been integrated with visualization and editing in Coot, and these restraints have been applied to both real-space refinement in Coot and reciprocal-space refinement in REFMAC.

  17. Tools for macromolecular model building and refinement into electron cryo-microscopy reconstructions

    International Nuclear Information System (INIS)

    Brown, Alan; Long, Fei; Nicholls, Robert A.; Toots, Jaan; Emsley, Paul; Murshudov, Garib

    2015-01-01

    A description is given of new tools to facilitate model building and refinement into electron cryo-microscopy reconstructions. The recent rapid development of single-particle electron cryo-microscopy (cryo-EM) now allows structures to be solved by this method at resolutions close to 3 Å. Here, a number of tools to facilitate the interpretation of EM reconstructions with stereochemically reasonable all-atom models are described. The BALBES database has been repurposed as a tool for identifying protein folds from density maps. Modifications to Coot, including new Jiggle Fit and morphing tools and improved handling of nucleic acids, enhance its functionality for interpreting EM maps. REFMAC has been modified for optimal fitting of atomic models into EM maps. As external structural information can enhance the reliability of the derived atomic models, stabilize refinement and reduce overfitting, ProSMART has been extended to generate interatomic distance restraints from nucleic acid reference structures, and a new tool, LIBG, has been developed to generate nucleic acid base-pair and parallel-plane restraints. Furthermore, restraint generation has been integrated with visualization and editing in Coot, and these restraints have been applied to both real-space refinement in Coot and reciprocal-space refinement in REFMAC

  18. Single-particle cryo-electron microscopy of Rift Valley fever virus

    International Nuclear Information System (INIS)

    Sherman, Michael B.; Freiberg, Alexander N.; Holbrook, Michael R.; Watowich, Stanley J.

    2009-01-01

    Rift Valley fever virus (RVFV; Bunyaviridae; Phlebovirus) is an emerging human and veterinary pathogen causing acute hepatitis in ruminants and has the potential to cause hemorrhagic fever in humans. We report a three-dimensional reconstruction of RVFV vaccine strain MP-12 (RVFV MP-12) by cryo-electron microcopy using icosahedral symmetry of individual virions. Although the genomic core of RVFV MP-12 is apparently poorly ordered, the glycoproteins on the virus surface are highly symmetric and arranged on a T = 12 icosahedral lattice. Our RVFV MP-12 structure allowed clear identification of inter-capsomer contacts and definition of possible glycoprotein arrangements within capsomers. This structure provides a detailed model for phleboviruses, opens new avenues for high-resolution structural studies of the bunyavirus family, and aids the design of antiviral diagnostics and effective subunit vaccines.

  19. Single-particle cryo-electron microscopy of Rift Valley fever virus.

    Science.gov (United States)

    Sherman, Michael B; Freiberg, Alexander N; Holbrook, Michael R; Watowich, Stanley J

    2009-04-25

    Rift Valley fever virus (RVFV; Bunyaviridae; Phlebovirus) is an emerging human and veterinary pathogen causing acute hepatitis in ruminants and has the potential to cause hemorrhagic fever in humans. We report a three-dimensional reconstruction of RVFV vaccine strain MP-12 (RVFV MP-12) by cryo-electron microcopy using icosahedral symmetry of individual virions. Although the genomic core of RVFV MP-12 is apparently poorly ordered, the glycoproteins on the virus surface are highly symmetric and arranged on a T=12 icosahedral lattice. Our RVFV MP-12 structure allowed clear identification of inter-capsomer contacts and definition of possible glycoprotein arrangements within capsomers. This structure provides a detailed model for phleboviruses, opens new avenues for high-resolution structural studies of the bunyavirus family, and aids the design of antiviral diagnostics and effective subunit vaccines.

  20. High-resolution scanning near-field EBIC microscopy: Application to the characterisation of a shallow ion implanted p+-n silicon junction

    International Nuclear Information System (INIS)

    Smaali, K.; Faure, J.; El Hdiy, A.; Troyon, M.

    2008-01-01

    High-resolution electron beam induced current (EBIC) analyses were carried out on a shallow ion implanted p + -n silicon junction in a scanning electron microscope (SEM) and a scanning probe microscope (SPM) hybrid system. With this scanning near-field EBIC microscope, a sample can be conventionally imaged by SEM, its local topography investigated by SPM and high-resolution EBIC image simultaneously obtained. It is shown that the EBIC imaging capabilities of this combined instrument allows the study of p-n junctions with a resolution of about 20 nm

  1. Cryo-scanning electron microscopy discloses differences in dehydration of frozen boar semen stored in large containers.

    Science.gov (United States)

    Ekwall, H

    2009-02-01

    In general, freezing in flat plastic polyethylene terephthalate (PET) bags (FlatPacks) at 50 degrees C/min gives better post-thaw viability, in terms of sperm motility and membrane integrity, than does freezing in plastic maxi-straws, probably owing to differences in cryobiology. To test the hypothesis that this better survival post-thaw relates to the degree of sperm dehydration during freezing, the present study investigated the structure of boar semen in a frozen state using cryo-scanning electron microscopy (cryo-SEM) to compare two different packages (FlatPacks and maxi-straws) for single artificial insemination (AI) doses, and three different freezing rates. The semen was split-sample frozen in maxi-straws or FlatPacks (both holding 5 ml) using 3% glycerol as cryoprotectant. Three freezing rates were applied from -5 degrees C to -100 degrees C, namely 2 degrees C/min, 50 degrees C/min and 1200 degrees C/min, the lattermost by plunging the samples into liquid nitrogen (LN(2)). The samples were thereafter fractured into LN(2) and larger areas of extra-cellular, unbound frozen water ('ice lakes') were measured to determine the degree of dehydration of the spermatozoa. These areas decreased in size with an increase in cooling rate, the differences in size being more dramatic for maxi-straws than for FlatPacks. Size of ice lakes was also influenced by location within package in relation to cooling rate, the central values being always smaller in maxi-straws than in Flatpacks (p < 0.05 at 2 degrees C/min and 50 degrees C/min) but not at 1200 degrees C/min, which suggested the FlatPack allows for more homogenous freezing of boar semen.

  2. Optimized cryo-focused ion beam sample preparation aimed at in situ structural studies of membrane proteins.

    Science.gov (United States)

    Schaffer, Miroslava; Mahamid, Julia; Engel, Benjamin D; Laugks, Tim; Baumeister, Wolfgang; Plitzko, Jürgen M

    2017-02-01

    While cryo-electron tomography (cryo-ET) can reveal biological structures in their native state within the cellular environment, it requires the production of high-quality frozen-hydrated sections that are thinner than 300nm. Sample requirements are even more stringent for the visualization of membrane-bound protein complexes within dense cellular regions. Focused ion beam (FIB) sample preparation for transmission electron microscopy (TEM) is a well-established technique in material science, but there are only few examples of biological samples exhibiting sufficient quality for high-resolution in situ investigation by cryo-ET. In this work, we present a comprehensive description of a cryo-sample preparation workflow incorporating additional conductive-coating procedures. These coating steps eliminate the adverse effects of sample charging on imaging with the Volta phase plate, allowing data acquisition with improved contrast. We discuss optimized FIB milling strategies adapted from material science and each critical step required to produce homogeneously thin, non-charging FIB lamellas that make large areas of unperturbed HeLa and Chlamydomonas cells accessible for cryo-ET at molecular resolution. Copyright © 2016 Elsevier Inc. All rights reserved.

  3. Neuroanatomy from Mesoscopic to Nanoscopic Scales: An Improved Method for the Observation of Semithin Sections by High-Resolution Scanning Electron Microscopy.

    Science.gov (United States)

    Rodríguez, José-Rodrigo; Turégano-López, Marta; DeFelipe, Javier; Merchán-Pérez, Angel

    2018-01-01

    Semithin sections are commonly used to examine large areas of tissue with an optical microscope, in order to locate and trim the regions that will later be studied with the electron microscope. Ideally, the observation of semithin sections would be from mesoscopic to nanoscopic scales directly, instead of using light microscopy and then electron microscopy (EM). Here we propose a method that makes it possible to obtain high-resolution scanning EM images of large areas of the brain in the millimeter to nanometer range. Since our method is compatible with light microscopy, it is also feasible to generate hybrid light and electron microscopic maps. Additionally, the same tissue blocks that have been used to obtain semithin sections can later be used, if necessary, for transmission EM, or for focused ion beam milling and scanning electron microscopy (FIB-SEM).

  4. FOLD-EM: automated fold recognition in medium- and low-resolution (4-15 Å) electron density maps.

    Science.gov (United States)

    Saha, Mitul; Morais, Marc C

    2012-12-15

    Owing to the size and complexity of large multi-component biological assemblies, the most tractable approach to determining their atomic structure is often to fit high-resolution radiographic or nuclear magnetic resonance structures of isolated components into lower resolution electron density maps of the larger assembly obtained using cryo-electron microscopy (cryo-EM). This hybrid approach to structure determination requires that an atomic resolution structure of each component, or a suitable homolog, is available. If neither is available, then the amount of structural information regarding that component is limited by the resolution of the cryo-EM map. However, even if a suitable homolog cannot be identified using sequence analysis, a search for structural homologs should still be performed because structural homology often persists throughout evolution even when sequence homology is undetectable, As macromolecules can often be described as a collection of independently folded domains, one way of searching for structural homologs would be to systematically fit representative domain structures from a protein domain database into the medium/low resolution cryo-EM map and return the best fits. Taken together, the best fitting non-overlapping structures would constitute a 'mosaic' backbone model of the assembly that could aid map interpretation and illuminate biological function. Using the computational principles of the Scale-Invariant Feature Transform (SIFT), we have developed FOLD-EM-a computational tool that can identify folded macromolecular domains in medium to low resolution (4-15 Å) electron density maps and return a model of the constituent polypeptides in a fully automated fashion. As a by-product, FOLD-EM can also do flexible multi-domain fitting that may provide insight into conformational changes that occur in macromolecular assemblies.

  5. Quantitative atomic resolution mapping using high-angle annular dark field scanning transmission electron microscopy

    International Nuclear Information System (INIS)

    Van Aert, S.; Verbeeck, J.; Erni, R.; Bals, S.; Luysberg, M.; Dyck, D. Van; Tendeloo, G. Van

    2009-01-01

    A model-based method is proposed to relatively quantify the chemical composition of atomic columns using high angle annular dark field (HAADF) scanning transmission electron microscopy (STEM) images. The method is based on a quantification of the total intensity of the scattered electrons for the individual atomic columns using statistical parameter estimation theory. In order to apply this theory, a model is required describing the image contrast of the HAADF STEM images. Therefore, a simple, effective incoherent model has been assumed which takes the probe intensity profile into account. The scattered intensities can then be estimated by fitting this model to an experimental HAADF STEM image. These estimates are used as a performance measure to distinguish between different atomic column types and to identify the nature of unknown columns with good accuracy and precision using statistical hypothesis testing. The reliability of the method is supported by means of simulated HAADF STEM images as well as a combination of experimental images and electron energy-loss spectra. It is experimentally shown that statistically meaningful information on the composition of individual columns can be obtained even if the difference in averaged atomic number Z is only 3. Using this method, quantitative mapping at atomic resolution using HAADF STEM images only has become possible without the need of simultaneously recorded electron energy loss spectra.

  6. Near-Atomic Resolution Structure of a Highly Neutralizing Fab Bound to Canine Parvovirus.

    Science.gov (United States)

    Organtini, Lindsey J; Lee, Hyunwook; Iketani, Sho; Huang, Kai; Ashley, Robert E; Makhov, Alexander M; Conway, James F; Parrish, Colin R; Hafenstein, Susan

    2016-11-01

    Canine parvovirus (CPV) is a highly contagious pathogen that causes severe disease in dogs and wildlife. Previously, a panel of neutralizing monoclonal antibodies (MAb) raised against CPV was characterized. An antibody fragment (Fab) of MAb E was found to neutralize the virus at low molar ratios. Using recent advances in cryo-electron microscopy (cryo-EM), we determined the structure of CPV in complex with Fab E to 4.1 Å resolution, which allowed de novo building of the Fab structure. The footprint identified was significantly different from the footprint obtained previously from models fitted into lower-resolution maps. Using single-chain variable fragments, we tested antibody residues that control capsid binding. The near-atomic structure also revealed that Fab binding had caused capsid destabilization in regions containing key residues conferring receptor binding and tropism, which suggests a mechanism for efficient virus neutralization by antibody. Furthermore, a general technical approach to solving the structures of small molecules is demonstrated, as binding the Fab to the capsid allowed us to determine the 50-kDa Fab structure by cryo-EM. Using cryo-electron microscopy and new direct electron detector technology, we have solved the 4 Å resolution structure of a Fab molecule bound to a picornavirus capsid. The Fab induced conformational changes in regions of the virus capsid that control receptor binding. The antibody footprint is markedly different from the previous one identified by using a 12 Å structure. This work emphasizes the need for a high-resolution structure to guide mutational analysis and cautions against relying on older low-resolution structures even though they were interpreted with the best methodology available at the time. Copyright © 2016, American Society for Microbiology. All Rights Reserved.

  7. Never at rest: insights into the conformational dynamics of ion channels from cryo-electron microscopy.

    Science.gov (United States)

    Lau, Carus; Hunter, Mark J; Stewart, Alastair; Perozo, Eduardo; Vandenberg, Jamie I

    2018-04-01

    The tightly regulated opening and closure of ion channels underlies the electrical signals that are vital for a wide range of physiological processes. Two decades ago the first atomic level view of ion channel structures led to a detailed understanding of ion selectivity and conduction. In recent years, spectacular developments in the field of cryo-electron microscopy have resulted in cryo-EM superseding crystallography as the technique of choice for determining near-atomic resolution structures of ion channels. Here, we will review the recent developments in cryo-EM and its specific application to the study of ion channel gating. We will highlight the advantages and disadvantages of the current technology and where the field is likely to head in the next few years. © 2018 The Authors. The Journal of Physiology © 2018 The Physiological Society.

  8. 3D structure of eukaryotic flagella/cilia by cryo-electron tomography.

    Science.gov (United States)

    Ishikawa, Takashi

    2013-01-01

    Flagella/cilia are motile organelles with more than 400 proteins. To understand the mechanism of such complex systems, we need methods to describe molecular arrange-ments and conformations three-dimensionally in vivo. Cryo-electron tomography enabled us such a 3D structural analysis. Our group has been working on 3D structure of flagella/cilia using this method and revealed highly ordered and beautifully organized molecular arrangement. 3D structure gave us insights into the mechanism to gener-ate bending motion with well defined waveforms. In this review, I summarize our recent structural studies on fla-gella/cilia by cryo-electron tomography, mainly focusing on dynein microtubule-based ATPase motor proteins and the radial spoke, a regulatory protein complex.

  9. Development of Scanning Ultrafast Electron Microscope Capability.

    Energy Technology Data Exchange (ETDEWEB)

    Collins, Kimberlee Chiyoko [Sandia National Lab. (SNL-CA), Livermore, CA (United States); Talin, Albert Alec [Sandia National Lab. (SNL-CA), Livermore, CA (United States); Chandler, David W. [Sandia National Lab. (SNL-CA), Livermore, CA (United States); Michael, Joseph R. [Sandia National Lab. (SNL-NM), Albuquerque, NM (United States)

    2016-11-01

    Modern semiconductor devices rely on the transport of minority charge carriers. Direct examination of minority carrier lifetimes in real devices with nanometer-scale features requires a measurement method with simultaneously high spatial and temporal resolutions. Achieving nanometer spatial resolutions at sub-nanosecond temporal resolution is possible with pump-probe methods that utilize electrons as probes. Recently, a stroboscopic scanning electron microscope was developed at Caltech, and used to study carrier transport across a Si p-n junction [ 1 , 2 , 3 ] . In this report, we detail our development of a prototype scanning ultrafast electron microscope system at Sandia National Laboratories based on the original Caltech design. This effort represents Sandia's first exploration into ultrafast electron microscopy.

  10. Local imaging of high mobility two-dimensional electron systems with virtual scanning tunneling microscopy

    Energy Technology Data Exchange (ETDEWEB)

    Pelliccione, M. [Department of Applied Physics, Stanford University, 348 Via Pueblo Mall, Stanford, California 94305 (United States); Stanford Institute for Materials and Energy Sciences, SLAC National Accelerator Laboratory, 2575 Sand Hill Road, Menlo Park, California 94025 (United States); Department of Physics, University of California, Santa Barbara, Santa Barbara, California 93106 (United States); Bartel, J.; Goldhaber-Gordon, D. [Stanford Institute for Materials and Energy Sciences, SLAC National Accelerator Laboratory, 2575 Sand Hill Road, Menlo Park, California 94025 (United States); Department of Physics, Stanford University, 382 Via Pueblo Mall, Stanford, California 94305 (United States); Sciambi, A. [Department of Applied Physics, Stanford University, 348 Via Pueblo Mall, Stanford, California 94305 (United States); Stanford Institute for Materials and Energy Sciences, SLAC National Accelerator Laboratory, 2575 Sand Hill Road, Menlo Park, California 94025 (United States); Pfeiffer, L. N.; West, K. W. [Department of Electrical Engineering, Princeton University, Princeton, New Jersey 08544 (United States)

    2014-11-03

    Correlated electron states in high mobility two-dimensional electron systems (2DESs), including charge density waves and microemulsion phases intermediate between a Fermi liquid and Wigner crystal, are predicted to exhibit complex local charge order. Existing experimental studies, however, have mainly probed these systems at micron to millimeter scales rather than directly mapping spatial organization. Scanning probes should be well-suited to study the spatial structure of these states, but high mobility 2DESs are found at buried semiconductor interfaces, beyond the reach of conventional scanning tunneling microscopy. Scanning techniques based on electrostatic coupling to the 2DES deliver important insights, but generally with resolution limited by the depth of the 2DES. In this letter, we present our progress in developing a technique called “virtual scanning tunneling microscopy” that allows local tunneling into a high mobility 2DES. Using a specially designed bilayer GaAs/AlGaAs heterostructure where the tunnel coupling between two separate 2DESs is tunable via electrostatic gating, combined with a scanning gate, we show that the local tunneling can be controlled with sub-250 nm resolution.

  11. Characterization of intact subcellular bodies in whole bacteria by cryo-electron tomography and spectroscopic imaging.

    Science.gov (United States)

    Comolli, L R; Kundmann, M; Downing, K H

    2006-07-01

    We illustrate the combined use of cryo-electron tomography and spectroscopic difference imaging in the study of subcellular structure and subcellular bodies in whole bacteria. We limited our goal and focus to bodies with a distinct elemental composition that was in a sufficiently high concentration to provide the necessary signal-to-noise level at the relatively large sample thicknesses of the intact cell. This combination proved very powerful, as demonstrated by the identification of a phosphorus-rich body in Caulobacter crescentus. We also confirmed the presence of a body rich in carbon, demonstrated that these two types of bodies are readily recognized and distinguished from each other, and provided, for the first time to our knowledge, structural information about them in their intact state. In addition, we also showed the presence of a similar type of phosphorus-rich body in Deinococcus grandis, a member of a completely unrelated bacteria genus. Cryo-electron microscopy and tomography allowed the study of the biogenesis and morphology of these bodies at resolutions better than 10 nm, whereas spectroscopic difference imaging provided a direct identification of their chemical composition.

  12. Advances and challenges in cryo ptychography at the Advanced Photon Source

    Energy Technology Data Exchange (ETDEWEB)

    Deng, J. [Applied Physics, Northwestern University, Evanston IL 60208 (United States); Vine, D. J.; Chen, S.; Vogt, S. [X-ray Science Division, Advanced Photon Source, Argonne National Laboratory, Argonne, IL 60439 (United States); Nashed, Y. S. G.; Peterka, T. [Mathematics and Computing Science Division, Argonne National Laboratory, Argonne, IL 60439 (United States); Jin, Q. [Department of Physics & Astronomy, Northwestern University, Evanston, IL 60208 (United States); Jacobsen, C. [X-ray Science Division, Advanced Photon Source, Argonne National Laboratory, Argonne, IL 60439 (United States); Department of Physics & Astronomy and Chemistry of Life Processes Institute, Northwestern University, Evanston, IL 60208 (United States)

    2016-01-28

    Ptychography has emerged as a nondestructive tool to quantitatively study extended samples at a high spatial resolution. In this manuscript, we report on recent developments from our team. We have combined cryo ptychography and fluorescence microscopy to provide simultaneous views of ultrastructure and elemental composition, we have developed multi-GPU parallel computation to speed up ptychographic reconstructions, and we have implemented fly-scan ptychography to allow for faster data acquisition. We conclude with a discussion of future challenges in high-resolution 3D ptychography.

  13. A comparative analysis of the sea ice freeboard from CryoSat. CryoVEx and IceBridge

    DEFF Research Database (Denmark)

    Kildegaard Rose, Stine; Connor, Laurence N.; Farrell, Sinead L.

    The CryoSat Validation Experiment (CryoVEx) has been conducted by ESA, to examine the uncertainties in the satellite measurement of e.g. sea ice thickness. In this study, we aim to estimate the sea ice freeboard from CryoSat-2, and compare it with the high-resolution Airborne Laser Scanner (ALS......) measurements collected along CryoSat-2 ground tracks from the CryoVEx 2012 campaign, together with NASA’s Operation IceBridge data. We will use the CryoSat SAR data level 1b (L1b) to discriminate the leads and from this, estimate the sea ice freeboard. Furthermore, we are looking at the CryoSat level 2 (L2...

  14. Attainment of 40.5 pm spatial resolution using 300 kV scanning transmission electron microscope equipped with fifth-order aberration corrector.

    Science.gov (United States)

    Morishita, Shigeyuki; Ishikawa, Ryo; Kohno, Yuji; Sawada, Hidetaka; Shibata, Naoya; Ikuhara, Yuichi

    2018-02-01

    The achievement of a fine electron probe for high-resolution imaging in scanning transmission electron microscopy requires technological developments, especially in electron optics. For this purpose, we developed a microscope with a fifth-order aberration corrector that operates at 300 kV. The contrast flat region in an experimental Ronchigram, which indicates the aberration-free angle, was expanded to 70 mrad. By using a probe with convergence angle of 40 mrad in the scanning transmission electron microscope at 300 kV, we attained the spatial resolution of 40.5 pm, which is the projected interatomic distance between Ga-Ga atomic columns of GaN observed along [212] direction.

  15. Monochromated scanning transmission electron microscopy

    International Nuclear Information System (INIS)

    Rechberger, W.; Kothleitner, G.; Hofer, F.

    2006-01-01

    Full text: Electron energy-loss spectroscopy (EELS) has developed into an established technique for chemical and structural analysis of thin specimens in the (scanning) transmission electron microscope (S)TEM. The energy resolution in EELS is largely limited by the stability of the high voltage supply, by the resolution of the spectrometer and by the energy spread of the source. To overcome this limitation a Wien filter monochromator was recently introduced with commercially available STEMs, offering the advantage to better resolve EELS fine structures, which contain valuable bonding information. The method of atomic resolution Z-contrast imaging within an STEM, utilizing a high-angle annular dark-field (HAADF) detector can perfectly complement the excellent energy resolution, since EELS spectra can be collected simultaneously. In combination with a monochromator microscope not only high spatial resolution images can be recorded but also high energy resolution EELS spectra are attainable. In this work we investigated the STEM performance of a 200 kV monochromated Tecnai F20 with a high resolution Gatan Imaging Filter (HR-GIF). (author)

  16. Helium ion microscopy and ultra-high-resolution scanning electron microscopy analysis of membrane-extracted cells reveals novel characteristics of the cytoskeleton of Giardia intestinalis.

    Science.gov (United States)

    Gadelha, Ana Paula Rocha; Benchimol, Marlene; de Souza, Wanderley

    2015-06-01

    Giardia intestinalis presents a complex microtubular cytoskeleton formed by specialized structures, such as the adhesive disk, four pairs of flagella, the funis and the median body. The ultrastructural organization of the Giardia cytoskeleton has been analyzed using different microscopic techniques, including high-resolution scanning electron microscopy. Recent advances in scanning microscopy technology have opened a new venue for the characterization of cellular structures and include scanning probe microscopy techniques such as ultra-high-resolution scanning electron microscopy (UHRSEM) and helium ion microscopy (HIM). Here, we studied the organization of the cytoskeleton of G. intestinalis trophozoites using UHRSEM and HIM in membrane-extracted cells. The results revealed a number of new cytoskeletal elements associated with the lateral crest and the dorsal surface of the parasite. The fine structure of the banded collar was also observed. The marginal plates were seen linked to a network of filaments, which were continuous with filaments parallel to the main cell axis. Cytoplasmic filaments that supported the internal structures were seen by the first time. Using anti-actin antibody, we observed a labeling in these filamentous structures. Taken together, these data revealed new surface characteristics of the cytoskeleton of G. intestinalis and may contribute to an improved understanding of the structural organization of trophozoites. Copyright © 2015 Elsevier Inc. All rights reserved.

  17. A design for a subminiature, low energy scanning electron microscope with atomic resolution

    International Nuclear Information System (INIS)

    Eastham, D. A.; Edmondson, P.; Greene, S.; Donnelly, S.; Olsson, E.; Svensson, K.; Bleloch, A.

    2009-01-01

    We describe a type of scanning electron microscope that works by directly imaging the electron field-emission sites on a nanotip. Electrons are extracted from the nanotip through a nanoscale aperture, accelerated in a high electric field, and focused to a spot using a microscale Einzel lens. If the whole microscope (accelerating section and lens) and the focal length are both restricted in size to below 10 μm, then computer simulations show that the effects of aberration are extremely small and it is possible to have a system with approximately unit magnification at electron energies as low as 300 eV. Thus a typical emission site of 1 nm diameter will produce an image of the same size, and an atomic emission site will give a resolution of 0.1-0.2 nm (1-2 A). Also, because the beam is not allowed to expand beyond 100 nm in diameter, the depth of field is large and the contribution to the beam spot size from chromatic aberrations is less than 0.02 nm (0.2 A) for 500 eV electrons. Since it is now entirely possible to make stable atomic sized emitters (nanopyramids), it is expected that this instrument will have atomic resolution. Furthermore the brightness of the beam is determined only by the field emission and can be up to 1x10 6 times larger than in a typical (high energy) electron microscope. The advantages of this low energy, bright-beam electron microscope with atomic resolution are described and include the possibility of it being used to rapidly sequence the human genome from a single strand of DNA as well as being able to identify atomic species directly from the elastic scattering of electrons

  18. 3D structure of eukaryotic flagella in a quiescent state revealed by cryo-electron tomography

    Science.gov (United States)

    Nicastro, Daniela; McIntosh, J. Richard; Baumeister, Wolfgang

    2005-01-01

    We have used cryo-electron tomography to investigate the 3D structure and macromolecular organization of intact, frozen-hydrated sea urchin sperm flagella in a quiescent state. The tomographic reconstructions provide information at a resolution better than 6 nm about the in situ arrangements of macromolecules that are key for flagellar motility. We have visualized the heptameric rings of the motor domains in the outer dynein arm complex and determined that they lie parallel to the plane that contains the axes of neighboring flagellar microtubules. Both the material associated with the central pair of microtubules and the radial spokes display a plane of symmetry that helps to explain the planar beat pattern of these flagella. Cryo-electron tomography has proven to be a powerful technique for helping us understand the relationships between flagellar structure and function and the design of macromolecular machines in situ. PMID:16246999

  19. High resolution techniques using scanning proton microprobe (SPM)

    International Nuclear Information System (INIS)

    Cholewa, M.; Saint, A.; Prawer, S.; Laird, J.S.; Legge, G.J.F.; Bardos, R.A.; Moorhead, G.F.; Taylor, G.N.; Stuart, S.A.; Howard, J.

    1994-01-01

    The very high resolution (down to 50 nm) achieved with low beam currents (fA) in a scanning ion microprobe have lead to many nondestructive techniques of microanalysis. This paper discusses recent developments and applications in the use of 3-D STIM (scanning transmission ion microscopy) Tomography, channeling STIM and IBIC (ion beam induced charge). (orig.)

  20. Cryo-EM studies of the structure and dynamics of vacuolar-type ATPases

    Science.gov (United States)

    Mazhab-Jafari, Mohammad T.; Rubinstein, John L.

    2016-01-01

    Electron cryomicroscopy (cryo-EM) has significantly advanced our understanding of molecular structure in biology. Recent innovations in both hardware and software have made cryo-EM a viable alternative for targets that are not amenable to x-ray crystallography or nuclear magnetic resonance (NMR) spectroscopy. Cryo-EM has even become the method of choice in some situations where x-ray crystallography and NMR spectroscopy are possible but where cryo-EM can determine structures at higher resolution or with less time or effort. Rotary adenosine triphosphatases (ATPases) are crucial to the maintenance of cellular homeostasis. These enzymes couple the synthesis or hydrolysis of adenosine triphosphate to the use or production of a transmembrane electrochemical ion gradient, respectively. However, the membrane-embedded nature and conformational heterogeneity of intact rotary ATPases have prevented their high-resolution structural analysis to date. Recent application of cryo-EM methods to the different types of rotary ATPase has led to sudden advances in understanding the structure and function of these enzymes, revealing significant conformational heterogeneity and characteristic transmembrane α helices that are highly tilted with respect to the membrane. In this Review, we will discuss what has been learned recently about rotary ATPase structure and function, with a particular focus on the vacuolar-type ATPases. PMID:27532044

  1. Method and apparatus for a high-resolution three dimensional confocal scanning transmission electron microscope

    Science.gov (United States)

    de Jonge, Niels [Oak Ridge, TN

    2010-08-17

    A confocal scanning transmission electron microscope which includes an electron illumination device providing an incident electron beam propagating in a direction defining a propagation axis, and a precision specimen scanning stage positioned along the propagation axis and movable in at least one direction transverse to the propagation axis. The precision specimen scanning stage is configured for positioning a specimen relative to the incident electron beam. A projector lens receives a transmitted electron beam transmitted through at least part of the specimen and focuses this transmitted beam onto an image plane, where the transmitted beam results from the specimen being illuminated by the incident electron beam. A detection system is placed approximately in the image plane.

  2. Parainfluenza virus type 5 (PIV-5) morphology revealed by cryo-electron microscopy.

    Science.gov (United States)

    Terrier, Olivier; Rolland, Jean-Paul; Rosa-Calatrava, Manuel; Lina, Bruno; Thomas, Daniel; Moules, Vincent

    2009-06-01

    The knowledge of parainfluenza type 5 (PIV-5) virion morphology is essentially based on the observation of negatively stained preparations in conventional transmission electron microscopy (CTEM). In this study, the ultrastructure of frozen-hydrated intact PIV-5 was examined by cryo-electron microscopy (cryo-EM). Cryo-EM revealed a majority of spherical virions (70%), with a lower pleiomorphy than originally observed in CTEM. Phospholipid bilayer thickness, spike length and glycoprotein spikes density were measured. About 2000 glycoprotein spikes were present in an average-sized spherical virion. Altogether, these data depict a more precise view of PIV-5 morphology.

  3. An electron beam linear scanning mode for industrial limited-angle nano-computed tomography

    Science.gov (United States)

    Wang, Chengxiang; Zeng, Li; Yu, Wei; Zhang, Lingli; Guo, Yumeng; Gong, Changcheng

    2018-01-01

    Nano-computed tomography (nano-CT), which utilizes X-rays to research the inner structure of some small objects and has been widely utilized in biomedical research, electronic technology, geology, material sciences, etc., is a high spatial resolution and non-destructive research technique. A traditional nano-CT scanning model with a very high mechanical precision and stability of object manipulator, which is difficult to reach when the scanned object is continuously rotated, is required for high resolution imaging. To reduce the scanning time and attain a stable and high resolution imaging in industrial non-destructive testing, we study an electron beam linear scanning mode of nano-CT system that can avoid mechanical vibration and object movement caused by the continuously rotated object. Furthermore, to further save the scanning time and study how small the scanning range could be considered with acceptable spatial resolution, an alternating iterative algorithm based on ℓ0 minimization is utilized to limited-angle nano-CT reconstruction problem with the electron beam linear scanning mode. The experimental results confirm the feasibility of the electron beam linear scanning mode of nano-CT system.

  4. Quantum ballistic transistor and low noise HEMT for cryo-electronics lower than 4.2 K

    International Nuclear Information System (INIS)

    Gremion, E.

    2008-01-01

    Next generations of cryo-detectors, widely used in physics of particles and physics of universe, will need in the future high-performance cryo-electronics less noisy and closer to the detector. Within this context, this work investigates properties of two dimensional electron gas GaAlAs/GaAs by studying two components, quantum point contact (QPC) and high electron mobility transistor (HEMT). Thanks to quantized conductance steps in QPC, we have realized a quantum ballistic transistor (voltage gain higher than 1), a new component useful for cryo-electronics thanks to its operating temperature and weak power consumption (about 1 nW). Moreover, the very low capacity of this component leads to promising performances for multiplexing low temperature bolometer dedicated to millimetric astronomy. The second study focused on HEMT with very high quality 2DEG. At 4.2 K, a voltage gain higher than 20 can be obtained with a very low power dissipation of less than 100 μW. Under the above experimental conditions, an equivalent input voltage noise of 1.2 nV/√(Hz) at 1 kHz and 0.12 nV/√(Hz) at 100 kHz has been reached. According to the Hooge formula, these noise performances are get by increasing gate capacity estimated to 60 pF. (author)

  5. Exploring an optimal wavelet-based filter for cryo-ET imaging.

    Science.gov (United States)

    Huang, Xinrui; Li, Sha; Gao, Song

    2018-02-07

    Cryo-electron tomography (cryo-ET) is one of the most advanced technologies for the in situ visualization of molecular machines by producing three-dimensional (3D) biological structures. However, cryo-ET imaging has two serious disadvantages-low dose and low image contrast-which result in high-resolution information being obscured by noise and image quality being degraded, and this causes errors in biological interpretation. The purpose of this research is to explore an optimal wavelet denoising technique to reduce noise in cryo-ET images. We perform tests using simulation data and design a filter using the optimum selected wavelet parameters (three-level decomposition, level-1 zeroed out, subband-dependent threshold, a soft-thresholding and spline-based discrete dyadic wavelet transform (DDWT)), which we call a modified wavelet shrinkage filter; this filter is suitable for noisy cryo-ET data. When testing using real cryo-ET experiment data, higher quality images and more accurate measures of a biological structure can be obtained with the modified wavelet shrinkage filter processing compared with conventional processing. Because the proposed method provides an inherent advantage when dealing with cryo-ET images, it can therefore extend the current state-of-the-art technology in assisting all aspects of cryo-ET studies: visualization, reconstruction, structural analysis, and interpretation.

  6. On the Progress of Scanning Transmission Electron Microscopy (STEM) Imaging in a Scanning Electron Microscope.

    Science.gov (United States)

    Sun, Cheng; Müller, Erich; Meffert, Matthias; Gerthsen, Dagmar

    2018-04-01

    Transmission electron microscopy (TEM) with low-energy electrons has been recognized as an important addition to the family of electron microscopies as it may avoid knock-on damage and increase the contrast of weakly scattering objects. Scanning electron microscopes (SEMs) are well suited for low-energy electron microscopy with maximum electron energies of 30 keV, but they are mainly used for topography imaging of bulk samples. Implementation of a scanning transmission electron microscopy (STEM) detector and a charge-coupled-device camera for the acquisition of on-axis transmission electron diffraction (TED) patterns, in combination with recent resolution improvements, make SEMs highly interesting for structure analysis of some electron-transparent specimens which are traditionally investigated by TEM. A new aspect is correlative SEM, STEM, and TED imaging from the same specimen region in a SEM which leads to a wealth of information. Simultaneous image acquisition gives information on surface topography, inner structure including crystal defects and qualitative material contrast. Lattice-fringe resolution is obtained in bright-field STEM imaging. The benefits of correlative SEM/STEM/TED imaging in a SEM are exemplified by structure analyses from representative sample classes such as nanoparticulates and bulk materials.

  7. High resolution scanning electron microscopy of rabbit corneal endothelium to show effects of UV-visible irradiation in the presence of chlorpromazine

    Energy Technology Data Exchange (ETDEWEB)

    Lea, P.J.; Hollenberg, M.J.; Menon, I.A.; Temkin, R.J.; Persad, S.D.; Basu, P.K. (Univ. of Toronto, Ontario (Canada))

    1989-01-01

    The ultrastructure of rabbit cornea endothelial cells was examined by scanning electron microscopy (SEM) in freeze-cleaved corneas using a Hitachi S-570 scanning electron microscope in the high resolution mode (HRSEM). In order to study phototoxic effects in vitro, rabbit corneas (experimental) were cultured as organ culture in the presence of 5 micrograms/ml chlorpromazine (CPZ) and irradiated. For comparison, control 1 corneas were not irradiated but incubated in the dark without CPZ in the medium; control 2 corneas were also kept in the dark but in the presence of CPZ; control 3 corneas were irradiated with no CPZ in the medium. Cellular damage was not seen in the three types of control corneas, but in the experimental corneas the endothelial cells showed extensive disruption of the cell membrane and some deterioration of the intracellular components. Our study confirmed that HRSEM is a satisfactory new technique for visualizing damage of the intracellular organelles of corneal endothelium.

  8. High resolution scanning electron microscopy of rabbit corneal endothelium to show effects of UV-visible irradiation in the presence of chlorpromazine

    International Nuclear Information System (INIS)

    Lea, P.J.; Hollenberg, M.J.; Menon, I.A.; Temkin, R.J.; Persad, S.D.; Basu, P.K.

    1989-01-01

    The ultrastructure of rabbit cornea endothelial cells was examined by scanning electron microscopy (SEM) in freeze-cleaved corneas using a Hitachi S-570 scanning electron microscope in the high resolution mode (HRSEM). In order to study phototoxic effects in vitro, rabbit corneas (experimental) were cultured as organ culture in the presence of 5 micrograms/ml chlorpromazine (CPZ) and irradiated. For comparison, control 1 corneas were not irradiated but incubated in the dark without CPZ in the medium; control 2 corneas were also kept in the dark but in the presence of CPZ; control 3 corneas were irradiated with no CPZ in the medium. Cellular damage was not seen in the three types of control corneas, but in the experimental corneas the endothelial cells showed extensive disruption of the cell membrane and some deterioration of the intracellular components. Our study confirmed that HRSEM is a satisfactory new technique for visualizing damage of the intracellular organelles of corneal endothelium

  9. Comparison of quantitative myocardial perfusion imaging CT to fluorescent microsphere-based flow from high-resolution cryo-images

    Science.gov (United States)

    Eck, Brendan L.; Fahmi, Rachid; Levi, Jacob; Fares, Anas; Wu, Hao; Li, Yuemeng; Vembar, Mani; Dhanantwari, Amar; Bezerra, Hiram G.; Wilson, David L.

    2016-03-01

    Myocardial perfusion imaging using CT (MPI-CT) has the potential to provide quantitative measures of myocardial blood flow (MBF) which can aid the diagnosis of coronary artery disease. We evaluated the quantitative accuracy of MPI-CT in a porcine model of balloon-induced LAD coronary artery ischemia guided by fractional flow reserve (FFR). We quantified MBF at baseline (FFR=1.0) and under moderate ischemia (FFR=0.7) using MPI-CT and compared to fluorescent microsphere-based MBF from high-resolution cryo-images. Dynamic, contrast-enhanced CT images were obtained using a spectral detector CT (Philips Healthcare). Projection-based mono-energetic images were reconstructed and processed to obtain MBF. Three MBF quantification approaches were evaluated: singular value decomposition (SVD) with fixed Tikhonov regularization (ThSVD), SVD with regularization determined by the L-Curve criterion (LSVD), and Johnson-Wilson parameter estimation (JW). The three approaches over-estimated MBF compared to cryo-images. JW produced the most accurate MBF, with average error 33.3+/-19.2mL/min/100g, whereas LSVD and ThSVD had greater over-estimation, 59.5+/-28.3mL/min/100g and 78.3+/-25.6 mL/min/100g, respectively. Relative blood flow as assessed by a flow ratio of LAD-to-remote myocardium was strongly correlated between JW and cryo-imaging, with R2=0.97, compared to R2=0.88 and 0.78 for LSVD and ThSVD, respectively. We assessed tissue impulse response functions (IRFs) from each approach for sources of error. While JW was constrained to physiologic solutions, both LSVD and ThSVD produced IRFs with non-physiologic properties due to noise. The L-curve provided noise-adaptive regularization but did not eliminate non-physiologic IRF properties or optimize for MBF accuracy. These findings suggest that model-based MPI-CT approaches may be more appropriate for quantitative MBF estimation and that cryo-imaging can support the development of MPI-CT by providing spatial distributions of MBF.

  10. OMNY—A tOMography Nano crYo stage

    Science.gov (United States)

    Holler, M.; Raabe, J.; Diaz, A.; Guizar-Sicairos, M.; Wepf, R.; Odstrcil, M.; Shaik, F. R.; Panneels, V.; Menzel, A.; Sarafimov, B.; Maag, S.; Wang, X.; Thominet, V.; Walther, H.; Lachat, T.; Vitins, M.; Bunk, O.

    2018-04-01

    For many scientific questions gaining three-dimensional insight into a specimen can provide valuable information. We here present an instrument called "tOMography Nano crYo (OMNY)," dedicated to high resolution 3D scanning x-ray microscopy at cryogenic conditions via hard X-ray ptychography. Ptychography is a lens-less imaging method requiring accurate sample positioning. In OMNY, this in achieved via dedicated laser interferometry and closed-loop position control reaching sub-10 nm positioning accuracy. Cryogenic sample conditions are maintained via conductive cooling. 90 K can be reached when using liquid nitrogen as coolant, and 10 K is possible with liquid helium. A cryogenic sample-change mechanism permits measurements of cryogenically fixed specimens. We compare images obtained with OMNY with older measurements performed using a nitrogen gas cryo-jet of stained, epoxy-embedded retina tissue and of frozen-hydrated Chlamydomonas cells.

  11. Ribbon scanning confocal for high-speed high-resolution volume imaging of brain.

    Directory of Open Access Journals (Sweden)

    Alan M Watson

    Full Text Available Whole-brain imaging is becoming a fundamental means of experimental insight; however, achieving subcellular resolution imagery in a reasonable time window has not been possible. We describe the first application of multicolor ribbon scanning confocal methods to collect high-resolution volume images of chemically cleared brains. We demonstrate that ribbon scanning collects images over ten times faster than conventional high speed confocal systems but with equivalent spectral and spatial resolution. Further, using this technology, we reconstruct large volumes of mouse brain infected with encephalitic alphaviruses and demonstrate that regions of the brain with abundant viral replication were inaccessible to vascular perfusion. This reveals that the destruction or collapse of large regions of brain micro vasculature may contribute to the severe disease caused by Venezuelan equine encephalitis virus. Visualization of this fundamental impact of infection would not be possible without sampling at subcellular resolution within large brain volumes.

  12. High resolution transmission electron microscopy and microdiffraction for radiation damage analysis

    International Nuclear Information System (INIS)

    Sinclair, R.

    1982-01-01

    High resolution TEM techniques have developed to quite a sophisticated level over the past few years. In addition TEM instruments with a scanning capability have become available commercially which permit in particular the formation of a small electron probe at the specimen. Thus direct resolution and microdiffraction investigations of thin specimens are now possible, neither of which have been employed to any great extent in the analysis of radiation damage. Some recent advances which are thought to be relevant to this specific area of research are highlighted

  13. Cryo-transmission electron tomography of native casein micelles from bovine milk

    Science.gov (United States)

    Trejo, R.; Dokland, T.; Jurat-Fuentes, J.; Harte, F.

    2013-01-01

    Caseins are the principal protein components in milk and an important ingredient in the food industry. In liquid milk, caseins are found as micelles of casein proteins and colloidal calcium nanoclusters. Casein micelles were isolated from raw skim milk by size exclusion chromatography and suspended in milk protein-free serum produced by ultrafiltration (molecular weight cut-off of 3 kDa) of raw skim milk. The micelles were imaged by cryo-electron microscopy and subjected to tomographic reconstruction methods to visualize the 3-dimensional and internal organization of native casein micelles. This provided new insights into the internal architecture of the casein micelle that had not been apparent from prior cryo-transmission electron microscopy studies. This analysis demonstrated the presence of water-filled cavities (~20 to 30 nm in diameter), channels (diameter greater than ~5 nm), and several hundred high-density nanoclusters (6 to 12 nm in diameter) within the interior of the micelles. No spherical protein submicellar structures were observed. PMID:22118067

  14. Single-particle cryo-electron microscopy of Rift Valley fever virus

    OpenAIRE

    Sherman, Michael B.; Freiberg, Alexander N.; Holbrook, Michael R.; Watowich, Stanley J.

    2009-01-01

    Rift Valley fever virus (RVFV; Bunyaviridae; Phlebovirus) is an emerging human veterinary pathogen causing acute hepatitis in ruminants and has the potential to Single-particle cryo-EM reconstruction of RVFV MP-12 hemorrhagic fever in humans. We report a three-dimensional reconstruction of RVFV vaccine strain MP-12 (RVFV MP-12) by cryo-electron microcopy using icosahedral symmetry of individual virions. Although the genomic core of RVFV MP-12 is apparently poorly ordered, the glycoproteins on...

  15. 3D Micro-topography of Transferred Laboratory and Natural Ice Crystal Surfaces Imaged by Cryo and Environmental Scanning Electron Microscopy

    Science.gov (United States)

    Magee, N. B.; Boaggio, K.; Bancroft, L.; Bandamede, M.

    2015-12-01

    Recent work has highlighted micro-scale roughness on the surfaces of ice crystals grown and imaged in-situ within the chambers of environmental scanning electron microscopes (ESEM). These observations appear to align with theoretical and satellite observations that suggest a prevalence of rough ice in cirrus clouds. However, the atmospheric application of the lab observations are indeterminate because the observations have been based only on crystals grown on substrates and in pure-water vapor environments. In this work, we present details and results from the development of a transfer technique which allows natural and lab-grown ice and snow crystals to be captured, preserved, and transferred into the ESEM for 3D imaging. Ice crystals were gathered from 1) natural snow, 2) a balloon-borne cirrus particle capture device, and 3) lab-grown ice crystals from a diffusion chamber. Ice crystals were captured in a pre-conditioned small-volume (~1 cm3) cryo-containment cell. The cell was then sealed closed and transferred to a specially-designed cryogenic dewer (filled with liquid nitrogen or crushed dry ice) for transport to a new Hitachi Field Emission, Variable Pressure SEM (SU-5000). The cryo-cell was then removed from the dewer and quickly placed onto the pre-conditioned cryo transfer stage attached to the ESEM (Quorum 3010T). Quantitative 3D topographical digital elevation models of ice surfaces are reported from SEM for the first time, including a variety of objective measures of statistical surface roughness. The surfaces of the transported crystals clearly exhibit signatures of mesoscopic roughening that are similar to examples of roughness seen in ESEM-grown crystals. For most transported crystals, the habits and crystal edges are more intricate that those observed for ice grown directly on substrates within the ESEM chamber. Portions of some crystals do appear smooth even at magnification greater than 1000x, a rare observation in our ESEM-grown crystals. The

  16. Integration of a high-NA light microscope in a scanning electron microscope.

    Science.gov (United States)

    Zonnevylle, A C; Van Tol, R F C; Liv, N; Narvaez, A C; Effting, A P J; Kruit, P; Hoogenboom, J P

    2013-10-01

    We present an integrated light-electron microscope in which an inverted high-NA objective lens is positioned inside a scanning electron microscope (SEM). The SEM objective lens and the light objective lens have a common axis and focal plane, allowing high-resolution optical microscopy and scanning electron microscopy on the same area of a sample simultaneously. Components for light illumination and detection can be mounted outside the vacuum, enabling flexibility in the construction of the light microscope. The light objective lens can be positioned underneath the SEM objective lens during operation for sub-10 μm alignment of the fields of view of the light and electron microscopes. We demonstrate in situ epifluorescence microscopy in the SEM with a numerical aperture of 1.4 using vacuum-compatible immersion oil. For a 40-nm-diameter fluorescent polymer nanoparticle, an intensity profile with a FWHM of 380 nm is measured whereas the SEM performance is uncompromised. The integrated instrument may offer new possibilities for correlative light and electron microscopy in the life sciences as well as in physics and chemistry. © 2013 The Authors Journal of Microscopy © 2013 Royal Microscopical Society.

  17. Synthesis and electroplating of high resolution insulated carbon nanotube scanning probes for imaging in liquid solutions.

    Science.gov (United States)

    Roberts, N A; Noh, J H; Lassiter, M G; Guo, S; Kalinin, S V; Rack, P D

    2012-04-13

    High resolution and isolated scanning probe microscopy (SPM) is in demand for continued development of energy storage and conversion systems involving chemical reactions at the nanoscale as well as an improved understanding of biological systems. Carbon nanotubes (CNTs) have large aspect ratios and, if leveraged properly, can be used to develop high resolution SPM probes. Isolation of SPM probes can be achieved by depositing a dielectric film and selectively etching at the apex of the probe. In this paper the fabrication of a high resolution and isolated SPM tip is demonstrated using electron beam induced etching of a dielectric film deposited onto an SPM tip with an attached CNT at the apex.

  18. Electron tomography of cryo-immobilized plant tissue: a novel approach to studying 3D macromolecular architecture of mature plant cell walls in situ.

    Directory of Open Access Journals (Sweden)

    Purbasha Sarkar

    Full Text Available Cost-effective production of lignocellulosic biofuel requires efficient breakdown of cell walls present in plant biomass to retrieve the wall polysaccharides for fermentation. In-depth knowledge of plant cell wall composition is therefore essential for improving the fuel production process. The precise spatial three-dimensional (3D organization of cellulose, hemicellulose, pectin and lignin within plant cell walls remains unclear to date since the microscopy techniques used so far have been limited to two-dimensional, topographic or low-resolution imaging, or required isolation or chemical extraction of the cell walls. In this paper we demonstrate that by cryo-immobilizing fresh tissue, then either cryo-sectioning or freeze-substituting and resin embedding, followed by cryo- or room temperature (RT electron tomography, respectively, we can visualize previously unseen details of plant cell wall architecture in 3D, at macromolecular resolution (∼ 2 nm, and in near-native state. Qualitative and quantitative analyses showed that wall organization of cryo-immobilized samples were preserved remarkably better than conventionally prepared samples that suffer substantial extraction. Lignin-less primary cell walls were well preserved in both self-pressurized rapidly frozen (SPRF, cryo-sectioned samples as well as high-pressure frozen, freeze-substituted and resin embedded (HPF-FS-resin samples. Lignin-rich secondary cell walls appeared featureless in HPF-FS-resin sections presumably due to poor stain penetration, but their macromolecular features could be visualized in unprecedented details in our cryo-sections. While cryo-tomography of vitreous tissue sections is currently proving to be instrumental in developing 3D models of lignin-rich secondary cell walls, here we confirm that the technically easier method of RT-tomography of HPF-FS-resin sections could be used immediately for routine study of low-lignin cell walls. As a proof of principle, we

  19. Re-scan confocal microscopy: scanning twice for better resolution.

    Science.gov (United States)

    De Luca, Giulia M R; Breedijk, Ronald M P; Brandt, Rick A J; Zeelenberg, Christiaan H C; de Jong, Babette E; Timmermans, Wendy; Azar, Leila Nahidi; Hoebe, Ron A; Stallinga, Sjoerd; Manders, Erik M M

    2013-01-01

    We present a new super-resolution technique, Re-scan Confocal Microscopy (RCM), based on standard confocal microscopy extended with an optical (re-scanning) unit that projects the image directly on a CCD-camera. This new microscope has improved lateral resolution and strongly improved sensitivity while maintaining the sectioning capability of a standard confocal microscope. This simple technology is typically useful for biological applications where the combination high-resolution and high-sensitivity is required.

  20. Efficient estimation of three-dimensional covariance and its application in the analysis of heterogeneous samples in cryo-electron microscopy.

    Science.gov (United States)

    Liao, Hstau Y; Hashem, Yaser; Frank, Joachim

    2015-06-02

    Single-particle cryogenic electron microscopy (cryo-EM) is a powerful tool for the study of macromolecular structures at high resolution. Classification allows multiple structural states to be extracted and reconstructed from the same sample. One classification approach is via the covariance matrix, which captures the correlation between every pair of voxels. Earlier approaches employ computing-intensive resampling and estimate only the eigenvectors of the matrix, which are then used in a separate fast classification step. We propose an iterative scheme to explicitly estimate the covariance matrix in its entirety. In our approach, the flexibility in choosing the solution domain allows us to examine a part of the molecule in greater detail. Three-dimensional covariance maps obtained in this way from experimental data (cryo-EM images of the eukaryotic pre-initiation complex) prove to be in excellent agreement with conclusions derived by using traditional approaches, revealing in addition the interdependencies of ligand bindings and structural changes. Copyright © 2015 Elsevier Ltd. All rights reserved.

  1. Cryo-transmission electron microscopy of Ag nanoparticles grown on an ionic liquid substrate

    KAUST Repository

    Anjum, Dalaver H.

    2010-07-01

    We report a novel method of growing silver nanostructures by cathodic sputtering onto an ionic liquid (IL) and our visualization by transmission cryo-electron microscopy to avoid beam-induced motion of the nanoparticles. By freezing the IL suspension and controlling electron dose, we can assess properties of particle size, morphology, crystallinity, and aggregation in situ and at high detail. We observed round silver nanoparticles with a well-defined diameter of 7.0 ± 1.5 nm that are faceted with crystalline cubic structures and ∼80% of the particles have multiply twinned faults. We also applied cryo-electron tomography to investigate the structure of the nanoparticles and to directly visualize the IL wetting around them. In addition to particles, we observed nanorods that appear to have assembled from individual nanoparticles. Reexamination of the samples after 4-5 days from initial preparation showed significant changes in morphology, and potential mechanisms for this are discussed. © 2010 Materials Research Society.

  2. High resolution helium ion scanning microscopy of the rat kidney.

    Directory of Open Access Journals (Sweden)

    William L Rice

    Full Text Available Helium ion scanning microscopy is a novel imaging technology with the potential to provide sub-nanometer resolution images of uncoated biological tissues. So far, however, it has been used mainly in materials science applications. Here, we took advantage of helium ion microscopy to explore the epithelium of the rat kidney with unsurpassed image quality and detail. In addition, we evaluated different tissue preparation methods for their ability to preserve tissue architecture. We found that high contrast, high resolution imaging of the renal tubule surface is possible with a relatively simple processing procedure that consists of transcardial perfusion with aldehyde fixatives, vibratome tissue sectioning, tissue dehydration with graded methanol solutions and careful critical point drying. Coupled with the helium ion system, fine details such as membrane texture and membranous nanoprojections on the glomerular podocytes were visualized, and pores within the filtration slit diaphragm could be seen in much greater detail than in previous scanning EM studies. In the collecting duct, the extensive and striking apical microplicae of the intercalated cells were imaged without the shrunken or distorted appearance that is typical with conventional sample processing and scanning electron microscopy. Membrane depressions visible on principal cells suggest possible endo- or exocytotic events, and central cilia on these cells were imaged with remarkable preservation and clarity. We also demonstrate the use of colloidal gold probes for highlighting specific cell-surface proteins and find that 15 nm gold labels are practical and easily distinguishable, indicating that external labels of various sizes can be used to detect multiple targets in the same tissue. We conclude that this technology represents a technical breakthrough in imaging the topographical ultrastructure of animal tissues. Its use in future studies should allow the study of fine cellular details

  3. Spin-polarized scanning electron microscopy

    International Nuclear Information System (INIS)

    Kohashi, Teruo

    2014-01-01

    Spin-Polarized Scanning Electron Microscopy (Spin SEM) is one way for observing magnetic domain structures taking advantage of the spin polarization of the secondary electrons emitted from a ferromagnetic sample. This principle brings us several excellent capabilities such as high-spatial resolution better than 10 nm, and analysis of magnetization direction in three dimensions. In this paper, the principle and the structure of the spin SEM is briefly introduced, and some examples of the spin SEM measurements are shown. (author)

  4. Direct observation and analysis of yolk-shell materials using low-voltage high-resolution scanning electron microscopy: Nanometal-particles encapsulated in metal-oxide, carbon, and polymer

    Energy Technology Data Exchange (ETDEWEB)

    Asahina, Shunsuke; Suga, Mitsuo; Takahashi, Hideyuki [JEOL Ltd., SM Business Unit, Tokyo (Japan); Young Jeong, Hu [Graduate School of EEWS, WCU/BK21+, KAIST, Daejeon 305-701 (Korea, Republic of); Galeano, Carolina; Schüth, Ferdi [Department of Heterogeneous Catalysis, Max-Planck-Institut für Kohlenforschung, Mülheim (Germany); Terasaki, Osamu, E-mail: terasaki@mmk.su.se, E-mail: terasaki@kaist.ac.kr [Graduate School of EEWS, WCU/BK21+, KAIST, Daejeon 305-701 (Korea, Republic of); Department of Materials and Environmental Chemistry, Berzelii Centre EXSELENT on Porous Materials, Stockholm University, SE-10691 Stockholm (Sweden)

    2014-11-01

    Nanometal particles show characteristic features in chemical and physical properties depending on their sizes and shapes. For keeping and further enhancing their features, the particles should be protected from coalescence or degradation. One approach is to encapsulate the nanometal particles inside pores with chemically inert or functional materials, such as carbon, polymer, and metal oxides, which contain mesopores to allow permeation of only chemicals not the nanometal particles. Recently developed low-voltage high-resolution scanning electron microscopy was applied to the study of structural, chemical, and electron state of both nanometal particles and encapsulating materials in yolk-shell materials of Au@C, Ru/Pt@C, Au@TiO{sub 2}, and Pt@Polymer. Progresses in the following categories were shown for the yolk-shell materials: (i) resolution of topographic image contrast by secondary electrons, of atomic-number contrast by back-scattered electrons, and of elemental mapping by X-ray energy dispersive spectroscopy; (ii) sample preparation for observing internal structures; and (iii) X-ray spectroscopy such as soft X-ray emission spectroscopy. Transmission electron microscopy was also used for characterization of Au@C.

  5. Combining image processing and modeling to generate traces of beta-strands from cryo-EM density images of beta-barrels.

    Science.gov (United States)

    Si, Dong; He, Jing

    2014-01-01

    Electron cryo-microscopy (Cryo-EM) technique produces 3-dimensional (3D) density images of proteins. When resolution of the images is not high enough to resolve the molecular details, it is challenging for image processing methods to enhance the molecular features. β-barrel is a particular structure feature that is formed by multiple β-strands in a barrel shape. There is no existing method to derive β-strands from the 3D image of a β-barrel at medium resolutions. We propose a new method, StrandRoller, to generate a small set of possible β-traces from the density images at medium resolutions of 5-10Å. StrandRoller has been tested using eleven β-barrel images simulated to 10Å resolution and one image isolated from the experimentally derived cryo-EM density image at 6.7Å resolution. StrandRoller was able to detect 81.84% of the β-strands with an overall 1.5Å 2-way distance between the detected and the observed β-traces, if the best of fifteen detections is considered. Our results suggest that it is possible to derive a small set of possible β-traces from the β-barrel cryo-EM image at medium resolutions even when no separation of the β-strands is visible in the images.

  6. The surface topography of the choroid plexus. Environmental, low and high vacuum scanning electron microscopy.

    Science.gov (United States)

    Mestres, Pedro; Pütz, Norbert; Garcia Gómez de Las Heras, Soledad; García Poblete, Eduardo; Morguet, Andrea; Laue, Michael

    2011-05-01

    Environmental scanning electron microscopy (ESEM) allows the examination of hydrated and dried specimens without a conductive metal coating which could be advantageous in the imaging of biological and medical objects. The aim of this study was to assess the performance and benefits of wet-mode and low vacuum ESEM in comparison to high vacuum scanning electron microscopy (SEM) using the choroid plexus of chicken embryos as a model, an organ of the brain involved in the formation of cerebrospinal fluid in vertebrates. Specimens were fixed with or without heavy metals and examined directly or after critical point drying with or without metal coating. For wet mode ESEM freshly excised specimens without any pre-treatment were also examined. Conventional high vacuum SEM revealed the characteristic morphology of the choroid plexus cells at a high resolution and served as reference. With low vacuum ESEM of dried but uncoated samples the structure appeared well preserved but charging was a problem. It could be reduced by a short beam dwell time and averaging of images or by using the backscattered electron detector instead of the gaseous secondary electron detector. However, resolution was lower than with conventional SEM. Wet mode imaging was only possible with tissue that had been stabilized by fixation. Not all surface details (e.g. microvilli) could be visualized and other structures, like the cilia, were deformed. In summary, ESEM is an additional option for the imaging of bio-medical samples but it is problematic with regard to resolution and sample stability during imaging. Copyright © 2011 Elsevier GmbH. All rights reserved.

  7. Massively parallel unsupervised single-particle cryo-EM data clustering via statistical manifold learning.

    Directory of Open Access Journals (Sweden)

    Jiayi Wu

    Full Text Available Structural heterogeneity in single-particle cryo-electron microscopy (cryo-EM data represents a major challenge for high-resolution structure determination. Unsupervised classification may serve as the first step in the assessment of structural heterogeneity. However, traditional algorithms for unsupervised classification, such as K-means clustering and maximum likelihood optimization, may classify images into wrong classes with decreasing signal-to-noise-ratio (SNR in the image data, yet demand increased computational costs. Overcoming these limitations requires further development of clustering algorithms for high-performance cryo-EM data processing. Here we introduce an unsupervised single-particle clustering algorithm derived from a statistical manifold learning framework called generative topographic mapping (GTM. We show that unsupervised GTM clustering improves classification accuracy by about 40% in the absence of input references for data with lower SNRs. Applications to several experimental datasets suggest that our algorithm can detect subtle structural differences among classes via a hierarchical clustering strategy. After code optimization over a high-performance computing (HPC environment, our software implementation was able to generate thousands of reference-free class averages within hours in a massively parallel fashion, which allows a significant improvement on ab initio 3D reconstruction and assists in the computational purification of homogeneous datasets for high-resolution visualization.

  8. Massively parallel unsupervised single-particle cryo-EM data clustering via statistical manifold learning.

    Science.gov (United States)

    Wu, Jiayi; Ma, Yong-Bei; Congdon, Charles; Brett, Bevin; Chen, Shuobing; Xu, Yaofang; Ouyang, Qi; Mao, Youdong

    2017-01-01

    Structural heterogeneity in single-particle cryo-electron microscopy (cryo-EM) data represents a major challenge for high-resolution structure determination. Unsupervised classification may serve as the first step in the assessment of structural heterogeneity. However, traditional algorithms for unsupervised classification, such as K-means clustering and maximum likelihood optimization, may classify images into wrong classes with decreasing signal-to-noise-ratio (SNR) in the image data, yet demand increased computational costs. Overcoming these limitations requires further development of clustering algorithms for high-performance cryo-EM data processing. Here we introduce an unsupervised single-particle clustering algorithm derived from a statistical manifold learning framework called generative topographic mapping (GTM). We show that unsupervised GTM clustering improves classification accuracy by about 40% in the absence of input references for data with lower SNRs. Applications to several experimental datasets suggest that our algorithm can detect subtle structural differences among classes via a hierarchical clustering strategy. After code optimization over a high-performance computing (HPC) environment, our software implementation was able to generate thousands of reference-free class averages within hours in a massively parallel fashion, which allows a significant improvement on ab initio 3D reconstruction and assists in the computational purification of homogeneous datasets for high-resolution visualization.

  9. Molecular dynamics-based refinement and validation for sub-5 Å cryo-electron microscopy maps.

    Science.gov (United States)

    Singharoy, Abhishek; Teo, Ivan; McGreevy, Ryan; Stone, John E; Zhao, Jianhua; Schulten, Klaus

    2016-07-07

    Two structure determination methods, based on the molecular dynamics flexible fitting (MDFF) paradigm, are presented that resolve sub-5 Å cryo-electron microscopy (EM) maps with either single structures or ensembles of such structures. The methods, denoted cascade MDFF and resolution exchange MDFF, sequentially re-refine a search model against a series of maps of progressively higher resolutions, which ends with the original experimental resolution. Application of sequential re-refinement enables MDFF to achieve a radius of convergence of ~25 Å demonstrated with the accurate modeling of β-galactosidase and TRPV1 proteins at 3.2 Å and 3.4 Å resolution, respectively. The MDFF refinements uniquely offer map-model validation and B-factor determination criteria based on the inherent dynamics of the macromolecules studied, captured by means of local root mean square fluctuations. The MDFF tools described are available to researchers through an easy-to-use and cost-effective cloud computing resource on Amazon Web Services.

  10. Gas chromatographic quadrupole time-of-flight full scan high resolution mass spectrometric screening of human urine in antidoping analysis

    NARCIS (Netherlands)

    Abushareeda, Wadha; Lyris, Emmanouil; Kraiem, Suhail; Wahaibi, Aisha Al; Alyazidi, Sameera; Dbes, Najib; Lommen, Arjen; Nielen, Michel; Horvatovich, Peter L.; Alsayrafi, Mohammed; Georgakopoulos, Costas

    2017-01-01

    This paper presents the development and validation of a high-resolution full scan (FS) electron impact ionization (EI) gas chromatography coupled to quadrupole Time-of-Flight mass spectrometry (GC/QTOF) platform for screening anabolic androgenic steroids (AAS) in human urine samples. The World

  11. Imaging endosomes and autophagosomes in whole mammalian cells using correlative cryo-fluorescence and cryo-soft X-ray microscopy (cryo-CLXM)

    International Nuclear Information System (INIS)

    Duke, Elizabeth M.H.; Razi, Minoo; Weston, Anne; Guttmann, Peter; Werner, Stephan; Henzler, Katja; Schneider, Gerd; Tooze, Sharon A.; Collinson, Lucy M.

    2014-01-01

    Cryo-soft X-ray tomography (cryo-SXT) is a powerful imaging technique that can extract ultrastructural information from whole, unstained mammalian cells as close to the living state as possible. Subcellular organelles including the nucleus, the Golgi apparatus and mitochondria have been identified by morphology alone, due to the similarity in contrast to transmission electron micrographs. In this study, we used cryo-SXT to image endosomes and autophagosomes, organelles that are particularly susceptible to chemical fixation artefacts during sample preparation for electron microscopy. We used two approaches to identify these compartments. For early and recycling endosomes, which are accessible to externally-loaded markers, we used an anti-transferrin receptor antibody conjugated to 10 nm gold particles. For autophagosomes, which are not accessible to externally-applied markers, we developed a correlative cryo-fluorescence and cryo-SXT workflow (cryo-CLXM) to localise GFP-LC3 and RFP-Atg9. We used a stand-alone cryo-fluorescence stage in the home laboratory to localise the cloned fluorophores, followed by cryo-soft X-ray tomography at the synchrotron to analyse cellular ultrastructure. We mapped the 3D ultrastructure of the endocytic and autophagic structures, and discovered clusters of omegasomes arising from ‘hotspots’ on the ER. Thus, immunogold markers and cryo-CLXM can be used to analyse cellular processes that are inaccessible using other imaging modalities. - Highlights: • We image whole, unstained mammalian cells using cryo-soft X-ray tomography. • Endosomes are identified using a gold marker for the transferrin receptor. • A new workflow for correlative cryo-fluorescence and cryo-SXT is used to locate early autophagosomes. • Interactions between endosomes, endoplasmic reticulum and forming autophagosomes are mapped in 3D. • Multiple omegasomes are shown to form at ‘hotspots’ on the endoplasmic reticulum

  12. Gas chromatographic quadrupole time-of-flight full scan high resolution mass spectrometric screening of human urine in antidoping analysis

    NARCIS (Netherlands)

    Abushareeda, Wadha; Lyris, Emmanouil; Kraiem, Suhail; Wahaibi, Aisha Al; Alyazidi, Sameera; Dbes, Najib; Lommen, Arjen; Nielen, Michel; Horvatovich, Peter L.; Alsayrafi, Mohammed; Georgakopoulos, Costas

    2017-01-01

    This paper presents the development and validation of a high-resolution full scan (FS) electron impact ionization (EI) gas chromatography coupled to quadrupole Time-of-Flight mass spectrometry (GC/QTOF) platform for screening anabolic androgenic steroids (AAS) in human urine samples. The World

  13. Automated transmission-mode scanning electron microscopy (tSEM for large volume analysis at nanoscale resolution.

    Directory of Open Access Journals (Sweden)

    Masaaki Kuwajima

    Full Text Available Transmission-mode scanning electron microscopy (tSEM on a field emission SEM platform was developed for efficient and cost-effective imaging of circuit-scale volumes from brain at nanoscale resolution. Image area was maximized while optimizing the resolution and dynamic range necessary for discriminating key subcellular structures, such as small axonal, dendritic and glial processes, synapses, smooth endoplasmic reticulum, vesicles, microtubules, polyribosomes, and endosomes which are critical for neuronal function. Individual image fields from the tSEM system were up to 4,295 µm(2 (65.54 µm per side at 2 nm pixel size, contrasting with image fields from a modern transmission electron microscope (TEM system, which were only 66.59 µm(2 (8.160 µm per side at the same pixel size. The tSEM produced outstanding images and had reduced distortion and drift relative to TEM. Automated stage and scan control in tSEM easily provided unattended serial section imaging and montaging. Lens and scan properties on both TEM and SEM platforms revealed no significant nonlinear distortions within a central field of ∼100 µm(2 and produced near-perfect image registration across serial sections using the computational elastic alignment tool in Fiji/TrakEM2 software, and reliable geometric measurements from RECONSTRUCT™ or Fiji/TrakEM2 software. Axial resolution limits the analysis of small structures contained within a section (∼45 nm. Since this new tSEM is non-destructive, objects within a section can be explored at finer axial resolution in TEM tomography with current methods. Future development of tSEM tomography promises thinner axial resolution producing nearly isotropic voxels and should provide within-section analyses of structures without changing platforms. Brain was the test system given our interest in synaptic connectivity and plasticity; however, the new tSEM system is readily applicable to other biological systems.

  14. Direct observation and analysis of york-shell materials using low-voltage high-resolution scanning electron microscopy: Nanometal-particles encapsulated in metal-oxide, carbon, and polymer

    Directory of Open Access Journals (Sweden)

    Shunsuke Asahina

    2014-11-01

    Full Text Available Nanometal particles show characteristic features in chemical and physical properties depending on their sizes and shapes. For keeping and further enhancing their features, the particles should be protected from coalescence or degradation. One approach is to encapsulate the nanometal particles inside pores with chemically inert or functional materials, such as carbon, polymer, and metal oxides, which contain mesopores to allow permeation of only chemicals not the nanometal particles. Recently developed low-voltage high-resolution scanning electron microscopy was applied to the study of structural, chemical, and electron state of both nanometal particles and encapsulating materials in york-shell materials of Au@C, Ru/Pt@C, Au@TiO2, and Pt@Polymer. Progresses in the following categories were shown for the york-shell materials: (i resolution of topographic image contrast by secondary electrons, of atomic-number contrast by back-scattered electrons, and of elemental mapping by X-ray energy dispersive spectroscopy; (ii sample preparation for observing internal structures; and (iii X-ray spectroscopy such as soft X-ray emission spectroscopy. Transmission electron microscopy was also used for characterization of Au@C.

  15. High resolution, high speed ultrahigh vacuum microscopy

    International Nuclear Information System (INIS)

    Poppa, Helmut

    2004-01-01

    The history and future of transmission electron microscopy (TEM) is discussed as it refers to the eventual development of instruments and techniques applicable to the real time in situ investigation of surface processes with high resolution. To reach this objective, it was necessary to transform conventional high resolution instruments so that an ultrahigh vacuum (UHV) environment at the sample site was created, that access to the sample by various in situ sample modification procedures was provided, and that in situ sample exchanges with other integrated surface analytical systems became possible. Furthermore, high resolution image acquisition systems had to be developed to take advantage of the high speed imaging capabilities of projection imaging microscopes. These changes to conventional electron microscopy and its uses were slowly realized in a few international laboratories over a period of almost 40 years by a relatively small number of researchers crucially interested in advancing the state of the art of electron microscopy and its applications to diverse areas of interest; often concentrating on the nucleation, growth, and properties of thin films on well defined material surfaces. A part of this review is dedicated to the recognition of the major contributions to surface and thin film science by these pioneers. Finally, some of the important current developments in aberration corrected electron optics and eventual adaptations to in situ UHV microscopy are discussed. As a result of all the path breaking developments that have led to today's highly sophisticated UHV-TEM systems, integrated fundamental studies are now possible that combine many traditional surface science approaches. Combined investigations to date have involved in situ and ex situ surface microscopies such as scanning tunneling microscopy/atomic force microscopy, scanning Auger microscopy, and photoemission electron microscopy, and area-integrating techniques such as x-ray photoelectron

  16. Evaluation of gas chromatography – electron ionization – full scan high resolution Orbitrap mass spectrometry for pesticide residue analysis

    International Nuclear Information System (INIS)

    Mol, Hans G.J.; Tienstra, Marc; Zomer, Paul

    2016-01-01

    Gas chromatography with electron ionization and full scan high resolution mass spectrometry with an Orbitrap mass analyzer (GC-EI-full scan Orbitrap HRMS) was evaluated for residue analysis. Pesticides in fruit and vegetables were taken as an example application. The relevant aspects for GC-MS based residue analysis, including the resolving power (15,000 to 120,000 FWHM at m/z 200), scan rate, dynamic range, selectivity, sensitivity, analyte identification, and utility of existing EI-libraries, are assessed and discussed in detail. The optimum acquisition conditions in full scan mode (m/z 50–500) were a resolving power of 60,000 and an automatic-gain-control target value of 3E6. These conditions provided (i) an optimum mass accuracy: within 2 ppm over a wide concentration range, with/without matrix, enabling the use of ±5 ppm mass extraction windows (ii) adequate scan speed: minimum 12 scans/peak, (iii) an intra-scan dynamic range sufficient to achieve LOD/LOQs ≤0.5 pg in fruit/vegetable matrices (corresponding to ≤0.5 μg kg"−"1) for most pesticides. EI-Orbitrap spectra were consistent over a very wide concentration range (5 orders) with good match values against NIST (EI-quadrupole) spectra. The applicability for quantitative residue analysis was verified by validation of 54 pesticides in three matrices (tomato, leek, orange) at 10 and 50 μg/kg. The method involved a QuEChERS-based extraction with a solvent switch into iso-octane, and 1 μL hot splitless injection into the GC-HRMS system. A recovery between 70 and 120% and a repeatability RSD <10% was obtained in most cases. Linearity was demonstrated for the range ≤5–250 μg kg"−"1. The pesticides could be identified according to the applicable EU criteria for GC-HRMS (SANTE/11945/2015). GC-EI-full scan Orbitrap HRMS was found to be highly suited for quantitative pesticide residue analysis. The potential of qualitative screening to extend the scope makes it an attractive alternative to

  17. Evaluation of gas chromatography – electron ionization – full scan high resolution Orbitrap mass spectrometry for pesticide residue analysis

    Energy Technology Data Exchange (ETDEWEB)

    Mol, Hans G.J., E-mail: hans.mol@wur.nl; Tienstra, Marc; Zomer, Paul

    2016-09-07

    Gas chromatography with electron ionization and full scan high resolution mass spectrometry with an Orbitrap mass analyzer (GC-EI-full scan Orbitrap HRMS) was evaluated for residue analysis. Pesticides in fruit and vegetables were taken as an example application. The relevant aspects for GC-MS based residue analysis, including the resolving power (15,000 to 120,000 FWHM at m/z 200), scan rate, dynamic range, selectivity, sensitivity, analyte identification, and utility of existing EI-libraries, are assessed and discussed in detail. The optimum acquisition conditions in full scan mode (m/z 50–500) were a resolving power of 60,000 and an automatic-gain-control target value of 3E6. These conditions provided (i) an optimum mass accuracy: within 2 ppm over a wide concentration range, with/without matrix, enabling the use of ±5 ppm mass extraction windows (ii) adequate scan speed: minimum 12 scans/peak, (iii) an intra-scan dynamic range sufficient to achieve LOD/LOQs ≤0.5 pg in fruit/vegetable matrices (corresponding to ≤0.5 μg kg{sup −1}) for most pesticides. EI-Orbitrap spectra were consistent over a very wide concentration range (5 orders) with good match values against NIST (EI-quadrupole) spectra. The applicability for quantitative residue analysis was verified by validation of 54 pesticides in three matrices (tomato, leek, orange) at 10 and 50 μg/kg. The method involved a QuEChERS-based extraction with a solvent switch into iso-octane, and 1 μL hot splitless injection into the GC-HRMS system. A recovery between 70 and 120% and a repeatability RSD <10% was obtained in most cases. Linearity was demonstrated for the range ≤5–250 μg kg{sup −1}. The pesticides could be identified according to the applicable EU criteria for GC-HRMS (SANTE/11945/2015). GC-EI-full scan Orbitrap HRMS was found to be highly suited for quantitative pesticide residue analysis. The potential of qualitative screening to extend the scope makes it an attractive

  18. Atomic Resolution Structures of Human Bufaviruses Determined by Cryo-Electron Microscopy

    Directory of Open Access Journals (Sweden)

    Maria Ilyas

    2018-01-01

    Full Text Available Bufavirus strain 1 (BuV1, a member of the Protoparvovirus genus of the Parvoviridae, was first isolated from fecal samples of children with acute diarrhea in Burkina Faso. Since this initial discovery, BuVs have been isolated in several countries, including Finland, the Netherlands, and Bhutan, in pediatric patients exhibiting similar symptoms. Towards their characterization, the structures of virus-like particles of BuV1, BuV2, and BuV3, the current known genotypes, have been determined by cryo-electron microscopy and image reconstruction to 2.84, 3.79, and 3.25 Å, respectively. The BuVs, 65–73% identical in amino acid sequence, conserve the major viral protein, VP2, structure and general capsid surface features of parvoviruses. These include a core β-barrel (βB-βI, α-helix A, and large surface loops inserted between these elements in VP2. The capsid contains depressions at the icosahedral 2-fold and around the 5-fold axes, and has three separated protrusions surrounding the 3-fold axes. Structure comparison among the BuVs and to available parvovirus structures revealed capsid surface variations and capsid 3-fold protrusions that depart from the single pinwheel arrangement of the animal protoparvoviruses. These structures provide a platform to begin the molecular characterization of these potentially pathogenic viruses.

  19. Electron beam excitation assisted optical microscope with ultra-high resolution.

    Science.gov (United States)

    Inami, Wataru; Nakajima, Kentaro; Miyakawa, Atsuo; Kawata, Yoshimasa

    2010-06-07

    We propose electron beam excitation assisted optical microscope, and demonstrated its resolution higher than 50 nm. In the microscope, a light source in a few nanometers size is excited by focused electron beam in a luminescent film. The microscope makes it possible to observe dynamic behavior of living biological specimens in various surroundings, such as air or liquids. Scan speed of the nanometric light source is faster than that in conventional near-field scanning optical microscopes. The microscope enables to observe optical constants such as absorption, refractive index, polarization, and their dynamic behavior on a nanometric scale. The microscope opens new microscopy applications in nano-technology and nano-science.

  20. Cryo-EM structure of Hepatitis C virus IRES bound to the human ribosome at 3.9-Å resolution.

    Science.gov (United States)

    Quade, Nick; Boehringer, Daniel; Leibundgut, Marc; van den Heuvel, Joop; Ban, Nenad

    2015-07-08

    Hepatitis C virus (HCV), a widespread human pathogen, is dependent on a highly structured 5'-untranslated region of its mRNA, referred to as internal ribosome entry site (IRES), for the translation of all of its proteins. The HCV IRES initiates translation by directly binding to the small ribosomal subunit (40S), circumventing the need for many eukaryotic translation initiation factors required for mRNA scanning. Here we present the cryo-EM structure of the human 40S ribosomal subunit in complex with the HCV IRES at 3.9 Å resolution, determined by focused refinement of an 80S ribosome-HCV IRES complex. The structure reveals the molecular details of the interactions between the IRES and the 40S, showing that expansion segment 7 (ES7) of the 18S rRNA acts as a central anchor point for the HCV IRES. The structural data rationalizes previous biochemical and genetic evidence regarding the initiation mechanism of the HCV and other related IRESs.

  1. High-resolution photoluminescence electro-modulation microscopy by scanning lock-in

    Science.gov (United States)

    Koopman, W.; Muccini, M.; Toffanin, S.

    2018-04-01

    Morphological inhomogeneities and structural defects in organic semiconductors crucially determine the charge accumulation and lateral transport in organic thin-film transistors. Photoluminescence Electro-Modulation (PLEM) microscopy is a laser-scanning microscopy technique that relies on the modulation of the thin-film fluorescence in the presence of charge-carriers to image the spatial distribution of charges within the active organic semiconductor. Here, we present a lock-in scheme based on a scanning beam approach for increasing the PLEM microscopy resolution and contrast. The charge density in the device is modulated by a sinusoidal electrical signal, phase-locked to the scanning beam of the excitation laser. The lock-in detection scheme is achieved by acquiring a series of images with different phases between the beam scan and the electrical modulation. Application of high resolution PLEM to an organic transistor in accumulation mode demonstrates its potential to image local variations in the charge accumulation. A diffraction-limited precision of sub-300 nm and a signal to noise ratio of 21.4 dB could be achieved.

  2. Transmission electron microscopy of amyloid fibrils.

    Science.gov (United States)

    Gras, Sally L; Waddington, Lynne J; Goldie, Kenneth N

    2011-01-01

    Transmission Electron Microscopy of negatively stained and cryo-prepared specimens allows amyloid fibrils to be visualised at high resolution in a dried or a hydrated state, and is an essential method for characterising the morphology of fibrils and pre-fibrillar species. We outline the key steps involved in the preparation and observation of samples using negative staining and cryo-electron preservation. We also discuss methods to measure fibril characteristics, such as fibril width, from electron micrographs.

  3. High resolution wind turbine wake measurements with a scanning lidar

    DEFF Research Database (Denmark)

    Herges, T. G.; Maniaci, D. C.; Naughton, B. T.

    2017-01-01

    High-resolution lidar wake measurements are part of an ongoing field campaign being conducted at the Scaled Wind Farm Technology facility by Sandia National Laboratories and the National Renewable Energy Laboratory using a customized scanning lidar from the Technical University of Denmark. One...

  4. Magnetic lens apparatus for a low-voltage high-resolution electron microscope

    Science.gov (United States)

    Crewe, Albert V.

    1996-01-01

    A lens apparatus in which a beam of charged particles of low accelerating voltage is brought to a focus by a magnetic field, the lens being situated behind the target position. The lens comprises an electrically-conducting coil arranged around the axis of the beam and a magnetic pole piece extending along the axis of the beam at least within the space surrounded by the coil. The lens apparatus comprises the sole focusing lens for high-resolution imaging in a low-voltage scanning electron microscope.

  5. Technical aspects: development, manufacture and installation of a cryo-cooled HTS coil system for high-resolution in-vivo imaging of the mouse at 1.5 T.

    Science.gov (United States)

    Ginefri, Jean-Christophe; Poirier-Quinot, Marie; Girard, Olivier; Darrasse, Luc

    2007-09-01

    Signal-to-noise ratio improvement is of major importance to achieve microscopic spatial resolution in magnetic resonance experiments. Magnetic resonance imaging of small animals is particularly concerned since it typically requires voxels of less than (100 microm)(3) to observe the small anatomical structures having size reduction by a factor of more than 10 as compared to human being. The signal-to-noise ratio can be increased by working at high static magnetic field strengths, but the biomedical interest of such high-field systems may be limited due to field-dependent contrast mechanisms and severe technological difficulties. An alternative approach that allows working in clinical imaging system is to improve the sensitivity of the radio-frequency receiver coil. This can be done using small cryogenically operated coils made either of copper or high-temperature superconducting material. We report the technological development of cryo-cooled superconducting coils for high-resolution imaging in a whole-body magnetic resonance scanner operating at 1.5 T. The technological background supporting this development is first addressed, including HTS coil design, simulation tools, cryogenic mean description and electrical characterization procedure. To illustrate the performances of superconducting coils for magnetic resonance imaging at intermediate field strength, in-vivo mouse images of various anatomic sites acquired with a 12 mm diameter cryo-cooled superconducting coil are presented.

  6. High Resolution Scanning Ion Microscopy

    NARCIS (Netherlands)

    Castaldo, V.

    2011-01-01

    The structure of the thesis is the following. The first chapter is an introduction to scanning microscopy, where the path that led to the Focused Ion Beam (FIB) is described and the main differences between electrons and ion beams are highlighted. Chapter 2 is what is normally referred to (which I

  7. Imaging system for creating 3D block-face cryo-images of whole mice

    Science.gov (United States)

    Roy, Debashish; Breen, Michael; Salvado, Olivier; Heinzel, Meredith; McKinley, Eliot; Wilson, David

    2006-03-01

    We developed a cryomicrotome/imaging system that provides high resolution, high sensitivity block-face images of whole mice or excised organs, and applied it to a variety of biological applications. With this cryo-imaging system, we sectioned cryo-preserved tissues at 2-40 μm thickness and acquired high resolution brightfield and fluorescence images with microscopic in-plane resolution (as good as 1.2 μm). Brightfield images of normal and pathological anatomy show exquisite detail, especially in the abdominal cavity. Multi-planar reformatting and 3D renderings allow one to interrogate 3D structures. In this report, we present brightfield images of mouse anatomy, as well as 3D renderings of organs. For BPK mice model of polycystic kidney disease, we compared brightfield cryo-images and kidney volumes to MRI. The color images provided greater contrast and resolution of cysts as compared to in vivo MRI. We note that color cryo-images are closer to what a researcher sees in dissection, making it easier for them to interpret image data. The combination of field of view, depth of field, ultra high resolution and color/fluorescence contrast enables cryo-image volumes to provide details that cannot be found through in vivo imaging or other ex vivo optical imaging approaches. We believe that this novel imaging system will have applications that include identification of mouse phenotypes, characterization of diseases like blood vessel disease, kidney disease, and cancer, assessment of drug and gene therapy delivery and efficacy and validation of other imaging modalities.

  8. Calibration And Validation Of CryoSat-2 Low Resolution Mode Data

    Science.gov (United States)

    Naeije, M.; Schrama, E.; Scharroo, R.

    2011-02-01

    Running ahead of the continuously growing need for operational use of sea level products, TUDelft started off the Radar Altimeter Database System RADS many years ago. This system attends to a global international sea- level service. It supports, on one hand, science, like studies on ocean circulation, El Nio, sea level change, and ice topography, and on the other hand (offshore) operations, like delivery of ocean current information, wind and wave statistics, ice detection and ice classification. At present, the database is used by a large scientific community throughout the world, and is daily maintained and developed by Altimetrics LLC, TUDelft and NOAA. It contains all historic altimeter data, and now has to be up- dated with the data from ESAs ice mission CryoSat-2, which was launched successfully in April 2010. These new data are important to augment the data set and by that to improve the estimates of sea level change and its contributors. For this the data have to be validated and calibrated, necessary corrections added and improved (including modelling of corrections that are not directly available from the CryoSat-2 platform), and the orbit ac- curacy verified and if possible the orbits brushed up. Subsequently, value-added ocean and ice products need to be developed in synergy with all the other satellite altimeter data. During the commissioning phase we primarily looked at the sanity of the available level-1b and level-2 Low Resolution Mode (LRM) data. Here, for the 2011 CryoSat Validation Workshop, we present the results of our calibration and validation of LRM L2 data by internal comparison of CryoSat-2 and external comparison with other satellites. We have established a range bias of 3.77 (measurement range too long) and a timing bias of 8.2ms (measurement range too late).

  9. Observation of Magnetic Induction Distribution by Scanning Interference Electron Microscopy

    Science.gov (United States)

    Takahashi, Yoshio; Yajima, Yusuke; Ichikawa, Masakazu; Kuroda, Katsuhiro

    1994-09-01

    A scanning interference electron microscope (SIEM) capable of observing magnetic induction distribution with high sensitivity and spatial resolution has been developed. The SIEM uses a pair of fine coherent scanning probes and detects their relative phase change by magnetic induction, giving raster images of microscopic magnetic distributions. Its performance has been demonstrated by observing magnetic induction distributed near the edge of a recorded magnetic storage medium. Obtained images are compared with corresponding images taken in the scanning Lorentz electron microscope mode using the same microscope, and the differences between them are discussed.

  10. Cryo-electron tomography investigation of serum albumin-camouflaged tobacco mosaic virus nanoparticles.

    Science.gov (United States)

    Gulati, Neetu M; Pitek, Andrzej S; Steinmetz, Nicole F; Stewart, Phoebe L

    2017-03-09

    Nanoparticles offer great potential in drug delivery and imaging, but shielding strategies are necessary to increase circulation time and performance. Structure-function studies are required to define the design rules to achieve effective shielding. With several formulations reaching clinical testing and approval, the ability to assess and detail nanoparticle formulations at the single particle level is becoming increasingly important. To address this need, we use cryo-electron tomography (cryo-ET) to investigate stealth-coated nanoparticles. As a model system, we studied the soft matter nanotubes formed by tobacco mosaic virus (TMV) coated with human serum albumin (SA) stealth proteins. Cryo-ET and subtomogram averaging allow for visualization of individual SA molecules and determination of their orientations relative to the TMV surface, and also for measurement of the surface coverage provided by added stealth proteins. This information fills a critical gap in the understanding of the structural morphology of stealth-coated nanoparticles, and therefore cryo-ET may play an important role in guiding the development of future nanoparticle-based therapeutics.

  11. Real-time underwater object detection based on an electrically scanned high-resolution sonar

    DEFF Research Database (Denmark)

    Henriksen, Lars

    1994-01-01

    The paper describes an approach to real time detection and tracking of underwater objects, using image sequences from an electrically scanned high-resolution sonar. The use of a high resolution sonar provides a good estimate of the location of the objects, but strains the computers on board, beca...

  12. Force scanning: a rapid, high-resolution approach for spatial mechanical property mapping

    International Nuclear Information System (INIS)

    Darling, E M

    2011-01-01

    Atomic force microscopy (AFM) can be used to co-localize mechanical properties and topographical features through property mapping techniques. The most common approach for testing biological materials at the microscale and nanoscale is force mapping, which involves taking individual force curves at discrete sites across a region of interest. The limitations of force mapping include long testing times and low resolution. While newer AFM methodologies, like modulated scanning and torsional oscillation, circumvent this problem, their adoption for biological materials has been limited. This could be due to their need for specialized software algorithms and/or hardware. The objective of this study is to develop a novel force scanning technique using AFM to rapidly capture high-resolution topographical images of soft biological materials while simultaneously quantifying their mechanical properties. Force scanning is a straightforward methodology applicable to a wide range of materials and testing environments, requiring no special modification to standard AFMs. Essentially, if a contact-mode image can be acquired, then force scanning can be used to produce a spatial modulus map. The current study first validates this technique using agarose gels, comparing results to ones achieved by the standard force mapping approach. Biologically relevant demonstrations are then presented for high-resolution modulus mapping of individual cells, cell-cell interfaces, and articular cartilage tissue.

  13. DeepPicker: A deep learning approach for fully automated particle picking in cryo-EM.

    Science.gov (United States)

    Wang, Feng; Gong, Huichao; Liu, Gaochao; Li, Meijing; Yan, Chuangye; Xia, Tian; Li, Xueming; Zeng, Jianyang

    2016-09-01

    Particle picking is a time-consuming step in single-particle analysis and often requires significant interventions from users, which has become a bottleneck for future automated electron cryo-microscopy (cryo-EM). Here we report a deep learning framework, called DeepPicker, to address this problem and fill the current gaps toward a fully automated cryo-EM pipeline. DeepPicker employs a novel cross-molecule training strategy to capture common features of particles from previously-analyzed micrographs, and thus does not require any human intervention during particle picking. Tests on the recently-published cryo-EM data of three complexes have demonstrated that our deep learning based scheme can successfully accomplish the human-level particle picking process and identify a sufficient number of particles that are comparable to those picked manually by human experts. These results indicate that DeepPicker can provide a practically useful tool to significantly reduce the time and manual effort spent in single-particle analysis and thus greatly facilitate high-resolution cryo-EM structure determination. DeepPicker is released as an open-source program, which can be downloaded from https://github.com/nejyeah/DeepPicker-python. Copyright © 2016 Elsevier Inc. All rights reserved.

  14. Austenitic Reversion of Cryo-rolled Ti-Stabilized Austenitic Stainless Steel: High-Resolution EBSD Investigation

    Science.gov (United States)

    Tiamiyu, A. A.; Odeshi, A. G.; Szpunar, J. A.

    2018-02-01

    In this study, AISI 321 austenitic stainless steel (ASS) was cryo-rolled and subsequently annealed at 650 and 800 °C to reverse BCC α'-martensite to FCC γ-austenite. The texture evolution associated with the reversion at the selected temperatures was investigated using high-resolution EBSD. After the reversion, TiC precipitates were observed to be more stable in 650 °C-annealed specimens than those reversed at 800 °C. {110} texture was mainly developed in specimens subjected to both annealing temperatures. However, specimens reversed at 650 °C have stronger texture than those annealed at 800 °C, even at the higher annealing time. The strong intensity of {110} texture component is attributed to the ability of AISI 321 ASS to memorize the crystallographic orientation of the deformed austenite, a phenomenon termed texture memory. The development of weaker texture in 800 °C-annealed specimens is attributed to the residual strain relief in grains, dissolution of grain boundary precipitates, and an increase in atomic migration along the grain boundaries. Based on the observed features of the reversed austenite grains and estimation from an existing model, it is suspected that the austenite reversion at 650 and 800 °C undergone diffusional and martensitic shear reversion, respectively.

  15. Emission sources in scanning electron microscopy

    International Nuclear Information System (INIS)

    Malkusch, W.

    1990-01-01

    Since the beginning of the commercial scanning electron microscopy, there are two kinds of emission sources generally used for generation of the electron beam. The first group covers the cathodes heated directly and indirectly (tungsten hair-needle cathodes and lanthanum hexaboride single crystals, LaB 6 cathode). The other group is the field emission cathodes. The advantages of the thermal sources are their low vacuum requirement and their high beam current which is necessary for the application of microanalysis units. Disadvantages are the short life and the low resolution. Advantages of the field emission cathode unambiguously are the possibilities of the very high resolution, especially in the case of low acceleration voltages. Disadvantages are the necessary ultra-high vacuum and the low beam current. An alternative source is the thermally induced ZrO/W field emission cathode which works stably as compared to the cold field emission and does not need periodic flashing for emitter tip cleaning. (orig.) [de

  16. Structure of the TRPV1 ion channel determined by electron cryo-microscopy.

    Science.gov (United States)

    Liao, Maofu; Cao, Erhu; Julius, David; Cheng, Yifan

    2013-12-05

    Transient receptor potential (TRP) channels are sensors for a wide range of cellular and environmental signals, but elucidating how these channels respond to physical and chemical stimuli has been hampered by a lack of detailed structural information. Here we exploit advances in electron cryo-microscopy to determine the structure of a mammalian TRP channel, TRPV1, at 3.4 Å resolution, breaking the side-chain resolution barrier for membrane proteins without crystallization. Like voltage-gated channels, TRPV1 exhibits four-fold symmetry around a central ion pathway formed by transmembrane segments 5-6 (S5-S6) and the intervening pore loop, which is flanked by S1-S4 voltage-sensor-like domains. TRPV1 has a wide extracellular 'mouth' with a short selectivity filter. The conserved 'TRP domain' interacts with the S4-S5 linker, consistent with its contribution to allosteric modulation. Subunit organization is facilitated by interactions among cytoplasmic domains, including amino-terminal ankyrin repeats. These observations provide a structural blueprint for understanding unique aspects of TRP channel function.

  17. Fine structure of granal thylakoid membrane organization using cryo electron tomography

    NARCIS (Netherlands)

    Kouril, Roman; Oostergetel, Gert T.; Boekema, Egbert J.

    The architecture of grana membranes from spinach chloroplasts was studied by cryo electron tomography. Tomographic reconstructions of ice-embedded isolated grana stacks enabled to resolve features of photosystem II (PSII) in the native membrane and to assign the absolute orientation of individual

  18. High-resolution monochromated electron energy-loss spectroscopy of organic photovoltaic materials.

    Science.gov (United States)

    Alexander, Jessica A; Scheltens, Frank J; Drummy, Lawrence F; Durstock, Michael F; Hage, Fredrik S; Ramasse, Quentin M; McComb, David W

    2017-09-01

    Advances in electron monochromator technology are providing opportunities for high energy resolution (10 - 200meV) electron energy-loss spectroscopy (EELS) to be performed in the scanning transmission electron microscope (STEM). The energy-loss near-edge structure in core-loss spectroscopy is often limited by core-hole lifetimes rather than the energy spread of the incident illumination. However, in the valence-loss region, the reduced width of the zero loss peak makes it possible to resolve clearly and unambiguously spectral features at very low energy-losses (photovoltaics (OPVs): poly(3-hexlythiophene) (P3HT), [6,6] phenyl-C 61 butyric acid methyl ester (PCBM), copper phthalocyanine (CuPc), and fullerene (C 60 ). Data was collected on two different monochromated instruments - a Nion UltraSTEM 100 MC 'HERMES' and a FEI Titan 3 60-300 Image-Corrected S/TEM - using energy resolutions (as defined by the zero loss peak full-width at half-maximum) of 35meV and 175meV, respectively. The data was acquired to allow deconvolution of plural scattering, and Kramers-Kronig analysis was utilized to extract the complex dielectric functions. The real and imaginary parts of the complex dielectric functions obtained from the two instruments were compared to evaluate if the enhanced resolution in the Nion provides new opto-electronic information for these organic materials. The differences between the spectra are discussed, and the implications for STEM-EELS studies of advanced materials are considered. Copyright © 2017 Elsevier B.V. All rights reserved.

  19. Phase contrast scanning transmission electron microscopy imaging of light and heavy atoms at the limit of contrast and resolution.

    Science.gov (United States)

    Yücelen, Emrah; Lazić, Ivan; Bosch, Eric G T

    2018-02-08

    Using state of the art scanning transmission electron microscopy (STEM) it is nowadays possible to directly image single atomic columns at sub-Å resolution. In standard (high angle) annular dark field STEM ((HA)ADF-STEM), however, light elements are usually invisible when imaged together with heavier elements in one image. Here we demonstrate the capability of the recently introduced Integrated Differential Phase Contrast STEM (iDPC-STEM) technique to image both light and heavy atoms in a thin sample at sub-Å resolution. We use the technique to resolve both the Gallium and Nitrogen dumbbells in a GaN crystal in [[Formula: see text

  20. High-Resolution Electronics: Spontaneous Patterning of High-Resolution Electronics via Parallel Vacuum Ultraviolet (Adv. Mater. 31/2016).

    Science.gov (United States)

    Liu, Xuying; Kanehara, Masayuki; Liu, Chuan; Sakamoto, Kenji; Yasuda, Takeshi; Takeya, Jun; Minari, Takeo

    2016-08-01

    On page 6568, T. Minari and co-workers describe spontaneous patterning based on the parallel vacuum ultraviolet (PVUV) technique, enabling the homogeneous integration of complex, high-resolution electronic circuits, even on large-scale, flexible, transparent substrates. Irradiation of PVUV to the hydrophobic polymer surface precisely renders the selected surface into highly wettable regions with sharply defined boundaries, which spontaneously guides a metal nanoparticle ink into a series of circuit lines and gaps with the widths down to a resolution of 1 μm. © 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  1. Correlating Fluorescence and High-Resolution Scanning Electron Microscopy (HRSEM) for the study of GABAA receptor clustering induced by inhibitory synaptic plasticity

    KAUST Repository

    Orlando, Marta

    2017-10-17

    Both excitatory and inhibitory synaptic contacts display activity dependent dynamic changes in their efficacy that are globally termed synaptic plasticity. Although the molecular mechanisms underlying glutamatergic synaptic plasticity have been extensively investigated and described, those responsible for inhibitory synaptic plasticity are only beginning to be unveiled. In this framework, the ultrastructural changes of the inhibitory synapses during plasticity have been poorly investigated. Here we combined confocal fluorescence microscopy (CFM) with high resolution scanning electron microscopy (HRSEM) to characterize the fine structural rearrangements of post-synaptic GABAA Receptors (GABAARs) at the nanometric scale during the induction of inhibitory long-term potentiation (iLTP). Additional electron tomography (ET) experiments on immunolabelled hippocampal neurons allowed the visualization of synaptic contacts and confirmed the reorganization of post-synaptic GABAAR clusters in response to chemical iLTP inducing protocol. Altogether, these approaches revealed that, following the induction of inhibitory synaptic potentiation, GABAAR clusters increase in size and number at the post-synaptic membrane with no other major structural changes of the pre- and post-synaptic elements.

  2. Correlating Fluorescence and High-Resolution Scanning Electron Microscopy (HRSEM) for the study of GABAA receptor clustering induced by inhibitory synaptic plasticity

    KAUST Repository

    Orlando, Marta; Ravasenga, Tiziana; Petrini, Enrica Maria; Falqui, Andrea; Marotta, Roberto; Barberis, Andrea

    2017-01-01

    Both excitatory and inhibitory synaptic contacts display activity dependent dynamic changes in their efficacy that are globally termed synaptic plasticity. Although the molecular mechanisms underlying glutamatergic synaptic plasticity have been extensively investigated and described, those responsible for inhibitory synaptic plasticity are only beginning to be unveiled. In this framework, the ultrastructural changes of the inhibitory synapses during plasticity have been poorly investigated. Here we combined confocal fluorescence microscopy (CFM) with high resolution scanning electron microscopy (HRSEM) to characterize the fine structural rearrangements of post-synaptic GABAA Receptors (GABAARs) at the nanometric scale during the induction of inhibitory long-term potentiation (iLTP). Additional electron tomography (ET) experiments on immunolabelled hippocampal neurons allowed the visualization of synaptic contacts and confirmed the reorganization of post-synaptic GABAAR clusters in response to chemical iLTP inducing protocol. Altogether, these approaches revealed that, following the induction of inhibitory synaptic potentiation, GABAAR clusters increase in size and number at the post-synaptic membrane with no other major structural changes of the pre- and post-synaptic elements.

  3. Correlating Fluorescence and High-Resolution Scanning Electron Microscopy (HRSEM) for the study of GABAA receptor clustering induced by inhibitory synaptic plasticity.

    Science.gov (United States)

    Orlando, Marta; Ravasenga, Tiziana; Petrini, Enrica Maria; Falqui, Andrea; Marotta, Roberto; Barberis, Andrea

    2017-10-23

    Both excitatory and inhibitory synaptic contacts display activity dependent dynamic changes in their efficacy that are globally termed synaptic plasticity. Although the molecular mechanisms underlying glutamatergic synaptic plasticity have been extensively investigated and described, those responsible for inhibitory synaptic plasticity are only beginning to be unveiled. In this framework, the ultrastructural changes of the inhibitory synapses during plasticity have been poorly investigated. Here we combined confocal fluorescence microscopy (CFM) with high resolution scanning electron microscopy (HRSEM) to characterize the fine structural rearrangements of post-synaptic GABA A Receptors (GABA A Rs) at the nanometric scale during the induction of inhibitory long-term potentiation (iLTP). Additional electron tomography (ET) experiments on immunolabelled hippocampal neurons allowed the visualization of synaptic contacts and confirmed the reorganization of post-synaptic GABA A R clusters in response to chemical iLTP inducing protocol. Altogether, these approaches revealed that, following the induction of inhibitory synaptic potentiation, GABA A R clusters increase in size and number at the post-synaptic membrane with no other major structural changes of the pre- and post-synaptic elements.

  4. A national facility for biological cryo-electron microscopy

    International Nuclear Information System (INIS)

    Saibil, Helen R.; Grünewald, Kay; Stuart, David I.

    2015-01-01

    This review provides a brief update on the use of cryo-electron microscopy for integrated structural biology, along with an overview of the plans for the UK national facility for electron microscopy being built at the Diamond synchrotron. Three-dimensional electron microscopy is an enormously powerful tool for structural biologists. It is now able to provide an understanding of the molecular machinery of cells, disease processes and the actions of pathogenic organisms from atomic detail through to the cellular context. However, cutting-edge research in this field requires very substantial resources for equipment, infrastructure and expertise. Here, a brief overview is provided of the plans for a UK national three-dimensional electron-microscopy facility for integrated structural biology to enable internationally leading research on the machinery of life. State-of-the-art equipment operated with expert support will be provided, optimized for both atomic-level single-particle analysis of purified macromolecules and complexes and for tomography of cell sections. The access to and organization of the facility will be modelled on the highly successful macromolecular crystallography (MX) synchrotron beamlines, and will be embedded at the Diamond Light Source, facilitating the development of user-friendly workflows providing near-real-time experimental feedback

  5. Electron-optical design parameters for a high-resolution electron monochromator

    International Nuclear Information System (INIS)

    Tanaka, H.; Huebner, R.H.

    1976-01-01

    Detailed design parameters of a new, high-resolution electron monochromator are presented. The design utilizes a hemispherical filter as the energy-dispersing element and combines both cylindrical and aperture electrostatic lenses to accelerate, decelerate, transport, and focus the electron beam from the cathode to the interaction region

  6. Human enamel structure studied by high resolution electron microscopy

    International Nuclear Information System (INIS)

    Wen, S.L.

    1989-01-01

    Human enamel structural features are characterized by high resolution electron microscopy. The human enamel consists of polycrystals with a structure similar to Ca10(PO4)6(OH)2. This article describes the structural features of human enamel crystal at atomic and nanometer level. Besides the structural description, a great number of high resolution images are included. Research into the carious process in human enamel is very important for human beings. This article firstly describes the initiation of caries in enamel crystal at atomic and unit-cell level and secondly describes the further steps of caries with structural and chemical demineralization. The demineralization in fact, is the origin of caries in human enamel. The remineralization of carious areas in human enamel has drawn more and more attention as its potential application is realized. This process has been revealed by high resolution electron microscopy in detail in this article. On the other hand, the radiation effects on the structure of human enamel are also characterized by high resolution electron microscopy. In order to reveal this phenomenon clearly, a great number of electron micrographs have been shown, and a physical mechanism is proposed. 26 references

  7. Novel low-dose imaging technique for characterizing atomic structures through scanning transmission electron microscope

    Science.gov (United States)

    Su, Chia-Ping; Syu, Wei-Jhe; Hsiao, Chien-Nan; Lai, Ping-Shan; Chen, Chien-Chun

    2017-08-01

    To investigate dislocations or heterostructures across interfaces is now of great interest to condensed matter and materials scientists. With the advances in aberration-corrected electron optics, the scanning transmission electron microscope has demonstrated its excellent capability of characterizing atomic structures within nanomaterials, and well-resolved atomic-resolution images can be obtained through long-exposure data acquisition. However, the sample drifting, carbon contamination, and radiation damage hinder further analysis, such as deriving three-dimensional (3D) structures from a series of images. In this study, a method for obtaining atomic-resolution images with significantly reduced exposure time was developed, using which an original high-resolution image with approximately one tenth the electron dose can be obtained by combining a fast-scan high-magnification image and a slow-scan low-magnification image. The feasibility of obtaining 3D atomic structures using the proposed approach was demonstrated through multislice simulation. Finally, the feasibility and accuracy of image restoration were experimentally verified. This general method cannot only apply to electron microscopy but also benefit to image radiation-sensitive materials using various light sources.

  8. Vibration compensated high-resolution scanning white-light Linnik-interferometer

    Science.gov (United States)

    Tereschenko, Stanislav; Lehmann, Peter; Gollor, Pascal; Kuehnhold, Peter

    2017-06-01

    We present a high-resolution Linnik scanning white-light interferometer (SWLI) with integrated distance measuring interferometer (DMI) for close-to-machine applications in the presence of environmental vibrations. The distance, measured by DMI during the depth-scan, is used for vibration compensation of SWLI signals. The reconstruction of the white-light interference signals takes place after measurement by reordering the captured images in accordance with their real positions obtained by the DMI and subsequent trigonometrical approximation. This system is the further development of our previously presented Michelson-interferometer. We are able to compensate for arbitrary vibrations with frequencies up to several kilohertz and amplitudes in the lower micrometer range. Completely distorted SWLI signals can be reconstructed and the surface topography can be obtained with high accuracy. We demonstrate the feasibility of the method by examples of practical measurements with and without vibrational disturbances.

  9. Primed for Discovery: Atomic-Resolution Cryo-EM Structure of a Reovirus Entry Intermediate

    Directory of Open Access Journals (Sweden)

    Shane D. Trask

    2010-06-01

    Full Text Available A recently solved structure of the aquareovirus virion (Zhang, X; Jin, L.; Fang, Q; Hui, W.H.; Zhou Z.H. 3.3 Å Cryo-EM Structure of a Nonenveloped Virus Reveals a Priming Mechanism for Cell Entry. Cell 2010, 141, 472-482 [1] provides new insights into the order of entry events, as well as confirming and refining several aspects of the entry mechanism, for aquareovirus and the related orthoreovirus. In particular, the structure provides evidence of a defined order for the progressive proteolytic cleavages of myristoylated penetration protein VP5 that prime the virion for membrane penetration. These observations reinforce the concept that, much like enveloped viruses, nonenveloped virions often undergo priming events that lead to a meta-stable state, preparing the virus for membrane penetration under the appropriate circumstances. In addition, this and other recent studies highlight the increasing power of electron cryomicroscopy to analyze large, geometrically regular structures, such as icosahedral viruses, at atomic resolution.

  10. Molecular architecture of axonemal microtubule doublets revealedby cryo-electron tomography

    Energy Technology Data Exchange (ETDEWEB)

    Sui, Haixin; Downing, Kenneth H.

    2006-05-22

    The axoneme, which forms the core of eukaryotic flagella and cilia, is one of the largest macromolecular machines with a structure that is largely conserved from protists to mammals. Microtubule doublets are structural components of axonemes containing a number of proteins besides tubulin, and are usually found in arrays of nine doublets arranged around two singlet microtubules. Coordinated sliding of adjacent doublets, which involves a host of other proteins in the axoneme, produces periodic beating movements of the axoneme. We have obtained a 3D density map of intact microtubule doublets using cryo-electron tomography and image averaging. Our map, with a resolution of about 3 nm, provides insights into locations of particular proteins within the doublets and the structural features of the doublets that define their mechanical properties. We identify likely candidates for several of these non-tubulin components of the doublets. This work offers novel insight on how tubulin protofilaments and accessory proteins attach together to form the doublets and provides a structural basis for understanding doublet function in axonemes.

  11. Electron tomography and cryo-SEM characterization reveals novel ultrastructural features of host-parasite interaction during Chlamydia abortus infection.

    Science.gov (United States)

    Wilkat, M; Herdoiza, E; Forsbach-Birk, V; Walther, P; Essig, A

    2014-08-01

    Chlamydia (C.) abortus is a widely spread pathogen among ruminants that can be transmitted to women during pregnancy leading to severe systemic infection with consecutive abortion. As a member of the Chlamydiaceae, C. abortus shares the characteristic feature of an obligate intracellular biphasic developmental cycle with two morphological forms including elementary bodies (EBs) and reticulate bodies (RBs). In contrast to other chlamydial species, C. abortus ultrastructure has not been investigated yet. To do so, samples were fixed by high-pressure freezing and processed by different electron microscopic methods. Freeze-substituted samples were analysed by transmission electron microscopy, scanning transmission electron microscopical tomography and immuno-electron microscopy, and freeze-fractured samples were analysed by cryo-scanning electron microscopy. Here, we present three ultrastructural features of C. abortus that have not been reported up to now. Firstly, the morphological evidence that C. abortus is equipped with the type three secretion system. Secondly, the accumulation and even coating of whole inclusion bodies by membrane complexes consisting of multiple closely adjacent membranes which seems to be a C. abortus specific feature. Thirdly, the formation of small vesicles in the periplasmic space of RBs in the second half of the developmental cycle. Concerning the time point of their formation and the fact that they harbour chlamydial components, these vesicles might be morphological correlates of an intermediate step during the process of redifferentiation of RBs into EBs. As this feature has also been shown for C. trachomatis and C. pneumoniae, it might be a common characteristic of the family of Chlamydiaceae.

  12. Field Emission Scanning Electron Microscope (FESEM) Facility in BTI

    International Nuclear Information System (INIS)

    Cik Rohaida Che Hak; Foo, C.T.; Nor Azillah Fatimah Othman

    2015-01-01

    Field Emission Scanning Electron Microscope (FE-SEM) provides ultra-high resolution imaging at low accelerating voltages and small working distances. The GeminisSEM 500, a new FESEM imaging facility will be installed soon in MTEC, BTI. It provides resolution of the images is as low as 0.6 nm at 15 kV and 1.2 nm at 1 kV, allowing examination of the top surface of nano powders, nano film and nano fiber in the wide range of applications such as mineralogy, ceramics, polymer, metallurgy, electronic devices, chemistry, physics and life sciences. This system is equipped with several detectors to detect various signals such as secondary electrons (SE) detector for topographic information and back-scattered electrons (BSE) detector for materials composition contrast. Energy dispersive x-ray spectroscopy (EDS) with detector energy resolution of < 129 eV and detection limit in the range of 1000-3000 ppm coupled with FE-SEM is used to determine the chemical composition of micro-features including boron (B) to uranium (U). Wavelength dispersive x-ray spectroscopy (WDS) which has detector resolution of 2-20 eV and detection limit of 30-300 ppm coupled with FE-SEM is used to detect elements that cannot be resolved with EDS. The ultra-high resolution imaging combined with the high sensitivity WDS helps to resolve the thorium and rare earth elemental analysis. (author)

  13. The propagation of high power CW scanning electron beam in air

    International Nuclear Information System (INIS)

    Korenev, Sergey; Korenev, Ivan

    2002-01-01

    The question of propagation of high power electron beam in air presents the scientific and applied interests. The high power (80 kW) CW electron accelerator 'Rhodotron' with kinetic energy of electrons 5 and 10 MeV was used in the experiments. The experimental results for propagation of scanning electron beams in air are presented and discussed

  14. Understanding the structure of nanocatalysts with high resolution scanning/transmission electron microscopy

    International Nuclear Information System (INIS)

    Francis, L D; Rivas, J; José-Yacamán, M

    2014-01-01

    Nanomaterials including nanoparticles, nanowires and nanotubes play an important role in heterogeneous catalysis. Thanks to the rapid improvement of the electron microscopic techniques and with the advent of aberration corrected electron microscopy as well as theoretical methodologies, the potential effects induced by nanocatalysts are better understood than before by unravelling their atomic structure. A brief introduction to advanced electron microscopic techniques namely aberration corrected scanning transmission electron microscopy (Cs-STEM) is presented and subsequently two examples of nanocatalysts are considered in the present review. The first example will focus on the study of bimetallic/core-shell nanoalloys. In heterogeneous catalysis, catalysts containing two or more metals might show significantly different catalytic properties compared to the parent metals and thus are widely utilized in several catalytic reactions. Atom-by-atom insights of the nanoalloy based catalysts ex: Au-Pd will be described in the present review using a combination of advanced electron microscopic and spectroscopic techniques. A related example on the understanding of bimetallic clusters by HAADF-STEM will also be presented in addition to nanoparticles. In the second case understanding the structure of transition metal chalcogenide based nanocatalysts by HRTEM and aberration corrected STEM, for the case of MoS 2 will be discussed. MoS 2 -based catalysts serve as model catalysts and are employed in the hydrodesulphurisations (HDS) reactions in the removal of sulphur from gasoline and related petrochemical products. They have been studied in various forms including nanowires, nanotubes and nanoplates. Their structure, atomic insights and as a consequence elucidation of their corresponding catalytic activity are thus important

  15. Computer simulation of high resolution transmission electron micrographs: theory and analysis

    International Nuclear Information System (INIS)

    Kilaas, R.

    1985-03-01

    Computer simulation of electron micrographs is an invaluable aid in their proper interpretation and in defining optimum conditions for obtaining images experimentally. Since modern instruments are capable of atomic resolution, simulation techniques employing high precision are required. This thesis makes contributions to four specific areas of this field. First, the validity of a new method for simulating high resolution electron microscope images has been critically examined. Second, three different methods for computing scattering amplitudes in High Resolution Transmission Electron Microscopy (HRTEM) have been investigated as to their ability to include upper Laue layer (ULL) interaction. Third, a new method for computing scattering amplitudes in high resolution transmission electron microscopy has been examined. Fourth, the effect of a surface layer of amorphous silicon dioxide on images of crystalline silicon has been investigated for a range of crystal thicknesses varying from zero to 2 1/2 times that of the surface layer

  16. Low resolution scans can provide a sufficiently accurate, cost- and time-effective alternative to high resolution scans for 3D shape analyses

    Directory of Open Access Journals (Sweden)

    Ariel E. Marcy

    2018-06-01

    Full Text Available Background Advances in 3D shape capture technology have made powerful shape analyses, such as geometric morphometrics, more feasible. While the highly accurate micro-computed tomography (µCT scanners have been the “gold standard,” recent improvements in 3D surface scanners may make this technology a faster, portable, and cost-effective alternative. Several studies have already compared the two devices but all use relatively large specimens such as human crania. Here we perform shape analyses on Australia’s smallest rodent to test whether a 3D scanner produces similar results to a µCT scanner. Methods We captured 19 delicate mouse (Pseudomys delicatulus crania with a µCT scanner and a 3D scanner for geometric morphometrics. We ran multiple Procrustes ANOVAs to test how variation due to scan device compared to other sources such as biologically relevant variation and operator error. We quantified operator error as levels of variation and repeatability. Further, we tested if the two devices performed differently at classifying individuals based on sexual dimorphism. Finally, we inspected scatterplots of principal component analysis (PCA scores for non-random patterns. Results In all Procrustes ANOVAs, regardless of factors included, differences between individuals contributed the most to total variation. The PCA plots reflect this in how the individuals are dispersed. Including only the symmetric component of shape increased the biological signal relative to variation due to device and due to error. 3D scans showed a higher level of operator error as evidenced by a greater spread of their replicates on the PCA, a higher level of multivariate variation, and a lower repeatability score. However, the 3D scan and µCT scan datasets performed identically in classifying individuals based on intra-specific patterns of sexual dimorphism. Discussion Compared to µCT scans, we find that even low resolution 3D scans of very small specimens are

  17. Cryo-scanning electron microscopy investigation of the Octopus Vulgaris arm structures for the design of an octopus-like arm artefact.

    Science.gov (United States)

    Minnocci, Antonio; Cianchetti, Matteo; Mazzolai, Barbara; Sebastiani, Luca; Laschi, Cecilia

    2015-12-01

    Octopus vulgaris is a cephalopod of the Octopodidae family. It has four pairs of arms and two rows of suckers which perform many functions, including bending and elongation. For this reason the octopus was chosen as model to develop a new generation of soft-body robots. In order to explain some of the fine structures of the octopus arm in relation to its specific ability, we examined the external and internal structures of O. vulgaris arms in a frozen-hydrated state using cryo-scanning electron microscopy. The arms showed skin with a very complex design that is useful to elongation, and a pore pattern distribution on their surface which is functional to cutaneous oxygen uptake. The analysis of freeze-fractured frozen-hydrated arm samples allowed us to describe the developmental differences in the relative proportion of the areas of axial nerve cord, intrinsic and extrinsic musculature, in relation to the growth of the arms and of the increase in functional capability. In the suckers, we analyzed the shedding mechanisms in the outer part of the infundibulum and described the outer and inner characteristics of the denticles, showing in detail their pore system, which is fundamental for their ability to explore the environment. These results are discussed by considering their possible application in the design of new octopus-like artefacts, which will be able to take advantage of some of these ultrastructure characteristics and achieve advanced bioinspired functionalities. © 2015 Wiley Periodicals, Inc.

  18. Cryo-transmission electron microscopy of Ag nanoparticles grown on an ionic liquid substrate

    KAUST Repository

    Anjum, Dalaver H.; Stiger, Rebecca M.; Finley, James J.; Conway, James F.

    2010-01-01

    We report a novel method of growing silver nanostructures by cathodic sputtering onto an ionic liquid (IL) and our visualization by transmission cryo-electron microscopy to avoid beam-induced motion of the nanoparticles. By freezing the IL

  19. In situ investigation of ion-induced dewetting of a thin iron-oxide film on silicon by high resolution scanning electron microscopy

    Energy Technology Data Exchange (ETDEWEB)

    Amirthapandian, S. [Institut fuer Halbleiteroptik und Funktionelle Grenzflaechen, Universitaet Stuttgart, 70569 Stuttgart (Germany); Material Physics Division, Materials Science Group, Indira Gandhi Centre for Atomic Research, Kalpakkam 603 102 (India); Schuchart, F.; Garmatter, D.; Bolse, W. [Institut fuer Halbleiteroptik und Funktionelle Grenzflaechen, Universitaet Stuttgart, 70569 Stuttgart (Germany)

    2012-11-15

    Using our new in situ high resolution scanning electron microscope, which is integrated into the UNILAC ion beamline at the Helmholtzzentrum fuer Schwerionenforschung (GSI) in Darmstadt, Germany, we investigated the swift heavy ion induced dewetting of a thin iron oxide layer on Si. Besides heterogeneous hole nucleation at defects and spontaneous (homogeneous) hole nucleation, we could clearly identify a dewetting mechanism, which is similar to the spinodal dewetting observed for liquid films. Instead of being due to capillary waves, it is based on a stress induced surface instability. The latter results in the formation of a wavy surface with constant dominant wave-length and increasing amplitude during ion irradiation. Dewetting sets in as soon as the wave-troughs reach the film-substrate interface. Inspection of the hole radii and rim shapes indicates that removal of the material from the hole area occurs mainly by plastic deformation at the inner boundary and ion induced viscous flow in the peripheral zone due to surface tension.

  20. Characterization of Emulsions of Fish Oil and Water by Cryo Scanning Electron Microscopy

    DEFF Research Database (Denmark)

    Jensen, Louise Helene Søgaard; Horn, Anna Frisenfeldt; Jacobsen, Charlotte

    Addition of fish oil to industrially prepared food products is attractive to the food industry because of the well-documented health effects of the omega 3 fatty acids in the fish oil [1]. Polyunsaturated Fatty Acids including omega 3 fatty acids are highly susceptible to lipid oxidation due...... to the many double bonds. Emulsions of fish oil in water are potential candidates for a delivery system of fish oil to food products. It has been suggested that oxidation of oil-in-water emulsions is initiated at the interface between oil and water. It has also been proposed that oxidation is to some extent...... is to characterize fish oil in water emulsions with respect to oil droplet size, distribution, and ultimately to view the structure and thickness of the interface layer. A freeze-fractured surface viewed at low temperatures under the scanning electron microscope is a promising strategy to reveal variations...

  1. a New Approach for Subway Tunnel Deformation Monitoring: High-Resolution Terrestrial Laser Scanning

    Science.gov (United States)

    Li, J.; Wan, Y.; Gao, X.

    2012-07-01

    With the improvement of the accuracy and efficiency of laser scanning technology, high-resolution terrestrial laser scanning (TLS) technology can obtain high precise points-cloud and density distribution and can be applied to high-precision deformation monitoring of subway tunnels and high-speed railway bridges and other fields. In this paper, a new approach using a points-cloud segmentation method based on vectors of neighbor points and surface fitting method based on moving least squares was proposed and applied to subway tunnel deformation monitoring in Tianjin combined with a new high-resolution terrestrial laser scanner (Riegl VZ-400). There were three main procedures. Firstly, a points-cloud consisted of several scanning was registered by linearized iterative least squares approach to improve the accuracy of registration, and several control points were acquired by total stations (TS) and then adjusted. Secondly, the registered points-cloud was resampled and segmented based on vectors of neighbor points to select suitable points. Thirdly, the selected points were used to fit the subway tunnel surface with moving least squares algorithm. Then a series of parallel sections obtained from temporal series of fitting tunnel surfaces were compared to analysis the deformation. Finally, the results of the approach in z direction were compared with the fiber optical displacement sensor approach and the results in x, y directions were compared with TS respectively, and comparison results showed the accuracy errors of x, y, z directions were respectively about 1.5 mm, 2 mm, 1 mm. Therefore the new approach using high-resolution TLS can meet the demand of subway tunnel deformation monitoring.

  2. CTF determination and correction in electron cryotomography

    International Nuclear Information System (INIS)

    Fernandez, J.J.; Li, S.; Crowther, R.A.

    2006-01-01

    Electron cryotomography (cryoET) has the potential to elucidate the structure of complex biological specimens at molecular resolution but technical and computational improvements are still needed. This work addresses the determination and correction of the contrast transfer function (CTF) of the electron microscope in cryoET. Our approach to CTF detection and defocus determination depends on strip-based periodogram averaging, extended throughout the tilt series to overcome the low contrast conditions found in cryoET. A method for CTF correction that deals with the defocus gradient in images of tilted specimens is also proposed. These approaches to CTF determination and correction have been applied here to several examples of cryoET of pleomorphic specimens and of single particles. CTF correction is essential for improving the resolution, particularly in those studies that combine cryoET with single particle averaging techniques

  3. Deceleration of probe beam by stage bias potential improves resolution of serial block-face scanning electron microscopic images.

    Science.gov (United States)

    Bouwer, James C; Deerinck, Thomas J; Bushong, Eric; Astakhov, Vadim; Ramachandra, Ranjan; Peltier, Steven T; Ellisman, Mark H

    2017-01-01

    Serial block-face scanning electron microscopy (SBEM) is quickly becoming an important imaging tool to explore three-dimensional biological structure across spatial scales. At probe-beam-electron energies of 2.0 keV or lower, the axial resolution should improve, because there is less primary electron penetration into the block face. More specifically, at these lower energies, the interaction volume is much smaller, and therefore, surface detail is more highly resolved. However, the backscattered electron yield for metal contrast agents and the backscattered electron detector sensitivity are both sub-optimal at these lower energies, thus negating the gain in axial resolution. We found that the application of a negative voltage (reversal potential) applied to a modified SBEM stage creates a tunable electric field at the sample. This field can be used to decrease the probe-beam-landing energy and, at the same time, alter the trajectory of the signal to increase the signal collected by the detector. With decelerated low landing-energy electrons, we observed that the probe-beam-electron-penetration depth was reduced to less than 30 nm in epoxy-embedded biological specimens. Concurrently, a large increase in recorded signal occurred due to the re-acceleration of BSEs in the bias field towards the objective pole piece where the detector is located. By tuning the bias field, we were able to manipulate the trajectories of the  primary and secondary electrons, enabling the spatial discrimination of these signals using an advanced ring-type BSE detector configuration or a standard monolithic BSE detector coupled with a blocking aperture.

  4. Refinement procedure for the image alignment in high-resolution electron tomography

    International Nuclear Information System (INIS)

    Houben, L.; Bar Sadan, M.

    2011-01-01

    High-resolution electron tomography from a tilt series of transmission electron microscopy images requires an accurate image alignment procedure in order to maximise the resolution of the tomogram. This is the case in particular for ultra-high resolution where even very small misalignments between individual images can dramatically reduce the fidelity of the resultant reconstruction. A tomographic-reconstruction based and marker-free method is proposed, which uses an iterative optimisation of the tomogram resolution. The method utilises a search algorithm that maximises the contrast in tomogram sub-volumes. Unlike conventional cross-correlation analysis it provides the required correlation over a large tilt angle separation and guarantees a consistent alignment of images for the full range of object tilt angles. An assessment based on experimental reconstructions shows that the marker-free procedure is competitive to the reference of marker-based procedures at lower resolution and yields sub-pixel accuracy even for simulated high-resolution data. -- Highlights: → Alignment procedure for electron tomography based on iterative tomogram contrast optimisation. → Marker-free, independent of object, little user interaction. → Accuracy competitive with fiducial marker methods and suited for high-resolution tomography.

  5. Healthy and diseased striated muscle studied by analytical scanning electron microscopy with special reference to fibre type

    International Nuclear Information System (INIS)

    Wroblewski, R.

    1982-01-01

    X-ray microanalytical investigations of striated muscles in the scanning electron microscope are reviewed. The main part of the studies was performed on cryosections cut with a conventional cryostat operating at -20 degrees C to -40 degrees C. The preparation procedure including different types of attachment of the sections to the specimen holder is described in detail. The elemental changes in muscle are related to the muscle fibre type as demonstrated by histochemical methods or to histochemically demonstrated inclusions in diseased muscles. This is of great importance, because muscle disorders are often characterised by selective involvement of different muscle fibre types. The preparation methods of muscle for analytical scanning electron microscopy and the obtained results are compared with studies performed on thin cryo and epoxy sections, analysed in the transmission and scanning-transmission electron microscope. It is evident that X-ray microanalysis performed on thick cryosections provide a quick survey of the elemental composition of whole cells, and should be followed in interesting cases by close examination on the organelle level studied in thin cryosections in the transmission and scanning-transmission electron microscope

  6. Time-resolved GISAXS and cryo-microscopy characterization of block copolymer membrane formation

    KAUST Repository

    Marques, Debora S.; Dorin, Rachel Mika; Wiesner, Ulrich B.; Smilgies, Detlef Matthias; Behzad, Ali Reza; Vainio, Ulla; Peinemann, Klaus-Viktor; Nunes, Suzana Pereira

    2014-01-01

    Time-resolved grazing-incidence small-angle X-ray scattering (GISAXS) and cryo-microscopy were used for the first time to understand the pore evolution by copolymer assembly, leading to the formation of isoporous membranes with exceptional porosity and regularity. The formation of copolymer micelle strings in solution (in DMF/DOX/THF and DMF/DOX) was confirmed by cryo field emission scanning electron microscopy (cryo-FESEM) with a distance of 72 nm between centers of micelles placed in different strings. SAXS measurement of block copolymer solutions in DMF/DOX indicated hexagonal assembly with micelle-to-micelle distance of 84-87 nm for 14-20 wt% copolymer solutions. GISAXS in-plane peaks were detected, revealing order close to hexagonal. The d-spacing corresponding to the first peak in this case was 100-130 nm (lattice constant 115-150 nm) for 17 wt% copolymer solutions evaporating up to 100 s. Time-resolved cryo-FESEM showed the formation of incipient pores on the film surface after 4 s copolymer solution casting with distances between void centers of 125 nm. © 2014 Elsevier Ltd. All rights reserved.

  7. Time-resolved GISAXS and cryo-microscopy characterization of block copolymer membrane formation

    KAUST Repository

    Marques, Debora S.

    2014-03-01

    Time-resolved grazing-incidence small-angle X-ray scattering (GISAXS) and cryo-microscopy were used for the first time to understand the pore evolution by copolymer assembly, leading to the formation of isoporous membranes with exceptional porosity and regularity. The formation of copolymer micelle strings in solution (in DMF/DOX/THF and DMF/DOX) was confirmed by cryo field emission scanning electron microscopy (cryo-FESEM) with a distance of 72 nm between centers of micelles placed in different strings. SAXS measurement of block copolymer solutions in DMF/DOX indicated hexagonal assembly with micelle-to-micelle distance of 84-87 nm for 14-20 wt% copolymer solutions. GISAXS in-plane peaks were detected, revealing order close to hexagonal. The d-spacing corresponding to the first peak in this case was 100-130 nm (lattice constant 115-150 nm) for 17 wt% copolymer solutions evaporating up to 100 s. Time-resolved cryo-FESEM showed the formation of incipient pores on the film surface after 4 s copolymer solution casting with distances between void centers of 125 nm. © 2014 Elsevier Ltd. All rights reserved.

  8. Transmission environmental scanning electron microscope with scintillation gaseous detection device

    International Nuclear Information System (INIS)

    Danilatos, Gerasimos; Kollia, Mary; Dracopoulos, Vassileios

    2015-01-01

    A transmission environmental scanning electron microscope with use of a scintillation gaseous detection device has been implemented. This corresponds to a transmission scanning electron microscope but with addition of a gaseous environment acting both as environmental and detection medium. A commercial type of low vacuum machine has been employed together with appropriate modifications to the detection configuration. This involves controlled screening of various emitted signals in conjunction with a scintillation gaseous detection device already provided with the machine for regular surface imaging. Dark field and bright field imaging has been obtained along with other detection conditions. With a progressive series of modifications and tests, the theory and practice of a novel type of microscopy is briefly shown now ushering further significant improvements and developments in electron microscopy as a whole. - Highlights: • Novel scanning transmission electron microscopy (STEM) with an environmental scanning electron microscope (ESEM) called TESEM. • Use of the gaseous detection device (GDD) in scintillation mode that allows high resolution bright and dark field imaging in the TESEM. • Novel approach towards a unification of both vacuum and environmental conditions in both bulk/surface and transmission mode of electron microscopy

  9. Structure and assembly of the mouse ASC inflammasome by combined NMR spectroscopy and cryo-electron microscopy

    Science.gov (United States)

    Sborgi, Lorenzo; Ravotti, Francesco; Dandey, Venkata P.; Dick, Mathias S.; Mazur, Adam; Reckel, Sina; Chami, Mohamed; Scherer, Sebastian; Huber, Matthias; Böckmann, Anja; Egelman, Edward H.; Stahlberg, Henning; Broz, Petr; Meier, Beat H.; Hiller, Sebastian

    2015-01-01

    Inflammasomes are multiprotein complexes that control the innate immune response by activating caspase-1, thus promoting the secretion of cytokines in response to invading pathogens and endogenous triggers. Assembly of inflammasomes is induced by activation of a receptor protein. Many inflammasome receptors require the adapter protein ASC [apoptosis-associated speck-like protein containing a caspase-recruitment domain (CARD)], which consists of two domains, the N-terminal pyrin domain (PYD) and the C-terminal CARD. Upon activation, ASC forms large oligomeric filaments, which facilitate procaspase-1 recruitment. Here, we characterize the structure and filament formation of mouse ASC in vitro at atomic resolution. Information from cryo-electron microscopy and solid-state NMR spectroscopy is combined in a single structure calculation to obtain the atomic-resolution structure of the ASC filament. Perturbations of NMR resonances upon filament formation monitor the specific binding interfaces of ASC-PYD association. Importantly, NMR experiments show the rigidity of the PYD forming the core of the filament as well as the high mobility of the CARD relative to this core. The findings are validated by structure-based mutagenesis experiments in cultured macrophages. The 3D structure of the mouse ASC-PYD filament is highly similar to the recently determined human ASC-PYD filament, suggesting evolutionary conservation of ASC-dependent inflammasome mechanisms. PMID:26464513

  10. A NEW APPROACH FOR SUBWAY TUNNEL DEFORMATION MONITORING: HIGH-RESOLUTION TERRESTRIAL LASER SCANNING

    Directory of Open Access Journals (Sweden)

    J. Li

    2012-07-01

    Full Text Available With the improvement of the accuracy and efficiency of laser scanning technology, high-resolution terrestrial laser scanning (TLS technology can obtain high precise points-cloud and density distribution and can be applied to high-precision deformation monitoring of subway tunnels and high-speed railway bridges and other fields. In this paper, a new approach using a points-cloud segmentation method based on vectors of neighbor points and surface fitting method based on moving least squares was proposed and applied to subway tunnel deformation monitoring in Tianjin combined with a new high-resolution terrestrial laser scanner (Riegl VZ-400. There were three main procedures. Firstly, a points-cloud consisted of several scanning was registered by linearized iterative least squares approach to improve the accuracy of registration, and several control points were acquired by total stations (TS and then adjusted. Secondly, the registered points-cloud was resampled and segmented based on vectors of neighbor points to select suitable points. Thirdly, the selected points were used to fit the subway tunnel surface with moving least squares algorithm. Then a series of parallel sections obtained from temporal series of fitting tunnel surfaces were compared to analysis the deformation. Finally, the results of the approach in z direction were compared with the fiber optical displacement sensor approach and the results in x, y directions were compared with TS respectively, and comparison results showed the accuracy errors of x, y, z directions were respectively about 1.5 mm, 2 mm, 1 mm. Therefore the new approach using high-resolution TLS can meet the demand of subway tunnel deformation monitoring.

  11. Cryo-EM Structure Determination Using Segmented Helical Image Reconstruction.

    Science.gov (United States)

    Fromm, S A; Sachse, C

    2016-01-01

    Treating helices as single-particle-like segments followed by helical image reconstruction has become the method of choice for high-resolution structure determination of well-ordered helical viruses as well as flexible filaments. In this review, we will illustrate how the combination of latest hardware developments with optimized image processing routines have led to a series of near-atomic resolution structures of helical assemblies. Originally, the treatment of helices as a sequence of segments followed by Fourier-Bessel reconstruction revealed the potential to determine near-atomic resolution structures from helical specimens. In the meantime, real-space image processing of helices in a stack of single particles was developed and enabled the structure determination of specimens that resisted classical Fourier helical reconstruction and also facilitated high-resolution structure determination. Despite the progress in real-space analysis, the combination of Fourier and real-space processing is still commonly used to better estimate the symmetry parameters as the imposition of the correct helical symmetry is essential for high-resolution structure determination. Recent hardware advancement by the introduction of direct electron detectors has significantly enhanced the image quality and together with improved image processing procedures has made segmented helical reconstruction a very productive cryo-EM structure determination method. © 2016 Elsevier Inc. All rights reserved.

  12. The cryo-electron microscopy structure of huntingtin

    Science.gov (United States)

    Guo, Qiang; Bin Huang; Cheng, Jingdong; Seefelder, Manuel; Engler, Tatjana; Pfeifer, Günter; Oeckl, Patrick; Otto, Markus; Moser, Franziska; Maurer, Melanie; Pautsch, Alexander; Baumeister, Wolfgang; Fernández-Busnadiego, Rubén; Kochanek, Stefan

    2018-03-01

    Huntingtin (HTT) is a large (348 kDa) protein that is essential for embryonic development and is involved in diverse cellular activities such as vesicular transport, endocytosis, autophagy and the regulation of transcription. Although an integrative understanding of the biological functions of HTT is lacking, the large number of identified HTT interactors suggests that it serves as a protein-protein interaction hub. Furthermore, Huntington’s disease is caused by a mutation in the HTT gene, resulting in a pathogenic expansion of a polyglutamine repeat at the amino terminus of HTT. However, only limited structural information regarding HTT is currently available. Here we use cryo-electron microscopy to determine the structure of full-length human HTT in a complex with HTT-associated protein 40 (HAP40; encoded by three F8A genes in humans) to an overall resolution of 4 Å. HTT is largely α-helical and consists of three major domains. The amino- and carboxy-terminal domains contain multiple HEAT (huntingtin, elongation factor 3, protein phosphatase 2A and lipid kinase TOR) repeats arranged in a solenoid fashion. These domains are connected by a smaller bridge domain containing different types of tandem repeats. HAP40 is also largely α-helical and has a tetratricopeptide repeat-like organization. HAP40 binds in a cleft and contacts the three HTT domains by hydrophobic and electrostatic interactions, thereby stabilizing the conformation of HTT. These data rationalize previous biochemical results and pave the way for improved understanding of the diverse cellular functions of HTT.

  13. Single particle and molecular assembly analysis of polyribosomes by single- and double-tilt cryo electron tomography

    Energy Technology Data Exchange (ETDEWEB)

    Myasnikov, Alexander G. [IGBMC (Institute of Genetics and of Molecular and Cellular Biology), Department of Integrative Structural Biology, Centre National de la Recherche Scientifique (CNRS) UMR 7104/ Institut National de la Santé de la Recherche Médicale INSERM U964/ Université de Strasbourg, 1 rue Laurent Fries, 67404 Illkirch (France); Afonina, Zhanna A. [Institute of Protein Research, Russian Academy of Sciences, 142290 Pushchino, Moscow Region (Russian Federation); Klaholz, Bruno P., E-mail: klaholz@igbmc.fr [IGBMC (Institute of Genetics and of Molecular and Cellular Biology), Department of Integrative Structural Biology, Centre National de la Recherche Scientifique (CNRS) UMR 7104/ Institut National de la Santé de la Recherche Médicale INSERM U964/ Université de Strasbourg, 1 rue Laurent Fries, 67404 Illkirch (France)

    2013-03-15

    Cryo electron tomography (cryo-ET) can provide cellular and molecular structural information on various biological samples. However, the detailed interpretation of tomograms reconstructed from single-tilt data tends to suffer from low signal-to-noise ratio and artefacts caused by some systematically missing angular views. While these can be overcome by sub-tomogram averaging, they remain limiting for the analysis of unique structures. Double-tilt ET can improve the tomogram quality by acquiring a second tilt series after an in-plane rotation, but its usage is not widespread yet because it is considered technically demanding and it is rarely used under cryo conditions. Here we show that double-tilt cryo-ET improves the quality of 3D reconstructions so significantly that even single particle analysis can be envisaged despite of the intrinsically low image contrast obtained from frozen-hydrated specimens. This is illustrated by the analysis of eukaryotic polyribosomes in which individual ribosomes were reconstructed using single-tilt, partial and full double-tilt geometries. The improved tomograms favour the faster convergence of iterative sub-tomogram averaging and allow a better 3D classification using multivariate statistical analysis. Our study of single particles and molecular assemblies within polysomes illustrates that the dual-axis approach is particularly useful for cryo applications of ET, both for unique objects and for structures that can be classified and averaged. - Highlights: ► Double-tilt cryo-ET improves 3D reconstructions thus making single particle analysis possible. ► Dual-axis cryo-ET data favour a faster convergence of iterative sub-tomogram averaging. ► Individual ribosomes were reconstructed from single-tilt, partial/ full double-tilt geometries. ► Double-tilt cryo-ET facilitates analysis of larger molecular assemblies such as in cell sections. ► Dual-axis cryo-ET is applicable to unique objects and to structures that can be

  14. Dynamic Low-Vacuum Scanning Electron Microscope Freeze Drying Observation for Fresh Water Algae

    International Nuclear Information System (INIS)

    Mohsen, H.T.; Ghaly, W.A.; Zahran, N.F.; Helal, A.I.

    2010-01-01

    A new perpetration method for serving in dynamic examinations of the fresh water algae is developed in connection with the Low-Vacuum Scanning Electron Microscope (LV-SEM) freeze drying technique. Specimens are collected from fresh water of Ismailia channel then transferred directly to freeze by liquid nitrogen and dried in the chamber of the scanning electron microscope in the low vacuum mode. Scanning electron micrographs revealed that the drying method presented the microstructure of algae. Dehydration in a graded ethanol series is not necessary in the new method. Dried algae specimen is observed in SEM high vacuum mode after conductive coating at higher resolution. Low-vacuum SEM freeze drying technique is a simple, time-saving and reproducible method for scanning electron microscopy that is applicable to various aquatic microorganisms covered with soft tissues.

  15. Scanning SQUID susceptometers with sub-micron spatial resolution

    Energy Technology Data Exchange (ETDEWEB)

    Kirtley, John R., E-mail: jkirtley@stanford.edu; Rosenberg, Aaron J.; Palmstrom, Johanna C.; Holland, Connor M.; Moler, Kathryn A. [Department of Applied Physics, Stanford University, Stanford, California 94305-4045 (United States); Paulius, Lisa [Department of Physics, Western Michigan University, Kalamazoo, Michigan 49008-5252 (United States); Spanton, Eric M. [Department of Physics, Stanford University, Stanford, California 94305-4045 (United States); Schiessl, Daniel [Attocube Systems AG, Königinstraße 11A, 80539 Munich (Germany); Jermain, Colin L.; Gibbons, Jonathan [Department of Physics, Cornell University, Cornell, Ithaca, New York 14853 (United States); Fung, Y.-K.K.; Gibson, Gerald W. [IBM Research Division, T. J. Watson Research Center, Yorktown Heights, New York 10598 (United States); Huber, Martin E. [Department of Physics, University of Colorado Denver, Denver, Colorado 80217-3364 (United States); Ralph, Daniel C. [Department of Physics, Cornell University, Cornell, Ithaca, New York 14853 (United States); Kavli Institute at Cornell, Ithaca, New York 14853 (United States); Ketchen, Mark B. [OcteVue, Hadley, Massachusetts 01035 (United States)

    2016-09-15

    Superconducting QUantum Interference Device (SQUID) microscopy has excellent magnetic field sensitivity, but suffers from modest spatial resolution when compared with other scanning probes. This spatial resolution is determined by both the size of the field sensitive area and the spacing between this area and the sample surface. In this paper we describe scanning SQUID susceptometers that achieve sub-micron spatial resolution while retaining a white noise floor flux sensitivity of ≈2μΦ{sub 0}/Hz{sup 1/2}. This high spatial resolution is accomplished by deep sub-micron feature sizes, well shielded pickup loops fabricated using a planarized process, and a deep etch step that minimizes the spacing between the sample surface and the SQUID pickup loop. We describe the design, modeling, fabrication, and testing of these sensors. Although sub-micron spatial resolution has been achieved previously in scanning SQUID sensors, our sensors not only achieve high spatial resolution but also have integrated modulation coils for flux feedback, integrated field coils for susceptibility measurements, and batch processing. They are therefore a generally applicable tool for imaging sample magnetization, currents, and susceptibilities with higher spatial resolution than previous susceptometers.

  16. Scanning SQUID susceptometers with sub-micron spatial resolution

    International Nuclear Information System (INIS)

    Kirtley, John R.; Rosenberg, Aaron J.; Palmstrom, Johanna C.; Holland, Connor M.; Moler, Kathryn A.; Paulius, Lisa; Spanton, Eric M.; Schiessl, Daniel; Jermain, Colin L.; Gibbons, Jonathan; Fung, Y.-K.K.; Gibson, Gerald W.; Huber, Martin E.; Ralph, Daniel C.; Ketchen, Mark B.

    2016-01-01

    Superconducting QUantum Interference Device (SQUID) microscopy has excellent magnetic field sensitivity, but suffers from modest spatial resolution when compared with other scanning probes. This spatial resolution is determined by both the size of the field sensitive area and the spacing between this area and the sample surface. In this paper we describe scanning SQUID susceptometers that achieve sub-micron spatial resolution while retaining a white noise floor flux sensitivity of ≈2μΦ_0/Hz"1"/"2. This high spatial resolution is accomplished by deep sub-micron feature sizes, well shielded pickup loops fabricated using a planarized process, and a deep etch step that minimizes the spacing between the sample surface and the SQUID pickup loop. We describe the design, modeling, fabrication, and testing of these sensors. Although sub-micron spatial resolution has been achieved previously in scanning SQUID sensors, our sensors not only achieve high spatial resolution but also have integrated modulation coils for flux feedback, integrated field coils for susceptibility measurements, and batch processing. They are therefore a generally applicable tool for imaging sample magnetization, currents, and susceptibilities with higher spatial resolution than previous susceptometers.

  17. Prediction of protein-protein interactions in dengue virus coat proteins guided by low resolution cryoEM structures

    Directory of Open Access Journals (Sweden)

    Srinivasan Narayanaswamy

    2010-06-01

    Full Text Available Abstract Background Dengue virus along with the other members of the flaviviridae family has reemerged as deadly human pathogens. Understanding the mechanistic details of these infections can be highly rewarding in developing effective antivirals. During maturation of the virus inside the host cell, the coat proteins E and M undergo conformational changes, altering the morphology of the viral coat. However, due to low resolution nature of the available 3-D structures of viral assemblies, the atomic details of these changes are still elusive. Results In the present analysis, starting from Cα positions of low resolution cryo electron microscopic structures the residue level details of protein-protein interaction interfaces of dengue virus coat proteins have been predicted. By comparing the preexisting structures of virus in different phases of life cycle, the changes taking place in these predicted protein-protein interaction interfaces were followed as a function of maturation process of the virus. Besides changing the current notion about the presence of only homodimers in the mature viral coat, the present analysis indicated presence of a proline-rich motif at the protein-protein interaction interface of the coat protein. Investigating the conservation status of these seemingly functionally crucial residues across other members of flaviviridae family enabled dissecting common mechanisms used for infections by these viruses. Conclusions Thus, using computational approach the present analysis has provided better insights into the preexisting low resolution structures of virus assemblies, the findings of which can be made use of in designing effective antivirals against these deadly human pathogens.

  18. The Enhancement of 3D Scans Depth Resolution Obtained by Confocal Scanning of Porous Materials

    Science.gov (United States)

    Martisek, Dalibor; Prochazkova, Jana

    2017-12-01

    The 3D reconstruction of simple structured materials using a confocal microscope is widely used in many different areas including civil engineering. Nonetheless, scans of porous materials such as concrete or cement paste are highly problematic. The well-known problem of these scans is low depth resolution in comparison to the horizontal and vertical resolution. The degradation of the image depth resolution is caused by systematic errors and especially by different random events. Our method is focused on the elimination of such random events, mainly the additive noise. We use an averaging method based on the Lindeberg-Lévy theorem that improves the final depth resolution to a level comparable with horizontal and vertical resolution. Moreover, using the least square method, we also precisely determine the limit value of a depth resolution. Therefore, we can continuously evaluate the difference between current resolution and the optimal one. This substantially simplifies the scanning process because the operator can easily determine the required number of scans.

  19. The Enhancement of 3D Scans Depth Resolution Obtained by Confocal Scanning of Porous Materials

    Directory of Open Access Journals (Sweden)

    Martisek Dalibor

    2017-12-01

    Full Text Available The 3D reconstruction of simple structured materials using a confocal microscope is widely used in many different areas including civil engineering. Nonetheless, scans of porous materials such as concrete or cement paste are highly problematic. The well-known problem of these scans is low depth resolution in comparison to the horizontal and vertical resolution. The degradation of the image depth resolution is caused by systematic errors and especially by different random events. Our method is focused on the elimination of such random events, mainly the additive noise. We use an averaging method based on the Lindeberg-Lévy theorem that improves the final depth resolution to a level comparable with horizontal and vertical resolution. Moreover, using the least square method, we also precisely determine the limit value of a depth resolution. Therefore, we can continuously evaluate the difference between current resolution and the optimal one. This substantially simplifies the scanning process because the operator can easily determine the required number of scans.

  20. Imaging of Norway spruce early somatic embryos with the ESEM, Cryo-SEM and laser scanning microscope

    Czech Academy of Sciences Publication Activity Database

    Neděla, Vilém; Hřib, Jiří; Havel, L.; Hudec, Jiří; Runštuk, Jiří

    2016-01-01

    Roč. 84, May 2016 (2016), s. 67-71 ISSN 0968-4328 R&D Projects: GA ČR(CZ) GA14-22777S Institutional support: RVO:68081731 Keywords : ESEM * Cryo-SEM * bright field/dark field microscopy * extracellular matrix * Picea abies * somatic embryogenesis Subject RIV: JA - Electronics ; Optoelectronics, Electrical Engineering Impact factor: 1.980, year: 2016

  1. Monitoring Snow and Land Ice Using Satellite data in the GMES Project CryoLand

    Science.gov (United States)

    Bippus, Gabriele; Nagler, Thomas

    2013-04-01

    The main objectives of the project "CryoLand - GMES Service Snow and Land Ice" are to develop, implement and validate services for snow, glaciers and lake and river ice products as a Downstream Service within the Global Monitoring for Environment and Security (GMES) program of the European Commission. CryoLand exploits Earth Observation data from current optical and microwave sensors and of the upcoming GMES Sentinel satellite family. The project prepares also the basis for the cryospheric component of the GMES Land Monitoring services. The CryoLand project team consists of 10 partner organisations from Austria, Finland, Norway, Sweden, Switzerland and Romania and is funded by the 7th Framework Program of the European Commission. The CryoLand baseline products for snow include fractional snow extent from optical satellite data, the extent of melting snow from SAR data, and coarse resolution snow water equivalent maps from passive microwave data. Experimental products include maps of snow surface wetness and temperature. The products range from large scale coverage at medium resolution to regional products with high resolution, in order to address a wide user community. Medium resolution optical data (e.g. MODIS, in the near future Sentinel-3) and SAR (ENVISAT ASAR, in the near future Sentinel-1) are the main sources of EO data for generating large scale products in near real time. For generation of regional products high resolution satellite data are used. Glacier products are based on high resolution optical (e.g. SPOT-5, in the near future Sentinel-2) and SAR (TerraSAR-X, in the near future Sentinel-1) data and include glacier outlines, mapping of glacier facies, glacier lakes and ice velocity. The glacier products are generated on users demand. Current test areas are located in the Alps, Norway, Greenland and the Himalayan Mountains. The lake and river ice products include ice extent and its temporal changes and snow extent on ice. The algorithms for these

  2. Dose-dependent high-resolution electron ptychography

    International Nuclear Information System (INIS)

    D'Alfonso, A. J.; Allen, L. J.; Sawada, H.; Kirkland, A. I.

    2016-01-01

    Recent reports of electron ptychography at atomic resolution have ushered in a new era of coherent diffractive imaging in the context of electron microscopy. We report and discuss electron ptychography under variable electron dose conditions, exploring the prospects of an approach which has considerable potential for imaging where low dose is needed

  3. Scanning magnetic tunnel junction microscope for high-resolution imaging of remanent magnetization fields

    Science.gov (United States)

    Lima, E. A.; Bruno, A. C.; Carvalho, H. R.; Weiss, B. P.

    2014-10-01

    Scanning magnetic microscopy is a new methodology for mapping magnetic fields with high spatial resolution and field sensitivity. An important goal has been to develop high-performance instruments that do not require cryogenic technology due to its high cost, complexity, and limitation on sensor-to-sample distance. Here we report the development of a low-cost scanning magnetic microscope based on commercial room-temperature magnetic tunnel junction (MTJ) sensors that typically achieves spatial resolution better than 7 µm. By comparing different bias and detection schemes, optimal performance was obtained when biasing the MTJ sensor with a modulated current at 1.0 kHz in a Wheatstone bridge configuration while using a lock-in amplifier in conjunction with a low-noise custom-made preamplifier. A precision horizontal (x-y) scanning stage comprising two coupled nanopositioners controls the position of the sample and a linear actuator adjusts the sensor-to-sample distance. We obtained magnetic field sensitivities better than 150 nT/Hz1/2 between 0.1 and 10 Hz, which is a critical frequency range for scanning magnetic microscopy. This corresponds to a magnetic moment sensitivity of 10-14 A m2, a factor of 100 better than achievable with typical commercial superconducting moment magnetometers. It also represents an improvement in sensitivity by a factor between 10 and 30 compared to similar scanning MTJ microscopes based on conventional bias-detection schemes. To demonstrate the capabilities of the instrument, two polished thin sections of representative geological samples were scanned along with a synthetic sample containing magnetic microparticles. The instrument is usable for a diversity of applications that require mapping of samples at room temperature to preserve magnetic properties or viability, including paleomagnetism and rock magnetism, nondestructive evaluation of materials, and biological assays.

  4. Scanning magnetic tunnel junction microscope for high-resolution imaging of remanent magnetization fields

    International Nuclear Information System (INIS)

    Lima, E A; Weiss, B P; Bruno, A C; Carvalho, H R

    2014-01-01

    Scanning magnetic microscopy is a new methodology for mapping magnetic fields with high spatial resolution and field sensitivity. An important goal has been to develop high-performance instruments that do not require cryogenic technology due to its high cost, complexity, and limitation on sensor-to-sample distance. Here we report the development of a low-cost scanning magnetic microscope based on commercial room-temperature magnetic tunnel junction (MTJ) sensors that typically achieves spatial resolution better than 7 µm. By comparing different bias and detection schemes, optimal performance was obtained when biasing the MTJ sensor with a modulated current at 1.0 kHz in a Wheatstone bridge configuration while using a lock-in amplifier in conjunction with a low-noise custom-made preamplifier. A precision horizontal (x–y) scanning stage comprising two coupled nanopositioners controls the position of the sample and a linear actuator adjusts the sensor-to-sample distance. We obtained magnetic field sensitivities better than 150 nT/Hz 1/2 between 0.1 and 10 Hz, which is a critical frequency range for scanning magnetic microscopy. This corresponds to a magnetic moment sensitivity of 10 –14  A m 2 , a factor of 100 better than achievable with typical commercial superconducting moment magnetometers. It also represents an improvement in sensitivity by a factor between 10 and 30 compared to similar scanning MTJ microscopes based on conventional bias-detection schemes. To demonstrate the capabilities of the instrument, two polished thin sections of representative geological samples were scanned along with a synthetic sample containing magnetic microparticles. The instrument is usable for a diversity of applications that require mapping of samples at room temperature to preserve magnetic properties or viability, including paleomagnetism and rock magnetism, nondestructive evaluation of materials, and biological assays. (paper)

  5. The neural elements in the lining of the ventricular-subventricular zone: making an old story new by high-resolution scanning electron microscopy

    Directory of Open Access Journals (Sweden)

    Carlos Alexandre Dos Santos Haemmerle

    2015-10-01

    Full Text Available The classical description of the neural elements that compose the lining of brain ventricles introduces us to the single layer of ependymal cells. However, new findings, especially in the lateral ventricle - the major niche for the generation of new neurons in the adult brain - have provided information about additional cell elements that influence the organization of this part of the ventricular system and produce important contributions to neurogenesis. To complement the cell neurochemistry findings, we present a three-dimensional in situ description that demonstrates the anatomical details of the different types of ciliated cells and the innervation of these elements. After processing adult rat brains for ultrastructural analysis by high-resolution scanning electron microscopy and transmission electron microscopy, we observed a heterogeneous pattern of cilia distribution at the different poles of the lateral ventricle surface. Furthermore, we describe the particular three-dimensional aspects of the ciliated cells of the lateral ventricle, in addition the fiber bundles and varicose axons surrounding these cells. Therefore, we provide a unique ultrastructural description of the three-dimensional in situ organization of the lateral ventricle surface, highlighting its innervation, to corroborate the available neurochemical and functional findings regarding the factors that regulate this neurogenic niche.

  6. Refinement procedure for the image alignment in high-resolution electron tomography.

    Science.gov (United States)

    Houben, L; Bar Sadan, M

    2011-01-01

    High-resolution electron tomography from a tilt series of transmission electron microscopy images requires an accurate image alignment procedure in order to maximise the resolution of the tomogram. This is the case in particular for ultra-high resolution where even very small misalignments between individual images can dramatically reduce the fidelity of the resultant reconstruction. A tomographic-reconstruction based and marker-free method is proposed, which uses an iterative optimisation of the tomogram resolution. The method utilises a search algorithm that maximises the contrast in tomogram sub-volumes. Unlike conventional cross-correlation analysis it provides the required correlation over a large tilt angle separation and guarantees a consistent alignment of images for the full range of object tilt angles. An assessment based on experimental reconstructions shows that the marker-free procedure is competitive to the reference of marker-based procedures at lower resolution and yields sub-pixel accuracy even for simulated high-resolution data. Copyright © 2011 Elsevier B.V. All rights reserved.

  7. Scanning photoelectron microscope for nanoscale three-dimensional spatial-resolved electron spectroscopy for chemical analysis.

    Science.gov (United States)

    Horiba, K; Nakamura, Y; Nagamura, N; Toyoda, S; Kumigashira, H; Oshima, M; Amemiya, K; Senba, Y; Ohashi, H

    2011-11-01

    In order to achieve nondestructive observation of the three-dimensional spatially resolved electronic structure of solids, we have developed a scanning photoelectron microscope system with the capability of depth profiling in electron spectroscopy for chemical analysis (ESCA). We call this system 3D nano-ESCA. For focusing the x-ray, a Fresnel zone plate with a diameter of 200 μm and an outermost zone width of 35 nm is used. In order to obtain the angular dependence of the photoelectron spectra for the depth-profile analysis without rotating the sample, we adopted a modified VG Scienta R3000 analyzer with an acceptance angle of 60° as a high-resolution angle-resolved electron spectrometer. The system has been installed at the University-of-Tokyo Materials Science Outstation beamline, BL07LSU, at SPring-8. From the results of the line-scan profiles of the poly-Si/high-k gate patterns, we achieved a total spatial resolution better than 70 nm. The capability of our system for pinpoint depth-profile analysis and high-resolution chemical state analysis is demonstrated. © 2011 American Institute of Physics

  8. A Simple Metric for Determining Resolution in Optical, Ion, and Electron Microscope Images.

    Science.gov (United States)

    Curtin, Alexandra E; Skinner, Ryan; Sanders, Aric W

    2015-06-01

    A resolution metric intended for resolution analysis of arbitrary spatially calibrated images is presented. By fitting a simple sigmoidal function to pixel intensities across slices of an image taken perpendicular to light-dark edges, the mean distance over which the light-dark transition occurs can be determined. A fixed multiple of this characteristic distance is then reported as the image resolution. The prefactor is determined by analysis of scanning transmission electron microscope high-angle annular dark field images of Si. This metric has been applied to optical, scanning electron microscope, and helium ion microscope images. This method provides quantitative feedback about image resolution, independent of the tool on which the data were collected. In addition, our analysis provides a nonarbitrary and self-consistent framework that any end user can utilize to evaluate the resolution of multiple microscopes from any vendor using the same metric.

  9. High-resolution observation by double-biprism electron holography

    International Nuclear Information System (INIS)

    Harada, Ken; Tonomura, Akira; Matsuda, Tsuyoshi; Akashi, Tetsuya; Togawa, Yoshihiko

    2004-01-01

    High-resolution electron holography has been achieved by using a double-biprism interferometer implemented on a 1 MV field emission electron microscope. The interferometer was installed behind the first magnifying lens to narrow carrier fringes and thus enabled complete separation of sideband Fourier spectrum from center band in reconstruction process. Holograms of Au fine particles and single-crystalline thin films with the finest fringe spacing of 4.2 pm were recorded and reconstructed. The overall holography system including the reconstruction process performed well for holograms in which carrier fringes had a spacing of around 10 pm. High-resolution lattice images of the amplitude and phase were clearly reconstructed without mixing of the center band and sideband information. Additionally, entire holograms were recorded without Fresnel fringes normally generated by the filament electrode of the biprism, and the holograms were thus reconstructed without the artifacts caused by Fresnel fringes

  10. A High Rigidity and Precision Scanning Tunneling Microscope with Decoupled XY and Z Scans.

    Science.gov (United States)

    Chen, Xu; Guo, Tengfei; Hou, Yubin; Zhang, Jing; Meng, Wenjie; Lu, Qingyou

    2017-01-01

    A new scan-head structure for the scanning tunneling microscope (STM) is proposed, featuring high scan precision and rigidity. The core structure consists of a piezoelectric tube scanner of quadrant type (for XY scans) coaxially housed in a piezoelectric tube with single inner and outer electrodes (for Z scan). They are fixed at one end (called common end). A hollow tantalum shaft is coaxially housed in the XY -scan tube and they are mutually fixed at both ends. When the XY scanner scans, its free end will bring the shaft to scan and the tip which is coaxially inserted in the shaft at the common end will scan a smaller area if the tip protrudes short enough from the common end. The decoupled XY and Z scans are desired for less image distortion and the mechanically reduced scan range has the superiority of reducing the impact of the background electronic noise on the scanner and enhancing the tip positioning precision. High quality atomic resolution images are also shown.

  11. A High Rigidity and Precision Scanning Tunneling Microscope with Decoupled XY and Z Scans

    Directory of Open Access Journals (Sweden)

    Xu Chen

    2017-01-01

    Full Text Available A new scan-head structure for the scanning tunneling microscope (STM is proposed, featuring high scan precision and rigidity. The core structure consists of a piezoelectric tube scanner of quadrant type (for XY scans coaxially housed in a piezoelectric tube with single inner and outer electrodes (for Z scan. They are fixed at one end (called common end. A hollow tantalum shaft is coaxially housed in the XY-scan tube and they are mutually fixed at both ends. When the XY scanner scans, its free end will bring the shaft to scan and the tip which is coaxially inserted in the shaft at the common end will scan a smaller area if the tip protrudes short enough from the common end. The decoupled XY and Z scans are desired for less image distortion and the mechanically reduced scan range has the superiority of reducing the impact of the background electronic noise on the scanner and enhancing the tip positioning precision. High quality atomic resolution images are also shown.

  12. Defining the limits and reliability of rigid-body fitting in cryo-EM maps using multi-scale image pyramids

    NARCIS (Netherlands)

    van Zundert, G. C P; Bonvin, A. M J J

    Cryo-electron microscopy provides fascinating structural insight into large macromolecular machines at increasing detail. Despite significant advances in the field, the resolution of the resulting three-dimensional images is still typically insufficient for . de novo model building. To bridge the

  13. Note: Electron energy spectroscopy mapping of surface with scanning tunneling microscope.

    Science.gov (United States)

    Li, Meng; Xu, Chunkai; Zhang, Panke; Li, Zhean; Chen, Xiangjun

    2016-08-01

    We report a novel scanning probe electron energy spectrometer (SPEES) which combines a double toroidal analyzer with a scanning tunneling microscope to achieve both topography imaging and electron energy spectroscopy mapping of surface in situ. The spatial resolution of spectroscopy mapping is determined to be better than 0.7 ± 0.2 μm at a tip sample distance of 7 μm. Meanwhile, the size of the field emission electron beam spot on the surface is also measured, and is about 3.6 ± 0.8 μm in diameter. This unambiguously demonstrates that the spatial resolution of SPEES technique can be much better than the size of the incident electron beam.

  14. Note: Electron energy spectroscopy mapping of surface with scanning tunneling microscope

    Energy Technology Data Exchange (ETDEWEB)

    Li, Meng; Xu, Chunkai, E-mail: xuck@ustc.edu.cn, E-mail: xjun@ustc.edu.cn; Zhang, Panke; Li, Zhean; Chen, Xiangjun, E-mail: xuck@ustc.edu.cn, E-mail: xjun@ustc.edu.cn [Hefei National Laboratory for Physical Science at Microscale and Department of Modern Physics, University of Science and Technology of China, Hefei 230026, China and Synergetic Innovation Center of Quantum Information and Quantum Physics, University of Science and Technology of China, Hefei 230026 (China)

    2016-08-15

    We report a novel scanning probe electron energy spectrometer (SPEES) which combines a double toroidal analyzer with a scanning tunneling microscope to achieve both topography imaging and electron energy spectroscopy mapping of surface in situ. The spatial resolution of spectroscopy mapping is determined to be better than 0.7 ± 0.2 μm at a tip sample distance of 7 μm. Meanwhile, the size of the field emission electron beam spot on the surface is also measured, and is about 3.6 ± 0.8 μm in diameter. This unambiguously demonstrates that the spatial resolution of SPEES technique can be much better than the size of the incident electron beam.

  15. Note: Electron energy spectroscopy mapping of surface with scanning tunneling microscope

    International Nuclear Information System (INIS)

    Li, Meng; Xu, Chunkai; Zhang, Panke; Li, Zhean; Chen, Xiangjun

    2016-01-01

    We report a novel scanning probe electron energy spectrometer (SPEES) which combines a double toroidal analyzer with a scanning tunneling microscope to achieve both topography imaging and electron energy spectroscopy mapping of surface in situ. The spatial resolution of spectroscopy mapping is determined to be better than 0.7 ± 0.2 μm at a tip sample distance of 7 μm. Meanwhile, the size of the field emission electron beam spot on the surface is also measured, and is about 3.6 ± 0.8 μm in diameter. This unambiguously demonstrates that the spatial resolution of SPEES technique can be much better than the size of the incident electron beam.

  16. Minimum detection limit and spatial resolution of thin-sample field-emission electron probe microanalysis

    International Nuclear Information System (INIS)

    Kubo, Yugo; Hamada, Kotaro; Urano, Akira

    2013-01-01

    The minimum detection limit and spatial resolution for a thinned semiconductor sample were determined by electron probe microanalysis (EPMA) using a Schottky field emission (FE) electron gun and wavelength dispersive X-ray spectrometry. Comparison of the FE-EPMA results with those obtained using energy dispersive X-ray spectrometry in conjunction with scanning transmission electron microscopy, confirmed that FE-EPMA is largely superior in terms of detection sensitivity. Thin-sample FE-EPMA is demonstrated as a very effective method for high resolution, high sensitivity analysis in a laboratory environment because a high probe current and high signal-to-noise ratio can be achieved. - Highlights: • Minimum detection limit and spatial resolution determined for FE-EPMA. • Detection sensitivity of FE-EPMA greatly superior to that of STEM-EDX. • Minimum detection limit and spatial resolution controllable by probe current

  17. Observations of localised dielectric excitations, secondary events and ionisation damage by scanning transmission electron microscopy

    International Nuclear Information System (INIS)

    Howie, A.

    1988-01-01

    In the scanning transmission electron microscope (STEM) a high intensity /approximately/0.5nm diameter, probe of 100 keV electrons is formed. This can be positioned to collect energy loss spectra from surfaces, interfaces, small spheres or other particles at controlled values of impact parameter or can be scanned across the object (usually a thin film) to produce high resolution images formed from a variety of signals - small angle or large angle (Z contrast) elastic scattering, inelastic scattering (both valence and core losses), secondary electron emission and x-ray or optical photon emission. The high spatial resolution achievable in a variety of simple structures raises many unsolved theoretical problems concerning the generation, propagation and decay of excitations in inhomogeneous media. These range from quite well posed problems in the mathematical physics of dielectric excitation to problems of plasmon propagation and rather more exotic and less well understood problems of radiation damage. 15 refs., 4 figs

  18. Cryo-SEM studies of latex/ceramic nanoparticle coating microstructure development.

    Science.gov (United States)

    Luo, Hui; Scriven, L E; Francis, Lorraine F

    2007-12-15

    Cryogenic scanning electron microscopy (cryo-SEM) was used to investigate microstructure development of composite coatings prepared from dispersions of antimony-doped tin oxide (ATO) nanoparticles (approximately 30 nm) or indium tin oxide (ITO) nanoparticles (approximately 40 nm) and latex particles (polydisperse, D(v): approximately 300 nm). Cryo-SEM images of ATO/latex dispersions as-frozen show small clusters of ATO and individual latex particles homogeneously distribute in a frozen water matrix. In contrast, cryo-SEM images of ITO/latex dispersions as-frozen show ITO particles adsorb onto latex particle surfaces. Electrostatic repulsion between negatively charged ATO and negatively charged latex particles stabilizes the ATO/latex dispersion, whereas in ITO/latex dispersion, positively charged ITO particles are attracted onto surfaces of negatively charged latex particles. These results are consistent with calculations of interaction potentials from past research. Cryo-SEM images of frozen and fractured coatings reveal that both ceramic nanoparticles and latex become more concentrated as drying proceeds; larger latex particles consolidate with ceramic nanoparticles in the interstitial spaces. With more drying, compaction flattens the latex-latex particle contacts and shrinks the voids between them. Thus, ceramic nanoparticles are forced to pack closely in the interstitial spaces, forming an interconnected network. Finally, latex particles partially coalesce at their flattened contacts, thereby yielding a coherent coating. The research reveals how nanoparticles segregate and interconnect among latex particles during drying.

  19. Simultaneous Confocal Scanning Laser Ophthalmoscopy Combined with High-Resolution Spectral-Domain Optical Coherence Tomography: A Review

    Directory of Open Access Journals (Sweden)

    Verônica Castro Lima

    2011-01-01

    Full Text Available We aimed to evaluate technical aspects and the clinical relevance of a simultaneous confocal scanning laser ophthalmoscope and a high-speed, high-resolution, spectral-domain optical coherence tomography (SDOCT device for retinal imaging. The principle of confocal scanning laser imaging provides a high resolution of retinal and choroidal vasculature with low light exposure. Enhanced contrast, details, and image sharpness are generated using confocality. The real-time SDOCT provides a new level of accuracy for assessment of the angiographic and morphological correlation. The combined system allows for simultaneous recordings of topographic and tomographic images with accurate correlation between them. Also it can provide simultaneous multimodal imaging of retinal pathologies, such as fluorescein and indocyanine green angiographies, infrared and blue reflectance (red-free images, fundus autofluorescence images, and OCT scans (Spectralis HRA + OCT; Heidelberg Engineering, Heidelberg, Germany. The combination of various macular diagnostic tools can lead to a better understanding and improved knowledge of macular diseases.

  20. Big data in cryoEM: automated collection, processing and accessibility of EM data.

    Science.gov (United States)

    Baldwin, Philip R; Tan, Yong Zi; Eng, Edward T; Rice, William J; Noble, Alex J; Negro, Carl J; Cianfrocco, Michael A; Potter, Clinton S; Carragher, Bridget

    2018-06-01

    The scope and complexity of cryogenic electron microscopy (cryoEM) data has greatly increased, and will continue to do so, due to recent and ongoing technical breakthroughs that have led to much improved resolutions for macromolecular structures solved using this method. This big data explosion includes single particle data as well as tomographic tilt series, both generally acquired as direct detector movies of ∼10-100 frames per image or per tilt-series. We provide a brief survey of the developments leading to the current status, and describe existing cryoEM pipelines, with an emphasis on the scope of data acquisition, methods for automation, and use of cloud storage and computing. Copyright © 2017 Elsevier Ltd. All rights reserved.

  1. Characterisation of radiation damage in perovskite using high angular resolution electron channeling x-ray spectroscopy (HARECXS)

    International Nuclear Information System (INIS)

    Smith, K.L.; Zaluzec, N.J.

    2002-01-01

    Full text: Predicting and/or modelling the occurrence of radiation damage induced defects and their effects on physical properties (eg. amorphisation induced swelling, electrical conductivity., optical response etc.) in ceramic phases requires knowledge of the displacement energies, E d , of cations and anions in those phases. In this study, High Angular Resolution Electron Channelling X-ray Spectroscopy (HARECXS) spectra were collected from perovskite (CaTiO 3 ) samples that had been exposed to high-energy electrons or high-energy heavy ions. Calculations based on experimental data were then used to indicate the E d of the cations in perovskite. The HARECXS measurements were conducted on a Philips EM 420T AEM (LaB6 source, operated at 120 kV) fitted with an EDAX ultra thin window Si(Li) detector. The specimen was first manually oriented to an appropriate zone axis. Then control of the relative orientation of the incident probe was accomplished via direct computer control of the beam tilt coils, Typical acquisition times for a complete two-dimensional scan were 18-24 hours, while one dimensional scans ranged from 1-5 hours. Our experiments established that: a) HARECXS can detect radiation damage in perovskite caused by either high energy heavy ions or high energy electrons, b) the HARECXS signature of perovskite shows a systematic change with ion dose, c) HARECXS detects damage in perovskite that has been irradiated with 900kV electrons and does not detect damage in perovskite that has been irradiated with 620kV electrons, indicating the existance of an electron irradiation damage threshold. Calculations based on the latter results indicate that the displacement energy, E d of calcium and titanium in perovskite lie between 50 and 85eV. Copyright (2002) Australian Society for Electron Microscopy Inc

  2. Scanning tunnel microscope with large vision field compatible with a scanning electron microscope

    International Nuclear Information System (INIS)

    Volodin, A.P.; Stepanyan, G.A.; Khajkin, M.S.; Ehdel'man, V.S.

    1989-01-01

    A scanning tunnel microscope (STM) with the 20μm vision field and 1nm resolution, designed to be compatible with a scanning electron microscope (SEM), is described. The sample scanning area is chosen within the 3x10mm limits with a 0.1-1μm step. The STM needle is moved automatically toward the sample surface from the maximum distance of 10mm until the tunneling current appears. Bimorphous elements of the KP-1 piezocorrector are used in the STM design. The device is installed on a table of SEM object holders

  3. High resolution STEM of quantum dots and quantum wires

    DEFF Research Database (Denmark)

    Kadkhodazadeh, Shima

    2013-01-01

    This article reviews the application of high resolution scanning transmission electron microscopy (STEM) to semiconductor quantum dots (QDs) and quantum wires (QWRs). Different imaging and analytical techniques in STEM are introduced and key examples of their application to QDs and QWRs...

  4. On the optical stability of high-resolution transmission electron microscopes

    International Nuclear Information System (INIS)

    Barthel, J.; Thust, A.

    2013-01-01

    In the recent two decades the technique of high-resolution transmission electron microscopy experienced an unprecedented progress through the introduction of hardware aberration correctors and by the improvement of the achievable resolution to the sub-Ångström level. The important aspect that aberration correction at a given resolution requires also a well defined amount of optical stability has received little attention so far. Therefore we investigate the qualification of a variety of high-resolution electron microscopes to maintain an aberration corrected optical state in terms of an optical lifetime. We develop a comprehensive statistical framework for the estimation of the optical lifetime and find remarkably low values between tens of seconds and a couple of minutes. Probability curves are introduced, which inform the operator about the chance to work still in the fully aberration corrected state. - Highlights: • We investigate the temporal stability of optical aberrations in HRTEM. • We develop a statistical framework for the estimation of optical lifetimes. • We introduce plots showing the success probability for aberration-free work. • Optical lifetimes in sub-Ångström electron microscopy are surprisingly low. • The success of aberration correction depends strongly on the optical stability

  5. Characterizing deformed ultrafine-grained and nanocrystalline materials using transmission Kikuchi diffraction in a scanning electron microscope

    International Nuclear Information System (INIS)

    Trimby, Patrick W.; Cao, Yang; Chen, Zibin; Han, Shuang; Hemker, Kevin J.; Lian, Jianshe; Liao, Xiaozhou; Rottmann, Paul; Samudrala, Saritha; Sun, Jingli; Wang, Jing Tao; Wheeler, John; Cairney, Julie M.

    2014-01-01

    Graphical abstract: -- Abstract: The recent development of transmission Kikuchi diffraction (TKD) in a scanning electron microscope enables fast, automated orientation mapping of electron transparent samples using standard electron backscatter diffraction (EBSD) hardware. TKD in a scanning electron microscope has significantly better spatial resolution than conventional EBSD, enabling routine characterization of nanocrystalline materials and allowing effective measurement of samples that have undergone severe plastic deformation. Combining TKD with energy dispersive X-ray spectroscopy (EDS) provides complementary chemical information, while a standard forescatter detector system below the EBSD detector can be used to generate dark field and oriented dark field images. Here we illustrate the application of this exciting new approach to a range of deformed, ultrafine grained and nanocrystalline samples, including duplex stainless steel, nanocrystalline copper and highly deformed titanium and nickel–cobalt. The results show that TKD combined with EDS is a highly effective and widely accessible tool for measuring key microstructural parameters at resolutions that are inaccessible using conventional EBSD

  6. Three-Dimensional scanning transmission electron microscopy of biological specimens

    KAUST Repository

    De Jonge, Niels

    2010-01-18

    A three-dimensional (3D) reconstruction of the cytoskeleton and a clathrin-coated pit in mammalian cells has been achieved from a focal-series of images recorded in an aberration-corrected scanning transmission electron microscope (STEM). The specimen was a metallic replica of the biological structure comprising Pt nanoparticles 2-3 nm in diameter, with a high stability under electron beam radiation. The 3D dataset was processed by an automated deconvolution procedure. The lateral resolution was 1.1 nm, set by pixel size. Particles differing by only 10 nm in vertical position were identified as separate objects with greater than 20% dip in contrast between them. We refer to this value as the axial resolution of the deconvolution or reconstruction, the ability to recognize two objects, which were unresolved in the original dataset. The resolution of the reconstruction is comparable to that achieved by tilt-series transmission electron microscopy. However, the focal-series method does not require mechanical tilting and is therefore much faster. 3D STEM images were also recorded of the Golgi ribbon in conventional thin sections containing 3T3 cells with a comparable axial resolution in the deconvolved dataset. © 2010 Microscopy Society of America.

  7. Electron Bio-Imaging Centre (eBIC): the UK national research facility for biological electron microscopy.

    Science.gov (United States)

    Clare, Daniel K; Siebert, C Alistair; Hecksel, Corey; Hagen, Christoph; Mordhorst, Valerie; Grange, Michael; Ashton, Alun W; Walsh, Martin A; Grünewald, Kay; Saibil, Helen R; Stuart, David I; Zhang, Peijun

    2017-06-01

    The recent resolution revolution in cryo-EM has led to a massive increase in demand for both time on high-end cryo-electron microscopes and access to cryo-electron microscopy expertise. In anticipation of this demand, eBIC was set up at Diamond Light Source in collaboration with Birkbeck College London and the University of Oxford, and funded by the Wellcome Trust, the UK Medical Research Council (MRC) and the Biotechnology and Biological Sciences Research Council (BBSRC) to provide access to high-end equipment through peer review. eBIC is currently in its start-up phase and began by offering time on a single FEI Titan Krios microscope equipped with the latest generation of direct electron detectors from two manufacturers. Here, the current status and modes of access for potential users of eBIC are outlined. In the first year of operation, 222 d of microscope time were delivered to external research groups, with 95 visits in total, of which 53 were from unique groups. The data collected have generated multiple high- to intermediate-resolution structures (2.8-8 Å), ten of which have been published. A second Krios microscope is now in operation, with two more due to come online in 2017. In the next phase of growth of eBIC, in addition to more microscope time, new data-collection strategies and sample-preparation techniques will be made available to external user groups. Finally, all raw data are archived, and a metadata catalogue and automated pipelines for data analysis are being developed.

  8. Practical considerations for high spatial and temporal resolution dynamic transmission electron microscopy

    Energy Technology Data Exchange (ETDEWEB)

    Armstrong, Michael R. [Materials Science and Technology Division, Chemistry and Materials Science Directorate, Lawrence Livermore National Laboratory, P.O. Box 808, L-356, Livermore, CA 94550 (United States)], E-mail: armstrong30@llnl.gov; Boyden, Ken [Materials Science and Technology Division, Chemistry and Materials Science Directorate, Lawrence Livermore National Laboratory, P.O. Box 808, L-356, Livermore, CA 94550 (United States); Browning, Nigel D. [Materials Science and Technology Division, Chemistry and Materials Science Directorate, Lawrence Livermore National Laboratory, P.O. Box 808, L-356, Livermore, CA 94550 (United States); Department of Chemical Engineering and Materials Science, University of California-Davis, One Shields Avenue, Davis, CA 95616 (United States); Campbell, Geoffrey H.; Colvin, Jeffrey D.; De Hope, William J.; Frank, Alan M. [Materials Science and Technology Division, Chemistry and Materials Science Directorate, Lawrence Livermore National Laboratory, P.O. Box 808, L-356, Livermore, CA 94550 (United States); Gibson, David J.; Hartemann, Fred [N Division, Physics and Advanced Technologies Directorate, Lawrence Livermore National Laboratory, P.O. Box 808, L-280, Livermore, CA 94550 (United States); Kim, Judy S. [Materials Science and Technology Division, Chemistry and Materials Science Directorate, Lawrence Livermore National Laboratory, P.O. Box 808, L-356, Livermore, CA 94550 (United States); Department of Chemical Engineering and Materials Science, University of California-Davis, One Shields Avenue, Davis, CA 95616 (United States); King, Wayne E.; La Grange, Thomas B.; Pyke, Ben J.; Reed, Bryan W.; Shuttlesworth, Richard M.; Stuart, Brent C.; Torralva, Ben R. [Materials Science and Technology Division, Chemistry and Materials Science Directorate, Lawrence Livermore National Laboratory, P.O. Box 808, L-356, Livermore, CA 94550 (United States)

    2007-04-15

    Although recent years have seen significant advances in the spatial resolution possible in the transmission electron microscope (TEM), the temporal resolution of most microscopes is limited to video rate at best. This lack of temporal resolution means that our understanding of dynamic processes in materials is extremely limited. High temporal resolution in the TEM can be achieved, however, by replacing the normal thermionic or field emission source with a photoemission source. In this case the temporal resolution is limited only by the ability to create a short pulse of photoexcited electrons in the source, and this can be as short as a few femtoseconds. The operation of the photo-emission source and the control of the subsequent pulse of electrons (containing as many as 5x10{sup 7} electrons) create significant challenges for a standard microscope column that is designed to operate with a single electron in the column at any one time. In this paper, the generation and control of electron pulses in the TEM to obtain a temporal resolution <10{sup -6} s will be described and the effect of the pulse duration and current density on the spatial resolution of the instrument will be examined. The potential of these levels of temporal and spatial resolution for the study of dynamic materials processes will also be discussed.

  9. Practical considerations for high spatial and temporal resolution dynamic transmission electron microscopy

    International Nuclear Information System (INIS)

    Armstrong, Michael R.; Boyden, Ken; Browning, Nigel D.; Campbell, Geoffrey H.; Colvin, Jeffrey D.; De Hope, William J.; Frank, Alan M.; Gibson, David J.; Hartemann, Fred; Kim, Judy S.; King, Wayne E.; La Grange, Thomas B.; Pyke, Ben J.; Reed, Bryan W.; Shuttlesworth, Richard M.; Stuart, Brent C.; Torralva, Ben R.

    2007-01-01

    Although recent years have seen significant advances in the spatial resolution possible in the transmission electron microscope (TEM), the temporal resolution of most microscopes is limited to video rate at best. This lack of temporal resolution means that our understanding of dynamic processes in materials is extremely limited. High temporal resolution in the TEM can be achieved, however, by replacing the normal thermionic or field emission source with a photoemission source. In this case the temporal resolution is limited only by the ability to create a short pulse of photoexcited electrons in the source, and this can be as short as a few femtoseconds. The operation of the photo-emission source and the control of the subsequent pulse of electrons (containing as many as 5x10 7 electrons) create significant challenges for a standard microscope column that is designed to operate with a single electron in the column at any one time. In this paper, the generation and control of electron pulses in the TEM to obtain a temporal resolution -6 s will be described and the effect of the pulse duration and current density on the spatial resolution of the instrument will be examined. The potential of these levels of temporal and spatial resolution for the study of dynamic materials processes will also be discussed

  10. APES: Acute Precipitating Electron Spectrometer - A High Time Resolution Monodirectional Magnetic Deflection Electron Spectrometer

    Science.gov (United States)

    Michell, R. G.; Samara, M.; Grubbs, G., II; Ogasawara, K.; Miller, G.; Trevino, J. A.; Webster, J.; Stange, J.

    2016-01-01

    We present a description of the Acute Precipitating Electron Spectrometer (APES) that was designed and built for the Ground-to-Rocket Electron Electrodynamics Correlative Experiment (GREECE) auroral sounding rocket mission. The purpose was to measure the precipitating electron spectrum with high time resolution, on the order of milliseconds. The trade-off made in order to achieve high time resolution was to limit the aperture to only one look direction. The energy selection was done by using a permanent magnet to separate the incoming electrons, such that the different energies would fall onto different regions of the microchannel plate and therefore be detected by different anodes. A rectangular microchannel plate (MCP) was used (15 mm x 100 mm), and there was a total of 50 discrete anodes under the MCP, each one 15 mm x 1.5 mm, with a 0.5 mm spacing between anodes. The target energy range of APES was 200 eV to 30 keV.

  11. Quantum ballistic transistor and low noise HEMT for cryo-electronics lower than 4.2 K; Transistor balistique quantique et HEMT bas-bruit pour la cryoelectronique inferieure a 4.2 K

    Energy Technology Data Exchange (ETDEWEB)

    Gremion, E

    2008-01-15

    Next generations of cryo-detectors, widely used in physics of particles and physics of universe, will need in the future high-performance cryo-electronics less noisy and closer to the detector. Within this context, this work investigates properties of two dimensional electron gas GaAlAs/GaAs by studying two components, quantum point contact (QPC) and high electron mobility transistor (HEMT). Thanks to quantized conductance steps in QPC, we have realized a quantum ballistic transistor (voltage gain higher than 1), a new component useful for cryo-electronics thanks to its operating temperature and weak power consumption (about 1 nW). Moreover, the very low capacity of this component leads to promising performances for multiplexing low temperature bolometer dedicated to millimetric astronomy. The second study focused on HEMT with very high quality 2DEG. At 4.2 K, a voltage gain higher than 20 can be obtained with a very low power dissipation of less than 100 {mu}W. Under the above experimental conditions, an equivalent input voltage noise of 1.2 nV/{radical}(Hz) at 1 kHz and 0.12 nV/{radical}(Hz) at 100 kHz has been reached. According to the Hooge formula, these noise performances are get by increasing gate capacity estimated to 60 pF. (author)

  12. Airborne campaigns for CryoSat pre-launch calibration and validation

    DEFF Research Database (Denmark)

    Hvidegaard, Sine Munk; Forsberg, René; Skourup, Henriette

    2010-01-01

    From 2003 to 2008 DTU Space together with ESA and several international partners carried out airborne and ground field campaigns in preparation for CryoSat validation; called CryoVEx: CryoSat Validation Experiments covering the main ice caps in Greenland, Canada and Svalbard and sea ice in the Ar......From 2003 to 2008 DTU Space together with ESA and several international partners carried out airborne and ground field campaigns in preparation for CryoSat validation; called CryoVEx: CryoSat Validation Experiments covering the main ice caps in Greenland, Canada and Svalbard and sea ice...... in the Arctic Ocean. The main goal of the airborne surveys was to acquire coincident scanning laser and CryoSat type radar elevation measurements of the surface; either sea ice or land ice. Selected lines have been surveyed along with detailed mapping of validation sites coordinated with insitu field work...... and helicopter electromagnetic surveying. This paper summarises the pre-launch campaigns and presents some of the result from the coincident measurement from airborne and ground observations....

  13. Structure Identification in High-Resolution Transmission Electron Microscopic Images

    DEFF Research Database (Denmark)

    Vestergaard, Jacob Schack; Kling, Jens; Dahl, Anders Bjorholm

    2014-01-01

    A connection between microscopic structure and macroscopic properties is expected for almost all material systems. High-resolution transmission electron microscopy is a technique offering insight into the atomic structure, but the analysis of large image series can be time consuming. The present ...

  14. HIGH RESOLUTION AIRBORNE LASER SCANNING AND HYPERSPECTRAL IMAGING WITH A SMALL UAV PLATFORM

    Directory of Open Access Journals (Sweden)

    M. Gallay

    2016-06-01

    Full Text Available The capabilities of unmanned airborne systems (UAS have become diverse with the recent development of lightweight remote sensing instruments. In this paper, we demonstrate our custom integration of the state-of-the-art technologies within an unmanned aerial platform capable of high-resolution and high-accuracy laser scanning, hyperspectral imaging, and photographic imaging. The technological solution comprises the latest development of a completely autonomous, unmanned helicopter by Aeroscout, the Scout B1-100 UAV helicopter. The helicopter is powered by a gasoline two-stroke engine and it allows for integrating 18 kg of a customized payload unit. The whole system is modular providing flexibility of payload options, which comprises the main advantage of the UAS. The UAS integrates two kinds of payloads which can be altered. Both payloads integrate a GPS/IMU with a dual GPS antenna configuration provided by OXTS for accurate navigation and position measurements during the data acquisition. The first payload comprises a VUX-1 laser scanner by RIEGL and a Sony A6000 E-Mount photo camera. The second payload for hyperspectral scanning integrates a push-broom imager AISA KESTREL 10 by SPECIM. The UAS was designed for research of various aspects of landscape dynamics (landslides, erosion, flooding, or phenology in high spectral and spatial resolution.

  15. High-resolution EELS investigation of the electronic structure of ilmenites

    NARCIS (Netherlands)

    Radtke, G.; Lazar, S.; Botton, G.A.

    2006-01-01

    The electronic structure of a series of compounds belonging to the ilmenite family is investigated using high resolution electron energy loss spectroscopy (EELS). The energy loss near edge structure (ELNES) of the O-K, Ti-L23 and transition metal L23 edges have been recorded in MnTiO3, FeTiO3,

  16. CryoEM at IUCrJ: a new era

    Directory of Open Access Journals (Sweden)

    Sriram Subramaniam

    2016-01-01

    Full Text Available In this overview, we briefly outline recent advances in electron cryomicroscopy (cryoEM and explain why the journal IUCrJ, published by the International Union of Crystallography, could provide a natural home for publications covering many present and future developments in the cryoEM field.

  17. Aberration-corrected scanning transmission electron microscopy of semiconductors

    International Nuclear Information System (INIS)

    Krivanek, O L; Dellby, N; Murfitt, M F

    2011-01-01

    The scanning transmission electron microscope (STEM) has been able to image individual heavy atoms in a light matrix for some time. It is now able to do much more: it can resolve individual atoms as light as boron in monolayer materials; image atomic columns as light as hydrogen, identify the chemical type of individual isolated atoms from the intensity of their annular dark field (ADF) image and by electron energy loss spectroscopy (EELS); and map elemental composition at atomic resolution by EELS and energy-dispersive X-ray spectroscopy (EDXS). It can even map electronic states, also by EELS, at atomic resolution. The instrumentation developments that have made this level of performance possible are reviewed, and examples of applications to semiconductors and oxides are shown.

  18. Applications of molecules as high-resolution, high-sensitivity threshold electron detectors

    International Nuclear Information System (INIS)

    Chutjian, A.

    1991-01-01

    The goal of the work under the contract entitled ''Applications of Molecules as High-Resolution, High-Sensitivity Threshold Electron Detectors'' (DoE IAA No. DE-AI01-83ER13093 Mod. A006) was to explore the electron attachment properties of a variety of molecules at electron energies not accessible by other experimental techniques. As a result of this work, not only was a large body of basic data measured on attachment cross sections and rate constants; but also extensive theoretical calculations were carried out to verify the underlying phenomenon of s-wave attachment. Important outgrowths of this week were also realized in other areas of research. The basic data have applications in fields such as combustion, soot reduction, rocket-exhaust modification, threshold photoelectron spectroscopy, and trace species detection

  19. Cryo-electron microscopy structure of the lysosomal calcium-permeable channel TRPML3.

    Science.gov (United States)

    Hirschi, Marscha; Herzik, Mark A; Wie, Jinhong; Suo, Yang; Borschel, William F; Ren, Dejian; Lander, Gabriel C; Lee, Seok-Yong

    2017-10-19

    The modulation of ion channel activity by lipids is increasingly recognized as a fundamental component of cellular signalling. The transient receptor potential mucolipin (TRPML) channel family belongs to the TRP superfamily and is composed of three members: TRPML1-TRPML3. TRPMLs are the major Ca 2+ -permeable channels on late endosomes and lysosomes (LEL). They regulate the release of Ca 2+ from organelles, which is important for various physiological processes, including organelle trafficking and fusion. Loss-of-function mutations in the MCOLN1 gene, which encodes TRPML1, cause the neurodegenerative lysosomal storage disorder mucolipidosis type IV, and a gain-of-function mutation (Ala419Pro) in TRPML3 gives rise to the varitint-waddler (Va) mouse phenotype. Notably, TRPML channels are activated by the low-abundance and LEL-enriched signalling lipid phosphatidylinositol-3,5-bisphosphate (PtdIns(3,5)P 2 ), whereas other phosphoinositides such as PtdIns(4,5)P 2 , which is enriched in plasma membranes, inhibit TRPMLs. Conserved basic residues at the N terminus of the channel are important for activation by PtdIns(3,5)P 2 and inhibition by PtdIns(4,5)P 2 . However, owing to a lack of structural information, the mechanism by which TRPML channels recognize PtdIns(3,5)P 2 and increase their Ca 2+ conductance remains unclear. Here we present the cryo-electron microscopy (cryo-EM) structure of a full-length TRPML3 channel from the common marmoset (Callithrix jacchus) at an overall resolution of 2.9 Å. Our structure reveals not only the molecular basis of ion conduction but also the unique architecture of TRPMLs, wherein the voltage sensor-like domain is linked to the pore via a cytosolic domain that we term the mucolipin domain. Combined with functional studies, these data suggest that the mucolipin domain is responsible for PtdIns(3,5)P 2 binding and subsequent channel activation, and that it acts as a 'gating pulley' for lipid-dependent TRPML gating.

  20. Scanning Auger Electron Microscope

    Data.gov (United States)

    Federal Laboratory Consortium — A JEOL model 7830F field emission source, scanning Auger microscope.Specifications / Capabilities:Ultra-high vacuum (UHV), electron gun range from 0.1 kV to 25 kV,...

  1. High-resolution electron microscopy and its applications.

    Science.gov (United States)

    Li, F H

    1987-12-01

    A review of research on high-resolution electron microscopy (HREM) carried out at the Institute of Physics, the Chinese Academy of Sciences, is presented. Apart from the direct observation of crystal and quasicrystal defects for some alloys, oxides, minerals, etc., and the structure determination for some minute crystals, an approximate image-contrast theory named pseudo-weak-phase object approximation (PWPOA), which shows the image contrast change with crystal thickness, is described. Within the framework of PWPOA, the image contrast of lithium ions in the crystal of R-Li2Ti3O7 has been observed. The usefulness of diffraction analysis techniques such as the direct method and Patterson method in HREM is discussed. Image deconvolution and resolution enhancement for weak-phase objects by use of the direct method are illustrated. In addition, preliminary results of image restoration for thick crystals are given.

  2. Construction of a high resolution electron beam profile monitor

    International Nuclear Information System (INIS)

    Norem, J.; Dawson, J.; Haberichter, W.; Novak, W.; Reed, L.; Yang, X.F.

    1993-01-01

    Bremsstrahlung from an electron beam on a heavy target can be used to image the beam profile using collimators and slits. The limiting resolution using this system is determined by Fresnel diffraction, and is ∼ √(λd/2), where λ is the photon wavelength and d is determined by the linear dimensions of the system. For linear colliders this resolution could be a few nm. The highest resolution requires detectors which see only high energy, (small λ), photons, and this is accomplished by converting photons to pairs, and detecting Cherenkov light in a nearly forward angle with a CCD detector or streak camera. Tests are planned at the Argonne APS and SLAC FFTB

  3. Annular dark field electron microscope images with better than 2 A resolution at 100 kV

    International Nuclear Information System (INIS)

    Shin, D.H.; Kirkland, E.J.; Silcox, J.

    1989-01-01

    High-resolution scanning transmission electron microscope (STEM) images in the annular dark field (ADF) imaging mode approaching the theoretical point-to-point resolution limit are presented. The ADF images were obtained from a high T c superconducting YBa 2 Cu 3 O 7-x thin-film specimen at 100 kV. The 1.9 A resolution lattice image, which is the smallest lattice spacing in the specimen, corresponds to the minimum resolvable spatial frequency with 5% contrast in the contrast transfer function for annular dark field, and is smaller than the resolution limit given by the Rayleigh criterion. This demonstrates that STEM ADF imaging can have a resolution approximately 40% better than that of the bright field conventional transmission electron microscope (CTEM) imaging at Scherzer condition

  4. Diagnosis of hearing impairment by high resolution CT scanning of inner ear anomalies

    International Nuclear Information System (INIS)

    Murata, Kiyotaka; Isono, Michio; Ohta, Fumihiko

    1988-01-01

    High resolution CT scanning of the temporal bone in our clinic has provided a more detailed radiological classification of inner ear anomalies than before. The statistical analysis of inner ear malformations based on the theory of quantification II has produced discriminant equations for the measurable diagnosis of hearing impairment and development of the inner ear. This analysis may make it possible to diagnose total and partial deafness on ipsi- and contralateral sides. (author)

  5. Long helical filaments are not seen encircling cells in electron cryotomograms of rod-shaped bacteria

    International Nuclear Information System (INIS)

    Swulius, Matthew T.; Chen, Songye; Jane Ding, H.; Li, Zhuo; Briegel, Ariane; Pilhofer, Martin; Tocheva, Elitza I.; Lybarger, Suzanne R.; Johnson, Tanya L.; Sandkvist, Maria; Jensen, Grant J.

    2011-01-01

    Highlights: → No long helical filaments are seen near or along rod-shaped bacterial inner membranes by electron cryo-tomography. → Electron cryo-tomography has the resolution to detect single filaments in vivo. -- Abstract: How rod-shaped bacteria form and maintain their shape is an important question in bacterial cell biology. Results from fluorescent light microscopy have led many to believe that the actin homolog MreB and a number of other proteins form long helical filaments along the inner membrane of the cell. Here we show using electron cryotomography of six different rod-shaped bacterial species, at macromolecular resolution, that no long (>80 nm) helical filaments exist near or along either surface of the inner membrane. We also use correlated cryo-fluorescent light microscopy (cryo-fLM) and electron cryo-tomography (ECT) to identify cytoplasmic bundles of MreB, showing that MreB filaments are detectable by ECT. In light of these results, the structure and function of MreB must be reconsidered: instead of acting as a large, rigid scaffold that localizes cell-wall synthetic machinery, moving MreB complexes may apply tension to growing peptidoglycan strands to ensure their orderly, linear insertion.

  6. The impact of irradiation induced specimen charging on microanalysis in a scanning electron microscope

    International Nuclear Information System (INIS)

    Stevens-Kalceff, M.A.

    2003-01-01

    Full text: It is necessary to assess and characterize the perturbing influences of experimental probes on the specimens under investigation. The significant influence of electron beam irradiation on poorly conducting materials has been assessed by a combination of specialized analytical scanning electron and scanning probe microscopy techniques including Cathodoluminescence Microanalysis and Kelvin Probe Microscopy. These techniques enable the defect structure and the residual charging of materials to be characterized at high spatial resolution. Cathodoluminescence is the non-incandescent emission of light resulting from the electron irradiation. CL microscopy and spectroscopy in a Scanning Electron Microscope (SEM) enables high spatial resolution and high sensitivity detection of defects in poorly conducting materials. Local variations in the distribution of defects can be non-destructively characterized with high spatial (lateral and depth) resolution by adjusting electron beam parameters to select the specimen micro-volume of interest. Kelvin Probe Microscopy (KPM) is a Scanning Probe Microscopy technique in which long-range Coulomb forces between a conductive atomic force probe and the specimen enable the surface potential to be characterized with high spatial resolution. A combination of Kelvin Probe Microscopy (KPM) and Cathodoluminescence (CL) microanalysis has been used to characterize ultra pure silicon dioxide exposed to electron irradiation in a Scanning Electron Microscope. Silicon dioxide is an excellent model specimen with which to investigate charging induced effects. It is a very poor electrical conductor, homogeneous and electron irradiation produces easily identifiable surface modification which enables irradiated regions to be easily and unambiguously located. A conductive grounded coating is typically applied to poorly conducting specimens prior to investigation in an SEM to prevent deflection of the electron beam and surface charging, however

  7. 4.0-A resolution cryo-EM structure of the mammalian chaperonin TRiC/CCT reveals its unique subunit arrangement.

    Science.gov (United States)

    Cong, Yao; Baker, Matthew L; Jakana, Joanita; Woolford, David; Miller, Erik J; Reissmann, Stefanie; Kumar, Ramya N; Redding-Johanson, Alyssa M; Batth, Tanveer S; Mukhopadhyay, Aindrila; Ludtke, Steven J; Frydman, Judith; Chiu, Wah

    2010-03-16

    The essential double-ring eukaryotic chaperonin TRiC/CCT (TCP1-ring complex or chaperonin containing TCP1) assists the folding of approximately 5-10% of the cellular proteome. Many TRiC substrates cannot be folded by other chaperonins from prokaryotes or archaea. These unique folding properties are likely linked to TRiC's unique heterooligomeric subunit organization, whereby each ring consists of eight different paralogous subunits in an arrangement that remains uncertain. Using single particle cryo-EM without imposing symmetry, we determined the mammalian TRiC structure at 4.7-A resolution. This revealed the existence of a 2-fold axis between its two rings resulting in two homotypic subunit interactions across the rings. A subsequent 2-fold symmetrized map yielded a 4.0-A resolution structure that evinces the densities of a large fraction of side chains, loops, and insertions. These features permitted unambiguous identification of all eight individual subunits, despite their sequence similarity. Independent biochemical near-neighbor analysis supports our cryo-EM derived TRiC subunit arrangement. We obtained a Calpha backbone model for each subunit from an initial homology model refined against the cryo-EM density. A subsequently optimized atomic model for a subunit showed approximately 95% of the main chain dihedral angles in the allowable regions of the Ramachandran plot. The determination of the TRiC subunit arrangement opens the way to understand its unique function and mechanism. In particular, an unevenly distributed positively charged wall lining the closed folding chamber of TRiC differs strikingly from that of prokaryotic and archaeal chaperonins. These interior surface chemical properties likely play an important role in TRiC's cellular substrate specificity.

  8. Longitudinal profile diagnostic scheme with subfemtosecond resolution for high-brightness electron beams

    Directory of Open Access Journals (Sweden)

    G. Andonian

    2011-07-01

    Full Text Available High-resolution measurement of the longitudinal profile of a relativistic electron beam is of utmost importance for linac based free-electron lasers and other advanced accelerator facilities that employ ultrashort bunches. In this paper, we investigate a novel scheme to measure ultrashort bunches (subpicosecond with exceptional temporal resolution (hundreds of attoseconds and dynamic range. The scheme employs two orthogonally oriented deflecting sections. The first imparts a short-wavelength (fast temporal resolution horizontal angular modulation on the beam, while the second imparts a long-wavelength (slow angular kick in the vertical dimension. Both modulations are observable on a standard downstream screen in the form of a streaked sinusoidal beam structure. We demonstrate, using scaled variables in a quasi-1D approximation, an expression for the temporal resolution of the scheme and apply it to a proof-of-concept experiment at the UCLA Neptune high-brightness injector facility. The scheme is also investigated for application at the SLAC NLCTA facility, where we show that the subfemtosecond resolution is sufficient to resolve the temporal structure of the beam used in the echo-enabled free-electron laser. We employ beam simulations to verify the effect for typical Neptune and NLCTA parameter sets and demonstrate the feasibility of the concept.

  9. Measurement needs guided by synthetic radar scans in high-resolution model output

    Science.gov (United States)

    Varble, A.; Nesbitt, S. W.; Borque, P.

    2017-12-01

    Microphysical and dynamical process interactions within deep convective clouds are not well understood, partly because measurement strategies often focus on statistics of cloud state rather than cloud processes. While processes cannot be directly measured, they can be inferred with sufficiently frequent and detailed scanning radar measurements focused on the life cycleof individual cloud regions. This is a primary goal of the 2018-19 DOE ARM Cloud, Aerosol, and Complex Terrain Interactions (CACTI) and NSF Remote sensing of Electrification, Lightning, And Mesoscale/microscale Processes with Adaptive Ground Observations (RELAMPAGO) field campaigns in central Argentina, where orographic deep convective initiation is frequent with some high-impact systems growing into the tallest and largest in the world. An array of fixed and mobile scanning multi-wavelength dual-polarization radars will be coupled with surface observations, sounding systems, multi-wavelength vertical profilers, and aircraft in situ measurements to characterize convective cloud life cycles and their relationship with environmental conditions. While detailed cloud processes are an observational target, the radar scan patterns that are most ideal for observing them are unclear. They depend on the locations and scales of key microphysical and dynamical processes operating within the cloud. High-resolution simulations of clouds, while imperfect, can provide information on these locations and scales that guide radar measurement needs. Radar locations are set in the model domain based on planned experiment locations, and simulatedorographic deep convective initiation and upscale growth are sampled using a number of different scans involving RHIs or PPIs with predefined elevation and azimuthal angles that approximately conform with radar range and beam width specifications. Each full scan pattern is applied to output atsingle model time steps with time step intervals that depend on the length of time

  10. Imaging and elemental mapping of biological specimens with a dual-EDS dedicated scanning transmission electron microscope

    Energy Technology Data Exchange (ETDEWEB)

    Wu, J.S., E-mail: jinsong-wu@northwestern.edu [Northwestern University Atomic and Nanoscale Characterization Experimental (NUANCE) Center, Northwestern University, Evanston, IL 60208 (United States); Department of Materials Science and Engineering, Northwestern University, Evanston, IL 60208 (United States); Kim, A.M. [Department of Obstetrics and Gynecology, Feinberg School of Medicine, Northwestern University, Chicago, IL 60611 (United States); Chemistry of Life Processes Institute, Northwestern University, Evanston, IL 60208 (United States); Bleher, R. [Department of Materials Science and Engineering, Northwestern University, Evanston, IL 60208 (United States); Chemistry of Life Processes Institute, Northwestern University, Evanston, IL 60208 (United States); Myers, B.D. [Northwestern University Atomic and Nanoscale Characterization Experimental (NUANCE) Center, Northwestern University, Evanston, IL 60208 (United States); Department of Materials Science and Engineering, Northwestern University, Evanston, IL 60208 (United States); Marvin, R.G. [Department of Chemistry, Northwestern University, Evanston, IL 60208 (United States); Chemistry of Life Processes Institute, Northwestern University, Evanston, IL 60208 (United States); Inada, H.; Nakamura, K. [Hitachi High-Technologies Corporation, Ibaraki 312-8504 (Japan); Zhang, X.F. [Hitachi High Technologies America, Inc., 5960 Inglewood Drive, Pleasanton, California 94588 (United States); Roth, E. [Department of Materials Science and Engineering, Northwestern University, Evanston, IL 60208 (United States); Chemistry of Life Processes Institute, Northwestern University, Evanston, IL 60208 (United States); Li, S.Y. [Northwestern University Atomic and Nanoscale Characterization Experimental (NUANCE) Center, Northwestern University, Evanston, IL 60208 (United States); and others

    2013-05-15

    A dedicated analytical scanning transmission electron microscope (STEM) with dual energy dispersive spectroscopy (EDS) detectors has been designed for complementary high performance imaging as well as high sensitivity elemental analysis and mapping of biological structures. The performance of this new design, based on a Hitachi HD-2300A model, was evaluated using a variety of biological specimens. With three imaging detectors, both the surface and internal structure of cells can be examined simultaneously. The whole-cell elemental mapping, especially of heavier metal species that have low cross-section for electron energy loss spectroscopy (EELS), can be faithfully obtained. Optimization of STEM imaging conditions is applied to thick sections as well as thin sections of biological cells under low-dose conditions at room and cryogenic temperatures. Such multimodal capabilities applied to soft/biological structures usher a new era for analytical studies in biological systems. - Highlights: ► Applications of STEM in characterization of biological samples are demonstrated. ► Elemental analyses are performed by dual EDS and EELS. ► Both the surface and internal structure of cells can be studied simultaneously. ► The imaging contrast in low-dose cryo-STEM has been analyzed.

  11. Imaging and elemental mapping of biological specimens with a dual-EDS dedicated scanning transmission electron microscope

    International Nuclear Information System (INIS)

    Wu, J.S.; Kim, A.M.; Bleher, R.; Myers, B.D.; Marvin, R.G.; Inada, H.; Nakamura, K.; Zhang, X.F.; Roth, E.; Li, S.Y.

    2013-01-01

    A dedicated analytical scanning transmission electron microscope (STEM) with dual energy dispersive spectroscopy (EDS) detectors has been designed for complementary high performance imaging as well as high sensitivity elemental analysis and mapping of biological structures. The performance of this new design, based on a Hitachi HD-2300A model, was evaluated using a variety of biological specimens. With three imaging detectors, both the surface and internal structure of cells can be examined simultaneously. The whole-cell elemental mapping, especially of heavier metal species that have low cross-section for electron energy loss spectroscopy (EELS), can be faithfully obtained. Optimization of STEM imaging conditions is applied to thick sections as well as thin sections of biological cells under low-dose conditions at room and cryogenic temperatures. Such multimodal capabilities applied to soft/biological structures usher a new era for analytical studies in biological systems. - Highlights: ► Applications of STEM in characterization of biological samples are demonstrated. ► Elemental analyses are performed by dual EDS and EELS. ► Both the surface and internal structure of cells can be studied simultaneously. ► The imaging contrast in low-dose cryo-STEM has been analyzed

  12. A fast cross-validation method for alignment of electron tomography images based on Beer-Lambert law

    Science.gov (United States)

    Yan, Rui; Edwards, Thomas J.; Pankratz, Logan M.; Kuhn, Richard J.; Lanman, Jason K.; Liu, Jun; Jiang, Wen

    2015-01-01

    In electron tomography, accurate alignment of tilt series is an essential step in attaining high-resolution 3D reconstructions. Nevertheless, quantitative assessment of alignment quality has remained a challenging issue, even though many alignment methods have been reported. Here, we report a fast and accurate method, tomoAlignEval, based on the Beer-Lambert law, for the evaluation of alignment quality. Our method is able to globally estimate the alignment accuracy by measuring the goodness of log-linear relationship of the beam intensity attenuations at different tilt angles. Extensive tests with experimental data demonstrated its robust performance with stained and cryo samples. Our method is not only significantly faster but also more sensitive than measurements of tomogram resolution using Fourier shell correlation method (FSCe/o). From these tests, we also conclude that while current alignment methods are sufficiently accurate for stained samples, inaccurate alignments remain a major limitation for high resolution cryo-electron tomography. PMID:26455556

  13. A fast cross-validation method for alignment of electron tomography images based on Beer-Lambert law.

    Science.gov (United States)

    Yan, Rui; Edwards, Thomas J; Pankratz, Logan M; Kuhn, Richard J; Lanman, Jason K; Liu, Jun; Jiang, Wen

    2015-11-01

    In electron tomography, accurate alignment of tilt series is an essential step in attaining high-resolution 3D reconstructions. Nevertheless, quantitative assessment of alignment quality has remained a challenging issue, even though many alignment methods have been reported. Here, we report a fast and accurate method, tomoAlignEval, based on the Beer-Lambert law, for the evaluation of alignment quality. Our method is able to globally estimate the alignment accuracy by measuring the goodness of log-linear relationship of the beam intensity attenuations at different tilt angles. Extensive tests with experimental data demonstrated its robust performance with stained and cryo samples. Our method is not only significantly faster but also more sensitive than measurements of tomogram resolution using Fourier shell correlation method (FSCe/o). From these tests, we also conclude that while current alignment methods are sufficiently accurate for stained samples, inaccurate alignments remain a major limitation for high resolution cryo-electron tomography. Copyright © 2015 Elsevier Inc. All rights reserved.

  14. Transmission/Scanning Transmission Electron Microscopy | Materials Science

    Science.gov (United States)

    crystallographic structure of a material. Amplitude-contrast images yield information about the chemistry and microstructure of a material and its defects. Phase-contrast imaging or high-resolution (HR) TEM imaging gives information about the microstructure of a material and its defects at an atomic resolution. With scanning

  15. High resolution RGB color line scan camera

    Science.gov (United States)

    Lynch, Theodore E.; Huettig, Fred

    1998-04-01

    A color line scan camera family which is available with either 6000, 8000 or 10000 pixels/color channel, utilizes off-the-shelf lenses, interfaces with currently available frame grabbers, includes on-board pixel by pixel offset correction, and is configurable and controllable via RS232 serial port for computer controlled or stand alone operation is described in this paper. This line scan camera is based on an available 8000 element monochrome line scan camera designed by AOA for OEM use. The new color version includes improvements such as better packaging and additional user features which make the camera easier to use. The heart of the camera is a tri-linear CCD sensor with on-chip color balancing for maximum accuracy and pinned photodiodes for low lag response. Each color channel is digitized to 12 bits and all three channels are multiplexed together so that the resulting camera output video is either a 12 or 8 bit data stream at a rate of up to 24Megpixels/sec. Conversion from 12 to 8 bit, or user-defined gamma, is accomplished by on board user-defined video look up tables. The camera has two user-selectable operating modes; lows speed, high sensitivity mode or high speed, reduced sensitivity mode. The intended uses of the camera include industrial inspection, digital archiving, document scanning, and graphic arts applications.

  16. Strain fields around dislocation arrays in a Σ9 silicon bicrystal measured by scanning transmission electron microscopy

    Science.gov (United States)

    Couillard, Martin; Radtke, Guillaume; Botton, Gianluigi A.

    2013-04-01

    Strain fields around grain boundary dislocations are measured by applying geometric phase analysis on atomic resolution images obtained from multiple fast acquisitions in scanning transmission electron microscopy. Maps of lattice distortions in silicon introduced by an array of pure edge dislocations located at a Σ9(122) grain boundary are compared with the predictions from isotropic elastic theory, and the atomic structure of dislocation cores is deduced from images displaying all the atomic columns. For strain measurements, reducing the acquisition time is found to significantly decrease the effects of instabilities on the high-resolution images. Contributions from scanning artefacts are also diminished by summing multiple images following a cross-correlation alignment procedure. Combined with the sub-Ångström resolution obtained with an aberration corrector, and the stable dedicated microscope's environment, therapid acquisition method provides the measurements of atomic displacements with accuracy below 10 pm. Finally, the advantages of combining strain measurements with the collection of various analytical signals in a scanning transmission electron microscope are discussed.

  17. Large-scale high-resolution scanning Hall probe microscope used for MgB2 filament characterization

    International Nuclear Information System (INIS)

    Cambel, V; Fedor, J; Gregusova, D; Kovac, P; Husek, I

    2005-01-01

    The scanning Hall probe microscope (SHPM) is an important imaging tool used for detailed studies of superconductors in basic science as well as in the industrial sector. It can be used for the studies of losses, current distribution, and effects at grain boundaries. However, only a few SHPMs for magnetic field imaging at temperatures below 77 K have been proposed up to now, most of them designed for small-area (∼10x10 μm 2 ) scanning. We present a large-scale low-temperature SHPM developed for imaging the entire magnetic field in close proximity to magnetic and superconducting samples at 4.2-300 K. The microscope combines a large scanned area and high spatial and magnetic field resolution. The instrument is designed as an insert of standard helium flowing cryostats. The Hall sensor scans an area up to 7 x 25 mm 2 in the whole temperature interval with a spatial resolution better than 5 μm. The presented system is used for the study of ex situ prepared MgB 2 filament. We show that external magnetic field induces local supercurrents in the MgB 2 , from which the critical current can be estimated. Moreover, it indicates the microstructure and space homogeneity of the superconductor

  18. Solving Research Tasks Using Desk top Scanning Electron Microscope Phenom ProX

    Directory of Open Access Journals (Sweden)

    Vertsanova, O.V.

    2014-03-01

    Full Text Available Phenom ProX — morden effective universal desktop Scanning Electron Microscope with integrated EDS system. Phenom-World helps customers to stay competitive in a world where critical dimensions are continuously getting smaller. All Phenom desktop systems give direct access to the high resolution and high-quality imaging and analysis required in a large variety of applications. They are affordable, flexible and a fast tool enabling engineers, technicians, researchers and educational professionals to investigate micron and submicron structures.

  19. Simultaneous correlative scanning electron and high-NA fluorescence microscopy.

    Directory of Open Access Journals (Sweden)

    Nalan Liv

    Full Text Available Correlative light and electron microscopy (CLEM is a unique method for investigating biological structure-function relations. With CLEM protein distributions visualized in fluorescence can be mapped onto the cellular ultrastructure measured with electron microscopy. Widespread application of correlative microscopy is hampered by elaborate experimental procedures related foremost to retrieving regions of interest in both modalities and/or compromises in integrated approaches. We present a novel approach to correlative microscopy, in which a high numerical aperture epi-fluorescence microscope and a scanning electron microscope illuminate the same area of a sample at the same time. This removes the need for retrieval of regions of interest leading to a drastic reduction of inspection times and the possibility for quantitative investigations of large areas and datasets with correlative microscopy. We demonstrate Simultaneous CLEM (SCLEM analyzing cell-cell connections and membrane protrusions in whole uncoated colon adenocarcinoma cell line cells stained for actin and cortactin with AlexaFluor488. SCLEM imaging of coverglass-mounted tissue sections with both electron-dense and fluorescence staining is also shown.

  20. Composition quantification of electron-transparent samples by backscattered electron imaging in scanning electron microscopy

    Energy Technology Data Exchange (ETDEWEB)

    Müller, E., E-mail: erich.mueller@kit.edu; Gerthsen, D.

    2017-02-15

    The contrast of backscattered electron (BSE) images in scanning electron microscopy (SEM) depends on material parameters which can be exploited for composition quantification if some information on the material system is available. As an example, the In-concentration in thin In{sub x}Ga{sub 1−x}As layers embedded in a GaAs matrix is analyzed in this work. The spatial resolution of the technique is improved by using thin electron-transparent specimens instead of bulk samples. Although the BSEs are detected in a comparably small angular range by an annular semiconductor detector, the image intensity can be evaluated to determine the composition and local thickness of the specimen. The measured intensities are calibrated within one single image to eliminate the influence of the detection and amplification system. Quantification is performed by comparison of experimental and calculated data. Instead of using time-consuming Monte-Carlo simulations, an analytical model is applied for BSE-intensity calculations which considers single electron scattering and electron diffusion. - Highlights: • Sample thickness and composition are quantified by backscattered electron imaging. • A thin sample is used to achieve spatial resolution of few nanometers. • Calculations are carried out with a time-saving electron diffusion model. • Small differences in atomic number and density detected at low electron energies.

  1. High-resolution dichroic imaging of magnetic flux distributions in superconductors with scanning x-ray microscopy

    Energy Technology Data Exchange (ETDEWEB)

    Ruoss, Stephen; Stahl, Claudia; Weigand, Markus; Schuetz, Gisela [Max-Planck-Institut fuer Intelligente Systeme, Stuttgart (Germany); Albrecht, Joachim [Research Institute for Innovative Surfaces, FINO, Aalen University (Germany)

    2015-07-01

    The penetration of magnetic flux into the high-temperature superconductor YBCO has been observed using a new high-resolution technique based on X-ray magnetic circular dichroism (XMCD). Superconductors coated with thin soft magnetic layers of CoFeB are observed in a scanning x-ray microscope providing cooling of the sample down to 83 K under the influence of external magnetic fields. Resulting electrical currents create an inhomogeneous magnetic field distribution above the superconductor which leads to a local reorientation of the ferromagnetic layer. X-ray absorption measurements with circular polarized radiation allows the analysis of the magnetic flux distribution in the superconductor via the ferromagnetic layer. In this work we present first images taken at 83K with high spatial resolution in the nanoscale.

  2. Elemental mapping in scanning transmission electron microscopy

    International Nuclear Information System (INIS)

    Allen, L J; D'Alfonso, A J; Lugg, N R; Findlay, S D; LeBeau, J M; Stemmer, S

    2010-01-01

    We discuss atomic resolution chemical mapping in scanning transmission electron microscopy (STEM) based on core-loss electron energy loss spectroscopy (EELS) and also on energy dispersive X-ray (EDX) imaging. Chemical mapping using EELS can yield counterintuitive results which, however, can be understood using first principles calculations. Experimental chemical maps based on EDX bear out the thesis that such maps are always likely to be directly interpretable. This can be explained in terms of the local nature of the effective optical potential for ionization under those imaging conditions. This is followed by an excursion into the complementary technique of elemental mapping using energy-filtered transmission electron microscopy (EFTEM) in a conventional transmission electron microscope. We will then consider the widely used technique of Z-contrast or high-angle annular dark field (HAADF) imaging, which is based on phonon excitation, where it has recently been shown that intensity variations can be placed on an absolute scale by normalizing the measured intensities to the incident beam. Results, showing excellent agreement between theory and experiment to within a few percent, are shown for Z-contrast imaging from a sample of PbWO 4 .

  3. Investigations and characterization of the microstructure of special ceramic materials using the high-resolution electron microscope

    International Nuclear Information System (INIS)

    Kirn, M.

    1979-01-01

    The possibilities to characterize phases and microstructures by direct lattice imaging are indicated in the following work. Ceramic materials are particularly suitable for this as these exhibit a high mechanical stability in the investigation in the transmission electron microscope. First of all the fundamentals of the high-resolution electron microscopy are introduced and the various resulting possibilities to characterize microstructures are presented. A report then follows on experimental observations on undisturbed crystals of special ceramics on a Si 3 N 4 basis. Furthermore, it is shown that the high-resolution electron microscope provides valuable contributions to the determination of structure, in particular of twin variants. Finally, revealing information on the structure of the interfaces was obtained with the help of high-resolution electron microscopy. (orig./IHOE) [de

  4. Recent advances in cryo-TEM imaging of soft lipid nanoparticles

    DEFF Research Database (Denmark)

    Helvig, Shen Yu; Mat Azmi, Intan Diana Binti; Moghimi, Seyed Moien

    2015-01-01

    Cryo-transmission electron microscopy (Cryo-TEM), and its technological variations thereof, have become a powerful tool for detailed morphological characterization and 3D tomography of soft lipid and polymeric nanoparticles as well as biological materials such as viruses and DNA without chemical...... fixation. Here, we review and discuss recent advances in Cryo-TEM analysis of lipid-based drug nanocarriers with particular emphasis on morphological and internal nanostructure characterization of lyotropic liquid crystalline nanoparticles such as cubosomes and hexosomes....

  5. Diagnostic accuracy of cone-beam computed tomography scans with high- and low-resolution modes for the detection of root perforations.

    Science.gov (United States)

    Shokri, Abbas; Eskandarloo, Amir; Norouzi, Marouf; Poorolajal, Jalal; Majidi, Gelareh; Aliyaly, Alireza

    2018-03-01

    This study compared the diagnostic accuracy of cone-beam computed tomography (CBCT) scans obtained with 2 CBCT systems with high- and low-resolution modes for the detection of root perforations in endodontically treated mandibular molars. The root canals of 72 mandibular molars were cleaned and shaped. Perforations measuring 0.2, 0.3, and 0.4 mm in diameter were created at the furcation area of 48 roots, simulating strip perforations, or on the external surfaces of 48 roots, simulating root perforations. Forty-eight roots remained intact (control group). The roots were filled using gutta-percha (Gapadent, Tianjin, China) and AH26 sealer (Dentsply Maillefer, Ballaigues, Switzerland). The CBCT scans were obtained using the NewTom 3G (QR srl, Verona, Italy) and Cranex 3D (Soredex, Helsinki, Finland) CBCT systems in high- and low-resolution modes, and were evaluated by 2 observers. The chi-square test was used to assess the nominal variables. In strip perforations, the accuracies of low- and high-resolution modes were 75% and 83% for NewTom 3G and 67% and 69% for Cranex 3D. In root perforations, the accuracies of low- and high-resolution modes were 79% and 83% for NewTom 3G and was 56% and 73% for Cranex 3D. The accuracy of the 2 CBCT systems was different for the detection of strip and root perforations. The Cranex 3D had non-significantly higher accuracy than the NewTom 3G. In both scanners, the high-resolution mode yielded significantly higher accuracy than the low-resolution mode. The diagnostic accuracy of CBCT scans was not affected by the perforation diameter.

  6. Microsphere-based super-resolution scanning optical microscope.

    Science.gov (United States)

    Huszka, Gergely; Yang, Hui; Gijs, Martin A M

    2017-06-26

    High-refractive index dielectric microspheres positioned within the field of view of a microscope objective in a dielectric medium can focus the light into a so-called photonic nanojet. A sample placed in such nanojet can be imaged by the objective with super-resolution, i.e. with a resolution beyond the classical diffraction limit. However, when imaging nanostructures on a substrate, the propagation distance of a light wave in the dielectric medium in between the substrate and the microsphere must be small enough to reveal the sample's nanometric features. Therefore, only the central part of an image obtained through a microsphere shows super-resolution details, which are typically ∼100 nm using white light (peak at λ = 600 nm). We have performed finite element simulations of the role of this critical distance in the super-resolution effect. Super-resolution imaging of a sample placed beneath the microsphere is only possible within a very restricted central area of ∼10 μm 2 , where the separation distance between the substrate and the microsphere surface is very small (∼1 μm). To generate super-resolution images over larger areas of the sample, we have fixed a microsphere on a frame attached to the microscope objective, which is automatically scanned over the sample in a step-by-step fashion. This generates a set of image tiles, which are subsequently stitched into a single super-resolution image (with resolution of λ/4-λ/5) of a sample area of up to ∼10 4 μm 2 . Scanning a standard optical microscope objective with microsphere therefore enables super-resolution microscopy over the complete field-of-view of the objective.

  7. Composition measurement in substitutionally disordered materials by atomic resolution energy dispersive X-ray spectroscopy in scanning transmission electron microscopy.

    Science.gov (United States)

    Chen, Z; Taplin, D J; Weyland, M; Allen, L J; Findlay, S D

    2017-05-01

    The increasing use of energy dispersive X-ray spectroscopy in atomic resolution scanning transmission electron microscopy invites the question of whether its success in precision composition determination at lower magnifications can be replicated in the atomic resolution regime. In this paper, we explore, through simulation, the prospects for composition measurement via the model system of Al x Ga 1-x As, discussing the approximations used in the modelling, the variability in the signal due to changes in configuration at constant composition, and the ability to distinguish between different compositions. Results are presented in such a way that the number of X-ray counts, and thus the expected variation due to counting statistics, can be gauged for a range of operating conditions. Copyright © 2016 Elsevier B.V. All rights reserved.

  8. Laboratory design for high-performance electron microscopy

    Energy Technology Data Exchange (ETDEWEB)

    O' Keefe, Michael A.; Turner, John H.; Hetherington, Crispin J.D.; Cullis, A.G.; Carragher, Bridget; Jenkins, Ron; Milgrim, Julie; Milligan,Ronald A.; Potter, Clinton S.; Allard, Lawrence F.; Blom, Douglas A.; Degenhardt, Lynn; Sides, William H.

    2004-04-23

    Proliferation of electron microscopes with field emission guns, imaging filters and hardware spherical aberration correctors (giving higher spatial and energy resolution) has resulted in the need to construct special laboratories. As resolutions improve, transmission electron microscopes (TEMs) and scanning transmission electron microscopes (STEMs) become more sensitive to ambient conditions. State-of-the-art electron microscopes require state-of-the-art environments, and this means careful design and implementation of microscope sites, from the microscope room to the building that surrounds it. Laboratories have been constructed to house high-sensitive instruments with resolutions ranging down to sub-Angstrom levels; we present the various design philosophies used for some of these laboratories and our experiences with them. Four facilities are described: the National Center for Electron Microscopy OAM Laboratory at LBNL; the FEGTEM Facility at the University of Sheffield; the Center for Integrative Molecular Biosciences at TSRI; and the Advanced Microscopy Laboratory at ORNL.

  9. Line-scanning confocal microscopy for high-resolution imaging of upconverting rare-earth-based contrast agents

    Science.gov (United States)

    Higgins, Laura M.; Zevon, Margot; Ganapathy, Vidya; Sheng, Yang; Tan, Mei Chee; Riman, Richard E.; Roth, Charles M.; Moghe, Prabhas V.; Pierce, Mark C.

    2015-01-01

    Abstract. Rare-earth (RE) doped nanocomposites emit visible luminescence when illuminated with continuous wave near-infrared light, making them appealing candidates for use as contrast agents in biomedical imaging. However, the emission lifetime of these materials is much longer than the pixel dwell times used in scanning intravital microscopy. To overcome this limitation, we have developed a line-scanning confocal microscope for high-resolution, optically sectioned imaging of samples labeled with RE-based nanomaterials. Instrument performance is quantified using calibrated test objects. NaYF4:Er,Yb nanocomposites are imaged in vitro, and in ex vivo tissue specimens, with direct comparison to point-scanning confocal microscopy. We demonstrate that the extended pixel dwell time of line-scanning confocal microscopy enables subcellular-level imaging of these nanomaterials while maintaining optical sectioning. The line-scanning approach thus enables microscopic imaging of this emerging class of contrast agents for preclinical studies, with the potential to be adapted for real-time in vivo imaging in the clinic. PMID:26603495

  10. Visualization of bacterial polysaccharides by scanning transmission electron microscopy.

    Science.gov (United States)

    Wolanski, B S; McAleer, W J; Hilleman, M R

    1983-04-01

    Highly purified capsular polysaccharides of Neisseria meningitidis groups A, B, and C have been visualized by high resolution Scanning Transmission Electron Microscopy (STEM). Spheroidal macromolecules approximately 200 A in diameter are characteristic of the Meningococcus A and C polysaccharides whereas filaments that are 400-600 A in length are found in Meningococcus B polysaccharide preparations. Filaments are occasionally found associated with the spheroidal Meningococcus A and C polysaccharides and it is proposed that these structures are composed of a long (1-4 microns) filament or filaments that are arranged in spheroidal molecules or micelles of high molecular weight. The Meningococcus B polysaccharide, by contrast, is a short flexuous filament or strand of relatively low molecular weight. A relationship between morphology and antigenicity is proposed.

  11. Zernike Phase Contrast Electron Cryo-Tomography Applied to Marine Cyanobacteria Infected with Cyanophages

    Science.gov (United States)

    Dai, Wei; Fu, Caroline; Khant, Htet A.; Ludtke, Steven J.; Schmid, Michael F.; Chiu, Wah

    2015-01-01

    Advances in electron cryo-tomography have provided a new opportunity to visualize the internal 3D structures of a bacterium. An electron microscope equipped with Zernike phase contrast optics produces images with dramatically increased contrast compared to images obtained by conventional electron microscopy. Here we describe a protocol to apply Zernike phase plate technology for acquiring electron tomographic tilt series of cyanophage-infected cyanobacterial cells embedded in ice, without staining or chemical fixation. We detail the procedures for aligning and assessing phase plates for data collection, and methods to obtain 3D structures of cyanophage assembly intermediates in the host, by subtomogram alignment, classification and averaging. Acquiring three to four tomographic tilt series takes approximately 12 h on a JEM2200FS electron microscope. We expect this time requirement to decrease substantially as the technique matures. Time required for annotation and subtomogram averaging varies widely depending on the project goals and data volume. PMID:25321408

  12. Electron beam fabrication and characterization of high-resolution magnetic force microscopy tips

    Science.gov (United States)

    Rührig, M.; Porthun, S.; Lodder, J. C.; McVitie, S.; Heyderman, L. J.; Johnston, A. B.; Chapman, J. N.

    1996-03-01

    The stray field, magnetic microstructure, and switching behavior of high-resolution electron beam fabricated thin film tips for magnetic force microscopy (MFM) are investigated with different imaging modes in a transmission electron microscope (TEM). As the tiny smooth carbon needles covered with a thermally evaporated magnetic thin film are transparent to the electron energies used in these TEMs it is possible to observe both the external stray field emanating from the tips as well as their internal domain structure. The experiments confirm the basic features of electron beam fabricated thin film tips concluded from various MFM observations using these tips. Only a weak but highly concentrated stray field is observed emanating from the immediate apex region of the tip, consistent with their capability for high resolution. It also supports the negligible perturbation of the magnetization sample due to the tip stray field observed in MFM experiments. Investigation of the magnetization distributions within the tips, as well as preliminary magnetizing experiments, confirm a preferred single domain state of the high aspect ratio tips. To exclude artefacts of the observation techniques both nonmagnetic tips and those supporting different magnetization states are used for comparison.

  13. Ultrafast terahertz scanning tunneling microscopy with atomic resolution

    DEFF Research Database (Denmark)

    Jelic, Vedran; Iwaszczuk, Krzysztof; Nguyen, Peter H.

    2016-01-01

    We demonstrate that ultrafast terahertz scanning tunneling microscopy (THz-STM) can probe single atoms on a silicon surface with simultaneous sub-nanometer and sub-picosecond spatio-temporal resolution. THz-STM is established as a new technique for exploring high-field non-equilibrium tunneling...

  14. Optical depth sectioning in the aberration-corrected scanning transmission and scanning confocal electron microscope

    International Nuclear Information System (INIS)

    Behan, G; Nellist, P D

    2008-01-01

    The use of spherical aberration correctors in the scanning transmission electron microscope (STEM) has the effect of reducing the depth of field of the microscope, making three-dimensional imaging of a specimen possible by optical sectioning. Depth resolution can be improved further by placing aberration correctors and lenses pre and post specimen to achieve an imaging mode known as scanning confocal electron microscopy (SCEM). We present the calculated incoherent point spread functions (PSF) and optical transfer functions (OTF) of a STEM and SCEM. The OTF for a STEM is shown to have a missing cone region which results in severe blurring along the optic axis, which can be especially severe for extended objects. We also present strategies for reconstruction of experimental data, such as three-dimensional deconvolution of the point spread function.

  15. Hi-Res scan mode in clinical MDCT systems: Experimental assessment of spatial resolution performance.

    Science.gov (United States)

    Cruz-Bastida, Juan P; Gomez-Cardona, Daniel; Li, Ke; Sun, Heyi; Hsieh, Jiang; Szczykutowicz, Timothy P; Chen, Guang-Hong

    2016-05-01

    The introduction of a High-Resolution (Hi-Res) scan mode and another associated option that combines Hi-Res mode with the so-called High Definition (HD) reconstruction kernels (referred to as a Hi-Res/HD mode in this paper) in some multi-detector CT (MDCT) systems offers new opportunities to increase spatial resolution for some clinical applications that demand high spatial resolution. The purpose of this work was to quantify the in-plane spatial resolution along both the radial direction and tangential direction for the Hi-Res and Hi-Res/HD scan modes at different off-center positions. A technique was introduced and validated to address the signal saturation problem encountered in the attempt to quantify spatial resolution for the Hi-Res and Hi-Res/HD scan modes. Using the proposed method, the modulation transfer functions (MTFs) of a 64-slice MDCT system (Discovery CT750 HD, GE Healthcare) equipped with both Hi-Res and Hi-Res/HD modes were measured using a metal bead at nine different off-centered positions (0-16 cm with a step size of 2 cm); at each position, both conventional scans and Hi-Res scans were performed. For each type of scan and position, 80 repeated acquisitions were performed to reduce noise induced uncertainties in the MTF measurements. A total of 15 reconstruction kernels, including eight conventional kernels and seven HD kernels, were used to reconstruct CT images of the bead. An ex vivo animal study consisting of a bone fracture model was performed to corroborate the MTF results, as the detection of this high-contrast and high frequency task is predominantly determined by spatial resolution. Images of this animal model generated by different scan modes and reconstruction kernels were qualitatively compared with the MTF results. At the centered position, the use of Hi-Res mode resulted in a slight improvement in the MTF; each HD kernel generated higher spatial resolution than its counterpart conventional kernel. However, the MTF along the

  16. Local thermal conductivity of polycrystalline AlN ceramics measured by scanning thermal microscopy and complementary scanning electron microscopy techniques

    International Nuclear Information System (INIS)

    Zhang Yue-Fei; Wang Li; Wei Bin; Ji Yuan; Han Xiao-Dong; Zhang Ze; Heiderhoff, R.; Geinzer, A. K.; Balk, L. J.

    2012-01-01

    The local thermal conductivity of polycrystalline aluminum nitride (AlN) ceramics is measured and imaged by using a scanning thermal microscope (SThM) and complementary scanning electron microscope (SEM) based techniques at room temperature. The quantitative thermal conductivity for the AlN sample is gained by using a SThM with a spatial resolution of sub-micrometer scale through using the 3ω method. A thermal conductivity of 308 W/m·K within grains corresponding to that of high-purity single crystal AlN is obtained. The slight differences in thermal conduction between the adjacent grains are found to result from crystallographic misorientations, as demonstrated in the electron backscattered diffraction. A much lower thermal conductivity at the grain boundary is due to impurities and defects enriched in these sites, as indicated by energy dispersive X-ray spectroscopy. (condensed matter: structural, mechanical, and thermal properties)

  17. Cryo-electron microscopy of vitrified SV40 minichromosomes: the liquid drop model.

    Science.gov (United States)

    Dubochet, J; Adrian, M; Schultz, P; Oudet, P

    1986-03-01

    The structure of SV40 minichromosomes has been studied by cryo-electron microscopy of vitrified thin layers of solution. In high-salt buffer (130 mM NaCl), freshly prepared minichromosomes are condensed into globules 30 nm or more in diameter. On the micrograph, they appear to be formed by the close packing of 10 nm granules which give rise to a 10 nm reflection in the optical diffractogram. The globules can adopt many different conformations. At high concentration, they fuse into a homogeneous 'sea' of closely packed 10 nm granules. In low-salt buffer (less than 10 mM NaCl), the globules open, first into 10 nm filaments, and then into nucleosome-strings. The 'liquid drop' model is proposed to explain the condensed structure of the minichromosome in high-salt buffer: nucleosomes stack specifically on top of one another, thus forming the 10 nm filaments. 10 nm filaments in turn, tend to aggregate laterally. Optimizing both these interactions results in the condensation of 10 nm filaments or portions thereof into a structure similar to that of a liquid. Some implications of this model for the structure of cellular chromatin are discussed.

  18. Nano-tomography of porous geological materials using focused ion beam-scanning electron microscopy

    NARCIS (Netherlands)

    Liu, Yang; King, Helen E.; van Huis, Marijn A.; Drury, Martyn R.; Plümper, Oliver

    2016-01-01

    Tomographic analysis using focused ion beam-scanning electron microscopy (FIB-SEM) provides three-dimensional information about solid materials with a resolution of a few nanometres and thus bridges the gap between X-ray and transmission electron microscopic tomography techniques. This contribution

  19. Improved cryo-resistors with low temperature coefficients

    International Nuclear Information System (INIS)

    Warnecke, P.; Braun, E.

    1989-01-01

    A new type of 10- and 12.9κΩ cryo-resistors operating in a liquid helium bath with small temperature coefficient of resistivity have been built. Details for the fabrication of these improved cryo-resistors are reported. Experimental evidence of their drift rates are on the order of a few parts in 10 9 per day. A reduction of the mean pressure of 98.7 kPa in the helium dewar to 86.1 kPa, corresponding to a temperature decrease from 4.19 to 4.07 Κ, did not change the resistance value by more than the experimental resolution of 4 parts in 10 8

  20. EMHP: an accurate automated hole masking algorithm for single-particle cryo-EM image processing.

    Science.gov (United States)

    Berndsen, Zachary; Bowman, Charles; Jang, Haerin; Ward, Andrew B

    2017-12-01

    The Electron Microscopy Hole Punch (EMHP) is a streamlined suite of tools for quick assessment, sorting and hole masking of electron micrographs. With recent advances in single-particle electron cryo-microscopy (cryo-EM) data processing allowing for the rapid determination of protein structures using a smaller computational footprint, we saw the need for a fast and simple tool for data pre-processing that could run independent of existing high-performance computing (HPC) infrastructures. EMHP provides a data preprocessing platform in a small package that requires minimal python dependencies to function. https://www.bitbucket.org/chazbot/emhp Apache 2.0 License. bowman@scripps.edu. Supplementary data are available at Bioinformatics online. © The Author(s) 2017. Published by Oxford University Press.

  1. High-resolution MRI of the labyrinth. Optimization of scan parameters with 3D-FSE

    International Nuclear Information System (INIS)

    Sakata, Motomichi; Harada, Kuniaki; Shirase, Ryuji; Kumagai, Akiko; Ogasawara, Masashi

    2005-01-01

    The aim of our study was to optimize the parameters of high-resolution MRI of the labyrinth with a 3D fast spin-echo (3D-FSE) sequence. We investigated repetition time (TR), echo time (TE), Matrix, field of view (FOV), and coil selection in terms of CNR (contrast-to-noise ratio) and SNR (signal-to-noise ratio) by comparing axial images and/or three-dimensional images. The optimal 3D-FSE sequence parameters were as follows: 1.5 Tesla MR unit (Signa LX, GE Medical Systems), 3D-FSE sequence, dual 3-inch surface coil, acquisition time=12.08 min, TR=5000 msec, TE=300 msec, 3 number of excitations (NEX), FOV=12 cm, matrix=256 x 256, slice thickness=0.5 mm/0.0 sp, echo train=64, bandwidth=±31.5 kHz. High-resolution MRI of the labyrinth using the optimized 3D-FSE sequence parameters permits visualization of important anatomic details (such as scala tympani and scala vestibuli), making it possible to determine inner ear anomalies and the patency of cochlear turns. To obtain excellent heavily T2-weighted axial and three-dimensional images in the labyrinth, high CNR, SNR, and spatial resolution are significant factors at the present time. Furthermore, it is important not only to optimize the scan parameters of 3D-FSE but also to select an appropriate coil for high-resolution MRI of the labyrinth. (author)

  2. Visualising reacting single atoms under controlled conditions: Advances in atomic resolution in situ Environmental (Scanning) Transmission Electron Microscopy (E(S)TEM)

    Science.gov (United States)

    Boyes, Edward D.; Gai, Pratibha L.

    2014-02-01

    Advances in atomic resolution Environmental (Scanning) Transmission Electron Microscopy (E(S)TEM) for probing gas-solid catalyst reactions in situ at the atomic level under controlled reaction conditions of gas environment and temperature are described. The recent development of the ESTEM extends the capability of the ETEM by providing the direct visualisation of single atoms and the atomic structure of selected solid state heterogeneous catalysts in their working states in real-time. Atomic resolution E(S)TEM provides a deeper understanding of the dynamic atomic processes at the surface of solids and their mechanisms of operation. The benefits of atomic resolution-E(S)TEM to science and technology include new knowledge leading to improved technological processes with substantial economic benefits, improved healthcare, reductions in energy needs and the management of environmental waste generation. xml:lang="fr"

  3. Dynamic Raman imaging system with high spatial and temporal resolution

    Science.gov (United States)

    Wang, Lei; Dai, Yinzhen; He, Hao; Lv, Ruiqi; Zong, Cheng; Ren, Bin

    2017-09-01

    There is an increasing need to study dynamic changing systems with significantly high spatial and temporal resolutions. In this work, we integrated point-scanning, line-scanning, and wide-field Raman imaging techniques into a single system. By using an Electron Multiplying CCD (EMCCD) with a high gain and high frame rate, we significantly reduced the time required for wide-field imaging, making it possible to monitor the electrochemical reactions in situ. The highest frame rate of EMCDD was ˜50 fps, and the Raman images for a specific Raman peak can be obtained by passing the signal from the sample through the Liquid Crystal Tunable Filter. The spatial resolutions of scanning imaging and wide-field imaging with a 100× objective (NA = 0.9) are 0.5 × 0.5 μm2 and 0.36 × 0.36 μm2, respectively. The system was used to study the surface plasmon resonance of Au nanorods, the surface-enhanced Raman scattering signal distribution for Au Nanoparticle aggregates, and dynamic Raman imaging of an electrochemical reacting system.

  4. Characterisation of colloidal drug delivery systems from the naked eye to Cryo-FESEM

    DEFF Research Database (Denmark)

    Krauel, Karen; Girvan, Liz; Hook, Sarah

    2007-01-01

    Poly(ethylcyanoacrylate) nanoparticles prepared by interfacial polymerisation on the basis of microemulsions were prepared in this study and both colloidal systems, nanoparticles and microemulsions, were analysed by visual observation and several microscopic techniques. Phase boundaries...... for the microemulsions of the two pseudoternary systems ethyloleate, polyoxyethylene 20 sorbitan mono-oleate/sorbitan monolaurate and water with and without butanol as a cosurfactant were determined by visual observation of the samples. Microemulsions containing liquid crystals were determined by polarisation light...... microscopy. Using freeze-fracture transmission electron microscopy and Cryo-field emission scanning electron microscopy the type of microemulsion (w/o droplet, bicontinuous, solution) was characterised. Nanoparticles prepared from the different types of microemulsion were additionally observed...

  5. Screening of plant toxins in food,feed and botanicals using full-scan high-resolution (Orbitrap) mass spectrometry

    NARCIS (Netherlands)

    Mol, J.G.J.; Dam, van R.C.J.; Zomer, P.; Mulder, P.P.J.

    2011-01-01

    A generic method based on LC with full-scan high-resolution (Orbitrap) mass spectrometry (MS) was systematically investigated for the simultaneous detection of a wide range of plant toxins in a variety of food and feed matrices. For a selection of 150 substances, representing various chemical

  6. Breaking Symmetry in Viral Icosahedral Capsids as Seen through the Lenses of X-ray Crystallography and Cryo-Electron Microscopy

    Directory of Open Access Journals (Sweden)

    Kristin N. Parent

    2018-02-01

    Full Text Available The majority of viruses on Earth form capsids built by multiple copies of one or more types of a coat protein arranged with 532 symmetry, generating an icosahedral shell. This highly repetitive structure is ideal to closely pack identical protein subunits and to enclose the nucleic acid genomes. However, the icosahedral capsid is not merely a passive cage but undergoes dynamic events to promote packaging, maturation and the transfer of the viral genome into the host. These essential processes are often mediated by proteinaceous complexes that interrupt the shell’s icosahedral symmetry, providing a gateway through the capsid. In this review, we take an inventory of molecular structures observed either internally, or at the 5-fold vertices of icosahedral DNA viruses that infect bacteria, archea and eukaryotes. Taking advantage of the recent revolution in cryo-electron microscopy (cryo-EM and building upon a wealth of crystallographic structures of individual components, we review the design principles of non-icosahedral structural components that interrupt icosahedral symmetry and discuss how these macromolecules play vital roles in genome packaging, ejection and host receptor-binding.

  7. Super-resolution biomolecular crystallography with low-resolution data.

    Science.gov (United States)

    Schröder, Gunnar F; Levitt, Michael; Brunger, Axel T

    2010-04-22

    X-ray diffraction plays a pivotal role in the understanding of biological systems by revealing atomic structures of proteins, nucleic acids and their complexes, with much recent interest in very large assemblies like the ribosome. As crystals of such large assemblies often diffract weakly (resolution worse than 4 A), we need methods that work at such low resolution. In macromolecular assemblies, some of the components may be known at high resolution, whereas others are unknown: current refinement methods fail as they require a high-resolution starting structure for the entire complex. Determining the structure of such complexes, which are often of key biological importance, should be possible in principle as the number of independent diffraction intensities at a resolution better than 5 A generally exceeds the number of degrees of freedom. Here we introduce a method that adds specific information from known homologous structures but allows global and local deformations of these homology models. Our approach uses the observation that local protein structure tends to be conserved as sequence and function evolve. Cross-validation with R(free) (the free R-factor) determines the optimum deformation and influence of the homology model. For test cases at 3.5-5 A resolution with known structures at high resolution, our method gives significant improvements over conventional refinement in the model as monitored by coordinate accuracy, the definition of secondary structure and the quality of electron density maps. For re-refinements of a representative set of 19 low-resolution crystal structures from the Protein Data Bank, we find similar improvements. Thus, a structure derived from low-resolution diffraction data can have quality similar to a high-resolution structure. Our method is applicable to the study of weakly diffracting crystals using X-ray micro-diffraction as well as data from new X-ray light sources. Use of homology information is not restricted to X

  8. Super-resolution for scanning light stimulation systems

    Energy Technology Data Exchange (ETDEWEB)

    Bitzer, L. A.; Neumann, K.; Benson, N., E-mail: niels.benson@uni-due.de; Schmechel, R. [Faculty of Engineering, NST and CENIDE, University of Duisburg-Essen, Bismarckstr. 81, 47057 Duisburg (Germany)

    2016-09-15

    Super-resolution (SR) is a technique used in digital image processing to overcome the resolution limitation of imaging systems. In this process, a single high resolution image is reconstructed from multiple low resolution images. SR is commonly used for CCD and CMOS (Complementary Metal-Oxide-Semiconductor) sensor images, as well as for medical applications, e.g., magnetic resonance imaging. Here, we demonstrate that super-resolution can be applied with scanning light stimulation (LS) systems, which are common to obtain space-resolved electro-optical parameters of a sample. For our purposes, the Projection Onto Convex Sets (POCS) was chosen and modified to suit the needs of LS systems. To demonstrate the SR adaption, an Optical Beam Induced Current (OBIC) LS system was used. The POCS algorithm was optimized by means of OBIC short circuit current measurements on a multicrystalline solar cell, resulting in a mean square error reduction of up to 61% and improved image quality.

  9. Self-sensing cantilevers with integrated conductive coaxial tips for high-resolution electrical scanning probe metrology

    International Nuclear Information System (INIS)

    Haemmerli, Alexandre J.; Pruitt, Beth L.; Harjee, Nahid; Koenig, Markus; Garcia, Andrei G. F.; Goldhaber-Gordon, David

    2015-01-01

    The lateral resolution of many electrical scanning probe techniques is limited by the spatial extent of the electrostatic potential profiles produced by their probes. Conventional unshielded conductive atomic force microscopy probes produce broad potential profiles. Shielded probes could offer higher resolution and easier data interpretation in the study of nanostructures. Electrical scanning probe techniques require a method of locating structures of interest, often by mapping surface topography. As the samples studied with these techniques are often photosensitive, the typical laser measurement of cantilever deflection can excite the sample, causing undesirable changes electrical properties. In this work, we present the design, fabrication, and characterization of probes that integrate coaxial tips for spatially sharp potential profiles with piezoresistors for self-contained, electrical displacement sensing. With the apex 100 nm above the sample surface, the electrostatic potential profile produced by our coaxial tips is more than 2 times narrower than that of unshielded tips with no long tails. In a scan bandwidth of 1 Hz–10 kHz, our probes have a displacement resolution of 2.9 Å at 293 K and 79 Å at 2 K, where the low-temperature performance is limited by amplifier noise. We show scanning gate microscopy images of a quantum point contact obtained with our probes, highlighting the improvement to lateral resolution resulting from the coaxial tip

  10. Reversible bronchial dilatation in children: comparison of serial high-resolution computer tomography scans of the lungs

    Energy Technology Data Exchange (ETDEWEB)

    Gaillard, E.A. E-mail: erol.gaillard@lwh-tr.nwest.nhs.uk; Carty, H.; Heaf, D.; Smyth, R.L

    2003-09-01

    Introduction: bronchiectasis is generally considered irreversible in the adult population, largely based on studies employing bronchography in cases with a significant clinical history. It is assumed, that the same is true for children. Few studies have examined the natural history of bronchiectasis in children and diagnostic criteria on high-resolution computer tomography of the lungs are derived from studies on adults. Frequently, bronchiectasis is reported in children in cases where localised bronchial dilatation is present, incorrectly labelling these children with an irreversible life-long condition. Objective: to evaluate changes in appearance of bronchial dilatation, unrelated to cystic fibrosis in children, as assessed by sequential high-resolution computer tomography (HRCT) of the lungs. Methods: the scans of 22 children with a radiological diagnosis of bronchiectasis, seen at Alder Hey Children's Hospital between 1994 and 2000, who had at least two CT scans of the lungs were reviewed by a single radiologist, who was blinded to the original report. Results: following a median scan interval of 21 months (range 2-43), bronchial dilatation resolved completely in six children and there was improvement in appearances in a further eight, with medical treatment alone. Discussion: a radiological diagnosis of bronchiectasis should be considered with caution in children as diagnostic criteria derived from studies in adults have not been validated in children and the condition is generally considered irreversible.

  11. CryoSat Ice Processor: High-Level Overview of Baseline-C Data and Quality-Control

    Science.gov (United States)

    Mannan, R.; Webb, E.; Hall, A.; Bouffard, J.; Femenias, P.; Parrinello, T.; Bouffard, J.; Brockley, D.; Baker, S.; Scagliola, M.; Urien, S.

    2016-08-01

    Since April 2015, the CryoSat ice products have been generated with the new Baseline-C Instrument Processing Facilities (IPFs). This represents a major upgrade to the CryoSat ice IPFs and is the baseline for the second CryoSat Reprocessing Campaign. Baseline- C introduces major evolutions with respect to Baseline- B, most notably the release of freeboard data within the L2 SAR products, following optimisation of the SAR retracker. Additional L2 improvements include a new Arctic Mean Sea Surface (MSS) in SAR; a new tuneable land ice retracker in LRM; and a new Digital Elevation Model (DEM) in SARIn. At L1B new attitude fields have been introduced and existing datation and range biases reduced. This paper provides a high level overview of the changes and evolutions implemented at Baseline-C in order to improve CryoSat L1B and L2 data characteristics and exploitation over polar regions. An overview of the main Quality Control (QC) activities performed on operational Baseline-C products is also presented.

  12. Non-Proportionality of Electron Response and Energy Resolution of Compton Electrons in Scintillators

    Science.gov (United States)

    Swiderski, L.; Marcinkowski, R.; Szawlowski, M.; Moszynski, M.; Czarnacki, W.; Syntfeld-Kazuch, A.; Szczesniak, T.; Pausch, G.; Plettner, C.; Roemer, K.

    2012-02-01

    Non-proportionality of light yield and energy resolution of Compton electrons in three scintillators (LaBr3:Ce, LYSO:Ce and CsI:Tl) were studied in a wide energy range from 10 keV up to 1 MeV. The experimental setup was comprised of a High Purity Germanium detector and tested scintillators coupled to a photomultiplier. Probing the non-proportionality and energy resolution curves at different energies was obtained by changing the position of various radioactive sources with respect to both detectors. The distance between both detectors and source was kept small to make use of Wide Angle Compton Coincidence (WACC) technique, which allowed us to scan large range of scattering angles simultaneously and obtain relatively high coincidence rate of 100 cps using weak sources of about 10 μCi activity. The results are compared with those obtained by direct irradiation of the tested scintillators with gamma-ray sources and fitting the full-energy peaks.

  13. Quantitative 3D Imaging by Cryo-EM (Shapes, membranes, and mapping of subcellular structures in whole bacteria)

    OpenAIRE

    Comolli, Luis R.; Downing, Kenneth H.; Fero, Michael J.; McAdams, Harley

    2006-01-01

    The shape of the bacteria cell wall during division and the deformation at the division plane are a function of the material properties of the cell wall, the growth rate, and the force due to the FtsZ ring. We have obtained 23 cryo-EM tomographic reconstructions through a time course following the cell division process in Caulobacter crescentus. We have also acquired over 300 images of the process by high resolution cryo-EM. The cell membranes have been segmented by an in-house developed util...

  14. Atomic-resolution transmission electron microscopy of electron beam–sensitive crystalline materials

    Science.gov (United States)

    Zhang, Daliang; Zhu, Yihan; Liu, Lingmei; Ying, Xiangrong; Hsiung, Chia-En; Sougrat, Rachid; Li, Kun; Han, Yu

    2018-02-01

    High-resolution imaging of electron beam–sensitive materials is one of the most difficult applications of transmission electron microscopy (TEM). The challenges are manifold, including the acquisition of images with extremely low beam doses, the time-constrained search for crystal zone axes, the precise image alignment, and the accurate determination of the defocus value. We develop a suite of methods to fulfill these requirements and acquire atomic-resolution TEM images of several metal organic frameworks that are generally recognized as highly sensitive to electron beams. The high image resolution allows us to identify individual metal atomic columns, various types of surface termination, and benzene rings in the organic linkers. We also apply our methods to other electron beam–sensitive materials, including the organic-inorganic hybrid perovskite CH3NH3PbBr3.

  15. Atomic-resolution transmission electron microscopy of electron beam–sensitive crystalline materials

    KAUST Repository

    Zhang, Daliang

    2018-01-18

    High-resolution imaging of electron beam-sensitive materials is one of the most difficult applications of transmission electron microscopy (TEM). The challenges are manifold, including the acquisition of images with extremely low beam doses, the time-constrained search for crystal zone axes, the precise image alignment, and the accurate determination of the defocus value. We develop a suite of methods to fulfill these requirements and acquire atomic-resolution TEM images of several metal organic frameworks that are generally recognized as highly sensitive to electron beams. The high image resolution allows us to identify individual metal atomic columns, various types of surface termination, and benzene rings in the organic linkers. We also apply our methods to other electron beam–sensitive materials, including the organic-inorganic hybrid perovskite CH3NH3PbBr3.

  16. Fast and sensitive rigid-body fitting into cryo-EM density maps with PowerFit

    NARCIS (Netherlands)

    C.p.van Zundert, Gydo; M.j.j. Bonvin, Alexandre

    2015-01-01

    Cryo-EM is a rapidly developing method to investigate the three dimensional structure of large macromolecular complexes. In spite of all the advances in the field, the resolution of most cryo-EM density maps is too low for de novo model building. Therefore, the data are often complemented by fitting

  17. Image quality of high-resolution CT with 16-channel multidetector-row CT. Comparison between helical scan and conventional step-shoot scan

    International Nuclear Information System (INIS)

    Sumikawa, Hiromitsu; Johkoh, Takeshi; Koyama, Mitsuhiro

    2005-01-01

    The aim of this study was to evaluate the image quality of high-resolution CT (HRCT) reconstructed from volumetric data with 16-channel multidetector-row CT (MDCT). Eleven autopsy lungs that were diagnosed histopathologically were scanned by 16-channel MDCT with the step-and-shoot scan mode and three helical scan modes. Each helical mode had each size of focal spot, pitch, and time of gantry rotation. HRCT images were reconstructed from the volumetric data with each helical mode and axial sequence data. Two observers evaluated the image quality and noted the most appropriate diagnosis for each imaging. Visualization of abnormal structures with one helical mode was equal to those with axial mode, whereas those with the other two helical modes were inferior to those with axial mode (Wilcoxon signed rank test; p<0.0001). There was no significant difference in diagnostic efficacy between modes. The image quality of HRCT with appropriate helical mode is equal to that with axial mode and diagnostic efficacy is equal among all modes. These results may indicate that sufficient HRCT images can be obtained by only one helical scan without the addition of conventional axial scans. (author)

  18. Precision crystal alignment for high-resolution electron microscope imaging

    International Nuclear Information System (INIS)

    Wood, G.J.; Beeching, M.J.

    1990-01-01

    One of the more difficult tasks involved in obtaining quality high-resolution electron micrographs is the precise alignment of a specimen into the required zone. The current accepted procedure, which involves changing to diffraction mode and searching for symmetric point diffraction pattern, is insensitive to small amounts of misalignment and at best qualitative. On-line analysis of the fourier space representation of the image, both for determining and correcting crystal tilt, is investigated. 8 refs., 42 figs

  19. Accurate virus quantitation using a Scanning Transmission Electron Microscopy (STEM) detector in a scanning electron microscope.

    Science.gov (United States)

    Blancett, Candace D; Fetterer, David P; Koistinen, Keith A; Morazzani, Elaine M; Monninger, Mitchell K; Piper, Ashley E; Kuehl, Kathleen A; Kearney, Brian J; Norris, Sarah L; Rossi, Cynthia A; Glass, Pamela J; Sun, Mei G

    2017-10-01

    A method for accurate quantitation of virus particles has long been sought, but a perfect method still eludes the scientific community. Electron Microscopy (EM) quantitation is a valuable technique because it provides direct morphology information and counts of all viral particles, whether or not they are infectious. In the past, EM negative stain quantitation methods have been cited as inaccurate, non-reproducible, and with detection limits that were too high to be useful. To improve accuracy and reproducibility, we have developed a method termed Scanning Transmission Electron Microscopy - Virus Quantitation (STEM-VQ), which simplifies sample preparation and uses a high throughput STEM detector in a Scanning Electron Microscope (SEM) coupled with commercially available software. In this paper, we demonstrate STEM-VQ with an alphavirus stock preparation to present the method's accuracy and reproducibility, including a comparison of STEM-VQ to viral plaque assay and the ViroCyt Virus Counter. Copyright © 2017 The Authors. Published by Elsevier B.V. All rights reserved.

  20. Deep learning-based subdivision approach for large scale macromolecules structure recovery from electron cryo tomograms.

    Science.gov (United States)

    Xu, Min; Chai, Xiaoqi; Muthakana, Hariank; Liang, Xiaodan; Yang, Ge; Zeev-Ben-Mordehai, Tzviya; Xing, Eric P

    2017-07-15

    Cellular Electron CryoTomography (CECT) enables 3D visualization of cellular organization at near-native state and in sub-molecular resolution, making it a powerful tool for analyzing structures of macromolecular complexes and their spatial organizations inside single cells. However, high degree of structural complexity together with practical imaging limitations makes the systematic de novo discovery of structures within cells challenging. It would likely require averaging and classifying millions of subtomograms potentially containing hundreds of highly heterogeneous structural classes. Although it is no longer difficult to acquire CECT data containing such amount of subtomograms due to advances in data acquisition automation, existing computational approaches have very limited scalability or discrimination ability, making them incapable of processing such amount of data. To complement existing approaches, in this article we propose a new approach for subdividing subtomograms into smaller but relatively homogeneous subsets. The structures in these subsets can then be separately recovered using existing computation intensive methods. Our approach is based on supervised structural feature extraction using deep learning, in combination with unsupervised clustering and reference-free classification. Our experiments show that, compared with existing unsupervised rotation invariant feature and pose-normalization based approaches, our new approach achieves significant improvements in both discrimination ability and scalability. More importantly, our new approach is able to discover new structural classes and recover structures that do not exist in training data. Source code freely available at http://www.cs.cmu.edu/∼mxu1/software . mxu1@cs.cmu.edu. Supplementary data are available at Bioinformatics online. © The Author 2017. Published by Oxford University Press. All rights reserved. For Permissions, please e-mail: journals.permissions@oup.com

  1. CryoVEx 2011-12 Airborne Campaigns for CryoSat Validation

    DEFF Research Database (Denmark)

    Skourup, Henriette; Hvidegaard, Sine Munk; Forsberg, René

    2013-01-01

    After the successful launch of CryoSat-2 in April 2010, the first direct validation campaign of the satellite was carried out in the April-May 2011. Part of this was repeated in Spring 2012. DTU Space has been involved in ESA’s CryoSat Validation Experiment (CryoVEx) with airborne activities since...... 2003. To validate the performance of the CryoSat-2 radar altimeter (SIRAL), the aircraft is equipped with an airborne version of the SIRAL altimeter (ASIRAS) together with a laser scanner. Of particular interest is to study the penetration depth of SIRAL into both land- and sea ice. This can be done...... of Alert and sea ice around Svalbard in the Fram Strait. Selected tracks were planned to match CryoSat-2 passes and a few of them were flown in formation flight with the AlfredWegener Institute (AWI) Polar- 5 carrying an EM induction sounder. The paper presents an overview of the 2011-12 airborne campaigns...

  2. ESA-NASA collaboration in support of CryoSat-2

    Science.gov (United States)

    Casal, T. G.; Davidson, M.; Schuettemeyer, D.; Perrera, A.; Armitage, T.; Bianchi, R.; Parrinello, T.; Fornari, M.; Skourup, H.

    2012-12-01

    In the framework of its Earth Observation Programmes the European Space Agency (ESA) carries out groundbased and airborne campaigns to support geophysical algorithm development, calibration/validation, simulation of future spaceborne earth observation missions, and applications development related to land, oceans and atmosphere. ESA has been conducting airborne and ground measurements campaigns since 1981 by deploying a broad range of active and passive instrumentation in both the optical and microwave regions of the electromagnetic spectrum such as lidars, limb/nadir sounding interferometers/spectrometers, high-resolution spectral imagers, advanced synthetic aperture radars, altimeters and radiometers. These campaigns take place inside and outside Europe in collaboration with national research organisations in the ESA member states as well as with international organisations harmonising European campaign activities. For the different activities a rich variety of datasets has been recorded, are archived and users can access campaign data through the EOPI web portal [http://eopi.esa.int]. CryoSat-2, ESA's third Earth Explorer, is Europe's first mission dedicated to monitoring Earth's ice fields. The satellite carries a sophisticated radar altimeter that can measure the thickness of sea ice down to centimetres and also monitor changes in ice sheets, particularly around the edges where icebergs are calved from the vast ice sheets that cover Greenland and Antarctica. On order to gather data to help ensure the accuracy of ESA's ice mission, in yet another remarkable collaborative effort, ESA and NASA met up over the Arctic Ocean in April 2012 to perform some carefully coordinated flights directly under CryoSat orbiting above. The aim of this large-scale campaign was to record sea-ice thickness and conditions of the ice exactly along the line traced by ESA's CryoSat satellite orbiting high above. A range of sensors installed on the different aircraft was used to gather

  3. High resolution and high voltage electron microscopy at the University of California, Berkeley

    International Nuclear Information System (INIS)

    Thomas, G.; Westmacott, K.H.

    1978-01-01

    Recent applications of high-resolution and high-voltage techniques at Berkely are described, using 100-kV TEMs and a standard 650-keV HVEM: grain boundary precipitation in Al--Zn, lattice imaging of grain boundaries in ceramics, steels, phase transitions and magnetic properties of ferrites, lattice defects, precipitation in Al--Si and behavior of interstitial dislocations under electron irradiation, effect of oxide films on loop formation in Al--Mg, and polytypism in magnesium Sialon. 13 refs. 12 figs

  4. Calibration of high-resolution electronic autocollimators with demanded low uncertainties using single reading head angle encoders

    International Nuclear Information System (INIS)

    Yandayan, Tanfer; Akgoz, S Asli; Asar, Muharrem

    2014-01-01

    Calibration of high-resolution electronic autocollimators is carried out in TUBITAK UME using an angle comparator to ensure direct traceability to the SI unit of plane angle, radian (rad). The device is a specially designed air-bearing rotary table fitted with a commercially available angular encoder utilizing a single reading head. It is shown that high-resolution electronic autocollimators in the large measurement range (e.g. ±1000 arcsec) can be calibrated with an expanded uncertainty of 0.035 arcsec (k = 2) in conventional dimensional laboratory conditions, applying good measurement strategy for single reading head angle encoders and taking simple but smart precautions. Description of the angle comparator is presented with various test results derived using different high-precision autocollimators, and a detailed uncertainty budget is given for the calibration of a high-resolution electronic autocollimator. (paper)

  5. Dysprosium disilicide nanostructures on silicon(001) studied by scanning tunneling microscopy and transmission electron microscopy

    International Nuclear Information System (INIS)

    Ye Gangfeng; Nogami, Jun; Crimp, Martin A.

    2006-01-01

    The microstructure of self-assembled dysprosium silicide nanostructures on silicon(001) has been studied by scanning tunneling microscopy and transmission electron microscopy. The studies focused on nanostructures that involve multiple atomic layers of the silicide. Cross-sectional high resolution transmission electron microscopy images and fast Fourier transform analysis showed that both hexagonal and orthorhombic/tetragonal silicide phases were present. Both the magnitude and the anisotropy of lattice mismatch between the silicide and the substrate play roles in the morphology and epitaxial growth of the nanostructures formed

  6. Sample preparation methods for scanning electron microscopy of homogenized Al-Mg-Si billets: A comparative study

    International Nuclear Information System (INIS)

    Österreicher, Johannes Albert; Kumar, Manoj; Schiffl, Andreas; Schwarz, Sabine; Hillebrand, Daniel; Bourret, Gilles Remi

    2016-01-01

    Characterization of Mg-Si precipitates is crucial for optimizing the homogenization heat treatment of Al-Mg-Si alloys. Although sample preparation is key for high quality scanning electron microscopy imaging, most common methods lead to dealloying of Mg-Si precipitates. In this article we systematically evaluate different sample preparation methods: mechanical polishing, etching with various reagents, and electropolishing using different electrolytes. We demonstrate that the use of a nitric acid and methanol electrolyte for electropolishing a homogenized Al-Mg-Si alloy prevents the dissolution of Mg-Si precipitates, resulting in micrographs of higher quality. This preparation method is investigated in depth and the obtained scanning electron microscopy images are compared with transmission electron micrographs: the shape and size of Mg-Si precipitates appear very similar in either method. The scanning electron micrographs allow proper identification and measurement of the Mg-Si phases including needles with lengths of roughly 200 nm. These needles are β″ precipitates as confirmed by high resolution transmission electron microscopy. - Highlights: •Secondary precipitation in homogenized 6xxx Al alloys is crucial for extrudability. •Existing sample preparation methods for SEM are improvable. •Electropolishing with nitric acid/methanol yields superior quality in SEM. •The obtained micrographs are compared to TEM micrographs.

  7. Sample preparation methods for scanning electron microscopy of homogenized Al-Mg-Si billets: A comparative study

    Energy Technology Data Exchange (ETDEWEB)

    Österreicher, Johannes Albert; Kumar, Manoj [LKR Light Metals Technologies Ranshofen, Austrian Institute of Technology, Postfach 26, 5282 Ranshofen (Austria); Schiffl, Andreas [Hammerer Aluminium Industries Extrusion GmbH, Lamprechtshausener Straße 69, 5282 Ranshofen (Austria); Schwarz, Sabine [University Service Centre for Transmission Electron Microscopy, Vienna University of Technology, Wiedner Hauptstr. 8-10, 1040 Wien (Austria); Hillebrand, Daniel [Hammerer Aluminium Industries Extrusion GmbH, Lamprechtshausener Straße 69, 5282 Ranshofen (Austria); Bourret, Gilles Remi, E-mail: gilles.bourret@sbg.ac.at [Department of Materials Science and Physics, University of Salzburg, Hellbrunner Straße 34, 5020 Salzburg (Austria)

    2016-12-15

    Characterization of Mg-Si precipitates is crucial for optimizing the homogenization heat treatment of Al-Mg-Si alloys. Although sample preparation is key for high quality scanning electron microscopy imaging, most common methods lead to dealloying of Mg-Si precipitates. In this article we systematically evaluate different sample preparation methods: mechanical polishing, etching with various reagents, and electropolishing using different electrolytes. We demonstrate that the use of a nitric acid and methanol electrolyte for electropolishing a homogenized Al-Mg-Si alloy prevents the dissolution of Mg-Si precipitates, resulting in micrographs of higher quality. This preparation method is investigated in depth and the obtained scanning electron microscopy images are compared with transmission electron micrographs: the shape and size of Mg-Si precipitates appear very similar in either method. The scanning electron micrographs allow proper identification and measurement of the Mg-Si phases including needles with lengths of roughly 200 nm. These needles are β″ precipitates as confirmed by high resolution transmission electron microscopy. - Highlights: •Secondary precipitation in homogenized 6xxx Al alloys is crucial for extrudability. •Existing sample preparation methods for SEM are improvable. •Electropolishing with nitric acid/methanol yields superior quality in SEM. •The obtained micrographs are compared to TEM micrographs.

  8. Scanning transmission electron microscopy: Albert Crewe's vision and beyond

    International Nuclear Information System (INIS)

    Krivanek, Ondrej L.; Chisholm, Matthew F.; Murfitt, Matthew F.; Dellby, Niklas

    2012-01-01

    Some four decades were needed to catch up with the vision that Albert Crewe and his group had for the scanning transmission electron microscope (STEM) in the nineteen sixties and seventies: attaining 0.5 Å resolution, and identifying single atoms spectroscopically. With these goals now attained, STEM developments are turning toward new directions, such as rapid atomic resolution imaging and exploring atomic bonding and electronic properties of samples at atomic resolution. The accomplishments and the future challenges are reviewed and illustrated with practical examples. -- Highlights: ► TV-rate STEM imaging of heavy atoms is demonstrated. ► DNA sequencing by STEM dark field imaging should be possible at a rate of 10 6 bases/s. ► Individual silicon atom impurities in graphene are imaged atom-by-atom. ► Single atoms of nitrogen and boron incorporated in graphene are imaged spectroscopically. ► Bonding of individual atoms can be probed by analyzing the fine structures of their EEL spectra.

  9. 'Crazy-Paving' Patterns on High-Resolution CT Scans in Patients with Pulmonary Complications after Hematopoietic Stem Cell Transplantation

    International Nuclear Information System (INIS)

    Marchiori, Edson; Escuissato, Dante L.; Gasparetto, Taisa Davaus; Considera, Daniela Peixoto; Franquet, Tomas

    2009-01-01

    To describe the pulmonary complications following hematopoietic stem cell transplantation (HSCT) that can present with a 'crazy-paving' pattern in high-resolution CT scans. Retrospective review of medical records from 2,537 patients who underwent HSCT. The 'crazy-paving' pattern consists of interlobular and intralobular septal thickening superimposed on an area of ground-glass attenuation on high-resolution CT scans. The CT scans were retrospectively reviewed by two radiologists, who reached final decisions by consensus. We identified 10 cases (2.02%), seven male and three female, with pulmonary complications following HSCT that presented with the 'crazy-paving' pattern. Seven (70%) patients had infectious pneumonia (adenovirus, herpes simplex, influenza virus, cytomegalovirus, respiratory syncytial virus, and toxoplasmosis), and three patients presented with non-infectious complications (idiopathic pneumonia syndrome and acute pulmonary edema). The 'crazy-paving' pattern was bilateral in all cases, with diffuse distribution in nine patients (90%), predominantly in the middle and inferior lung regions in seven patients (70%), and involving the anterior and posterior regions of the lungs in nine patients (90%). The 'crazy-paving' pattern is rare in HSCT recipients with pulmonary complications and is associated with infectious complications more commonly than non-infectious conditions

  10. Construction of a four tip scanning tunneling microscope/scanning electron microscope combination and conductivity measurements of silicide nanowires

    International Nuclear Information System (INIS)

    Zubkov, Evgeniy

    2013-01-01

    In this work the combination of a four-tip scanning tunneling microscope with a scanning electron microscope is presented. By means of this apparatus it is possible to perform the conductivity measurements on the in-situ prepared nanostructures in ultra-high vacuum. With the aid of a scanning electron microscope (SEM), it becomes possible to position the tunneling tips of the four-tip scanning tunneling microscope (STM), so that an arrangement for a four-point probe measurement on nanostructures can be obtained. The STM head was built according to the novel coaxial Beetle concept. This concept allows on the one hand, a very compact arrangement of the components of the STM and on the other hand, the new-built STM head has a good mechanical stability, in order to achieve atomic resolution with all four STM units. The atomic resolution of the STM units was confirmed by scanning a Si(111)-7 x 7 surface. The thermal drift during the STM operation, as well as the resonant frequencies of the mechanical structure of the STM head, were determined. The scanning electron microscope allows the precise and safe navigation of the tunneling tips on the sample surface. Multi tip spectroscopy with up to four STM units can be performed synchronously. To demonstrate the capabilities of the new-built apparatus the conductivity measurements were carried out on metallic yttrium silicide nanowires. The nanowires were prepared by the in-situ deposition of yttrium on a heated Si(110) sample surface. Current-voltage curves were recorded on the nanowires and on the wetting layer in-between. The curves indicate an existence of the Schottky barrier between the yttrium silicide nanowires and the silicon bulk. By means of the two-tip measurements with a gate, the insulating property of the Schottky barrier has been confirmed. Using this Schottky barrier, it is possible to limit the current to the nanowire and to prevent it from flowing through the silicon bulk. A four-tip resistance measurement

  11. High resolution electron microscopy and electron diffraction of YBa2Cu3O(7-x)

    International Nuclear Information System (INIS)

    Krakow, W.; Shaw, T.M.

    1988-01-01

    Experimental high resolution electron micrographs and computer simulation experiments have been used to evaluate the visibility of the atomic constituents of YBa 2 Cu 3 O(7-x). In practice, the detection of oxygen has not been possible in contradiction to that predicted by modelling of perfect crystalline material. Preliminary computer experiments of the electron diffraction patterns when oxygen vacancies are introduced on the Cu-O sheets separating Ba layers show the diffuse streaks characteristic of short range ordering. 7 references

  12. Design and performance of a spin-polarized electron energy loss spectrometer with high momentum resolution

    Energy Technology Data Exchange (ETDEWEB)

    Vasilyev, D.; Kirschner, J. [Max-Planck-Institut für Mikrostrukturphysik, Weinberg 2, 06120 Halle (Germany)

    2016-08-15

    We describe a new “complete” spin-polarized electron energy loss spectrometer comprising a spin-polarized primary electron source, an imaging electron analyzer, and a spin analyzer of the “spin-polarizing mirror” type. Unlike previous instruments, we have a high momentum resolution of less than 0.04 Å{sup −1}, at an energy resolution of 90-130 meV. Unlike all previous studies which reported rather broad featureless data in both energy and angle dependence, we find richly structured spectra depending sensitively on small changes of the primary energy, the kinetic energy after scattering, and of the angle of incidence. The key factor is the momentum resolution.

  13. Practical Approaches to Mitigation of Specimen Charging in High-Resolution Transmission Electron Microscopy

    Directory of Open Access Journals (Sweden)

    Young-Min Kim

    2010-09-01

    Full Text Available Specimen charging that is associated with the electron bombardment on the sample is a practical hindrance to high-resolution transmission electron microscopy (HRTEM analysis because it causes a severe loss of resolution in either diffraction or image data. Conductive thin film deposition on an insulating specimen has been proposed as an effective approach to the mitigation of the specimen charging; however, this method is generally not useful in HRTEM imaging of materials because the deposited film induces another artifact in the HRTEM image contrast. In this study, we propose practical methods to mitigate the specimen charging that takes place during the HRTEM of materials. For bulk-type specimens prepared by either an ion-thinning or focused-ion beam (FIB process, a plasma cleaning treatment is significantly effective in eliminating the charging phenomenon. In the case of low-dimensional nanomaterials such as nanowires and nanoparticles, the plasma cleaning is not feasible; however, the charging effect can be effectively eliminated by adjusting the electron illumination condition. The proposed methods facilitate a decrease in the buildup of specimen charging, thereby enhancing the quality of high-resolution images significantly.

  14. Development of a high-resolution electron-beam profile monitor using Fresnel zone plates

    International Nuclear Information System (INIS)

    Nakamura, Norio; Sakai, Hiroshi; Muto, Toshiya; Hayano, Hitoshi

    2004-01-01

    We present a high-resolution and real-time beam profile monitor using Fresnel zone plates (FZPs) developed in the KEK-ATF damping ring. The monitor system has an X-ray imaging optics with two FZPs. In this monitor, the synchrotron radiation from the electron beam at the bending magnet is monochromatized by a crystal monochromator and the transverse electron beam image is twenty-times magnified by the two FZPs and detected on an X-ray CCD camera. The expected spatial resolution for the selected photon energy of 3.235 keV is less than 1 μm. With the beam profile monitor, we succeeded in obtaining a clear electron-beam image and measuring the extremely small beam size less than 10 μm. It is greatly expected that the beam profile monitor will be used in high-brilliance light sources and low-emittance accelerators. (author)

  15. Spatial Localization of the Ebola Virus Glycoprotein Mucin-Like Domain Determined by Cryo-Electron Tomography

    OpenAIRE

    Tran, Erin E. H.; Simmons, James A.; Bartesaghi, Alberto; Shoemaker, Charles J.; Nelson, Elizabeth; White, Judith M.; Subramaniam, Sriram

    2014-01-01

    The Ebola virus glycoprotein mucin-like domain (MLD) is implicated in Ebola virus cell entry and immune evasion. Using cryo-electron tomography of Ebola virus-like particles, we determined a three-dimensional structure for the full-length glycoprotein in a near-native state and compared it to that of a glycoprotein lacking the MLD. Our results, which show that the MLD is located at the apex and the sides of each glycoprotein monomer, provide a structural template for analysis of MLD function.

  16. Energy-weighted dynamical scattering simulations of electron diffraction modalities in the scanning electron microscope.

    Science.gov (United States)

    Pascal, Elena; Singh, Saransh; Callahan, Patrick G; Hourahine, Ben; Trager-Cowan, Carol; Graef, Marc De

    2018-04-01

    Transmission Kikuchi diffraction (TKD) has been gaining momentum as a high resolution alternative to electron back-scattered diffraction (EBSD), adding to the existing electron diffraction modalities in the scanning electron microscope (SEM). The image simulation of any of these measurement techniques requires an energy dependent diffraction model for which, in turn, knowledge of electron energies and diffraction distances distributions is required. We identify the sample-detector geometry and the effect of inelastic events on the diffracting electron beam as the important factors to be considered when predicting these distributions. However, tractable models taking into account inelastic scattering explicitly are lacking. In this study, we expand the Monte Carlo (MC) energy-weighting dynamical simulations models used for EBSD [1] and ECP [2] to the TKD case. We show that the foil thickness in TKD can be used as a means of energy filtering and compare band sharpness in the different modalities. The current model is shown to correctly predict TKD patterns and, through the dictionary indexing approach, to produce higher quality indexed TKD maps than conventional Hough transform approach, especially close to grain boundaries. Copyright © 2018 The Authors. Published by Elsevier B.V. All rights reserved.

  17. Efficient creation of electron vortex beams for high resolution STEM imaging.

    Science.gov (United States)

    Béché, A; Juchtmans, R; Verbeeck, J

    2017-07-01

    The recent discovery of electron vortex beams carrying quantised angular momentum in the TEM has led to an active field of research, exploring a variety of potential applications including the possibility of mapping magnetic states at the atomic scale. A prerequisite for this is the availability of atomic sized electron vortex beams at high beam current and mode purity. In this paper we present recent progress showing that by making use of the Aharonov-Bohm effect near the tip of a long single domain ferromagnetic Nickel needle, a very efficient aperture for the production of electron vortex beams can be realised. The aperture transmits more than 99% of all electrons and provides a vortex mode purity of up to 92%. Placing this aperture in the condenser plane of a state of the art Cs corrected microscope allows us to demonstrate atomic resolution HAADF STEM images with spatial resolution better than 1 Angström, in agreement with theoretical expectations and only slightly inferior to the performance of a non-vortex probe on the same instrument. Copyright © 2016 Elsevier B.V. All rights reserved.

  18. Challenges of microtome‐based serial block‐face scanning electron microscopy in neuroscience

    Science.gov (United States)

    WANNER, A. A.; KIRSCHMANN, M. A.

    2015-01-01

    Summary Serial block‐face scanning electron microscopy (SBEM) is becoming increasingly popular for a wide range of applications in many disciplines from biology to material sciences. This review focuses on applications for circuit reconstruction in neuroscience, which is one of the major driving forces advancing SBEM. Neuronal circuit reconstruction poses exceptional challenges to volume EM in terms of resolution, field of view, acquisition time and sample preparation. Mapping the connections between neurons in the brain is crucial for understanding information flow and information processing in the brain. However, information on the connectivity between hundreds or even thousands of neurons densely packed in neuronal microcircuits is still largely missing. Volume EM techniques such as serial section TEM, automated tape‐collecting ultramicrotome, focused ion‐beam scanning electron microscopy and SBEM (microtome serial block‐face scanning electron microscopy) are the techniques that provide sufficient resolution to resolve ultrastructural details such as synapses and provides sufficient field of view for dense reconstruction of neuronal circuits. While volume EM techniques are advancing, they are generating large data sets on the terabyte scale that require new image processing workflows and analysis tools. In this review, we present the recent advances in SBEM for circuit reconstruction in neuroscience and an overview of existing image processing and analysis pipelines. PMID:25907464

  19. Polystyrene negative resist for high-resolution electron beam lithography

    Directory of Open Access Journals (Sweden)

    Ma Siqi

    2011-01-01

    Full Text Available Abstract We studied the exposure behavior of low molecular weight polystyrene as a negative tone electron beam lithography (EBL resist, with the goal of finding the ultimate achievable resolution. It demonstrated fairly well-defined patterning of a 20-nm period line array and a 15-nm period dot array, which are the densest patterns ever achieved using organic EBL resists. Such dense patterns can be achieved both at 20 and 5 keV beam energies using different developers. In addition to its ultra-high resolution capability, polystyrene is a simple and low-cost resist with easy process control and practically unlimited shelf life. It is also considerably more resistant to dry etching than PMMA. With a low sensitivity, it would find applications where negative resist is desired and throughput is not a major concern.

  20. Miniature scanning electron microscope for investigation of the interior surface of a superconducting Nb radiofrequency accelerating cavity

    International Nuclear Information System (INIS)

    Mathewson, A.G.; Grillot, A.

    1982-01-01

    A miniature scanning electron microscope with an electron beam diameter approx.1 μm has been constructed for high resolution examination at room temperature of the interior surface of a superconducting Nb radiofrequency accelerating cavity. Various objects and surface structures were observed, some of which could be correlated with lossy regions or ''hot spots'' detected previously on the outside surface during cavity operation at < or =4.2 K by a chain of carbon resistors. No internal surface features were observed which could conclusively be correlated with field emitting electron sources

  1. Preparation and Loading Process of Single Crystalline Samples into a Gas Environmental Cell Holder for In Situ Atomic Resolution Scanning Transmission Electron Microscopic Observation.

    Science.gov (United States)

    Straubinger, Rainer; Beyer, Andreas; Volz, Kerstin

    2016-06-01

    A reproducible way to transfer a single crystalline sample into a gas environmental cell holder for in situ transmission electron microscopic (TEM) analysis is shown in this study. As in situ holders have only single-tilt capability, it is necessary to prepare the sample precisely along a specific zone axis. This can be achieved by a very accurate focused ion beam lift-out preparation. We show a step-by-step procedure to prepare the sample and transfer it into the gas environmental cell. The sample material is a GaP/Ga(NAsP)/GaP multi-quantum well structure on Si. Scanning TEM observations prove that it is possible to achieve atomic resolution at very high temperatures in a nitrogen environment of 100,000 Pa.

  2. A low cost high resolution pattern generator for electron-beam lithography

    International Nuclear Information System (INIS)

    Pennelli, G.; D'Angelo, F.; Piotto, M.; Barillaro, G.; Pellegrini, B.

    2003-01-01

    A simple, very low cost pattern generator for electron-beam lithography is presented. When it is applied to a scanning electron microscope, the system allows a high precision positioning of the beam for lithography of very small structures. Patterns are generated by a suitable software implemented on a personal computer, by using very simple functions, allowing an easy development of new writing strategies for a great adaptability to different user necessities. Hardware solutions, as optocouplers and battery supply, have been implemented for reduction of noise and disturbs on the voltages controlling the positioning of the beam

  3. A compact sub-Kelvin ultrahigh vacuum scanning tunneling microscope with high energy resolution and high stability.

    Science.gov (United States)

    Zhang, L; Miyamachi, T; Tomanić, T; Dehm, R; Wulfhekel, W

    2011-10-01

    We designed a scanning tunneling microscope working at sub-Kelvin temperatures in ultrahigh vacuum (UHV) in order to study the magnetic properties on the nanoscale. An entirely homebuilt three-stage cryostat is used to cool down the microscope head. The first stage is cooled with liquid nitrogen, the second stage with liquid (4)He. The third stage uses a closed-cycle Joule-Thomson refrigerator of a cooling power of 1 mW. A base temperature of 930 mK at the microscope head was achieved using expansion of (4)He, which can be reduced to ≈400 mK when using (3)He. The cryostat has a low liquid helium consumption of only 38 ml/h and standing times of up to 280 h. The fast cooling down of the samples (3 h) guarantees high sample throughput. Test experiments with a superconducting tip show a high energy resolution of 0.3 meV when performing scanning tunneling spectroscopy. The vertical stability of the tunnel junction is well below 1 pm (peak to peak) and the electric noise floor of tunneling current is about 6fA/√Hz. Atomic resolution with a tunneling current of 1 pA and 1 mV was achieved on Au(111). The lateral drift of the microscope at stable temperature is below 20 pm/h. A superconducting spilt-coil magnet allows to apply an out-of-plane magnetic field of up to 3 T at the sample surface. The flux vortices of a Nb(110) sample were clearly resolved in a map of differential conductance at 1.1 K and a magnetic field of 0.21 T. The setup is designed for in situ preparation of tip and samples under UHV condition.

  4. Unsupervised Cryo-EM Data Clustering through Adaptively Constrained K-Means Algorithm.

    Science.gov (United States)

    Xu, Yaofang; Wu, Jiayi; Yin, Chang-Cheng; Mao, Youdong

    2016-01-01

    In single-particle cryo-electron microscopy (cryo-EM), K-means clustering algorithm is widely used in unsupervised 2D classification of projection images of biological macromolecules. 3D ab initio reconstruction requires accurate unsupervised classification in order to separate molecular projections of distinct orientations. Due to background noise in single-particle images and uncertainty of molecular orientations, traditional K-means clustering algorithm may classify images into wrong classes and produce classes with a large variation in membership. Overcoming these limitations requires further development on clustering algorithms for cryo-EM data analysis. We propose a novel unsupervised data clustering method building upon the traditional K-means algorithm. By introducing an adaptive constraint term in the objective function, our algorithm not only avoids a large variation in class sizes but also produces more accurate data clustering. Applications of this approach to both simulated and experimental cryo-EM data demonstrate that our algorithm is a significantly improved alterative to the traditional K-means algorithm in single-particle cryo-EM analysis.

  5. Exploring semiconductor quantum dots and wires by high resolution electron microscopy

    Energy Technology Data Exchange (ETDEWEB)

    Molina, S I [Departamento de Ciencia de los Materiales e Ing Metalurgica y Q. Inorganica, F. de Ciencias, Universidad de Cadiz, Campus Rio San Pedro. 11510 Puerto Real (Cadiz) (Spain); Galindo, P L [Departamento de Lenguajes y Sistemas Informaticos, CASEM, Universidad de Cadiz, Campus Rio San Pedro. 11510 Puerto Real (Cadiz) (Spain); Gonzalez, L; Ripalda, J M [Instituto de Microelectronica de Madrid (CNM, CSIC), Isaac Newton 8, 28760 Tres Cantos, Madrid (Spain); Varela, M; Pennycook, S J, E-mail: sergio.molina@uca.e [Materials Science and Technology Division, Oak Ridge National Laboratory, Oak Ridge TN 37831 (United States)

    2010-02-01

    We review in this communication our contribution to the structural characterisation of semiconductor quantum dots and wires by high resolution electron microscopy, both in phase-contrast and Z-contrast modes. We show how these techniques contribute to predict the preferential sites of nucleation of these nanostructures, and also determine the compositional distribution in 1D and 0D nanostructures. The results presented here were produced in the framework of the European Network of Excellence entitled {sup S}elf-Assembled semiconductor Nanostructures for new Devices in photonics and Electronics (SANDiE){sup .}

  6. GraDeR: Membrane Protein Complex Preparation for Single-Particle Cryo-EM.

    Science.gov (United States)

    Hauer, Florian; Gerle, Christoph; Fischer, Niels; Oshima, Atsunori; Shinzawa-Itoh, Kyoko; Shimada, Satoru; Yokoyama, Ken; Fujiyoshi, Yoshinori; Stark, Holger

    2015-09-01

    We developed a method, named GraDeR, which substantially improves the preparation of membrane protein complexes for structure determination by single-particle cryo-electron microscopy (cryo-EM). In GraDeR, glycerol gradient centrifugation is used for the mild removal of free detergent monomers and micelles from lauryl maltose-neopentyl glycol detergent stabilized membrane complexes, resulting in monodisperse and stable complexes to which standard processes for water-soluble complexes can be applied. We demonstrate the applicability of the method on three different membrane complexes, including the mammalian FoF1 ATP synthase. For this highly dynamic and fragile rotary motor, we show that GraDeR allows visualizing the asymmetry of the F1 domain, which matches the ground state structure of the isolated domain. Therefore, the present cryo-EM structure of FoF1 ATP synthase provides direct structural evidence for Boyer's binding change mechanism in the context of the intact enzyme. Copyright © 2015 Elsevier Ltd. All rights reserved.

  7. Improved image quality and diagnostic potential using ultra-high-resolution computed tomography of the lung with small scan FOV: A prospective study.

    Directory of Open Access Journals (Sweden)

    Huiyuan Zhu

    Full Text Available The aim of this study was to assess whether CT imaging using an ultra-high-resolution CT (UHRCT scan with a small scan field of view (FOV provides higher image quality and helps to reduce the follow-up period compared with a conventional high-resolution CT (CHRCT scan. We identified patients with at least one pulmonary nodule at our hospital from July 2015 to November 2015. CHRCT and UHRCT scans were conducted in all enrolled patients. Three experienced radiologists evaluated the image quality using a 5-point score and made diagnoses. The paired images were displayed side by side in a random manner and annotations of scan information were removed. The following parameters including image quality, diagnostic confidence of radiologists, follow-up recommendations and diagnostic accuracy were assessed. A total of 52 patients (62 nodules were included in this study. UHRCT scan provides a better image quality regarding the margin of nodules and solid internal component compared to that of CHRCT (P 0.05. These findings suggest that the UHRCT prototype scanner provides a better image quality of subsolid nodules compared to CHRCT and contributes significantly to reduce the patients' follow-up period.

  8. Effects of display resolution and size on primary diagnosis of chest images using a high-resolution electronic work station

    International Nuclear Information System (INIS)

    Fuhrman, C.R.; Cooperstein, L.A.; Herron, J.; Good, W.F.; Good, B.; Gur, D.; Maitz, G.; Tabor, E.; Hoy, R.J.

    1987-01-01

    To evaluate the acceptability of electronically displayed planar images, the authors have a high-resolution work station. This system utilizes a high-resolution film digitizer (100-micro resolution) interfaced to a mainframe computer and two high-resolution (2,048 X 2,048) display devices (Azuray). In a clinically simulated multiobserver blind study (19 cases and five observers) a prodetermined series of reading sessions is stored on magnetic disk and is transferred to the displays while the preceding set of images is being reviewed. Images can be linearly processed on the fly into 2,000 X 2,000 full resolution, 1,000 X 1,000 minified display, or 1,000 X 1,000 interpolated for full-size display. Results of the study indicate that radiologists accept but do not like significant minification (more than X2), and they rate 2,000 X 2,000 images as having better diagnostic quality than 1,000 X 1,000 images

  9. High Resolution Trichromatic Road Surface Scanning with a Line Scan Camera and Light Emitting Diode Lighting for Road-Kill Detection

    Directory of Open Access Journals (Sweden)

    Gil Lopes

    2016-04-01

    Full Text Available This paper presents a road surface scanning system that operates with a trichromatic line scan camera with light emitting diode (LED lighting achieving road surface resolution under a millimeter. It was part of a project named Roadkills—Intelligent systems for surveying mortality of amphibians in Portuguese roads, sponsored by the Portuguese Science and Technology Foundation. A trailer was developed in order to accommodate the complete system with standalone power generation, computer image capture and recording, controlled lighting to operate day or night without disturbance, incremental encoder with 5000 pulses per revolution attached to one of the trailer wheels, under a meter Global Positioning System (GPS localization, easy to utilize with any vehicle with a trailer towing system and focused on a complete low cost solution. The paper describes the system architecture of the developed prototype, its calibration procedure, the performed experimentation and some obtained results, along with a discussion and comparison with existing systems. Sustained operating trailer speeds of up to 30 km/h are achievable without loss of quality at 4096 pixels’ image width (1 m width of road surface with 250 µm/pixel resolution. Higher scanning speeds can be achieved by lowering the image resolution (120 km/h with 1 mm/pixel. Computer vision algorithms are under development to operate on the captured images in order to automatically detect road-kills of amphibians.

  10. High Resolution Trichromatic Road Surface Scanning with a Line Scan Camera and Light Emitting Diode Lighting for Road-Kill Detection.

    Science.gov (United States)

    Lopes, Gil; Ribeiro, A Fernando; Sillero, Neftalí; Gonçalves-Seco, Luís; Silva, Cristiano; Franch, Marc; Trigueiros, Paulo

    2016-04-19

    This paper presents a road surface scanning system that operates with a trichromatic line scan camera with light emitting diode (LED) lighting achieving road surface resolution under a millimeter. It was part of a project named Roadkills-Intelligent systems for surveying mortality of amphibians in Portuguese roads, sponsored by the Portuguese Science and Technology Foundation. A trailer was developed in order to accommodate the complete system with standalone power generation, computer image capture and recording, controlled lighting to operate day or night without disturbance, incremental encoder with 5000 pulses per revolution attached to one of the trailer wheels, under a meter Global Positioning System (GPS) localization, easy to utilize with any vehicle with a trailer towing system and focused on a complete low cost solution. The paper describes the system architecture of the developed prototype, its calibration procedure, the performed experimentation and some obtained results, along with a discussion and comparison with existing systems. Sustained operating trailer speeds of up to 30 km/h are achievable without loss of quality at 4096 pixels' image width (1 m width of road surface) with 250 µm/pixel resolution. Higher scanning speeds can be achieved by lowering the image resolution (120 km/h with 1 mm/pixel). Computer vision algorithms are under development to operate on the captured images in order to automatically detect road-kills of amphibians.

  11. Environmental high resolution electron microscopy and applications to chemical science

    OpenAIRE

    Boyes, Edward; Gai, Pratibha

    2017-01-01

    An environmental cell high resolution electron microscope (EHREM) has been developed for in situ studies of dynamic chemical reactions on the atomic scale. It allows access to metastable intermediate phases of catalysts and to sequences of reversible microstructural and chemical development associated with the activation, deactivation and poisoning of a catalyst. Materials transported through air can be restored or recreated and samples damaged, e.g. by dehydration, by the usual vacuum enviro...

  12. Analysis of improvement in performance and design parameters for enhancing resolution in an atmospheric scanning electron microscope.

    Science.gov (United States)

    Yoon, Yeo Hun; Kim, Seung Jae; Kim, Dong Hwan

    2015-12-01

    The scanning electron microscope is used in various fields to go beyond diffraction limits of the optical microscope. However, the electron pathway should be conducted in a vacuum so as not to scatter electrons. The pretreatment of the sample is needed for use in the vacuum. To directly observe large and fully hydrophilic samples without pretreatment, the atmospheric scanning electron microscope (ASEM) is needed. We developed an electron filter unit and an electron detector unit for implementation of the ASEM. The key of the electron filter unit is that electrons are transmitted while air molecules remain untransmitted through the unit. The electron detector unit collected the backscattered electrons. We conducted experiments using the selected materials with Havar foil, carbon film and SiN film. © The Author 2015. Published by Oxford University Press on behalf of The Japanese Society of Microscopy. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

  13. Advanced radiographic scanning, enhancement and electronic data storage

    International Nuclear Information System (INIS)

    Savoie, C.; Rivest, D.

    2003-01-01

    It is a well-known fact that radiographs deteriorate with time. Substantial cost is attributed to cataloguing and storage. To eliminate deterioration issues and save time retrieving radiographs, laser scanning techniques were developed in conjunction with viewing and enhancement software. This will allow radiographs to be successfully scanned and stored electronically for future reference. Todays radiographic laser scanners are capable Qf capturing images with an optical density of up to 4.1 at 256 grey levels and resolutions up to 4096 pixels per line. An industrial software interface was developed for the nondestructive testing industry so that, certain parameters such as scan resolution, number of scans, file format and location to be saved could be adjusted as needed. Once the radiographs have been scanned, the tiff images are stored, or retrieved into Radiance software (developed by Rivest Technologies Inc.), which will help to properly interpret the radiographs. Radiance was developed to allow the user to quickly view the radiographs correctness or enhance its defects for comparison and future evaluation. Radiance also allows the user to zoom, measure and annotate areas of interest. Physical cost associated with cataloguing, storing and retrieving radiographs can be eliminated. You can now successfully retrieve and view your radiographs from CD media or dedicated hard drive at will. For continuous searches and/or field access, dedicated hard drives controlled by a server would be the media of choice. All scanned radiographs will be archived to CD media (CD-R). Laser scanning with a proper acquisition interface and easy to use viewing software will permit a qualified user to identify areas of interest and share this information with his/her colleagues via e-mail or web data access. (author)

  14. (CryoTransmission Electron Microscopy of Phospholipid Model Membranes Interacting with Amphiphilic and Polyphilic Molecules

    Directory of Open Access Journals (Sweden)

    Annette Meister

    2017-10-01

    Full Text Available Lipid membranes can incorporate amphiphilic or polyphilic molecules leading to specific functionalities and to adaptable properties of the lipid bilayer host. The insertion of guest molecules into membranes frequently induces changes in the shape of the lipid matrix that can be visualized by transmission electron microscopy (TEM techniques. Here, we review the use of stained and vitrified specimens in (cryoTEM to characterize the morphology of amphiphilic and polyphilic molecules upon insertion into phospholipid model membranes. Special emphasis is placed on the impact of novel synthetic amphiphilic and polyphilic bolalipids and polymers on membrane integrity and shape stability.

  15. Preliminary gravity recovery from CryoSat-2 data in the Baffin Bay

    DEFF Research Database (Denmark)

    Stenseng, Lars; Andersen, Ole Baltazar

    2012-01-01

    recovery of geophysical signal was achieved from measurements averaged to 5 Hz, rather than the simulated 20 Hz full resolution data. A first validation of these theoretical modelling results with respect to resolution and noise is presented using various CryoSat-2 data, and evaluated against conventional...

  16. Sparsity-Based Super Resolution for SEM Images.

    Science.gov (United States)

    Tsiper, Shahar; Dicker, Or; Kaizerman, Idan; Zohar, Zeev; Segev, Mordechai; Eldar, Yonina C

    2017-09-13

    The scanning electron microscope (SEM) is an electron microscope that produces an image of a sample by scanning it with a focused beam of electrons. The electrons interact with the atoms in the sample, which emit secondary electrons that contain information about the surface topography and composition. The sample is scanned by the electron beam point by point, until an image of the surface is formed. Since its invention in 1942, the capabilities of SEMs have become paramount in the discovery and understanding of the nanometer world, and today it is extensively used for both research and in industry. In principle, SEMs can achieve resolution better than one nanometer. However, for many applications, working at subnanometer resolution implies an exceedingly large number of scanning points. For exactly this reason, the SEM diagnostics of microelectronic chips is performed either at high resolution (HR) over a small area or at low resolution (LR) while capturing a larger portion of the chip. Here, we employ sparse coding and dictionary learning to algorithmically enhance low-resolution SEM images of microelectronic chips-up to the level of the HR images acquired by slow SEM scans, while considerably reducing the noise. Our methodology consists of two steps: an offline stage of learning a joint dictionary from a sequence of LR and HR images of the same region in the chip, followed by a fast-online super-resolution step where the resolution of a new LR image is enhanced. We provide several examples with typical chips used in the microelectronics industry, as well as a statistical study on arbitrary images with characteristic structural features. Conceptually, our method works well when the images have similar characteristics, as microelectronics chips do. This work demonstrates that employing sparsity concepts can greatly improve the performance of SEM, thereby considerably increasing the scanning throughput without compromising on analysis quality and resolution.

  17. A new topology of the HK97-like fold revealed in Bordetella bacteriophage by cryoEM at 3.5 Å resolution

    Science.gov (United States)

    Zhang, Xing; Guo, Huatao; Jin, Lei; Czornyj, Elizabeth; Hodes, Asher; Hui, Wong H; Nieh, Angela W; Miller, Jeff F; Zhou, Z Hong

    2013-01-01

    Bacteriophage BPP-1 infects and kills Bordetella species that cause whooping cough. Its diversity-generating retroelement (DGR) provides a naturally occurring phage-display system, but engineering efforts are hampered without atomic structures. Here, we report a cryo electron microscopy structure of the BPP-1 head at 3.5 Å resolution. Our atomic model shows two of the three protein folds representing major viral lineages: jellyroll for its cement protein (CP) and HK97-like (‘Johnson’) for its major capsid protein (MCP). Strikingly, the fold topology of MCP is permuted non-circularly from the Johnson fold topology previously seen in viral and cellular proteins. We illustrate that the new topology is likely the only feasible alternative of the old topology. β-sheet augmentation and electrostatic interactions contribute to the formation of non-covalent chainmail in BPP-1, unlike covalent inter-protein linkages of the HK97 chainmail. Despite these complex interactions, the termini of both CP and MCP are ideally positioned for DGR-based phage-display engineering. DOI: http://dx.doi.org/10.7554/eLife.01299.001 PMID:24347545

  18. Scanning electron microscopy

    Energy Technology Data Exchange (ETDEWEB)

    Cox, B. [Atomic Energy of Canada Limited, Chalk River, Ontario (Canada)

    1970-05-15

    The JSM-11 scanning electron microscope at CRNL has been used extensively for topographical studies of oxidized metals, fracture surfaces, entomological and biological specimens. A non-dispersive X-ray attachment permits the microanalysis of the surface features. Techniques for the production of electron channeling patterns have been developed. (author)

  19. ESA CryoVEx 2014 - Airborne ASIRAS radar and laser scanner measurements during 2014 CryoVEx campaign in the Arctic

    DEFF Research Database (Denmark)

    Hvidegaard, S. M.; Nielsen, J. E.; Sørensen, L. Sandberg

    the penetration depth of the ASIRAS radar. An opportunity site on the Greenland Ice Sheet was surveyed near Jakobshavn Isbræ. No other ground experiments were coordinated with the CryoVEx campaing on the Greenland Ice Sheet. The CryoVEx 2014 campaign was a success and the processed data is of high quality......This report outlines the airborne field operations with the ESA airborne Ku‐band interferometric radar (ASIRAS), coincident airborne laser scanner (ALS) and vertical photography to acquire data over sea‐ and land ice along validation sites and CryoSat‐2 ground tracks. The airborne campaign...... in the Beaufort Sea lead by US office of Naval Research (ONR) and north of Greenland as a dedicated ESA CryoVEx initiative. In addition, selected CryoSat‐2 ground tracks were under‐flown in the Lincoln Sea from CFS Alert, North of Greenland and Svalbard from St. Nord and Longyearbyen. Several of the flights...

  20. Dark-field imaging based on post-processed electron backscatter diffraction patterns of bulk crystalline materials in a scanning electron microscope.

    Science.gov (United States)

    Brodusch, Nicolas; Demers, Hendrix; Gauvin, Raynald

    2015-01-01

    Dark-field (DF) images were acquired in the scanning electron microscope with an offline procedure based on electron backscatter diffraction (EBSD) patterns (EBSPs). These EBSD-DF images were generated by selecting a particular reflection on the electron backscatter diffraction pattern and by reporting the intensity of one or several pixels around this point at each pixel of the EBSD-DF image. Unlike previous studies, the diffraction information of the sample is the basis of the final image contrast with a pixel scale resolution at the EBSP providing DF imaging in the scanning electron microscope. The offline facility of this technique permits the selection of any diffraction condition available in the diffraction pattern and displaying the corresponding image. The high number of diffraction-based images available allows a better monitoring of deformation structures compared to electron channeling contrast imaging (ECCI) which is generally limited to a few images of the same area. This technique was applied to steel and iron specimens and showed its high capability in describing more rigorously the deformation structures around micro-hardness indents. Due to the offline relation between the reference EBSP and the EBSD-DF images, this new technique will undoubtedly greatly improve our knowledge of deformation mechanism and help to improve our understanding of the ECCI contrast mechanisms. Copyright © 2014 Elsevier B.V. All rights reserved.

  1. Carbon K-shell excitation in small molecules by high-resolution electron impact

    International Nuclear Information System (INIS)

    Tronc, M.; King, G.C.; Read, F.H.

    1979-01-01

    The excitation of 1s carbon electrons has been observed in C0, CH 4 , CF4, C0 2 , COS, C 2 H 2 and C 2 H 4 by means of the electron energy-loss technique with high resolution (70 meV in the 300 eV excitation energy range) and at an incident electron energy of 1.5 keV. The energies, widths and vibrational structures of excited states corresponding to the promotion of 1s carbon electrons to unoccupied valence and Rydberg orbitals have been obtained. The validity of the equivalent-core model, and the role of resonances caused by potential barriers, are discussed. (author)

  2. High-resolution electron-beam patternable nanocomposite containing metal nanoparticles for plasmonics

    International Nuclear Information System (INIS)

    Abargues, R; Marques-Hueso, J; Canet-Ferrer, J; Pedrueza, E; Valdes, J L; Jimenez, E; MartInez-Pastor, J P

    2008-01-01

    Polymer nanocomposites containing noble metal nanoparticles are promising materials for plasmonic applications. In this paper, we report on a high-resolution negative-tone nanocomposite resist based on poly(vinyl alcohol) where silver nanoparticles and nanopatterns are simultaneously generated by electron-beam lithography. Our results indicate nanostructures with a relatively high concentration of nanoparticles and, consequently, an electromagnetic coupling among the nanoparticles. Therefore, the patternable nanocomposite described in this work may be a suitable material for future plasmonic circuitry

  3. Single-view volumetric PIV via high-resolution scanning, isotropic voxel restructuring and 3D least-squares matching (3D-LSM)

    International Nuclear Information System (INIS)

    Brücker, C; Hess, D; Kitzhofer, J

    2013-01-01

    Scanning PIV as introduced by Brücker (1995 Exp. Fluids 19 255–63, 1996a Appl. Sci. Res. 56 157–79) has been successfully applied in the last 20 years to different flow problems where the frame rate was sufficient to ensure a ‘frozen’ field condition. The limited number of parallel planes however leads typically to an under-sampling in the scan direction in depth; therefore, the spatial resolution in depth is typically considerably lower than the spatial resolution in the plane of the laser sheet (depth resolution = scan shift Δz ≫ pixel unit in object space). In addition, a partial volume averaging effect due to the thickness of the light sheet must be taken into account. Herein, the method is further developed using a high-resolution scanning in combination with a Gaussian regression technique to achieve an isotropic representation of the tracer particles in a voxel-based volume reconstruction with cuboidal voxels. This eliminates the partial volume averaging effect due to light sheet thickness and leads to comparable spatial resolution of the particle field reconstructions in x-, y- and z-axes. In addition, advantage of voxel-based processing with estimations of translation, rotation and shear/strain is taken by using a 3D least-squares matching method, well suited for reconstruction of grey-level pattern fields. The method is discussed in this paper and used to investigate the ring vortex instability at Re = 2500 within a measurement volume of roughly 75 × 75 × 50 mm 3 with a spatial resolution of 100 µm/voxel (750 × 750 × 500 voxel elements). The volume has been scanned with a number of 100 light sheets and scan rates of 10 kHz. The results show the growth of the Tsai–Widnall azimuthal instabilities accompanied with a precession of the axis of the vortex ring. Prior to breakdown, secondary instabilities evolve along the core with streamwise oriented striations. The front stagnation point's streamwise distance to the core starts to decrease

  4. Spatial localization of the Ebola virus glycoprotein mucin-like domain determined by cryo-electron tomography.

    Science.gov (United States)

    Tran, Erin E H; Simmons, James A; Bartesaghi, Alberto; Shoemaker, Charles J; Nelson, Elizabeth; White, Judith M; Subramaniam, Sriram

    2014-09-01

    The Ebola virus glycoprotein mucin-like domain (MLD) is implicated in Ebola virus cell entry and immune evasion. Using cryo-electron tomography of Ebola virus-like particles, we determined a three-dimensional structure for the full-length glycoprotein in a near-native state and compared it to that of a glycoprotein lacking the MLD. Our results, which show that the MLD is located at the apex and the sides of each glycoprotein monomer, provide a structural template for analysis of MLD function. Copyright © 2014, American Society for Microbiology. All Rights Reserved.

  5. Cryo-STEM-EDX spectroscopy for the characterisation of nanoparticles in cell culture media

    Science.gov (United States)

    Ilett, M.; Bamiduro, F.; Matar, O.; Brown, A.; Brydson, R.; Hondow, N.

    2017-09-01

    We present a study of barium titanate nanoparticles dispersed in cell culture media. Scanning transmission electron microscopy combined with energy dispersive X-ray spectroscopy was undertaken on samples prepared using both conventional drop casting and also plunge freezing and examination under cryogenic conditions. This showed that drying artefacts occurred during conventional sample preparation, whereby some salt components of the cell culture media accumulated around the barium titanate nanoparticles; these were removed using the cryogenic route. Importantly, the formation of a calcium and phosphorus rich coating around the barium titanate nanoparticles was retained under cryo-conditions, highlighting that significant interactions do occur between nanomaterials and biological media.

  6. Towards the low-dose characterization of beam sensitive nanostructures via implementation of sparse image acquisition in scanning transmission electron microscopy

    International Nuclear Information System (INIS)

    Hwang, Sunghwan; Han, Chang Wan; Ortalan, Volkan; Venkatakrishnan, Singanallur V; Bouman, Charles A

    2017-01-01

    Scanning transmission electron microscopy (STEM) has been successfully utilized to investigate atomic structure and chemistry of materials with atomic resolution. However, STEM’s focused electron probe with a high current density causes the electron beam damages including radiolysis and knock-on damage when the focused probe is exposed onto the electron-beam sensitive materials. Therefore, it is highly desirable to decrease the electron dose used in STEM for the investigation of biological/organic molecules, soft materials and nanomaterials in general. With the recent emergence of novel sparse signal processing theories, such as compressive sensing and model-based iterative reconstruction, possibilities of operating STEM under a sparse acquisition scheme to reduce the electron dose have been opened up. In this paper, we report our recent approach to implement a sparse acquisition in STEM mode executed by a random sparse-scan and a signal processing algorithm called model-based iterative reconstruction (MBIR). In this method, a small portion, such as 5% of randomly chosen unit sampling areas (i.e. electron probe positions), which corresponds to pixels of a STEM image, within the region of interest (ROI) of the specimen are scanned with an electron probe to obtain a sparse image. Sparse images are then reconstructed using the MBIR inpainting algorithm to produce an image of the specimen at the original resolution that is consistent with an image obtained using conventional scanning methods. Experimental results for down to 5% sampling show consistency with the full STEM image acquired by the conventional scanning method. Although, practical limitations of the conventional STEM instruments, such as internal delays of the STEM control electronics and the continuous electron gun emission, currently hinder to achieve the full potential of the sparse acquisition STEM in realizing the low dose imaging condition required for the investigation of beam-sensitive materials

  7. Nanoscale Energy-Filtered Scanning Confocal Electron Microscopy Using a Double-Aberration-Corrected Transmission Electron Microscope

    International Nuclear Information System (INIS)

    Wang Peng; Behan, Gavin; Kirkland, Angus I.; Nellist, Peter D.; Takeguchi, Masaki; Hashimoto, Ayako; Mitsuishi, Kazutaka; Shimojo, Masayuki

    2010-01-01

    We demonstrate that a transmission electron microscope fitted with two spherical-aberration correctors can be operated as an energy-filtered scanning confocal electron microscope. A method for establishing this mode is described and initial results showing 3D chemical mapping with nanoscale sensitivity to height and thickness changes in a carbon film are presented. Importantly, uncorrected chromatic aberration does not limit the depth resolution of this technique and moreover performs an energy-filtering role, which is explained in terms of a combined depth and energy-loss response function.

  8. Development of an environmental high-voltage electron microscope for reaction science.

    Science.gov (United States)

    Tanaka, Nobuo; Usukura, Jiro; Kusunoki, Michiko; Saito, Yahachi; Sasaki, Katuhiro; Tanji, Takayoshi; Muto, Shunsuke; Arai, Shigeo

    2013-02-01

    Environmental transmission electron microscopy and ultra-high resolution electron microscopic observation using aberration correctors have recently emerged as topics of great interest. The former method is an extension of the so-called in situ electron microscopy that has been performed since the 1970s. Current research in this area has been focusing on dynamic observation with atomic resolution under gaseous atmospheres and in liquids. Since 2007, Nagoya University has been developing a new 1-MV high voltage (scanning) transmission electron microscope that can be used to observe nanomaterials under conditions that include the presence of gases, liquids and illuminating lights, and it can be also used to perform mechanical operations to nanometre-sized areas as well as electron tomography and elemental analysis by electron energy loss spectroscopy. The new instrument has been used to image and analyse various types of samples including biological ones.

  9. Implementation and performance of an optical motion tracking system for high resolution brain PET imaging

    Science.gov (United States)

    Lopresti, B. J.; Russo, A.; Jones, W. F.; Fisher, T.; Crouch, D. G.; Altenburger, D. E.; Townsend, D. W.

    1999-12-01

    Head motion during PET scanning is widely regarded as a source of image degradation and resolution loss. Recent improvements in the spatial resolution of state-of-the-art tomographs may be compromised by patient motion during scanning, as these high resolution data will be increasingly susceptible to smaller movements of the head. The authors have developed an opto-electronic motion tracking system based on commercially-available technology that is capable of very accurate real-time measurements of the position and orientation of the patient's head. These positions are transformed to the reference frame of the PET scanner, and could potentially be used to provide motion correction of list-mode emission data on an event-by-event basis.

  10. Atomic resolution three-dimensional electron diffraction microscopy

    International Nuclear Information System (INIS)

    Miao Jianwei; Ohsuna, Tetsu; Terasaki, Osamu; Hodgson, Keith O.; O'Keefe, Michael A.

    2002-01-01

    We report the development of a novel form of diffraction-based 3D microscopy to overcome resolution barriers inherent in high-resolution electron microscopy and tomography. By combining coherent electron diffraction with the oversampling phasing method, we show that the 3D structure of a nanocrystal can be determined ab initio at a resolution of 1 Angstrom from 29 simulated noisy diffraction patterns. This new form of microscopy can be used to image the 3D structures of nanocrystals and noncrystalline samples, with resolution limited only by the quality of sample diffraction

  11. Novel method of simultaneous multiple immunogold localization on resin sections in high resolution scanning electron microscopy

    Czech Academy of Sciences Publication Activity Database

    Nebesářová, Jana; Wandrol, P.; Vancová, Marie

    2016-01-01

    Roč. 12, č. 1 (2016), s. 105-517 ISSN 1549-9634 R&D Projects: GA TA ČR(CZ) TE01020118 Institutional support: RVO:60077344 Keywords : multiple immunolabeling * gold nanoparticles * high resolution SEM * STEM imaging * BSE imaging Subject RIV: EA - Cell Biology Impact factor: 5.720, year: 2016

  12. Atom-counting in High Resolution Electron Microscopy:TEM or STEM - That's the question.

    Science.gov (United States)

    Gonnissen, J; De Backer, A; den Dekker, A J; Sijbers, J; Van Aert, S

    2017-03-01

    In this work, a recently developed quantitative approach based on the principles of detection theory is used in order to determine the possibilities and limitations of High Resolution Scanning Transmission Electron Microscopy (HR STEM) and HR TEM for atom-counting. So far, HR STEM has been shown to be an appropriate imaging mode to count the number of atoms in a projected atomic column. Recently, it has been demonstrated that HR TEM, when using negative spherical aberration imaging, is suitable for atom-counting as well. The capabilities of both imaging techniques are investigated and compared using the probability of error as a criterion. It is shown that for the same incoming electron dose, HR STEM outperforms HR TEM under common practice standards, i.e. when the decision is based on the probability function of the peak intensities in HR TEM and of the scattering cross-sections in HR STEM. If the atom-counting decision is based on the joint probability function of the image pixel values, the dependence of all image pixel intensities as a function of thickness should be known accurately. Under this assumption, the probability of error may decrease significantly for atom-counting in HR TEM and may, in theory, become lower as compared to HR STEM under the predicted optimal experimental settings. However, the commonly used standard for atom-counting in HR STEM leads to a high performance and has been shown to work in practice. Copyright © 2017 Elsevier B.V. All rights reserved.

  13. Removing Contamination-Induced Reconstruction Artifacts from Cryo-electron Tomograms

    Science.gov (United States)

    Fernandez, Jose-Jesus; Laugks, Ulrike; Schaffer, Miroslava; Bäuerlein, Felix J.B.; Khoshouei, Maryam; Baumeister, Wolfgang; Lucic, Vladan

    2016-01-01

    Imaging of fully hydrated, vitrified biological samples by electron tomography yields structural information about cellular protein complexes in situ. Here we present a computational procedure that removes artifacts of three-dimensional reconstruction caused by contamination present in samples during imaging by electron microscopy. Applying the procedure to phantom data and electron tomograms of cellular samples significantly improved the resolution and the interpretability of tomograms. Artifacts caused by surface contamination associated with thinning by focused ion beam, as well as those arising from gold fiducial markers and from common, lower contrast contamination, could be removed. Our procedure is widely applicable and is especially suited for applications that strive to reach a higher resolution and involve the use of recently developed, state-of-the-art instrumentation. PMID:26743046

  14. High resolution electron attachment to molecules of atmospheric relevance

    International Nuclear Information System (INIS)

    Senn, G.

    2000-10-01

    This Ph.D. thesis is divided into three parts. The first is an introduction into the field of electron attachment. In the second part the experimental apparatus is described, and in the third part the results are presented. In the present thesis molecules were chosen for our investigations that are not only of academic interest but that also play an important role for applications or even the life on this planet. All the molecules studied in this work are of atmospheric relevance. NO, and OClO, are involved in the ozone depletion of the stratosphere. The D-layer of the ionosphere is an upper boundary of the ozone layer, therefore the interaction of the electrons from the D-layer with O 3 might play an important role for the chemistry in that part of the atmosphere. Especially the interaction of slow electrons (that will be present in the D-layer in large numbers) with ozone was emphasized in the present study. The production of O - and O 2 - by dissociative electron attachment to ozone was measured for incident electron energies between 0 and 10eV. A previously unobserved sharp structure was discovered in the formation of O - ions for electrons with zero kinetic energy. This additional cross section peak has important consequences for the role of ozone in the anion formation process in the ionosphere. Since OClO is a night time reservoir for chlorine atoms (Cl) and chlorine monoxide (ClO) both of which play a critical role in the depletion of the stratospheric ozone, we have studied negative ion formation following electron impact (0-10eV) to OClO. Despite its atmospheric relevance the mechanism of dissociative electron attachment (DEA) to NO is still a matter of controversy. DEA was studied at high energy resolution and with a kinetic-energy analysis of the O - fragment in two independent crossed electron-molecular-beam experiments. The DEA cross section exhibits a vertical onset near 7.45eV that corresponds to the energy threshold of the DEA channel O - ( 2 P

  15. Acquisition parameters optimization of a transmission electron forward scatter diffraction system in a cold-field emission scanning electron microscope for nanomaterials characterization.

    Science.gov (United States)

    Brodusch, Nicolas; Demers, Hendrix; Trudeau, Michel; Gauvin, Raynald

    2013-01-01

    Transmission electron forward scatter diffraction (t-EFSD) is a new technique providing crystallographic information with high resolution on thin specimens by using a conventional electron backscatter diffraction (EBSD) system in a scanning electron microscope. In this study, the impact of tilt angle, working distance, and detector distance on the Kikuchi pattern quality were investigated in a cold-field emission scanning electron microscope (CFE-SEM). We demonstrated that t-EFSD is applicable for tilt angles ranging from -20° to -40°. Working distance (WD) should be optimized for each material by choosing the WD for which the EBSD camera screen illumination is the highest, as the number of detected electrons on the screen is directly dependent on the scattering angle. To take advantage of the best performances of the CFE-SEM, the EBSD camera should be close to the sample and oriented towards the bottom to increase forward scattered electron collection efficiency. However, specimen chamber cluttering and beam/mechanical drift are important limitations in the CFE-SEM used in this work. Finally, the importance of t-EFSD in materials science characterization was illustrated through three examples of phase identification and orientation mapping. © Wiley Periodicals, Inc.

  16. A new high resolution tidal model in the arctic ocean

    DEFF Research Database (Denmark)

    Cancet, M.; Andersen, Ole Baltazar; Lyard, F.

    The Arctic Ocean is a challenging region for tidal modeling, because of its complex and not well-documented bathymetry, together combined with the intermittent presence of sea ice and the fact that the in situ tidal observations are rather scarce at such high latitudes. As a consequence, the accu......The Arctic Ocean is a challenging region for tidal modeling, because of its complex and not well-documented bathymetry, together combined with the intermittent presence of sea ice and the fact that the in situ tidal observations are rather scarce at such high latitudes. As a consequence......, the accuracy of the global tidal models decreases by several centimeters in the Polar Regions. In particular, it has a large impact on the quality of the satellite altimeter sea surface heights in these regions (ERS1/2, Envisat, CryoSat-2, SARAL/AltiKa and the future Sentinel-3 mission). Better knowledge......-growing maritime and industrial activities in this region. NOVELTIS and DTU Space have developed a regional, high-resolution tidal atlas in the Arctic Ocean, in the framework of the CryoSat Plus for Ocean (CP4O) ESA project. In particular, this atlas benefits from the assimilation of the most complete satellite...

  17. Fabrication of thin TEM sample of ionic liquid for high-resolution ELNES measurements

    Energy Technology Data Exchange (ETDEWEB)

    Miyata, Tomohiro, E-mail: tomo-m@iis.u-tokyo.ac.jp; Mizoguchi, Teruyasu, E-mail: teru@iis.u-tokyo.ac.jp

    2017-07-15

    Investigation of the local structure, ionic and molecular behavior, and chemical reactions at high spatial resolutions in liquids has become increasingly important. Improvements in these areas help to develop efficient batteries and improve organic syntheses. Transmission electron microscopy (TEM) and scanning-TEM (STEM) have excellent spatial resolution, and the electron energy-loss near edge structure (ELNES) measured by the accompanied electron energy-loss spectroscopy (EELS) is effective to analyze the liquid local structure owing to reflecting the electronic density of states. In this study, we fabricate a liquid-layer-only sample with thickness of single to tens nanometers using an ionic liquid. Because the liquid film has a thickness much less than the inelastic mean free path (IMFP) of the electron beam, the fine structure of the C-K edge electron energy loss near edge structure (ELNES) can be measured with sufficient resolution to allow meaningful analysis. The ELNES spectrum from the thin liquid film has been interpreted using first principles ELNES calculations. - Highlights: • A fabrication method of thin liquid film samples for STEM-EELS observations is proposed. • The thickness of the fabricated thin liquid film is about 10 nm. • An ELNES is measured from the thin liquid with a high energy resolution. • The peaks of the ELNES are interpreted using first principles calculations.

  18. Improved image quality and diagnostic potential using ultra-high-resolution computed tomography of the lung with small scan FOV: A prospective study.

    Science.gov (United States)

    Zhu, Huiyuan; Zhang, Lian; Wang, Yali; Hamal, Preeti; You, Xiaofang; Mao, Haixia; Li, Fei; Sun, Xiwen

    2017-01-01

    The aim of this study was to assess whether CT imaging using an ultra-high-resolution CT (UHRCT) scan with a small scan field of view (FOV) provides higher image quality and helps to reduce the follow-up period compared with a conventional high-resolution CT (CHRCT) scan. We identified patients with at least one pulmonary nodule at our hospital from July 2015 to November 2015. CHRCT and UHRCT scans were conducted in all enrolled patients. Three experienced radiologists evaluated the image quality using a 5-point score and made diagnoses. The paired images were displayed side by side in a random manner and annotations of scan information were removed. The following parameters including image quality, diagnostic confidence of radiologists, follow-up recommendations and diagnostic accuracy were assessed. A total of 52 patients (62 nodules) were included in this study. UHRCT scan provides a better image quality regarding the margin of nodules and solid internal component compared to that of CHRCT (P images than of CHRCT images (Pimages (P 0.05). These findings suggest that the UHRCT prototype scanner provides a better image quality of subsolid nodules compared to CHRCT and contributes significantly to reduce the patients' follow-up period.

  19. High-resolution electron collision spectroscopy with multicharged ions in merged beams

    Energy Technology Data Exchange (ETDEWEB)

    Lestinsky, M.

    2007-04-18

    The Heidelberg ion storage ring Tsr is currently the only ring equipped with two independent devices for the collinear merging of a cold electron beam with stored ions. This greatly improves the potential of electron-ion collision experiments, as the ion beam can be cooled with one electron beam, while the other one is used as a dedicated target for energy-resolved electron collision processes, such as recombination. The work describes the implementation of this system for rst electron collision spectroscopy experiments. A detection system has been realized including an ion detector and specroscopic beam-control software and instrumentation. Moreover, in order to improve the spectroscopic resolution systematical studies of intrinsic relaxation processes in the electron beam have been carried out. These include the dependence on the electron beam density, the magnetic guiding eld strength, and the acceleration geometry. The recombination measurements on low-lying resonances in lithiumlike Sc{sup 18+} yield a high-precision measurement of the 2s-2p{sub 3/2} transition energy in this system. Operation of the two-electron-beam setup at high collision energy ({approx}1000 eV) is established using resonances of hydrogenlike Mg{sup 11+}, while the unique possibility of modifying the beam-merging geometry con rms its importance for the electron-ion recombination rate at lowest relative energy, as demonstrated on F{sup 6+}. (orig.)

  20. High-resolution electron collision spectroscopy with multicharged ions in merged beams

    International Nuclear Information System (INIS)

    Lestinsky, M.

    2007-01-01

    The Heidelberg ion storage ring Tsr is currently the only ring equipped with two independent devices for the collinear merging of a cold electron beam with stored ions. This greatly improves the potential of electron-ion collision experiments, as the ion beam can be cooled with one electron beam, while the other one is used as a dedicated target for energy-resolved electron collision processes, such as recombination. The work describes the implementation of this system for rst electron collision spectroscopy experiments. A detection system has been realized including an ion detector and specroscopic beam-control software and instrumentation. Moreover, in order to improve the spectroscopic resolution systematical studies of intrinsic relaxation processes in the electron beam have been carried out. These include the dependence on the electron beam density, the magnetic guiding eld strength, and the acceleration geometry. The recombination measurements on low-lying resonances in lithiumlike Sc 18+ yield a high-precision measurement of the 2s-2p 3/2 transition energy in this system. Operation of the two-electron-beam setup at high collision energy (∼1000 eV) is established using resonances of hydrogenlike Mg 11+ , while the unique possibility of modifying the beam-merging geometry con rms its importance for the electron-ion recombination rate at lowest relative energy, as demonstrated on F 6+ . (orig.)

  1. Improved methods for high resolution electron microscopy

    Energy Technology Data Exchange (ETDEWEB)

    Taylor, J.R.

    1987-04-01

    Existing methods of making support films for high resolution transmission electron microscopy are investigated and novel methods are developed. Existing methods of fabricating fenestrated, metal reinforced specimen supports (microgrids) are evaluated for their potential to reduce beam induced movement of monolamellar crystals of C/sub 44/H/sub 90/ paraffin supported on thin carbon films. Improved methods of producing hydrophobic carbon films by vacuum evaporation, and improved methods of depositing well ordered monolamellar paraffin crystals on carbon films are developed. A novel technique for vacuum evaporation of metals is described which is used to reinforce microgrids. A technique is also developed to bond thin carbon films to microgrids with a polymer bonding agent. Unique biochemical methods are described to accomplish site specific covalent modification of membrane proteins. Protocols are given which covalently convert the carboxy terminus of papain cleaved bacteriorhodopsin to a free thiol. 53 refs., 19 figs., 1 tab.

  2. Thermal Design and Performance results of the first High-Beta Cryo-module for HIE-ISOLDE at CERN

    International Nuclear Information System (INIS)

    Valdarno, L; Delruelle, N; Leclercq, Y; Parma, V; Vandoni, G; Williams, L

    2015-01-01

    The High Energy and Intensity HIE-ISOLDE is a facility under construction at CERN whose target is ultimately, after the installation of six cryo-modules, to produce radioactive ion beams at 10MeV/u maximum energy in order to significantly expand the nuclear physics programme carried out by REX-ISOLDE. Since thermal control is essential to the performance of the whole cryo-module, a combination of a passive (materials, coatings, and surface finishes) and active (cryogenic loops, heaters) control has been designed to keep the cryostat operating within the allowable thermal budget. A numerical model based on Finite Element has been developed in order to generate a faithful global mapping of temperatures and heat fluxes inside the cryo-module. The numerical model, combined with the experimental results of the first test campaign, will serve as an optimization tool for the future cryo-modules in terms of improvement in the global and specific heat loads management. (paper)

  3. Thermal Design and Performance results of the first High-Beta Cryo-module for HIE-ISOLDE at CERN

    CERN Document Server

    Valdarno, L; Leclercq, Y; Parma, V; Vandoni, G; Williams, L

    2015-01-01

    The High Energy and Intensity HIE-ISOLDE is a facility under construction at CERN whose target is ultimately, after the installation of six cryo-modules, to produce radioactive ion beams at 10MeV/u maximum energy in order to significantly expand the nuclear physics programme carried out by REX-ISOLDE. Since thermal control is essential to the performance of the whole cryo-module, a combination of a passive (materials, coatings, and surface finishes) and active (cryogenic loops, heaters) control has been designed to keep the cryostat operating within the allowable thermal budget. A numerical model based on Finite Element has been developed in order to generate a faithful global mapping of temperatures and heat fluxes inside the cryo-module. The numerical model, combined with the experimental results of the first test campaign, will serve as an optimization tool for the future cryo-modules in terms of improvement in the global and specific heat loads management.

  4. Analysis of self-organized In(Ga)As quantum structures with the scanning transmission electron microscope

    International Nuclear Information System (INIS)

    Sauerwald, Andres

    2008-01-01

    Aim of this thesis was to apply the analytical methods of the scanning transmission electron microscopy to the study of self-organized In(Ga)As quantum structures. With the imaging methods Z contrast and bright field (position resolutions in the subnanometer range) and especially with the possibilities of the quantitative chemical EELS analysis of the scanning transmission electron microscope (STEM) fundamental questions concerning morphology and chemical properties of self-organized quantum structures should be answered. By the high position resolution of the STEM among others essentail morphological and structural parameters in the growth behaviour of ''dot in a well'' (DWell) structures and of vertically correlated quantum dots (QDs) could be analyzed. For the optimization of DWell structures samples were studied, the nominal InAs-QD growth position was directedly varied within the embedding InGaAs quantum wells. The STEM offers in connection with the EELS method a large potential for the chemical analysis of quantum structures. Studied was a sample series of self-organized InGaAs/GaAs structures on GaAs substrate, the stress of which was changed by varying the Ga content of the INGaAs material between 2.4 % and 4.3 % [de

  5. Not all trees sleep the same - High temporal resolution terrestrial laser scanning shows differences in nocturnal plant movement

    DEFF Research Database (Denmark)

    Zlinszky, András; Barfod, Anders; Molnár, Bence

    2017-01-01

    Circadian leaf movements are widely known in plants, but nocturnal movement of tree branches were only recently discovered by using terrestrial laser scanning (TLS), a high resolution three-dimensional surveying technique. TLS uses a pulsed laser emitted in a regular scan pattern for rapid...... surveyed a series of 18 full scans over a 12-h night period to measure nocturnal changes in shape simultaneously for an experimental setup of 22 plants representing different species. Resulting point clouds were evaluated by comparing changes in height percentiles of laser scanning points belonging...... to the canopy. Changes in crown shape were observed for all studied trees, but clearly distinguishable sleep movements are apparently rare. Ambient light conditions were continuously dark between sunset (7:30 p.m.) and sunrise (6:00 a.m.), but most changes in movement direction occurred during this period, thus...

  6. Cryo-electron microscopy and three-dimensional reconstructions of hepatitis C virus particles

    International Nuclear Information System (INIS)

    Yu Xuekui; Qiao Ming; Atanasov, Ivo; Hu Zongyi; Kato, Takanobu; Liang, T. Jake; Zhou, Z. Hong

    2007-01-01

    The structural details of hepatitis C virus (HCV) have been elusive because of the lack of a robust tissue culture system for producing an adequate amount of virions from infectious sources for in-depth three-dimensional (3D) structural analysis. Using both negative-stain and cryo-electron microscopy (cryoEM), we show that HCV virions isolated from cell culture have a rather uniform size of 500 A in diameter and that recombinantly expressed HCV-like particles (HCV-LPs) have similar morphologic, biophysical and antigenic features in spite of the varying sizes of the particles. 3D reconstructions were obtained from HCV-LPs with the same size as the HCV virions in the presence and absence of monoclonal antibodies bound to the E1 glycoprotein. The 3D reconstruction of HCV-LP reveals a multilayered architecture, with smooth outer-layer densities arranged in a 'fishbone' configuration. Reconstruction of the particles in complex with anti-E1 antibodies shows that sites of the E1 epitope are exposed and surround the 5-, 3- and 2-fold axes. The binding pattern of the anti-E1 antibody and the fitting of the structure of the dengue virus E glycoprotein into our 3D reconstructions further suggest that the HCV-LP E1 and E2 proteins form a tetramer (or dimer of heterodimers) that corresponds morphologically and functionally to the flavivirus E homodimer. This first 3D structural analysis of HCV particles offers important insights into the elusive mechanisms of HCV assembly and maturation

  7. Aberration-corrected scanning transmission electron microscopy for complex transition metal oxides

    Science.gov (United States)

    Qing-Hua, Zhang; Dong-Dong, Xiao; Lin, Gu

    2016-06-01

    Lattice, charge, orbital, and spin are the four fundamental degrees of freedom in condensed matter, of which the interactive coupling derives tremendous novel physical phenomena, such as high-temperature superconductivity (high-T c SC) and colossal magnetoresistance (CMR) in strongly correlated electronic system. Direct experimental observation of these freedoms is essential to understanding the structure-property relationship and the physics behind it, and also indispensable for designing new materials and devices. Scanning transmission electron microscopy (STEM) integrating multiple techniques of structure imaging and spectrum analysis, is a comprehensive platform for providing structural, chemical and electronic information of materials with a high spatial resolution. Benefiting from the development of aberration correctors, STEM has taken a big breakthrough towards sub-angstrom resolution in last decade and always steps forward to improve the capability of material characterization; many improvements have been achieved in recent years, thereby giving an in-depth insight into material research. Here, we present a brief review of the recent advances of STEM by some representative examples of perovskite transition metal oxides; atomic-scale mapping of ferroelectric polarization, octahedral distortions and rotations, valence state, coordination and spin ordering are presented. We expect that this brief introduction about the current capability of STEM could facilitate the understanding of the relationship between functional properties and these fundamental degrees of freedom in complex oxides. Project supported by the National Key Basic Research Project, China (Grant No. 2014CB921002), the Strategic Priority Research Program of Chinese Academy of Sciences (Grant No. XDB07030200), and the National Natural Science Foundation of China (Grant Nos. 51522212 and 51421002).

  8. Label-free visualization of ultrastructural features of artificial synapses via cryo-EM.

    Science.gov (United States)

    Gopalakrishnan, Gopakumar; Yam, Patricia T; Madwar, Carolin; Bostina, Mihnea; Rouiller, Isabelle; Colman, David R; Lennox, R Bruce

    2011-12-21

    The ultrastructural details of presynapses formed between artificial substrates of submicrometer silica beads and hippocampal neurons are visualized via cryo-electron microscopy (cryo-EM). The silica beads are derivatized by poly-d-lysine or lipid bilayers. Molecular features known to exist at presynapses are clearly present at these artificial synapses, as visualized by cryo-EM. Key synaptic features such as the membrane contact area at synaptic junctions, the presynaptic bouton containing presynaptic vesicles, as well as microtubular structures can be identified. This is the first report of the direct, label-free observation of ultrastructural details of artificial synapses.

  9. CryoEM structure of the human SLC4A4 sodium-coupled acid-base transporter NBCe1.

    Science.gov (United States)

    Huynh, Kevin W; Jiang, Jiansen; Abuladze, Natalia; Tsirulnikov, Kirill; Kao, Liyo; Shao, Xuesi; Newman, Debra; Azimov, Rustam; Pushkin, Alexander; Zhou, Z Hong; Kurtz, Ira

    2018-03-02

    Na + -coupled acid-base transporters play essential roles in human biology. Their dysfunction has been linked to cancer, heart, and brain disease. High-resolution structures of mammalian Na + -coupled acid-base transporters are not available. The sodium-bicarbonate cotransporter NBCe1 functions in multiple organs and its mutations cause blindness, abnormal growth and blood chemistry, migraines, and impaired cognitive function. Here, we have determined the structure of the membrane domain dimer of human NBCe1 at 3.9 Å resolution by cryo electron microscopy. Our atomic model and functional mutagenesis revealed the ion accessibility pathway and the ion coordination site, the latter containing residues involved in human disease-causing mutations. We identified a small number of residues within the ion coordination site whose modification transformed NBCe1 into an anion exchanger. Our data suggest that symporters and exchangers utilize comparable transport machinery and that subtle differences in their substrate-binding regions have very significant effects on their transport mode.

  10. High-resolution line-scan analysis of resin-embedded sediments using laser ablation-inductively coupled plasma-mass spectrometry (LA-ICP-MS)

    NARCIS (Netherlands)

    Hennekam, R.; Jilbert, T.; de Lange, G.J.; Reichart, G.J.

    2015-01-01

    Laser ablation-inductively coupled plasma-mass spectrometry (LA-ICP-MS) line-scanning is a promising technique for producing high-resolution (µm-scale) geochemical records on resin-embedded sediments. However, this approach has not yet been thoroughly tested on sediment samples of known elemental

  11. In situ fatigue loading stage inside scanning electron microscope

    Science.gov (United States)

    Telesman, Jack; Kantzos, Peter; Brewer, David

    1988-01-01

    A fatigue loading stage inside a scanning electron microscopy (SEM) was developed. The stage allows dynamic and static high-magnification and high-resolution viewing of the fatigue crack initiation and crack propagation processes. The loading stage is controlled by a closed-loop servohydraulic system. Maximum load is 1000 lb (4450 N) with test frequencies ranging up to 30 Hz. The stage accommodates specimens up to 2 inches (50 mm) in length and tolerates substantial specimen translation to view the propagating crack. At room temperature, acceptable working resolution is obtainable for magnifications ranging up to 10,000X. The system is equipped with a high-temperature setup designed for temperatures up to 2000 F (1100 C). The signal can be videotaped for further analysis of the pertinent fatigue damage mechanisms. The design allows for quick and easy interchange and conversion of the SEM from a loading stage configuration to its normal operational configuration and vice versa. Tests are performed entirely in the in-situ mode. In contrast to other designs, the NASA design has greatly extended the life of the loading stage by not exposing the bellows to cyclic loading. The loading stage was used to investigate the fatigue crack growth mechanisms in the (100)-oriented PWA 1480 single-crystal, nickel-based supperalloy. The high-magnification observations revealed the details of the crack growth processes.

  12. A Small Crack Length Evaluation Technique by Electronic Scanning

    International Nuclear Information System (INIS)

    Cho, Yong Sang; Kim, Jae Hoon

    2009-01-01

    The results of crack evaluation by conventional UT(Ultrasonic Test)is highly depend on the inspector's experience or knowledge of ultrasound. Phased array UT system and its application methods for small crack length evaluation will be a good alternative method which overcome present UT weakness. This study was aimed at checking the accuracy of crack length evaluation method by electronic scanning and discuss about characteristics of electronic scanning for crack length evaluation. Especially ultrasonic phased array with electronic scan technique was used in carrying out both sizing and detect ability of crack as its length changes. The response of ultrasonic phased array was analyzed to obtain the special method of determining crack length without moving the transducer and detectability of crack minimal length and depth from the material. A method of crack length determining by electronic scanning for the small crack is very real method which has it's accuracy and verify the effectiveness of method compared to a conventional crack length determination

  13. Cryo-EM structures of the human endolysosomal TRPML3 channel in three distinct states.

    Science.gov (United States)

    Zhou, Xiaoyuan; Li, Minghui; Su, Deyuan; Jia, Qi; Li, Huan; Li, Xueming; Yang, Jian

    2017-12-01

    TRPML3 channels are mainly localized to endolysosomes and play a critical role in the endocytic pathway. Their dysfunction causes deafness and pigmentation defects in mice. TRPML3 activity is inhibited by low endolysosomal pH. Here we present cryo-electron microscopy (cryo-EM) structures of human TRPML3 in the closed, agonist-activated, and low-pH-inhibited states, with resolutions of 4.06, 3.62, and 4.65 Å, respectively. The agonist ML-SA1 lodges between S5 and S6 and opens an S6 gate. A polycystin-mucolipin domain (PMD) forms a luminal cap. S1 extends into this cap, forming a 'gating rod' that connects directly to a luminal pore loop, which undergoes dramatic conformational changes in response to low pH. S2 extends intracellularly and interacts with several intracellular regions to form a 'gating knob'. These unique structural features, combined with the results of electrophysiological studies, indicate a new mechanism by which luminal pH and other physiological modulators such as PIP 2 regulate TRPML3 by changing S1 and S2 conformations.

  14. 3D imaging by serial block face scanning electron microscopy for materials science using ultramicrotomy.

    Science.gov (United States)

    Hashimoto, Teruo; Thompson, George E; Zhou, Xiaorong; Withers, Philip J

    2016-04-01

    Mechanical serial block face scanning electron microscopy (SBFSEM) has emerged as a means of obtaining three dimensional (3D) electron images over volumes much larger than possible by focused ion beam (FIB) serial sectioning and at higher spatial resolution than achievable with conventional X-ray computed tomography (CT). Such high resolution 3D electron images can be employed for precisely determining the shape, volume fraction, distribution and connectivity of important microstructural features. While soft (fixed or frozen) biological samples are particularly well suited for nanoscale sectioning using an ultramicrotome, the technique can also produce excellent 3D images at electron microscope resolution in a time and resource-efficient manner for engineering materials. Currently, a lack of appreciation of the capabilities of ultramicrotomy and the operational challenges associated with minimising artefacts for different materials is limiting its wider application to engineering materials. Consequently, this paper outlines the current state of the art for SBFSEM examining in detail how damage is introduced during slicing and highlighting strategies for minimising such damage. A particular focus of the study is the acquisition of 3D images for a variety of metallic and coated systems. Copyright © 2016 The Authors. Published by Elsevier B.V. All rights reserved.

  15. High-resolution line-scan analysis of resin-embedded sediments using laser ablation-inductively coupled plasma-mass spectrometry (LA-ICP-MS)

    NARCIS (Netherlands)

    Hennekam, Rick; Jilbert, Tom; Mason, Paul R D; de Lange, Gert J.; Reichart, Gert Jan

    2015-01-01

    Laser ablation-inductively coupled plasma-mass spectrometry (LA-ICP-MS) line-scanning is a promising technique for producing high-resolution (μm-scale) geochemical records on resin-embedded sediments. However, this approach has not yet been thoroughly tested on sediment samples of known elemental

  16. Design and development of a very high resolution thermal imager

    Science.gov (United States)

    Kuerbitz, Gunther; Duchateau, Ruediger

    1998-10-01

    The design goal of this project was to develop a thermal imaging system with ultimate geometrical resolution without sacrificing thermal sensitivity. It was necessary to fulfil the criteria for a future advanced video standard. This video standard is the so-called HDTV standard (HDTV High Definition TeleVision). The thermal imaging system is a parallel scanning system working in the 7...11 micrometer spectral region. The detector for that system has to have 576 X n (n number of TDI stages) detector elements taking into account a twofold interlace. It must be carefully optimized in terms of range performance and size of optics entrance pupil as well as producibility and yield. This was done in strong interaction with the detector manufacturer. The 16:9 aspect ratio of the HDTV standard together with the high number of 1920 pixels/line impose high demands on the scanner design in terms of scan efficiency and linearity. As an advanced second generation thermal imager the system has an internal thermal reference. The electronics is fully digitized and comprises circuits for Non Uniformity Correction (NUC), scan conversion, electronic zoom, auto gain and level, edge enhancement, up/down and left/right reversion etc. It can be completely remote-controlled via a serial interface.

  17. Indigenous development of scanning electron microscope

    International Nuclear Information System (INIS)

    Ambastha, K.P.; Chaudhari, Y.V.; Pal, Suvadip; Tikaria, Amit; Pious, Lizy; Dubey, B.P.; Chadda, V.K.

    2009-01-01

    Scanning electron microscope (SEM) is a precision instrument and plays very important role in scientific studies. Bhabha Atomic Research Centre has taken up the job of development of SEM indigenously. Standard and commercially available components like computer, high voltage power supply, detectors etc. shall be procured from market. Focusing and scanning coils, vacuum chamber, specimen stage, control hardware and software etc. shall be developed at BARC with the help of Indian industry. Procurement, design and fabrication of various parts of SEM are in progress. (author)

  18. Development of forage cryo minced fish technology

    Directory of Open Access Journals (Sweden)

    Titova S. A.

    2017-11-01

    Full Text Available The technology of grinding the frozen fish raw material using the useful model of a piston-type extrusion plant with cold working bodies for fodder minced fish (cryo minced fish feed by the method of cryoextrusion has been developed. The possibility of replacing the standard manufacturing operations (defrosting and subsequent grinding of raw materials to the one (cryoextrusion has been substantiated. The optimal shape of holes of the die for forcing the raw material has been determined in the form of "hourglass", the hole diameter is 7 mm, the length of the process of pushing is 40 s. It has been established that the finished product (cryo minced fish feed has a homogeneous grinding, juicy, crumbly texture, the temperature in the thickness of the product is equal to the temperature of the raw material before processing (–18 °C. The technological scheme of producing the minced fish feed by the method of cryoextrusion has been developed. Researching the chemical composition of fish and feed of mince produced by the traditional and developed methods it has been established that protein content in the cryo minced fish feed remains unchanged. The amino acid composition of cryo minced fish feed has been determined, the amount of essential amino acids is 365.6 mg/g of protein. In the proteins of the cryo minced fish feed methionine has been contained in the minimum quantity (10.0 mg/g protein and in the maximum – serine (155.3 mg/g protein. In the protein of the cryo minced fish feed the only limiting amino acid is methionine. The coefficient of rationality of the cryo minced fish feed whiting is equal to 0.33. The product is not toxic, is not subject to contamination with dangerous microorganisms, has high biological value, contains necessary in animal nutrition mineral elements, the energy value per 100 g of the product is 87 kcal. The combination of the processes of cryoextrusion and lyophilization allows to obtain a new food product with high

  19. CIRS High-Resolution Thermal Scans and the Structure of Saturn's B Ring

    Science.gov (United States)

    Brooks, S. M.; Spilker, L. J.; Showalter, M.; Pilorz, S.; Edgington, S. G.

    2017-12-01

    The flyby of Titan on November 29, 2016, sent the Cassini spacecraft on a trajectory that would take it within 10,000 kilometers of Saturn's F ring multiple times before a subsequent Titan encounter on April 22, 2017, would send it on ballistic trajectory carrying it between Saturn's cloud tops and the planet's D ring for several flybys. This geometry has proven beneficial for high-resolution studies of the rings, not just because of Cassini's proximity to the rings, but also because of the spacecraft's high elevation angle above the rings, which reduces the foreshortening that tends to degrade resolution in the ring plane. We will report on several observations of Saturn's main rings at the high spatial resolutions enabled by the end-of-mission geometry, particulary the B ring, with the Composite Infrared Spectrometer onboard Cassini during the F-ring and proximal orbits. CIRS' three infrared detectors cover a combined spectral range of 10 to 1400 cm-1 (1 mm down to 7 microns). We focus on data from Focal Plane 1, which covers the 10 to 600 cm-1 range (1 mm to 16 microns). The apodized spectral resolution of the instrument can be varied from 15 cm-1 to 0.5 cm-1 (Flasar et al. 2004). FP1's wavelength range makes it well-suited to sensing thermal emission from objects at temperatures typical of Saturn's rings. Correlating ring optical depth with temperatures retrieved from scans of the face of the rings exposed to direct solar illumination (the lit face) and the opposite (unlit) face suggests differences in ring structure or particle transport between the lit and unlit sides of the rings in different regions of the B ring. Lit side temperatures in the core of the B ring range between 82 and 87 K; temperatures on the unlit side of the core vary from 66 K up to 74 K. Ferrari and Reffet (2013) and Pilorz et al. (2015) published thorough analyses of the thermal throughput across this optically thick ring. We will discuss these recent CIRS rings observations and their

  20. Quantitative detection of gold nanoparticles on individual, unstained cancer cells by Scanning Electron Microscopy

    NARCIS (Netherlands)

    Hartsuiker, Liesbeth; van Es, Peter; Petersen, Wilhelmina; van Leeuwen, Ton; Terstappen, Leonardus Wendelinus Mathias Marie; Otto, Cornelis

    2011-01-01

    Gold nanoparticles are rapidly emerging for use in biomedical applications. Characterization of the interaction and delivery of nanoparticles to cells through microscopy is important. Scanning electron microscopes have the intrinsic resolution to visualize gold nanoparticles on cells. A novel sample

  1. Quantitative detection of gold nanoparticles on individual, unstained cancer cells by scanning electron microscopy

    NARCIS (Netherlands)

    Hartsuiker, L.; van Es, P.; Petersen, W.; van Leeuwen, T. G.; Terstappen, L. W. M. M.; Otto, C.

    2011-01-01

    Gold nanoparticles are rapidly emerging for use in biomedical applications. Characterization of the interaction and delivery of nanoparticles to cells through microscopy is important. Scanning electron microscopes have the intrinsic resolution to visualize gold nanoparticles on cells. A novel sample

  2. Human Skin Barrier Structure and Function Analyzed by Cryo-EM and Molecular Dynamics Simulation.

    Science.gov (United States)

    Lundborg, Magnus; Narangifard, Ali; Wennberg, Christian L; Lindahl, Erik; Daneholt, Bertil; Norlén, Lars

    2018-04-24

    In the present study we have analyzed the molecular structure and function of the human skin's permeability barrier using molecular dynamics simulation validated against cryo-electron microscopy data from near native skin. The skin's barrier capacity is located to an intercellular lipid structure embedding the cells of the superficial most layer of skin - the stratum corneum. According to the splayed bilayer model (Iwai et al., 2012) the lipid structure is organized as stacked bilayers of ceramides in a splayed chain conformation with cholesterol associated with the ceramide sphingoid moiety and free fatty acids associated with the ceramide fatty acid moiety. However, knowledge about the lipid structure's detailed molecular organization, and the roles of its different lipid constituents, remains circumstantial. Starting from a molecular dynamics model based on the splayed bilayer model, we have, by stepwise structural and compositional modifications, arrived at a thermodynamically stable molecular dynamics model expressing simulated electron microscopy patterns matching original cryo-electron microscopy patterns from skin extremely closely. Strikingly, the closer the individual molecular dynamics models' lipid composition was to that reported in human stratum corneum, the better was the match between the models' simulated electron microscopy patterns and the original cryo-electron microscopy patterns. Moreover, the closest-matching model's calculated water permeability and thermotropic behaviour were found compatible with that of human skin. The new model may facilitate more advanced physics-based skin permeability predictions of drugs and toxicants. The proposed procedure for molecular dynamics based analysis of cellular cryo-electron microscopy data might be applied to other biomolecular systems. Copyright © 2018. Published by Elsevier Inc.

  3. Compact design of a transmission electron microscope-scanning tunneling microscope holder with three-dimensional coarse motion

    International Nuclear Information System (INIS)

    Svensson, K.; Jompol, Y.; Olin, H.; Olsson, E.

    2003-01-01

    A scanning tunneling microscope (STM) with a compact, three-dimensional, inertial slider design is presented. Inertial sliding of the STM tip, in three dimensions, enables coarse motion and scanning using only one piezoelectric tube. Using the same electronics both for scanning and inertial sliding, step lengths of less than 5% of the piezo range were achieved. The compact design, less than 1 cm3 in volume, ensures a low mechanical noise level and enables us to fit the STM into the sample holder of a transmission electron microscope (TEM), while maintaining atomic scale resolution in both STM and TEM imaging

  4. Heparin binding sites on Ross River virus revealed by electron cryo-microscopy

    International Nuclear Information System (INIS)

    Zhang Wei; Heil, Marintha; Kuhn, Richard J.; Baker, Timothy S.

    2005-01-01

    Cell surface glycosaminoglycans play important roles in cell adhesion and viral entry. Laboratory strains of two alphaviruses, Sindbis and Semliki Forest virus, have been shown to utilize heparan sulfate as an attachment receptor, whereas Ross River virus (RRV) does not significantly interact with it. However, a single amino acid substitution at residue 218 in the RRV E2 glycoprotein adapts the virus to heparan sulfate binding and expands the host range of the virus into chicken embryo fibroblasts. Structures of the RRV mutant, E2 N218R, and its complex with heparin were determined through the use of electron cryo-microscopy and image reconstruction methods. Heparin was found to bind at the distal end of the RRV spikes, in a region of the E2 glycoprotein that has been previously implicated in cell-receptor recognition and antibody binding

  5. High-resolution compact Johann crystal spectrometer with the Livermore electron beam ion trap

    International Nuclear Information System (INIS)

    Robbins, D.L.; Chen, H.; Beiersdorfer, P.; Faenov, A.Ya.; Pikuz, T.A.; May, M.J.; Dunn, J.; Smith, A.J.

    2004-01-01

    A compact high-resolution (λ/Δλ≅10 000) spherically bent crystal spectrometer in the Johann geometry was recently installed and tested on the Lawrence Livermore National Laboratory SuperEBIT electron beam ion trap. The curvature of the mica (002) crystal grating allows for higher collection efficiency compared to the flat and cylindrically bent crystal spectrometers commonly used on the Livermore electron beam ion traps. The spectrometer's Johann configuration enables orientation of its dispersion plane to be parallel to the electron beam propagation. Used in concert with a crystal spectrometer, whose dispersion plane is perpendicular to the electron beam propagation, the polarization of x-ray emission lines can be measured

  6. An atomic model of brome mosaic virus using direct electron detection and real-space optimization

    Science.gov (United States)

    Wang, Zhao; Hryc, Corey F.; Bammes, Benjamin; Afonine, Pavel V.; Jakana, Joanita; Chen, Dong-Hua; Liu, Xiangan; Baker, Matthew L.; Kao, Cheng; Ludtke, Steven J.; Schmid, Michael F.; Adams, Paul D.; Chiu, Wah

    2014-09-01

    Advances in electron cryo-microscopy have enabled structure determination of macromolecules at near-atomic resolution. However, structure determination, even using de novo methods, remains susceptible to model bias and overfitting. Here we describe a complete workflow for data acquisition, image processing, all-atom modelling and validation of brome mosaic virus, an RNA virus. Data were collected with a direct electron detector in integrating mode and an exposure beyond the traditional radiation damage limit. The final density map has a resolution of 3.8 Å as assessed by two independent data sets and maps. We used the map to derive an all-atom model with a newly implemented real-space optimization protocol. The validity of the model was verified by its match with the density map and a previous model from X-ray crystallography, as well as the internal consistency of models from independent maps. This study demonstrates a practical approach to obtain a rigorously validated atomic resolution electron cryo-microscopy structure.

  7. 3D membrane segmentation and quantification of intact thick cells using cryo soft X-ray transmission microscopy: A pilot study

    Science.gov (United States)

    Klementieva, Oxana; Werner, Stephan; Guttmann, Peter; Pratsch, Christoph; Cladera, Josep

    2017-01-01

    Structural analysis of biological membranes is important for understanding cell and sub-cellular organelle function as well as their interaction with the surrounding environment. Imaging of whole cells in three dimension at high spatial resolution remains a significant challenge, particularly for thick cells. Cryo-transmission soft X-ray microscopy (cryo-TXM) has recently gained popularity to image, in 3D, intact thick cells (∼10μm) with details of sub-cellular architecture and organization in near-native state. This paper reports a new tool to segment and quantify structural changes of biological membranes in 3D from cryo-TXM images by tracking an initial 2D contour along the third axis of the microscope, through a multi-scale ridge detection followed by an active contours-based model, with a subsequent refinement along the other two axes. A quantitative metric that assesses the grayscale profiles perpendicular to the membrane surfaces is introduced and shown to be linearly related to the membrane thickness. Our methodology has been validated on synthetic phantoms using realistic microscope properties and structure dimensions, as well as on real cryo-TXM data. Results demonstrate the validity of our algorithms for cryo-TXM data analysis. PMID:28376110

  8. MRI isotropic resolution reconstruction from two orthogonal scans

    Science.gov (United States)

    Tamez-Pena, Jose G.; Totterman, Saara; Parker, Kevin J.

    2001-07-01

    An algorithm for the reconstructions of ISO-resolution volumetric MR data sets from two standard orthogonal MR scans having anisotropic resolution has been developed. The reconstruction algorithm starts by registering a pair of orthogonal volumetric MR data sets. The registration is done by maximizing the correlation between the gradient magnitude using a simple translation-rotation model in a multi-resolution approach. Then algorithm assumes that the individual voxels on the MR data are an average of the magnetic resonance properties of an elongated imaging volume. Then, the process is modeled as the projection of MR properties into a single sensor. This model allows the derivation of a set of linear equations that can be used to recover the MR properties of every single voxel in the SO-resolution volume given only two orthogonal MR scans. Projections on convex sets (POCS) was used to solve the set of linear equations. Experimental results show the advantage of having a ISO-resolution reconstructions for the visualization and analysis of small and thin muscular structures.

  9. Atmospheric scanning electron microscope observes cells and tissues in open medium through silicon nitride film.

    Science.gov (United States)

    Nishiyama, Hidetoshi; Suga, Mitsuo; Ogura, Toshihiko; Maruyama, Yuusuke; Koizumi, Mitsuru; Mio, Kazuhiro; Kitamura, Shinichi; Sato, Chikara

    2010-03-01

    Direct observation of subcellular structures and their characterization is essential for understanding their physiological functions. To observe them in open environment, we have developed an inverted scanning electron microscope with a detachable, open-culture dish, capable of 8 nm resolution, and combined with a fluorescence microscope quasi-simultaneously observing the same area from the top. For scanning electron microscopy from the bottom, a silicon nitride film window in the base of the dish maintains a vacuum between electron gun and open sample dish while allowing electrons to pass through. Electrons are backscattered from the sample and captured by a detector under the dish. Cells cultured on the open dish can be externally manipulated under optical microscopy, fixed, and observed using scanning electron microscopy. Once fine structures have been revealed by scanning electron microscopy, their component proteins may be identified by comparison with separately prepared fluorescence-labeled optical microscopic images of the candidate proteins, with their heavy-metal-labeled or stained ASEM images. Furthermore, cell nuclei in a tissue block stained with platinum-blue were successfully observed without thin-sectioning, which suggests the applicability of this inverted scanning electron microscope to cancer diagnosis. This microscope visualizes mesoscopic-scale structures, and is also applicable to non-bioscience fields including polymer chemistry. (c) 2010 Elsevier Inc. All rights reserved.

  10. High-resolution patterning of graphene by screen printing with a silicon stencil for highly flexible printed electronics.

    Science.gov (United States)

    Hyun, Woo Jin; Secor, Ethan B; Hersam, Mark C; Frisbie, C Daniel; Francis, Lorraine F

    2015-01-07

    High-resolution screen printing of pristine graphene is introduced for the rapid fabrication of conductive lines on flexible substrates. Well-defined silicon stencils and viscosity-controlled inks facilitate the preparation of high-quality graphene patterns as narrow as 40 μm. This strategy provides an efficient method to produce highly flexible graphene electrodes for printed electronics. © 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  11. Gas-phase synthesis of magnesium nanoparticles : A high-resolution transmission electron microscopy study

    NARCIS (Netherlands)

    Kooi, B.J.; Palasantzas, G.; de Hosson, J.T.M.

    2006-01-01

    Magnesium nanoparticles with size above 10 nm, prepared by gas-phase syntheses, were investigated by high-resolution transmission electron microscopy. The dominant particle shape is a hexagonal prism terminated by Mg(0002) and Mg{1010} facets. Oxidation of Mg yields a MgO shell (similar to 3 nm

  12. Atmospheric scanning electron microscope for correlative microscopy.

    Science.gov (United States)

    Morrison, Ian E G; Dennison, Clare L; Nishiyama, Hidetoshi; Suga, Mitsuo; Sato, Chikara; Yarwood, Andrew; O'Toole, Peter J

    2012-01-01

    The JEOL ClairScope is the first truly correlative scanning electron and optical microscope. An inverted scanning electron microscope (SEM) column allows electron images of wet samples to be obtained in ambient conditions in a biological culture dish, via a silicon nitride film window in the base. A standard inverted optical microscope positioned above the dish holder can be used to take reflected light and epifluorescence images of the same sample, under atmospheric conditions that permit biochemical modifications. For SEM, the open dish allows successive staining operations to be performed without moving the holder. The standard optical color camera used for fluorescence imaging can be exchanged for a high-sensitivity monochrome camera to detect low-intensity fluorescence signals, and also cathodoluminescence emission from nanophosphor particles. If these particles are applied to the sample at a suitable density, they can greatly assist the task of perfecting the correlation between the optical and electron images. Copyright © 2012 Elsevier Inc. All rights reserved.

  13. Design and construction of a high-stability, low-noise power supply for use with high-resolution electron energy loss spectrometers

    International Nuclear Information System (INIS)

    Katz, J.E.; Davies, P.W.; Crowell, J.E.; Somorjai, G.A.

    1982-01-01

    The design and construction of a high-stability, low-noise power supply which provides potentials for the lens and analyzer elements of a 127 0 Ehrhardt-type high-resolution electron energy loss spectrometer (HREELS) is described. The supply incorporates a filament emission-control circuit and facilities for measuring electron beam current at each spectrometer element, thus facilitating optimal tuning of the spectrometer. Spectra obtained using this supply are shown to have a four-fold improvement in signal-to-noise ratio and a higher resolution of the vibrational loss features when compared with spectra taken using a previously existing supply based on passive potential divider networks

  14. Sub-micron resolution selected area electron channeling patterns.

    Science.gov (United States)

    Guyon, J; Mansour, H; Gey, N; Crimp, M A; Chalal, S; Maloufi, N

    2015-02-01

    Collection of selected area channeling patterns (SACPs) on a high resolution FEG-SEM is essential to carry out quantitative electron channeling contrast imaging (ECCI) studies, as it facilitates accurate determination of the crystal plane normal with respect to the incident beam direction and thus allows control the electron channeling conditions. Unfortunately commercial SACP modes developed in the past were limited in spatial resolution and are often no longer offered. In this contribution we present a novel approach for collecting high resolution SACPs (HR-SACPs) developed on a Gemini column. This HR-SACP technique combines the first demonstrated sub-micron spatial resolution with high angular accuracy of about 0.1°, at a convenient working distance of 10mm. This innovative approach integrates the use of aperture alignment coils to rock the beam with a digitally calibrated beam shift procedure to ensure the rocking beam is maintained on a point of interest. Moreover a new methodology to accurately measure SACP spatial resolution is proposed. While column considerations limit the rocking angle to 4°, this range is adequate to index the HR-SACP in conjunction with the pattern simulated from the approximate orientation deduced by EBSD. This new technique facilitates Accurate ECCI (A-ECCI) studies from very fine grained and/or highly strained materials. It offers also new insights for developing HR-SACP modes on new generation high-resolution electron columns. Copyright © 2014 Elsevier B.V. All rights reserved.

  15. Cryo-immunogold electron microscopy for prions: toward identification of a conversion site.

    Science.gov (United States)

    Godsave, Susan F; Wille, Holger; Kujala, Pekka; Latawiec, Diane; DeArmond, Stephen J; Serban, Ana; Prusiner, Stanley B; Peters, Peter J

    2008-11-19

    Prion diseases are caused by accumulation of an abnormally folded isoform (PrP(Sc)) of the cellular prion protein (PrP(C)). The subcellular distribution of PrP(Sc) and the site of its formation in brain are still unclear. We performed quantitative cryo-immunogold electron microscopy on hippocampal sections from mice infected with the Rocky Mountain Laboratory strain of prions. Two antibodies were used: R2, which recognizes both PrP(C) and PrP(Sc); and F4-31, which only detects PrP(C) in undenatured sections. At a late subclinical stage of prion infection, both PrP(C) and PrP(Sc) were detected principally on neuronal plasma membranes and on vesicles resembling early endocytic or recycling vesicles in the neuropil. The R2 labeling was approximately six times higher in the infected than the uninfected hippocampus and gold clusters were only evident in infected tissue. The biggest increase in labeling density (24-fold) was found on the early/recycling endosome-like vesicles of small-diameter neurites, suggesting these as possible sites of conversion. Trypsin digestion of infected hippocampal sections resulted in a reduction in R2 labeling of >85%, which suggests that a high proportion of PrP(Sc) may be oligomeric, protease-sensitive PrP(Sc).

  16. Determining the resolution of scanning microwave impedance microscopy using atomic-precision buried donor structures

    Science.gov (United States)

    Scrymgeour, D. A.; Baca, A.; Fishgrab, K.; Simonson, R. J.; Marshall, M.; Bussmann, E.; Nakakura, C. Y.; Anderson, M.; Misra, S.

    2017-11-01

    To quantify the resolution limits of scanning microwave impedance microscopy (sMIM), we created scanning tunneling microscope (STM)-patterned donor nanostructures in silicon composed of 10 nm lines of highly conductive silicon buried under a protective top cap of silicon, and imaged them with sMIM. This dopant pattern is an ideal test of the resolution and sensitivity of the sMIM technique, as it is made with nm-resolution and offers minimal complications from topography convolution. It has been determined that typical sMIM tips can resolve lines down to ∼80 nm spacing, while resolution is independent of tip geometry as extreme tip wear does not change the resolving power, contrary to traditional scanning capacitance microscopy (SCM). Going forward, sMIM is an ideal technique for qualifying buried patterned devices, potentially allowing for quantitative post-fabrication characterization of donor structures, which may be an important tool for the study of atomic-scale transistors and state of the art quantum computation schemes.

  17. High-resolution transmission electron microscopy and energetics of flattened carbon nonoshells

    International Nuclear Information System (INIS)

    Bourgeois, L.N.; Bursill, L.A.

    1998-01-01

    When examined under a high-resolution transmission electron microscope, carbon soot produced alongside buckytubes in an arc-discharge is found to contain a small percentage of flattened carbon shells. These objects are shown to be small graphite flakes which eliminated their dangling bonds by terminating their edges with highly curved junctions. Ideal models for these structures are presented, and their energy estimated. The calculations show that the establishment of highly curved junctions is energetically favourable for a graphite flake in an inert atmosphere. Flattened shells also appear more stable than their 'inflated' counterparts (fullerene 'onions' and buckytubes) when the shell dimensions obey specific criteria.(authors)

  18. Retrofit implementation of Zernike phase plate imaging for cryo-TEM.

    Science.gov (United States)

    Marko, Michael; Leith, Ardean; Hsieh, Chyongere; Danev, Radostin

    2011-05-01

    In-focus phase-plate imaging is particularly beneficial for cryo-TEM because it offers a substantial overall increase in image contrast, without an electron dose penalty, and it simplifies image interpretation. We show how phase-plate cryo-TEM can be implemented with an appropriate existing TEM, and provide a basic practical introduction to use of thin-film (carbon) phase plates. We point out potential pitfalls of phase-plate operation, and discuss solutions. We provide information on evaluating a particular TEM for its suitability. Copyright © 2011 Elsevier Inc. All rights reserved.

  19. Electron Temperatures in W51 Complex from High Resolution, Low ...

    Indian Academy of Sciences (India)

    2001-03-09

    Mar 9, 2001 ... kinematics and local thermodynamic equilibrium (LTE) electron temperature (Te) of this region. H109α RRL observations by Wilsonet al.(1970) with a resolution of 4 and by Pankonin et al. (1979) with a resolution of 2.6 show that Te ∼ 6000 K in G49.5-0.4. Lower frequency observations for H137β and ...

  20. High-resolution CT of the lungs: Anatomic-pathologic correlation

    International Nuclear Information System (INIS)

    Stein, M.G.; Webb, W.R.; Finkbeiner, W.; Gamsu, G.

    1986-01-01

    The interpretation of thin-section (1.5-mm), high-resolution CT scans of the lungs has been limited by lack of direct radiologic and pathologic correlation. The author scanned fresh inflated isolated lungs from ten healthy and five diseased subjects using thin-section, high-resolution techniques. The lungs were then fixed by inflation with endobronchial Formalin. Gough sections (1 mm thick) were obtained at the same levels as the CT scans. In healthy subjects, secondary lobules were identified by the presence of visible interlobular septa and central arterioles. In some patients with disease, septal thickening was visible. In patients with honeycombing cystic areas of destroyed lung were seen, along with areas of fibrosis. Emphysema was well evaluated. Thin-section, high-resolution CT can define lung architecture and may resolve mild changes of the interstitium

  1. 'Crazy-Paving' Patterns on High-Resolution CT Scans in Patients with Pulmonary Complications after Hematopoietic Stem Cell Transplantation

    Energy Technology Data Exchange (ETDEWEB)

    Marchiori, Edson; Escuissato, Dante L.; Gasparetto, Taisa Davaus; Considera, Daniela Peixoto [Federal University, Sao Paulo (Brazil); Franquet, Tomas [Hospital de Sant Pau, Universitat Autonoma de Barcelona, Barcelona (Spain)

    2009-02-15

    To describe the pulmonary complications following hematopoietic stem cell transplantation (HSCT) that can present with a 'crazy-paving' pattern in high-resolution CT scans. Retrospective review of medical records from 2,537 patients who underwent HSCT. The 'crazy-paving' pattern consists of interlobular and intralobular septal thickening superimposed on an area of ground-glass attenuation on high-resolution CT scans. The CT scans were retrospectively reviewed by two radiologists, who reached final decisions by consensus. We identified 10 cases (2.02%), seven male and three female, with pulmonary complications following HSCT that presented with the 'crazy-paving' pattern. Seven (70%) patients had infectious pneumonia (adenovirus, herpes simplex, influenza virus, cytomegalovirus, respiratory syncytial virus, and toxoplasmosis), and three patients presented with non-infectious complications (idiopathic pneumonia syndrome and acute pulmonary edema). The 'crazy-paving' pattern was bilateral in all cases, with diffuse distribution in nine patients (90%), predominantly in the middle and inferior lung regions in seven patients (70%), and involving the anterior and posterior regions of the lungs in nine patients (90%). The 'crazy-paving' pattern is rare in HSCT recipients with pulmonary complications and is associated with infectious complications more commonly than non-infectious conditions.

  2. Angle selective backscattered electron contrast in the low-voltage scanning electron microscope: Simulation and experiment for polymers

    Energy Technology Data Exchange (ETDEWEB)

    Wan, Q., E-mail: qwan2@sheffield.ac.uk [Department of Material Science and Engineering, University of Sheffield, Western Bank, Sheffield S10 2TN (United Kingdom); Masters, R.C. [Department of Material Science and Engineering, University of Sheffield, Western Bank, Sheffield S10 2TN (United Kingdom); Lidzey, D. [Department of Physics and Astronomy, University of Sheffield, Western Bank, Sheffield S10 2TN (United Kingdom); Abrams, K.J. [Department of Material Science and Engineering, University of Sheffield, Western Bank, Sheffield S10 2TN (United Kingdom); Dapor, M. [European Centre for Theoretical Studies in Nuclear Physics and Related Areas (ECT-FBK) and Trento Institute for Fundamental Physics and Applications (TIFPA-INFN), via Sommarive 18, I-38123 Trento (Italy); Plenderleith, R.A. [Department of Material Science and Engineering, University of Sheffield, Western Bank, Sheffield S10 2TN (United Kingdom); Rimmer, S. [Department of Chemistry, University of Sheffield, Western Bank, Sheffield S10 2TN (United Kingdom); Claeyssens, F.; Rodenburg, C. [Department of Material Science and Engineering, University of Sheffield, Western Bank, Sheffield S10 2TN (United Kingdom)

    2016-12-15

    Recently developed detectors can deliver high resolution and high contrast images of nanostructured carbon based materials in low voltage scanning electron microscopes (LVSEM) with beam deceleration. Monte Carlo Simulations are also used to predict under which exact imaging conditions purely compositional contrast can be obtained and optimised. This allows the prediction of the electron signal intensity in angle selective conditions for back-scattered electron (BSE) imaging in LVSEM and compares it to experimental signals. Angle selective detection with a concentric back scattered (CBS) detector is considered in the model in the absence and presence of a deceleration field, respectively. The validity of the model prediction for both cases was tested experimentally for amorphous C and Cu and applied to complex nanostructured carbon based materials, namely a Poly(N-isopropylacrylamide)/Poly(ethylene glycol) Diacrylate (PNIPAM/PEGDA) semi-interpenetration network (IPN) and a Poly(3-hexylthiophene-2,5-diyl) (P3HT) film, to map nano-scale composition and crystallinity distribution by avoiding experimental imaging conditions that lead to a mixed topographical and compositional contrast - Highlights: • An optimised model for nano-scale analysis of beam sensitive materials by LVSEM. • Simulation and separation of composition and topography in a CBS detector. • Selective angle backscattered electron collection for mapping of polymers.

  3. Angle selective backscattered electron contrast in the low-voltage scanning electron microscope: Simulation and experiment for polymers

    International Nuclear Information System (INIS)

    Wan, Q.; Masters, R.C.; Lidzey, D.; Abrams, K.J.; Dapor, M.; Plenderleith, R.A.; Rimmer, S.; Claeyssens, F.; Rodenburg, C.

    2016-01-01

    Recently developed detectors can deliver high resolution and high contrast images of nanostructured carbon based materials in low voltage scanning electron microscopes (LVSEM) with beam deceleration. Monte Carlo Simulations are also used to predict under which exact imaging conditions purely compositional contrast can be obtained and optimised. This allows the prediction of the electron signal intensity in angle selective conditions for back-scattered electron (BSE) imaging in LVSEM and compares it to experimental signals. Angle selective detection with a concentric back scattered (CBS) detector is considered in the model in the absence and presence of a deceleration field, respectively. The validity of the model prediction for both cases was tested experimentally for amorphous C and Cu and applied to complex nanostructured carbon based materials, namely a Poly(N-isopropylacrylamide)/Poly(ethylene glycol) Diacrylate (PNIPAM/PEGDA) semi-interpenetration network (IPN) and a Poly(3-hexylthiophene-2,5-diyl) (P3HT) film, to map nano-scale composition and crystallinity distribution by avoiding experimental imaging conditions that lead to a mixed topographical and compositional contrast - Highlights: • An optimised model for nano-scale analysis of beam sensitive materials by LVSEM. • Simulation and separation of composition and topography in a CBS detector. • Selective angle backscattered electron collection for mapping of polymers.

  4. Cryo-EM structures of the 80S ribosomes from human parasites Trichomonas vaginalis and Toxoplasma gondii

    Institute of Scientific and Technical Information of China (English)

    Zhifei Li; Qiang Guo; Lvqin Zheng; Yongsheng Ji; Yi-Ting Xie; De-Hua Lai; Zhao-Rong Lun; Xun Suo; Ning Gao

    2017-01-01

    As an indispensable molecular machine universal in all living organisms,the ribosome has been selected by evolution to be the natural target of many antibiotics and small-molecule inhibitors.High-resolution structures of pathogen ribosomes are crucial for understanding the general and unique aspects of translation control in disease-causing microbes.With cryo-electron microscopy technique,we have determined structures of the cytosolic ribosomes from two human parasites,Trichomonas vaginalis and Toxoplasma gondii,at resolution of 3.2-3.4,(A).Although the ribosomal proteins from both pathogens are typical members of eukaryotic families,with a co-evolution pattern between certain species-specific insertions/extensions and neighboring ribosomal RNA (rRNA) expansion segments,the sizes of their rRNAs are sharply different.Very interestingly,rRNAs of T.vaginalis are in size comparable to prokaryotic counterparts,with nearly all the eukaryote-specific rRNA expansion segments missing.These structures facilitate the dissection of evolution path for ribosomal proteins and RNAs,and may aid in design of novel translation inhibitors.

  5. A scanning tunneling microscope with a scanning range from hundreds of micrometers down to nanometer resolution.

    Science.gov (United States)

    Kalkan, Fatih; Zaum, Christopher; Morgenstern, Karina

    2012-10-01

    A beetle type stage and a flexure scanning stage are combined to form a two stages scanning tunneling microscope (STM). It operates at room temperature in ultrahigh vacuum and is capable of scanning areas up to 300 μm × 450 μm down to resolution on the nanometer scale. This multi-scale STM has been designed and constructed in order to investigate prestructured metallic or semiconducting micro- and nano-structures in real space from atomic-sized structures up to the large-scale environment. The principle of the instrument is demonstrated on two different systems. Gallium nitride based micropillars demonstrate scan areas up to hundreds of micrometers; a Au(111) surface demonstrates nanometer resolution.

  6. Detecting and locating light atoms from high-resolution STEM images : The quest for a single optimal design

    NARCIS (Netherlands)

    Gonnissen, J; De Backer, A; den Dekker, A.J.; Sijbers, J.; Van Aert, S.

    2016-01-01

    In the present paper, the optimal detector design is investigated for both detecting and locating light atoms from high resolution scanning transmission electron microscopy (HR STEM) images. The principles of detection theory are used to quantify the probability of error for the detection of

  7. Understanding deformation with high angular resolution electron backscatter diffraction (HR-EBSD)

    Science.gov (United States)

    Britton, T. B.; Hickey, J. L. R.

    2018-01-01

    High angular resolution electron backscatter diffraction (HR-EBSD) affords an increase in angular resolution, as compared to ‘conventional’ Hough transform based EBSD, of two orders of magnitude, enabling measurements of relative misorientations of 1 x 10-4 rads (~ 0.006°) and changes in (deviatoric) lattice strain with a precision of 1 x 10-4. This is achieved through direct comparison of two or more diffraction patterns using sophisticated cross-correlation based image analysis routines. Image shifts between zone axes in the two-correlated diffraction pattern are measured with sub-pixel precision and this realises the ability to measure changes in interplanar angles and lattice orientation with a high degree of sensitivity. These shifts are linked to strains and lattice rotations through simple geometry. In this manuscript, we outline the basis of the technique and two case studies that highlight its potential to tackle real materials science challenges, such as deformation patterning in polycrystalline alloys.

  8. Forensic Scanning Electron Microscope

    Science.gov (United States)

    Keeley, R. H.

    1983-03-01

    The scanning electron microscope equipped with an x-ray spectrometer is a versatile instrument which has many uses in the investigation of crime and preparation of scientific evidence for the courts. Major applications include microscopy and analysis of very small fragments of paint, glass and other materials which may link an individual with a scene of crime, identification of firearms residues and examination of questioned documents. Although simultaneous observation and chemical analysis of the sample is the most important feature of the instrument, other modes of operation such as cathodoluminescence spectrometry, backscattered electron imaging and direct x-ray excitation are also exploited. Marks on two bullets or cartridge cases can be compared directly by sequential scanning with a single beam or electronic linkage of two instruments. Particles of primer residue deposited on the skin and clothing when a gun is fired can be collected on adhesive tape and identified by their morphology and elemental composition. It is also possible to differentiate between the primer residues of different types of ammunition. Bullets may be identified from the small fragments left behind as they pass through the body tissues. In the examination of questioned documents the scanning electron microscope is used to establish the order in which two intersecting ink lines were written and to detect traces of chemical markers added to the security inks on official documents.

  9. Investigation into scanning tunnelling luminescence microscopy

    International Nuclear Information System (INIS)

    Manson-Smith, S.K.

    2001-01-01

    This work reports on the development of a scanning tunnelling luminescence (STL) microscope and its application to the study of Ill-nitride semiconductor materials used in the production of light emitting devices. STL microscopy is a technique which uses the high resolution topographic imaging capabilities of the scanning tunnelling microscope (STM) to generate high resolution luminescence images. The STM tunnelling current acts as a highly localised source of electrons (or holes) which generates luminescence in certain materials. Light generated at the STM tunnelling junction is collected concurrently with the height variation of the tunnelling probe as it is scanned across a sample surface, producing simultaneous topographic and luminescence images. Due to the very localised excitation source, high resolution luminescence images can be obtained. Spectroscopic resolution can be obtained by using filters. Additionally, the variation of luminescence intensity with tunnel current and with bias voltage can provide information on recombination processes and material properties. The design and construction of a scanning tunnelling luminescence microscope is described in detail. Operating under ambient conditions, the microscope has several novel features, including a new type of miniature inertial slider-based approach motor, large solid-angle light collection optical arrangement and a tip-height regulation system which requires the minimum of operator input. (author)

  10. Advances in imaging and electron physics the scanning transmission electron microscope

    CERN Document Server

    Hawkes, Peter W

    2009-01-01

    Advances in Imaging and Electron Physics merges two long-running serials--Advances in Electronics and Electron Physics and Advances in Optical and Electron Microscopy. This series features extended articles on the physics of electron devices (especially semiconductor devices), particle optics at high and low energies, microlithography, image science and digital image processing, electromagnetic wave propagation, electron microscopy, and the computing methods used in all these domains.  This particular volume presents several timely articles on the scanning transmission electron microscope. Updated with contributions from leading international scholars and industry experts Discusses hot topic areas and presents current and future research trends Provides an invaluable reference and guide for physicists, engineers and mathematicians.

  11. Robust w-Estimators for Cryo-EM Class Means

    Science.gov (United States)

    Huang, Chenxi; Tagare, Hemant D.

    2016-01-01

    A critical step in cryogenic electron microscopy (cryo-EM) image analysis is to calculate the average of all images aligned to a projection direction. This average, called the “class mean”, improves the signal-to-noise ratio in single particle reconstruction (SPR). The averaging step is often compromised because of outlier images of ice, contaminants, and particle fragments. Outlier detection and rejection in the majority of current cryo-EM methods is done using cross-correlation with a manually determined threshold. Empirical assessment shows that the performance of these methods is very sensitive to the threshold. This paper proposes an alternative: a “w-estimator” of the average image, which is robust to outliers and which does not use a threshold. Various properties of the estimator, such as consistency and influence function are investigated. An extension of the estimator to images with different contrast transfer functions (CTFs) is also provided. Experiments with simulated and real cryo-EM images show that the proposed estimator performs quite well in the presence of outliers. PMID:26841397

  12. An ASIC implementation of digital front-end electronics for a high resolution PET scanner

    International Nuclear Information System (INIS)

    Newport, D.F.; Young, J.W.

    1993-01-01

    AN Application Specific Integrated Circuit (ASIC) has been designed and fabricated which implements many of the current functions found in the digital front-end electronics for a high resolution Positron Emission Tomography (PET) scanner. The ASIC performs crystal selection, energy qualification, time correction, and event counting functions for block technology high resolution PET scanners. Digitized x and y position, event energy, and time information are used by the ASIC to determine block crystal number, qualify the event based on energy, and correct the event time. In addition, event counting and block dead time calculations are performed for system dead time corrections. A loadable sequencer for controlling the analog front-end electronics is also implemented. The ASIC is implemented in a 37,000 gate, 1.0 micron CMOS gate-array and is capable of handling 4 million events/second while reducing parts count, cost, and power consumption over current board-level designs

  13. High energy resolution and first time-dependent positron annihilation induced Auger electron spectroscopty

    Energy Technology Data Exchange (ETDEWEB)

    Mayer, Jakob

    2010-04-03

    It was the aim of this thesis to improve the existing positron annihilation induced Auger spectrometer at the highly intense positron source NEPOMUC (NEutron induced POsitron source MUniCh) in several ways: Firstly, the measurement time for a single spectrum should be reduced from typically 12 h to roughly 1 h or even less. Secondly, the energy resolution, which amounted to {delta}E/E{approx}10%, should be increased by at least one order of magnitude in order to make high resolution positron annihilation induced Auger spectroscopy (PAES)-measurements of Auger transitions possible and thus deliver more information about the nature of the Auger process. In order to achieve these objectives, the PAES spectrometer was equipped with a new electron energy analyzer. For its ideal operation all other components of the Auger analysis chamber had to be adapted. Particularly the sample manipulation and the positron beam guidance had to be renewed. Simulations with SIMION {sup registered} ensured the optimal positron lens parameters. After the adjustment of the new analyzer and its components, first measurements illustrated the improved performance of the PAES setup: Firstly, the measurement time for short overview measurements was reduced from 3 h to 420 s. The measurement time for more detailed Auger spectra was shortened from 12 h to 80 min. Secondly, even with the reduced measurement time, the signal to noise ratio was also enhanced by one order of magnitude. Finally, the energy resolution was improved to {delta}E/E < 1. The exceptional surface sensitivity and elemental selectivity of PAES was demonstrated in measurements of Pd and Fe, both coated with Cu layers of varying thickness. PAES showed that with 0.96 monolayer of Cu on Fe, more than 55% of the detected Auger electrons stem from Cu. In the case of the Cu coated Pd sample 0.96 monolayer of Cu resulted in a Cu Auger fraction of more than 30% with PAES and less than 5% with electron induced Auger spectroscopy

  14. Scanning electron microscope

    International Nuclear Information System (INIS)

    Anon.

    1980-01-01

    The principle underlying the design of the scanning electron microscope (SEM), the design and functioning of SEM are described. Its applications in the areas of microcircuitry and materials science are outlined. The development of SEM in India is reviewed. (M.G.B.)

  15. High resolution electron microscopy and electron diffraction of YBa/sub 2/Cu/sub 3/O/sub 7-x/

    International Nuclear Information System (INIS)

    Krakow, W.; Shaw, T.M.

    1988-01-01

    Experimental high resolution electron micrographs and computer simulation experiments have been used to evaluate the visibility of the atomic constituents of YBa/sub 2/Cu/sub 3/O/sub 7-x/. In practice, the detection of oxygen has not been possible in contradiction to that predicted by modelling of perfect crystalline material. Preliminary computer experiments of the electron diffraction patterns when oxygen vacancies are introduced on the Cu-O sheets separating Ba layers show the diffuse streaks characteristic of short range ordering

  16. A comparative analysis of the cryo-compression and cryo-adsorption hydrogen storage methods

    Energy Technology Data Exchange (ETDEWEB)

    Petitpas, G [Lawrence Livermore National Lab. (LLNL), Livermore, CA (United States); Benard, P [Universite du Quebec a Trois-Rivieres (Canada); Klebanoff, L E [Sandia National Lab. (SNL-CA), Livermore, CA (United States); Xiao, J [Universite du Quebec a Trois-Rivieres (Canada); Aceves, S M [Lawrence Livermore National Lab. (LLNL), Livermore, CA (United States)

    2014-07-01

    While conventional low-pressure LH₂ dewars have existed for decades, advanced methods of cryogenic hydrogen storage have recently been developed. These advanced methods are cryo-compression and cryo-adsorption hydrogen storage, which operate best in the temperature range 30–100 K. We present a comparative analysis of both approaches for cryogenic hydrogen storage, examining how pressure and/or sorbent materials are used to effectively increase onboard H₂ density and dormancy. We start by reviewing some basic aspects of LH₂ properties and conventional means of storing it. From there we describe the cryo-compression and cryo-adsorption hydrogen storage methods, and then explore the relationship between them, clarifying the materials science and physics of the two approaches in trying to solve the same hydrogen storage task (~5–8 kg H₂, typical of light duty vehicles). Assuming that the balance of plant and the available volume for the storage system in the vehicle are identical for both approaches, the comparison focuses on how the respective storage capacities, vessel weight and dormancy vary as a function of temperature, pressure and type of cryo-adsorption material (especially, powder MOF-5 and MIL-101). By performing a comparative analysis, we clarify the science of each approach individually, identify the regimes where the attributes of each can be maximized, elucidate the properties of these systems during refueling, and probe the possible benefits of a combined “hybrid” system with both cryo-adsorption and cryo-compression phenomena operating at the same time. In addition the relationships found between onboard H₂ capacity, pressure vessel and/or sorbent mass and dormancy as a function of rated pressure, type of sorbent material and fueling conditions are useful as general designing guidelines in future engineering efforts using these two hydrogen storage approaches.

  17. CryoSat SIRAL Calibration and Performance

    Science.gov (United States)

    Fornari, Marco; Scagliola, Michele; Tagliani, Nicolas; Parrinello, Tommaso

    2013-04-01

    The main payload of CryoSat is a Ku band pulse-width limited radar altimeter, called SIRAL (Synthetic interferometric radar altimeter), that transmits pulses at a high pulse repetition frequency thus making the received echoes phase coherent and suitable for azimuth processing. This allows to reach an along track resolution of about 250 meters which is a significant improvement over traditional pulse-width limited altimeters. Due to the fact that SIRAL is a phase coherent pulse-width limited radar altimeter, a proper calibration approach has been developed, including both an internal and external calibration. The internal calibration monitors the instrument impulse response and the transfer function, like traditional altimeters. In addition to that, the interferometer requires a special calibration developed ad hoc for SIRAL. The external calibration is performed with the use of a ground transponder, located in Svalbard, which receives SIRAL signal and sends the echo back to the satellite. Internal calibration data are processed on ground by the CryoSat Instrument Processing Facility (IPF1) and then applied to the science data. By April 2013, almost 3 years of calibration data will be available, which will be shown in this poster. The external calibration (transponder) data are processed and analyzed independently from the operational chain. The use of an external transponder has been very useful to determine instrument performance and for the tuning of the on-ground processor. This poster presents the transponder results in terms of range noise and datation error.

  18. CryoSat Land Ice Product Validation within the CryoVal-LI project

    DEFF Research Database (Denmark)

    Sørensen, Louise Sandberg; Baker, Steven; Csatho, Bea

    The main objective of the ESA funded CryoVal-LI project has been to identify and quantify the error sources for the CryoSat-2 mission over land ice. This has been undertaken through the careful documentation of the possible error sources, the identification of suitable validation sites...

  19. Electronically-Scanned Pressure Sensors

    Science.gov (United States)

    Coe, C. F.; Parra, G. T.; Kauffman, R. C.

    1984-01-01

    Sensors not pneumatically switched. Electronic pressure-transducer scanning system constructed in modular form. Pressure transducer modules and analog to digital converter module small enough to fit within cavities of average-sized wind-tunnel models. All switching done electronically. Temperature controlled environment maintained within sensor modules so accuracy maintained while ambient temperature varies.

  20. Electron dose dependence of signal-to-noise ratio, atom contrast and resolution in transmission electron microscope images

    International Nuclear Information System (INIS)

    Lee, Z.; Rose, H.; Lehtinen, O.; Biskupek, J.; Kaiser, U.

    2014-01-01

    In order to achieve the highest resolution in aberration-corrected (AC) high-resolution transmission electron microscopy (HRTEM) images, high electron doses are required which only a few samples can withstand. In this paper we perform dose-dependent AC-HRTEM image calculations, and study the dependence of the signal-to-noise ratio, atom contrast and resolution on electron dose and sampling. We introduce dose-dependent contrast, which can be used to evaluate the visibility of objects under different dose conditions. Based on our calculations, we determine optimum samplings for high and low electron dose imaging conditions. - Highlights: • The definition of dose-dependent atom contrast is introduced. • The dependence of the signal-to-noise ratio, atom contrast and specimen resolution on electron dose and sampling is explored. • The optimum sampling can be determined according to different dose conditions

  1. Opto-mechano-electrical tripling in ZnO nanowires probed by photocurrent spectroscopy in a high-resolution transmission electron microscope

    Energy Technology Data Exchange (ETDEWEB)

    Zhang, C.; Golberg, D., E-mail: xuzhi@iphy.ac.cn, E-mail: golberg.dmitri@nims.go.jp [International Center for Materials Nanoarchitectonics (MANA), National Institute for Materials Science (NIMS), Namiki 1-1, Tsukuba, Ibaraki 3050044 (Japan); Graduate School of Pure and Applied Sciences, University of Tsukuba, Tennodai 1, Tsukuba, Ibaraki 3058577 (Japan); Xu, Z., E-mail: xuzhi@iphy.ac.cn, E-mail: golberg.dmitri@nims.go.jp [Beijing National Laboratory for Condensed Matter Physics, Institute of Physics, Chinese Academy of Sciences, Beijing 100190 (China); Kvashnin, D. G. [National University of Science and Technology, MISIS, Leninskiy Prospect 4, Moscow 119049 (Russian Federation); Tang, D.-M.; Xue, Y. M.; Bando, Y. [International Center for Materials Nanoarchitectonics (MANA), National Institute for Materials Science (NIMS), Namiki 1-1, Tsukuba, Ibaraki 3050044 (Japan); Sorokin, P. B. [National University of Science and Technology, MISIS, Leninskiy Prospect 4, Moscow 119049 (Russian Federation); Moscow Institute of Physics and Technology, Institutsky Lane 9, Dolgoprudny 141700 (Russian Federation)

    2015-08-31

    Photocurrent spectroscopy of individual free-standing ZnO nanowires inside a high-resolution transmission electron microscope (TEM) is reported. By using specially designed optical in situ TEM system capable of scanning tunneling microscopy probing paired with light illumination, opto-mechano-electrical tripling phenomenon in ZnO nanowires is demonstrated. Splitting of photocurrent spectra at around 3.3 eV under in situ TEM bending of ZnO nanowires directly corresponds to nanowire deformation and appearance of expanded and compressed nanowire sides. Theoretical simulation of a bent ZnO nanowire has an excellent agreement with the experimental data. The splitting effect could be explained by a change in the valence band structure of ZnO nanowires due to a lattice strain. The strain-induced splitting provides important clues for future flexible piezo-phototronics.

  2. High spatial resolution upgrade of the electron cyclotron emission radiometer for the DIII-D tokamak.

    Science.gov (United States)

    Truong, D D; Austin, M E

    2014-11-01

    The 40-channel DIII-D electron cyclotron emission (ECE) radiometer provides measurements of Te(r,t) at the tokamak midplane from optically thick, second harmonic X-mode emission over a frequency range of 83-130 GHz. The frequency spacing of the radiometer's channels results in a spatial resolution of ∼1-3 cm, depending on local magnetic field and electron temperature. A new high resolution subsystem has been added to the DIII-D ECE radiometer to make sub-centimeter (0.6-0.8 cm) resolution Te measurements. The high resolution subsystem branches off from the regular channels' IF bands and consists of a microwave switch to toggle between IF bands, a switched filter bank for frequency selectivity, an adjustable local oscillator and mixer for further frequency down-conversion, and a set of eight microwave filters in the 2-4 GHz range. Higher spatial resolution is achieved through the use of a narrower (200 MHz) filter bandwidth and closer spacing between the filters' center frequencies (250 MHz). This configuration allows for full coverage of the 83-130 GHz frequency range in 2 GHz bands. Depending on the local magnetic field, this translates into a "zoomed-in" analysis of a ∼2-4 cm radial region. Expected uses of these channels include mapping the spatial dependence of Alfven eigenmodes, geodesic acoustic modes, and externally applied magnetic perturbations. Initial Te measurements, which demonstrate that the desired resolution is achieved, are presented.

  3. A common structure for the potexviruses

    Energy Technology Data Exchange (ETDEWEB)

    Kendall, Amy; Bian, Wen; Maris, Alexander; Azzo, Caitlin; Groom, Joseph [Department of Biological Sciences and Center for Structural Biology, Vanderbilt University, Nashville, TN 37235 (United States); Williams, Dewight; Shi, Jian; Stewart, Phoebe L. [Department of Molecular Physiology and Biophysics and Center for Structural Biology, Vanderbilt University, Nashville, TN 37232 (United States); Wall, Joseph S. [Biology Department, Brookhaven National Laboratory, Upton, NY 11973 (United States); Stubbs, Gerald, E-mail: gerald.stubbs@vanderbilt.edu [Department of Biological Sciences and Center for Structural Biology, Vanderbilt University, Nashville, TN 37235 (United States)

    2013-02-05

    We have used fiber diffraction, cryo-electron microscopy, and scanning transmission electron microscopy to confirm the symmetry of three potexviruses, potato virus X, papaya mosaic virus, and narcissus mosaic virus, and to determine their low-resolution structures. All three viruses have slightly less than nine subunits per turn of the viral helix. Our data strongly support the view that all potexviruses have approximately the same symmetry. The structures are dominated by a large domain at high radius in the virion, with a smaller domain, which includes the putative RNA-binding site, extending to low radius.

  4. System and method for compressive scanning electron microscopy

    Science.gov (United States)

    Reed, Bryan W

    2015-01-13

    A scanning transmission electron microscopy (STEM) system is disclosed. The system may make use of an electron beam scanning system configured to generate a plurality of electron beam scans over substantially an entire sample, with each scan varying in electron-illumination intensity over a course of the scan. A signal acquisition system may be used for obtaining at least one of an image, a diffraction pattern, or a spectrum from the scans, the image, diffraction pattern, or spectrum representing only information from at least one of a select subplurality or linear combination of all pixel locations comprising the image. A dataset may be produced from the information. A subsystem may be used for mathematically analyzing the dataset to predict actual information that would have been produced by each pixel location of the image.

  5. Minimal exposure technique in the Cambridge University 600kV high resolution electron microscope

    International Nuclear Information System (INIS)

    Fryer, J.R.; Cleaver, J.R.A.; Smith, D.J.

    1980-01-01

    Radiation damage due to the incident electron beam imposes a fundamental limitation on the information obtainable by electron microscopy about organic materials; it is desirable therefore that exposure of the specimen to the electron beam should be restricted to the actual period during which the image is being recorded. A description is given of methods employed in the observation of the organic aromatic hydrocarbons quaterrylene, ovalene and coronene with the Cambridge University 600kV high resolution electron microscope (HREM). In particular, the condenser-objective mode of operation of this microscope lends itself to the use of an area-defining aperture below the second condenser lens conjugate with the specimen. Furthermore, operation at the higher accelerating voltage of this instrument could be anticipated to reduce the rate of damage, depending on the dominant beam-specimen interaction, whilst the increased width of the first broad band of the contrast transfer function of this microscope at the optimum defocus may overcome the reported resolution limitation of current 100kV microscopes for the observation of related materials. (author)

  6. The role of symmetry in the theory of inelastic high-energy electron scattering and its application to atomic-resolution core-loss imaging.

    Science.gov (United States)

    Dwyer, C

    2015-04-01

    The inelastic scattering of a high-energy electron in a solid constitutes a bipartite quantum system with an intrinsically large number of excitations, posing a considerable challenge for theorists. It is demonstrated how and why the utilization of symmetries, or approximate symmetries, can lead to significant improvements in both the description of the scattering physics and the efficiency of numerical computations. These ideas are explored thoroughly for the case of core-loss excitations, where it is shown that the coupled angular momentum basis leads to dramatic improvements over the bases employed in previous work. The resulting gains in efficiency are demonstrated explicitly for K-, L- and M-shell excitations, including such excitations in the context of atomic-resolution imaging in the scanning transmission electron microscope. The utilization of other symmetries is also discussed. Copyright © 2014 Elsevier B.V. All rights reserved.

  7. IN-SITU EXPERIMENTS OF VACUUM DISCHARGE USING SCANNING ELECTRON MICROSCOPES

    CERN Document Server

    Muranaka, T; Leifer, K; Ziemann, V

    2011-01-01

    The fundamental understanding of vacuum discharge mechanisms and induced surface damage is indispensable for the CLIC feasibility study. We have been conducting dc sparc experiments inside a Scanning Electron Microscope (SEM) at Uppsala university in order to investigate localized breakdown phenomena. By using a SEM, we achieve the resolution of the electron probe in the few-nm range, which is of great advantage as the surface roughness of the polished accelerating structures is in the same scale. The high accelerating field of 1 GV/m is realized by biasing an electrode with 1 kV set above the sample with a gap of sub μm. Furthermore, a second SEM equipped with a Focused Ion Beam (FIB) is used to modify the topography of sample surfaces thus the geometrical dependence of field emissions and vacuum discharges can be studied. The FIB can be used for the surface damage analysis as well. We have demonstrated subsurface damage observations by using FIB to sputter a rectangular recess into the sample in the breakd...

  8. Monte Carlo simulation for scanning technique with scattering foil free electron beam: A proof of concept study.

    Directory of Open Access Journals (Sweden)

    Wonmo Sung

    Full Text Available This study investigated the potential of a newly proposed scattering foil free (SFF electron beam scanning technique for the treatment of skin cancer on the irregular patient surfaces using Monte Carlo (MC simulation. After benchmarking of the MC simulations, we removed the scattering foil to generate SFF electron beams. Cylindrical and spherical phantoms with 1 cm boluses were generated and the target volume was defined from the surface to 5 mm depth. The SFF scanning technique with 6 MeV electrons was simulated using those phantoms. For comparison, volumetric modulated arc therapy (VMAT plans were also generated with two full arcs and 6 MV photon beams. When the scanning resolution resulted in a larger separation between beams than the field size, the plan qualities were worsened. In the cylindrical phantom with a radius of 10 cm, the conformity indices, homogeneity indices and body mean doses of the SFF plans (scanning resolution = 1° vs. VMAT plans were 1.04 vs. 1.54, 1.10 vs. 1.12 and 5 Gy vs. 14 Gy, respectively. Those of the spherical phantom were 1.04 vs. 1.83, 1.08 vs. 1.09 and 7 Gy vs. 26 Gy, respectively. The proposed SFF plans showed superior dose distributions compared to the VMAT plans.

  9. Monte Carlo simulation for scanning technique with scattering foil free electron beam: A proof of concept study.

    Science.gov (United States)

    Sung, Wonmo; Park, Jong In; Kim, Jung-In; Carlson, Joel; Ye, Sung-Joon; Park, Jong Min

    2017-01-01

    This study investigated the potential of a newly proposed scattering foil free (SFF) electron beam scanning technique for the treatment of skin cancer on the irregular patient surfaces using Monte Carlo (MC) simulation. After benchmarking of the MC simulations, we removed the scattering foil to generate SFF electron beams. Cylindrical and spherical phantoms with 1 cm boluses were generated and the target volume was defined from the surface to 5 mm depth. The SFF scanning technique with 6 MeV electrons was simulated using those phantoms. For comparison, volumetric modulated arc therapy (VMAT) plans were also generated with two full arcs and 6 MV photon beams. When the scanning resolution resulted in a larger separation between beams than the field size, the plan qualities were worsened. In the cylindrical phantom with a radius of 10 cm, the conformity indices, homogeneity indices and body mean doses of the SFF plans (scanning resolution = 1°) vs. VMAT plans were 1.04 vs. 1.54, 1.10 vs. 1.12 and 5 Gy vs. 14 Gy, respectively. Those of the spherical phantom were 1.04 vs. 1.83, 1.08 vs. 1.09 and 7 Gy vs. 26 Gy, respectively. The proposed SFF plans showed superior dose distributions compared to the VMAT plans.

  10. Comparison of high resolution computed tomography and pulmonary function tests in diagnosis of mild emphysema

    International Nuclear Information System (INIS)

    Kuwano, Kazuyoshi; Matsuba, Kenichi; Ikeda, Togo

    1989-01-01

    To assess the ability of high resolution CT scan and pulmonary function tests in detecting and grading mild emphysema, we correlated the high resolution CT scan and pulmonary function tests with the pathologic grade of emphysema and the destructive index of lung specimens from 42 patients undergoing thoracotomy for solitary pulmonary nodules. Using the high resolution CT scan, we could identify the pathologic grade of mild and moderate emphysema. By measuring diffusing capacity per unit alveolar gas volume (DLco/VA), it seemed to be possible to detect the mildest degree of alveolar destruction assessed by the destructive index, which was not detected by high resolution CT scan. The reason for these results seemed to be that we assessed the severity of emphysema by detecting the air space enlargement on high resolution CT scan images caused by the destruction of alveolar walls, which were detectable by measuring DLco/VA. We conclude that it is possible to detect mild emphysema using the combination of high resolution CT scan and pulmomary function tests. (author)

  11. CryoSat SAR/SARin Level1b products: assessment of BaselineC and improvements towards BaselineD

    Science.gov (United States)

    Scagliola, Michele; Fornari, Marco; Bouffard, Jerome; Parrinello, Tommaso

    2017-04-01

    CryoSat was launched on the 8th April 2010 and is the first European ice mission dedicated to the monitoring of precise changes in the thickness of polar ice sheets and floating sea ice. Cryosat carries an innovative radar altimeter called the Synthetic Aperture Interferometric Altimeter (SIRAL), that transmits pulses at a high pulse repetition frequency thus making the received echoes phase coherent and suitable for azimuth processing. This allows to reach a significantly improved along track resolution with respect to traditional pulse-width limited altimeters. CryoSat is the first altimetry mission operating in SAR mode and continuous improvements in the Level1 Instrument Processing Facility (IPF1) are being identified, tested and validated in order to improve the quality of the Level1b products. The current IPF, Baseline C, was released in operation in April 2015 and the second CryoSat reprocessing campaign was jointly initiated, taking benefit of the upgrade implemented in the IPF1 processing chain but also of some specific configurations for the calibration corrections. In particular, the CryoSat Level1b BaselineC products generated in the framework of the second reprocessing campaign include refined information for what concerns the mispointing angles and the calibration corrections. This poster will thus detail thus the evolutions that are currently planned for the CryoSat BaselineD SAR/SARin Level1b products and the corresponding quality improvements that are expected.

  12. Cryo-CMOS Circuits and Systems for Quantum Computing Applications

    NARCIS (Netherlands)

    Patra, B; Incandela, R.M.; van Dijk, J.P.G.; Homulle, H.A.R.; Song, Lin; Shahmohammadi, M.; Staszewski, R.B.; Vladimirescu, A.; Babaie, M.; Sebastiano, F.; Charbon, E.E.E.

    2018-01-01

    A fault-tolerant quantum computer with millions of quantum bits (qubits) requires massive yet very precise control electronics for the manipulation and readout of individual qubits. CMOS operating at cryogenic temperatures down to 4 K (cryo-CMOS) allows for closer system integration, thus promising

  13. Cryo-imaging of fluorescently labeled single cells in a mouse

    Science.gov (United States)

    Steyer, Grant J.; Roy, Debashish; Salvado, Olivier; Stone, Meredith E.; Wilson, David L.

    2009-02-01

    We developed a cryo-imaging system to provide single-cell detection of fluorescently labeled cells in mouse, with particular applicability to stem cells and metastatic cancer. The Case cryoimaging system consists of a fluorescence microscope, robotic imaging positioner, customized cryostat, PC-based control system, and visualization/analysis software. The system alternates between sectioning (10-40 μm) and imaging, collecting color brightfield and fluorescent blockface image volumes >60GB. In mouse experiments, we imaged quantum-dot labeled stem cells, GFP-labeled cancer and stem cells, and cell-size fluorescent microspheres. To remove subsurface fluorescence, we used a simplified model of light-tissue interaction whereby the next image was scaled, blurred, and subtracted from the current image. We estimated scaling and blurring parameters by minimizing entropy of subtracted images. Tissue specific attenuation parameters were found [uT : heart (267 +/- 47.6 μm), liver (218 +/- 27.1 μm), brain (161 +/- 27.4 μm)] to be within the range of estimates in the literature. "Next image" processing removed subsurface fluorescence equally well across multiple tissues (brain, kidney, liver, adipose tissue, etc.), and analysis of 200 microsphere images in the brain gave 97+/-2% reduction of subsurface fluorescence. Fluorescent signals were determined to arise from single cells based upon geometric and integrated intensity measurements. Next image processing greatly improved axial resolution, enabled high quality 3D volume renderings, and improved enumeration of single cells with connected component analysis by up to 24%. Analysis of image volumes identified metastatic cancer sites, found homing of stem cells to injury sites, and showed microsphere distribution correlated with blood flow patterns. We developed and evaluated cryo-imaging to provide single-cell detection of fluorescently labeled cells in mouse. Our cryo-imaging system provides extreme (>60GB), micron

  14. Correlative Cryo-Tem Cryo-Stxm and Cryo-Shxm Investigation of Selenium Bioreduction in a Contaminated Aquifer

    Science.gov (United States)

    Fakra, S.; Luef, B.; Tyliszczak, T.; Castelle, C. J.; Mullin, S. W.; Hug, L. A.; Williams, K. H.; Marcus, M.; Banfield, J. F.

    2015-12-01

    Accurate mapping of the composition and ultrastructure of minerals and cells is key to understanding biogeochemical process in contaminated environments. Here we developed two apparatus that allow correlation of cryogenic transmission electron microscopy (TEM), synchrotron hard X-ray microprobe (SHXM) and scanning transmission X-ray microscopy (STXM) datasets. These cryogenic methods enabled precise determination of the distribution, valence state and structure of selenium in intact biofilms sampled during a biostimulation experiment in a contaminated aquifer near Rifle, CO, USA. Results were replicated in the laboratory via anaerobic selenate-reducing enrichment cultures. 16S rRNA analyses of field-derived biofilm indicated the dominance of Betaproteobacteria from the Comamonadaceae family, and uncultivated members of the Simplicispira genus. The major product in field and culture-derived biofilms consists of ~25-300 nm red amorphous Se0 aggregates of colloidal nanoparticles. Correlative analyses of the cultures provided direct evidence for microbial dissimilatory reduction of Se(VI) to Se(IV) to Se0. X-ray diffraction and Se K-edge extended X-ray absorption fine structure spectroscopy revealed red amorphous Se0 with a first shell Se-Se interatomic distance of 2.339 ± 0.003 Å. STXM showed that these aggregates are strongly associated with a protein-rich biofilm matrix containing acidic polysaccharides. From Rifle groundwater, we isolated a strain that shares 98.9% 16S rRNA gene sequence identity with Dechloromonas aromatica RCB and grows anaerobically by oxidizing acetate and reducing selenate. We refer to this isolate as Dechloromonas selenatis strain RGW99. 3D cryo-electron tomography showed that Se0 particles do not form inside the cytoplasm but rather originate in the cell membrane. The end product of selenate reduction by D. selenatis is 240 ± 66 nm diameter red amorphous Se0 colloidal aggregates. This product was found to be stable for months. Overall

  15. Millisecond resolution electron fluxes from the Cluster satellites: Calibrated EDI ambient electron data

    Science.gov (United States)

    Förster, Matthias; Rashev, Mikhail; Haaland, Stein

    2017-04-01

    The Electron Drift Instrument (EDI) onboard Cluster can measure 500 eV and 1 keV electron fluxes with high time resolution during passive operation phases in its Ambient Electron (AE) mode. Data from this mode is available in the Cluster Science Archive since October 2004 with a cadence of 16 Hz in the normal mode or 128 Hz for burst mode telemetry intervals. The fluxes are recorded at pitch angles of 0, 90, and 180 degrees. This paper describes the calibration and validation of these measurements. The high resolution AE data allow precise temporal and spatial diagnostics of magnetospheric boundaries and will be used for case studies and statistical studies of low energy electron fluxes in the near-Earth space. We show examples of applications.

  16. CryoSat Level1b SAR/SARin: quality improvements towards BaselineC

    Science.gov (United States)

    Scagliola, Michele; Fornari, Marco; Bouzinac, Catherine; Tagliani, Nicolas; Parrinello, Tommaso

    2014-05-01

    CryoSat was launched on the 8th April 2010 and it is the first European ice mission dedicated to monitoring precise changes in the thickness of polar ice sheets and floating sea ice over a 3-year period. Cryosat carries an innovative radar altimeter called the Synthetic Aperture Interferometric Altimeter (SIRAL), that transmits pulses at a high pulse repetition frequency thus making the received echoes phase coherent and suitable for azimuth processing. This allows to reach a significantly improved along track resolution with respect to traditional pulse-width limited altimeters. CryoSat is the first altimetry mission operating in SAR mode and continuous improvement in the Level1 Instrument Processing Facility (IPF1) are being identified, tested and validated in order to improve the quality of the Level1b products. Towards the release of the BaselineC of the CryoSat Level1b SAR/SARin products, that is expected during 2014, several improvements have been identified: • a datation bias of about -0.5195 ms will be corrected • a range bias of about -0.6730 m will be corrected • the waveform length in the Level1b product will be doubled with respect to BaselineB • improved processing for 1Hz echoes to have sharper waveforms • surface sample stack weighting to filter out the single look echoes acquired at highest look angle, that results in a sharpening of the 20Hz waveforms This poster details the main improvements that are foreseen to be included in the CryoSat Level1b SAR/SARin products in BaselineC.

  17. Cryo-electron microscopy and single molecule fluorescent microscopy detect CD4 receptor induced HIV size expansion prior to cell entry

    International Nuclear Information System (INIS)

    Pham, Son; Tabarin, Thibault; Garvey, Megan; Pade, Corinna; Rossy, Jérémie; Monaghan, Paul; Hyatt, Alex; Böcking, Till; Leis, Andrew; Gaus, Katharina; Mak, Johnson

    2015-01-01

    Viruses are often thought to have static structure, and they only remodel after the viruses have entered target cells. Here, we detected a size expansion of virus particles prior to viral entry using cryo-electron microscopy (cryo-EM) and single molecule fluorescence imaging. HIV expanded both under cell-free conditions with soluble receptor CD4 (sCD4) targeting the CD4 binding site on the HIV-1 envelope protein (Env) and when HIV binds to receptor on cellular membrane. We have shown that the HIV Env is needed to facilitate receptor induced virus size expansions, showing that the ‘lynchpin’ for size expansion is highly specific. We demonstrate that the size expansion required maturation of HIV and an internal capsid core with wild type stability, suggesting that different HIV compartments are linked and are involved in remodelling. Our work reveals a previously unknown event in HIV entry, and we propose that this pre-entry priming process enables HIV particles to facilitate the subsequent steps in infection. - Highlights: • Cell free viruses are able to receive external trigger that leads to apparent size expansion. • Virus envelope and CD4 receptor engagement is the lynchpin of virus size expansion. • Internal capsid organisation can influence receptor mediated virus size expansion. • Pre-existing virus-associated lipid membrane in cell free virus can accommodate the receptor mediated virus size expansion.

  18. Cryo-electron microscopy and single molecule fluorescent microscopy detect CD4 receptor induced HIV size expansion prior to cell entry

    Energy Technology Data Exchange (ETDEWEB)

    Pham, Son [Deakin University, Victoria 3216 (Australia); CSIRO Australian Animal Health Laboratory, Victoria 3220 (Australia); Tabarin, Thibault [ARC Centre of Excellence in Advanced Molecular Imaging, University of New South Wales, New South Wales 3220 (Australia); Garvey, Megan; Pade, Corinna [Deakin University, Victoria 3216 (Australia); CSIRO Australian Animal Health Laboratory, Victoria 3220 (Australia); Rossy, Jérémie [ARC Centre of Excellence in Advanced Molecular Imaging, University of New South Wales, New South Wales 3220 (Australia); Monaghan, Paul; Hyatt, Alex [CSIRO Australian Animal Health Laboratory, Victoria 3220 (Australia); Böcking, Till [ARC Centre of Excellence in Advanced Molecular Imaging, University of New South Wales, New South Wales 3220 (Australia); Leis, Andrew [CSIRO Australian Animal Health Laboratory, Victoria 3220 (Australia); Gaus, Katharina, E-mail: k.gaus@unsw.edu.au [ARC Centre of Excellence in Advanced Molecular Imaging, University of New South Wales, New South Wales 3220 (Australia); Mak, Johnson, E-mail: j.mak@deakin.edu.au [Deakin University, Victoria 3216 (Australia); CSIRO Australian Animal Health Laboratory, Victoria 3220 (Australia)

    2015-12-15

    Viruses are often thought to have static structure, and they only remodel after the viruses have entered target cells. Here, we detected a size expansion of virus particles prior to viral entry using cryo-electron microscopy (cryo-EM) and single molecule fluorescence imaging. HIV expanded both under cell-free conditions with soluble receptor CD4 (sCD4) targeting the CD4 binding site on the HIV-1 envelope protein (Env) and when HIV binds to receptor on cellular membrane. We have shown that the HIV Env is needed to facilitate receptor induced virus size expansions, showing that the ‘lynchpin’ for size expansion is highly specific. We demonstrate that the size expansion required maturation of HIV and an internal capsid core with wild type stability, suggesting that different HIV compartments are linked and are involved in remodelling. Our work reveals a previously unknown event in HIV entry, and we propose that this pre-entry priming process enables HIV particles to facilitate the subsequent steps in infection. - Highlights: • Cell free viruses are able to receive external trigger that leads to apparent size expansion. • Virus envelope and CD4 receptor engagement is the lynchpin of virus size expansion. • Internal capsid organisation can influence receptor mediated virus size expansion. • Pre-existing virus-associated lipid membrane in cell free virus can accommodate the receptor mediated virus size expansion.

  19. High-resolution electron microscopic evidence for the filamentous structure of the cyst wall in Giardia muris and Giardia duodenalis.

    Science.gov (United States)

    Erlandsen, S L; Bemrick, W J; Pawley, J

    1989-10-01

    High-resolution morphological studies of the cyst wall of Giardia spp. were performed using low-voltage scanning electron microscopy (LVSEM) and transmission electron microscopy (TEM). The cyst wall was composed of membranous and filamentous layers. The membranous layer consisted of an inner and an outer cyst membrane separated by a thin layer of cytoplasm. The filamentous layer contained individual filaments that ranged from 7 to 20 nm in diameter when measured by LVSEM, formed a dense meshwork with branches or interconnections, and were occasionally arranged on the surface in whorled patterns. Cysts of Giardia muris from mice, Giardia duodenalis from dogs, pigs, voles, beavers, muskrats, and humans, and Giardia psittaci from a bird (parakeet), possessed an essentially identical wall composed of filaments. Inducement of excystation in viable Giardia cysts produced a dramatic increase in the interfilament spacing over an entire cyst, but none was observed in heat-killed or chemically fixed control cysts. These results demonstrated that the cyst wall of Giardia spp. was composed of a complex arrangement of filaments, presumably formed during the process of encystment.

  20. High resolution melting for mutation scanning of TP53 exons 5–8

    International Nuclear Information System (INIS)

    Krypuy, Michael; Dobrovic, Alexander; Ahmed, Ahmed Ashour; Etemadmoghadam, Dariush; Hyland, Sarah J; Australian Ovarian Cancer Study Group; Fazio, Anna de; Fox, Stephen B; Brenton, James D; Bowtell, David D

    2007-01-01

    p53 is commonly inactivated by mutations in the DNA-binding domain in a wide range of cancers. As mutant p53 often influences response to therapy, effective and rapid methods to scan for mutations in TP53 are likely to be of clinical value. We therefore evaluated the use of high resolution melting (HRM) as a rapid mutation scanning tool for TP53 in tumour samples. We designed PCR amplicons for HRM mutation scanning of TP53 exons 5 to 8 and tested them with DNA from cell lines hemizygous or homozygous for known mutations. We assessed the sensitivity of each PCR amplicon using dilutions of cell line DNA in normal wild-type DNA. We then performed a blinded assessment on ovarian tumour DNA samples that had been previously sequenced for mutations in TP53 to assess the sensitivity and positive predictive value of the HRM technique. We also performed HRM analysis on breast tumour DNA samples with unknown TP53 mutation status. One cell line mutation was not readily observed when exon 5 was amplified. As exon 5 contained multiple melting domains, we divided the exon into two amplicons for further screening. Sequence changes were also introduced into some of the primers to improve the melting characteristics of the amplicon. Aberrant HRM curves indicative of TP53 mutations were observed for each of the samples in the ovarian tumour DNA panel. Comparison of the HRM results with the sequencing results revealed that each mutation was detected by HRM in the correct exon. For the breast tumour panel, we detected seven aberrant melt profiles by HRM and subsequent sequencing confirmed the presence of these and no other mutations in the predicted exons. HRM is an effective technique for simple and rapid scanning of TP53 mutations that can markedly reduce the amount of sequencing required in mutational studies of TP53

  1. High-Resolution Measurements of Low-Energy Conversion Electrons

    CERN Multimedia

    Gizon, A; Putaux, J

    2002-01-01

    Measurements of low-energy internal conversion electrons have been performed with high energy resolution in some N = 105 odd and odd-odd nuclei using a semi-circular spectrograph associated to a specific tape transport system. These experiments aimed to answer the following questions~: \\begin{itemize} \\item Do M3 isomeric transitions exist in $^{183}$Pt and $^{181}$Os, isotones of $^{184}$Au~? \\item Are the neutron configurations proposed to describe the isomeric and ground states of $^{184}$Au right or wrong~? \\item Does it exist an isomeric state in $^{182}$Ir, isotone of $^{181}$Os, $^{183}$Pt and $^{184}$Au~? \\item What are the spin and parity values of the excited states of $^{182}$Ir~? \\end{itemize} In $^{183}$Pt, the 35.0 keV M3 isomeric transition has been clearly observed and the reduced transition probability has been determined. The deduced hindrance factor is close to that observed in the neighbouring odd-odd $^{184}$Au nucleus. This confirms the neutron configurations previously proposed for the ...

  2. Imaging collagen type I fibrillogenesis with high spatiotemporal resolution

    International Nuclear Information System (INIS)

    Stamov, Dimitar R; Stock, Erik; Franz, Clemens M; Jähnke, Torsten; Haschke, Heiko

    2015-01-01

    Fibrillar collagens, such as collagen type I, belong to the most abundant extracellular matrix proteins and they have received much attention over the last five decades due to their large interactome, complex hierarchical structure and high mechanical stability. Nevertheless, the collagen self-assembly process is still incompletely understood. Determining the real-time kinetics of collagen type I formation is therefore pivotal for better understanding of collagen type I structure and function, but visualising the dynamic self-assembly process of collagen I on the molecular scale requires imaging techniques offering high spatiotemporal resolution. Fast and high-speed scanning atomic force microscopes (AFM) provide the means to study such processes on the timescale of seconds under near-physiological conditions. In this study we have applied fast AFM tip scanning to study the assembly kinetics of fibrillar collagen type I nanomatrices with a temporal resolution reaching eight seconds for a frame size of 500 nm. By modifying the buffer composition and pH value, the kinetics of collagen fibrillogenesis can be adjusted for optimal analysis by fast AFM scanning. We furthermore show that amplitude-modulation imaging can be successfully applied to extract additional structural information from collagen samples even at high scan rates. Fast AFM scanning with controlled amplitude modulation therefore provides a versatile platform for studying dynamic collagen self-assembly processes at high resolution. - Highlights: • Continuous non-invasive time-lapse investigation of collagen I fibrillogenesis in situ. • Imaging of collagen I self-assembly with high spatiotemporal resolution. • Application of setpoint modulation to study the hierarchical structure of collagen I. • Observing real-time formation of the D-banding pattern in collagen I

  3. Imaging collagen type I fibrillogenesis with high spatiotemporal resolution

    Energy Technology Data Exchange (ETDEWEB)

    Stamov, Dimitar R, E-mail: stamov@jpk.com [JPK Instruments AG, Bouchéstrasse 12, 12435 Berlin (Germany); Stock, Erik [JPK Instruments AG, Bouchéstrasse 12, 12435 Berlin (Germany); Franz, Clemens M [DFG-Center for Functional Nanostructures (CFN), Karlsruhe Institute of Technology (KIT), Wolfgang-Gaede-Strasse 1a, 76131 Karlsruhe (Germany); Jähnke, Torsten; Haschke, Heiko [JPK Instruments AG, Bouchéstrasse 12, 12435 Berlin (Germany)

    2015-02-15

    Fibrillar collagens, such as collagen type I, belong to the most abundant extracellular matrix proteins and they have received much attention over the last five decades due to their large interactome, complex hierarchical structure and high mechanical stability. Nevertheless, the collagen self-assembly process is still incompletely understood. Determining the real-time kinetics of collagen type I formation is therefore pivotal for better understanding of collagen type I structure and function, but visualising the dynamic self-assembly process of collagen I on the molecular scale requires imaging techniques offering high spatiotemporal resolution. Fast and high-speed scanning atomic force microscopes (AFM) provide the means to study such processes on the timescale of seconds under near-physiological conditions. In this study we have applied fast AFM tip scanning to study the assembly kinetics of fibrillar collagen type I nanomatrices with a temporal resolution reaching eight seconds for a frame size of 500 nm. By modifying the buffer composition and pH value, the kinetics of collagen fibrillogenesis can be adjusted for optimal analysis by fast AFM scanning. We furthermore show that amplitude-modulation imaging can be successfully applied to extract additional structural information from collagen samples even at high scan rates. Fast AFM scanning with controlled amplitude modulation therefore provides a versatile platform for studying dynamic collagen self-assembly processes at high resolution. - Highlights: • Continuous non-invasive time-lapse investigation of collagen I fibrillogenesis in situ. • Imaging of collagen I self-assembly with high spatiotemporal resolution. • Application of setpoint modulation to study the hierarchical structure of collagen I. • Observing real-time formation of the D-banding pattern in collagen I.

  4. Can Low-Resolution Airborne Laser Scanning Data Be Used to Model Stream Rating Curves?

    Directory of Open Access Journals (Sweden)

    Steve W. Lyon

    2015-03-01

    Full Text Available This pilot study explores the potential of using low-resolution (0.2 points/m2 airborne laser scanning (ALS-derived elevation data to model stream rating curves. Rating curves, which allow the functional translation of stream water depth into discharge, making them integral to water resource monitoring efforts, were modeled using a physics-based approach that captures basic geometric measurements to establish flow resistance due to implicit channel roughness. We tested synthetically thinned high-resolution (more than 2 points/m2 ALS data as a proxy for low-resolution data at a point density equivalent to that obtained within most national-scale ALS strategies. Our results show that the errors incurred due to the effect of low-resolution versus high-resolution ALS data were less than those due to flow measurement and empirical rating curve fitting uncertainties. As such, although there likely are scale and technical limitations to consider, it is theoretically possible to generate rating curves in a river network from ALS data of the resolution anticipated within national-scale ALS schemes (at least for rivers with relatively simple geometries. This is promising, since generating rating curves from ALS scans would greatly enhance our ability to monitor streamflow by simplifying the overall effort required.

  5. Can low-resolution airborne laser scanning data be used to model stream rating curves?

    Science.gov (United States)

    Lyon, Steve; Nathanson, Marcus; Lam, Norris; Dahlke, Helen; Rutzinger, Martin; Kean, Jason W.; Laudon, Hjalmar

    2015-01-01

    This pilot study explores the potential of using low-resolution (0.2 points/m2) airborne laser scanning (ALS)-derived elevation data to model stream rating curves. Rating curves, which allow the functional translation of stream water depth into discharge, making them integral to water resource monitoring efforts, were modeled using a physics-based approach that captures basic geometric measurements to establish flow resistance due to implicit channel roughness. We tested synthetically thinned high-resolution (more than 2 points/m2) ALS data as a proxy for low-resolution data at a point density equivalent to that obtained within most national-scale ALS strategies. Our results show that the errors incurred due to the effect of low-resolution versus high-resolution ALS data were less than those due to flow measurement and empirical rating curve fitting uncertainties. As such, although there likely are scale and technical limitations to consider, it is theoretically possible to generate rating curves in a river network from ALS data of the resolution anticipated within national-scale ALS schemes (at least for rivers with relatively simple geometries). This is promising, since generating rating curves from ALS scans would greatly enhance our ability to monitor streamflow by simplifying the overall effort required.

  6. High-Resolution Detector For X-Ray Diffraction

    Science.gov (United States)

    Carter, Daniel C.; Withrow, William K.; Pusey, Marc L.; Yost, Vaughn H.

    1988-01-01

    Proposed x-ray-sensitive imaging detector offers superior spatial resolution, counting-rate capacity, and dynamic range. Instrument based on laser-stimulated luminescence and reusable x-ray-sensitive film. Detector scans x-ray film line by line. Extracts latent image in film and simultaneously erases film for reuse. Used primarily for protein crystallography. Principle adapted to imaging detectors for electron microscopy and fluorescence spectroscopy and general use in astronomy, engineering, and medicine.

  7. The measurement and calculation of the X-ray spatial resolution obtained in the analytical electron microscope

    International Nuclear Information System (INIS)

    Michael, J.R.; Williams, D.B.

    1990-01-01

    The X-ray microanalytical spatial resolution is determined experimentally in various analytical electron microscopes by measuring the degradation of an atomically discrete composition profile across an interphase interface in a thin-foil of Ni-Cr-Fe. The experimental spatial resolutions are then compared with calculated values. The calculated spatial resolutions are obtained by the mathematical convolution of the electron probe size with an assumed beam-broadening distribution and the single-scattering model of beam broadening. The probe size is measured directly from an image of the probe in a TEM/SETEM and indirectly from dark-field signal changes resulting from scanning the probe across the edge of an MgO crystal in a dedicated STEM. This study demonstrates the applicability of the convolution technique to the calculation of the microanalytical spatial resolution obtained in the analytical electron microscope. It is demonstrated that, contrary to popular opinion, the electron probe size has a major impact on the measured spatial resolution in foils < 150 nm thick. (author)

  8. Immunogold scanning electron microscopy can reveal the polysaccharide architecture of xylem cell walls

    Science.gov (United States)

    Sun, Yuliang; Juzenas, Kevin

    2017-01-01

    Abstract Immunofluorescence microscopy (IFM) and immunogold transmission electron microscopy (TEM) are the two main techniques commonly used to detect polysaccharides in plant cell walls. Both are important in localizing cell wall polysaccharides, but both have major limitations, such as low resolution in IFM and restricted sample size for immunogold TEM. In this study, we have developed a robust technique that combines immunocytochemistry with scanning electron microscopy (SEM) to study cell wall polysaccharide architecture in xylem cells at high resolution over large areas of sample. Using multiple cell wall monoclonal antibodies (mAbs), this immunogold SEM technique reliably localized groups of hemicellulosic and pectic polysaccharides in the cell walls of five different xylem structures (vessel elements, fibers, axial and ray parenchyma cells, and tyloses). This demonstrates its important advantages over the other two methods for studying cell wall polysaccharide composition and distribution in these structures. In addition, it can show the three-dimensional distribution of a polysaccharide group in the vessel lateral wall and the polysaccharide components in the cell wall of developing tyloses. This technique, therefore, should be valuable for understanding the cell wall polysaccharide composition, architecture and functions of diverse cell types. PMID:28398585

  9. High spatial resolution upgrade of the electron cyclotron emission radiometer for the DIII-D tokamak

    Energy Technology Data Exchange (ETDEWEB)

    Truong, D. D., E-mail: dtruong@wisc.edu [Department of Engineering Physics, University of Wisconsin-Madison, Madison, Wisconsin 53706 (United States); Austin, M. E. [Institute for Fusion Studies, University of Texas, Austin, Texas, 78712 (United States)

    2014-11-15

    The 40-channel DIII-D electron cyclotron emission (ECE) radiometer provides measurements of T{sub e}(r,t) at the tokamak midplane from optically thick, second harmonic X-mode emission over a frequency range of 83–130 GHz. The frequency spacing of the radiometer's channels results in a spatial resolution of ∼1–3 cm, depending on local magnetic field and electron temperature. A new high resolution subsystem has been added to the DIII-D ECE radiometer to make sub-centimeter (0.6–0.8 cm) resolution T{sub e} measurements. The high resolution subsystem branches off from the regular channels’ IF bands and consists of a microwave switch to toggle between IF bands, a switched filter bank for frequency selectivity, an adjustable local oscillator and mixer for further frequency down-conversion, and a set of eight microwave filters in the 2–4 GHz range. Higher spatial resolution is achieved through the use of a narrower (200 MHz) filter bandwidth and closer spacing between the filters’ center frequencies (250 MHz). This configuration allows for full coverage of the 83–130 GHz frequency range in 2 GHz bands. Depending on the local magnetic field, this translates into a “zoomed-in” analysis of a ∼2–4 cm radial region. Expected uses of these channels include mapping the spatial dependence of Alfven eigenmodes, geodesic acoustic modes, and externally applied magnetic perturbations. Initial T{sub e} measurements, which demonstrate that the desired resolution is achieved, are presented.

  10. ESA CryoVEx 2012

    DEFF Research Database (Denmark)

    Skourup, Henriette; Einarsson, Indriði; Forsberg, René

    Institute (AWI) with fixed‐wing airplane (Polar‐5, Basler BT‐67). The airborne systems are described in detail, together with campaign implementation plan, data processing and data quality analysis. The CryoVEx 2012 campaign was a success and the processed data is of high quality. The data set includes 16...

  11. Fabrication of all diamond scanning probes for nanoscale magnetometry

    OpenAIRE

    Appel Patrick; Neu Elke; Ganzhorn Marc; Barfuss Arne; Batzer Marietta; Gratz Micha; Tschoepe Andreas; Maletinsky Patrick

    2016-01-01

    The electronic spin of the nitrogen vacancy (NV) center in diamond forms an atomically sized, highly sensitive sensor for magnetic fields. To harness the full potential of individual NV centers for sensing with high sensitivity and nanoscale spatial resolution, NV centers have to be incorporated into scanning probe structures enabling controlled scanning in close proximity to the sample surface. Here, we present an optimized procedure to fabricate single-crystal, all-diamond scanning probes s...

  12. The performance and potentials of the CryoSat-2 SAR and SARIn modes for lake level estimation

    DEFF Research Database (Denmark)

    Nielsen, Karina; Stenseng, Lars; Andersen, Ole Baltazar

    2017-01-01

    Over the last few decades, satellite altimetry has proven to be valuable for monitoring lake levels. With the new generation of altimetry missions, CryoSat-2 and Sentinel-3, which operate in Synthetic Aperture Radar (SAR) and SAR Interferometric (SARIn) modes, the footprint size is reduced...... to approximately 300 m in the along-track direction. Here, the performance of these new modes is investigated in terms of uncertainty of the estimated water level from CryoSat-2 data and the agreement with in situ data. The data quality is compared to conventional low resolution mode (LRM) altimetry products from...... of that of the Envisat results. Generally, the CryoSat-2 lake levels also show a better agreement with the in situ data. The lower uncertainty of the CryoSat-2 results entails a more detailed description of water level variations....

  13. Graphene-enabled electron microscopy and correlated super-resolution microscopy of wet cells.

    Science.gov (United States)

    Wojcik, Michal; Hauser, Margaret; Li, Wan; Moon, Seonah; Xu, Ke

    2015-06-11

    The application of electron microscopy to hydrated biological samples has been limited by high-vacuum operating conditions. Traditional methods utilize harsh and laborious sample dehydration procedures, often leading to structural artefacts and creating difficulties for correlating results with high-resolution fluorescence microscopy. Here, we utilize graphene, a single-atom-thick carbon meshwork, as the thinnest possible impermeable and conductive membrane to protect animal cells from vacuum, thus enabling high-resolution electron microscopy of wet and untreated whole cells with exceptional ease. Our approach further allows for facile correlative super-resolution and electron microscopy of wet cells directly on the culturing substrate. In particular, individual cytoskeletal actin filaments are resolved in hydrated samples through electron microscopy and well correlated with super-resolution results.

  14. Review: two-photon scanning systems for clinical high resolution in vivo tissue imaging

    Science.gov (United States)

    König, K.; Müller, J.; Höfer, M.; Müller, C.; Weinigel, M.; Bückle, R.; Elsner, P.; Kaatz, M.; Messerschmidt, B.

    2008-02-01

    The femtosecond laser multiphoton tomograph DermaInspect as well as high NA two-photon GRIN microendoscopes for in vivo tomography of human skin have been used to detect malignant melanoma as well as to study the diffusion and intradermal accumulation of topically applied cosmetical and pharmaceutical components. So far, more than 500 patients and volunteers in Europe, Australia, and Asia have been investigated with this unique tomograph. Near infrared 80 MHz picojoule femtosecond laser pulses were employed to excite endogenous fluorophores such as NAD(P)H, flavoproteins, melanin, and elastin as well as fluorescent components of a variety of ointments via a twophoton excitation process. In addition, collagen has been imaged by second harmonic generation. Using a two-PMT detection system, the ratio of elastin to collagen was determined during optical sectioning. A high submicron spatial resolution and 50 picosecond temporal resolution was achieved using galvoscan mirrors and piezodriven focusing optics as well as a time-correlated single photon counting module with a fast microchannel plate detector and fast photomultipliers. Individual intratissue cells, mitochondria, melanosomes, and the morphology of the nuclei as well as extracellular matrix elements could be clearly visualized due to molecular imaging and the calculation of fluorescence lifetime images. Nanoparticles and intratissue drugs have been detected non-invasively, in situ and over a period of up to 3 months. In addition, hydration effects and UV effects were studied by monitoring modifications of cellular morphology and autofluorescence. The system was used to observe the diffusion through the stratum corneum and the accumulation and release of functionalized nanoparticles along hair shafts and epidermal ridges. The DermaInspect been also employed to gain information on skin age and wound healing in patients with ulcers. Novel developments include a galvo/piezo-scan driven flexible articulated arm as

  15. Reprint of: Atmospheric scanning electron microscope observes cells and tissues in open medium through silicon nitride film.

    Science.gov (United States)

    Nishiyama, Hidetoshi; Suga, Mitsuo; Ogura, Toshihiko; Maruyama, Yuusuke; Koizumi, Mitsuru; Mio, Kazuhiro; Kitamura, Shinichi; Sato, Chikara

    2010-11-01

    Direct observation of subcellular structures and their characterization is essential for understanding their physiological functions. To observe them in open environment, we have developed an inverted scanning electron microscope with a detachable, open-culture dish, capable of 8 nm resolution, and combined with a fluorescence microscope quasi-simultaneously observing the same area from the top. For scanning electron microscopy from the bottom, a silicon nitride film window in the base of the dish maintains a vacuum between electron gun and open sample dish while allowing electrons to pass through. Electrons are backscattered from the sample and captured by a detector under the dish. Cells cultured on the open dish can be externally manipulated under optical microscopy, fixed, and observed using scanning electron microscopy. Once fine structures have been revealed by scanning electron microscopy, their component proteins may be identified by comparison with separately prepared fluorescence-labeled optical microscopic images of the candidate proteins, with their heavy-metal-labeled or stained ASEM images. Furthermore, cell nuclei in a tissue block stained with platinum-blue were successfully observed without thin-sectioning, which suggests the applicability of this inverted scanning electron microscope to cancer diagnosis. This microscope visualizes mesoscopic-scale structures, and is also applicable to non-bioscience fields including polymer chemistry. Copyright © 2010 Elsevier Inc. All rights reserved.

  16. Gas-phase synthesis of magnesium nanoparticles: A high-resolution transmission electron microscopy study

    International Nuclear Information System (INIS)

    Kooi, B. J.; Palasantzas, G.; De Hosson, J. Th. M.

    2006-01-01

    Magnesium nanoparticles with size above 10 nm, prepared by gas-phase syntheses, were investigated by high-resolution transmission electron microscopy. The dominant particle shape is a hexagonal prism terminated by Mg(0002) and Mg(1010) facets. Oxidation of Mg yields a MgO shell (∼3 nm thick), which has an orientation relation with the Mg. Inhomogeneous facet oxidation influences their growth kinetics resulting in a relatively broad size and shape distribution. Faceted voids between Mg and MgO shells indicate a fast outward diffusion of Mg and vacancy rearrangement into voids. The faceting of polar (220) planes is assisted by electron irradiation

  17. Can X-ray spectrum imaging replace backscattered electrons for compositional contrast in the scanning electron microscope?

    Science.gov (United States)

    Newbury, Dale E; Ritchie, Nicholas W M

    2011-01-01

    The high throughput of the silicon drift detector energy dispersive X-ray spectrometer (SDD-EDS) enables X-ray spectrum imaging (XSI) in the scanning electron microscope to be performed in frame times of 10-100 s, the typical time needed to record a high-quality backscattered electron (BSE) image. These short-duration XSIs can reveal all elements, except H, He, and Li, present as major constituents, defined as 0.1 mass fraction (10 wt%) or higher, as well as minor constituents in the range 0.01-0.1 mass fraction, depending on the particular composition and possible interferences. Although BSEs have a greater abundance by a factor of 100 compared with characteristic X-rays, the strong compositional contrast in element-specific X-ray maps enables XSI mapping to compete with BSE imaging to reveal compositional features. Differences in the fraction of the interaction volume sampled by the BSE and X-ray signals lead to more delocalization of the X-ray signal at abrupt compositional boundaries, resulting in poorer spatial resolution. Improved resolution in X-ray elemental maps occurs for the case of a small feature composed of intermediate to high atomic number elements embedded in a matrix of lower atomic number elements. XSI imaging strongly complements BSE imaging, and the SDD-EDS technology enables an efficient combined BSE-XSI measurement strategy that maximizes the compositional information. If 10 s or more are available for the measurement of an area of interest, the analyst should always record the combined BSE-XSI information to gain the advantages of both measures of compositional contrast. Copyright © 2011 Wiley Periodicals, Inc.

  18. Contact detection for nanomanipulation in a scanning electron microscope.

    Science.gov (United States)

    Ru, Changhai; To, Steve

    2012-07-01

    Nanomanipulation systems require accurate knowledge of the end-effector position in all three spatial coordinates, XYZ, for reliable manipulation of nanostructures. Although the images acquired by a scanning electron microscope (SEM) provide high resolution XY information, the lack of depth information in the Z-direction makes 3D nanomanipulation time-consuming. Existing approaches for contact detection of end-effectors inside SEM typically utilize fragile touch sensors that are difficult to integrate into a nanomanipulation system. This paper presents a method for determining the contact between an end-effector and a target surface during nanomanipulation inside SEM, purely based on the processing of SEM images. A depth-from-focus method is used in the fast approach of the end-effector to the substrate, followed by fine contact detection. Experimental results demonstrate that the contact detection approach is capable of achieving an accuracy of 21.5 nm at 50,000× magnification while inducing little end-effector damage. Copyright © 2012 Elsevier B.V. All rights reserved.

  19. Electron spin resonance scanning tunneling microscope

    International Nuclear Information System (INIS)

    Guo Yang; Li Jianmei; Lu Xinghua

    2015-01-01

    It is highly expected that the future informatics will be based on the spins of individual electrons. The development of elementary information unit will eventually leads to novel single-molecule or single-atom devices based on electron spins; the quantum computer in the future can be constructed with single electron spins as the basic quantum bits. However, it is still a great challenge in detection and manipulation of a single electron spin, as well as its coherence and entanglement. As an ideal experimental tool for such tasks, the development of electron spin resonance scanning tunneling microscope (ESR-STM) has attracted great attention for decades. This paper briefly introduces the basic concept of ESR-STM. The development history of this instrument and recent progresses are reviewed. The underlying mechanism is explored and summarized. The challenges and possible solutions are discussed. Finally, the prospect of future direction and applications are presented. (authors)

  20. CryoSat-2 altimetry derived Arctic bathymetry map: first results and validation

    Science.gov (United States)

    Andersen, O. B.; Abulaitijiang, A.; Cancet, M.; Knudsen, P.

    2017-12-01

    The Technical University of Denmark (DTU), DTU Space has been developing high quality high resolution gravity fields including the new highly accurate CryoSat-2 radar altimetry satellite data which extends the global coverage of altimetry data up to latitude 88°. With its exceptional Synthetic Aperture Radar (SAR) mode being operating throughout the Arctic Ocean, leads, i.e., the ocean surface heights, is used to retrieve the sea surface height with centimeter-level range precision. Combined with the long repeat cycle ( 369 days), i.e., dense cross-track coverage, the high-resolution Arctic marine gravity can be modelled using the CryoSat-2 altimetry. Further, the polar gap can be filled by the available ArcGP product, thus yielding the complete map of the Arctic bathymetry map. In this presentation, we will make use of the most recent DTU17 marine gravity, to derive the arctic bathymetry map using inversion based on best available hydrographic maps. Through the support of ESA a recent evaluation of existing hydrographic models of the Arctic Ocean Bathymetry models (RTOPO, GEBCO, IBCAO etc) and various inconsistencies have been identified and means to rectify these inconsistencies have been taken prior to perform the inversion using altimetry. Simultaneously DTU Space has been placing great effort on the Arctic data screening, filtering, and de-noising using various altimetry retracking solutions and classifications. All the pre-processing contributed to the fine modelling of Actic gravity map. Thereafter, the arctic marine gravity grids will eventually be translated (downward continuation operation) to a new altimetry enhanced Arctic bathymetry map using appropriate band-pass filtering.

  1. High resolution CT in cystic fibrosis--the contribution of expiratory scans

    International Nuclear Information System (INIS)

    Dorloechter, Ludger; Nes, Harald; Fluge, Gjermund; Rosendahl, Karen

    2003-01-01

    Introduction: The use of high-resolution computed tomography (HRCT) is well accepted as an accurate method for evaluation of lung parenchyma in cystic fibrosis (CF). Several scoring methods exist and, in common, all are based on HRCT findings during inspiration alone. Objective: To examine whether expiratory HRCT scans could add information about the degree of mosaic perfusion in patients with CF. Methods and patients: Pulmonary HRCT was performed in 17 CF patients (median age of 12 years) with 1-mm thin sections and 10-mm intervals during inspiration, followed by 1-mm thin sections with 20-mm intervals during expiration. HRCT was scored by using a modified Bhalla method. Results: The mean HRCT score was 8.2. Out of 17 patients, 11 (65%) demonstrated a pathological mosaic perfusion in expiration, while only three patients showed mosaic perfusion in inspiration. The degree of expiratory mosaic perfusion was graded as severe in nine patients and moderate in two patients. There was a significant correlation between our modified HRCT score and lung function, as measured by forced expiratory volume in 1 s (FEV1% predicted, P<0.01). Conclusion: Mosaic perfusion in expiration was a common pathological HRCT finding in our study group. The clinical significance of this finding needs further evaluation

  2. Development of a dielectric barrier discharge (DBD) cryo-microplasma: generation and diagnostics

    International Nuclear Information System (INIS)

    Ishihara, Daisuke; Noma, Yuri; Stauss, Sven; Sai, Masaki; Tomai, Takaaki; Terashima, Kazuo

    2008-01-01

    We developed a cryo-microplasma, which can continuously control gas temperature below room temperature and below the freezing point of water. To develop the cryo-microplasma, we first developed an atmospheric-pressure low-temperature microplasma that can suppress the increase in its gas temperature. Helium gas was employed, which was generated in open air. The average estimated electron density and temperature were 10 8 -10 9 cm -3 and 4-5 eV, respectively, independent of the applied voltage. Then, helium gas, which was the working gas of the atmospheric-pressure low-temperature microplasma, was cooled by liquid nitrogen to generate an atmospheric-pressure cryo-microplasma in open air. We observed the generation of frost around the quartz tube in which the plasma was generated and an increase in atomic oxygen emission by optical emission spectroscopy. Finally, to avoid the generation of frost, a cryo-microplasma was generated in a reactor chamber separated from open air. Helium, nitrogen and oxygen were employed as working gases. Using thermocouples and by estimation from the nitrogen rotational temperature, we verified that the gas temperature of the cryo-microplasma was much lower (T g ∼ 180-300 K) than that of the conventional atmospheric-pressure low-temperature plasma (above 300 K).

  3. Development of a dielectric barrier discharge (DBD) cryo-microplasma: generation and diagnostics

    Science.gov (United States)

    Ishihara, Daisuke; Noma, Yuri; Stauss, Sven; Sai, Masaki; Tomai, Takaaki; Terashima, Kazuo

    2008-08-01

    We developed a cryo-microplasma, which can continuously control gas temperature below room temperature and below the freezing point of water. To develop the cryo-microplasma, we first developed an atmospheric-pressure low-temperature microplasma that can suppress the increase in its gas temperature. Helium gas was employed, which was generated in open air. The average estimated electron density and temperature were 108-109 cm-3 and 4-5 eV, respectively, independent of the applied voltage. Then, helium gas, which was the working gas of the atmospheric-pressure low-temperature microplasma, was cooled by liquid nitrogen to generate an atmospheric-pressure cryo-microplasma in open air. We observed the generation of frost around the quartz tube in which the plasma was generated and an increase in atomic oxygen emission by optical emission spectroscopy. Finally, to avoid the generation of frost, a cryo-microplasma was generated in a reactor chamber separated from open air. Helium, nitrogen and oxygen were employed as working gases. Using thermocouples and by estimation from the nitrogen rotational temperature, we verified that the gas temperature of the cryo-microplasma was much lower (Tg ≈ 180-300 K) than that of the conventional atmospheric-pressure low-temperature plasma (above 300 K).

  4. A high resolution electron microscopy investigation of curvature in multilayer graphite sheets

    International Nuclear Information System (INIS)

    Wang Zhenxia; Hu Jun; Wang Wenmin; Yu Guoqing

    1998-01-01

    Here the authors report a carbon sample generated by ultrasonic wave high oriented pyrolytic graphite (HOPG) in ethanol, water or ethanol-water mixed solution. High resolution transmission electron microscopy (HRTEM) revealed many multilayer graphite sheets with a total curved angle that is multiples of θ 0 (= 30 degree C). Close examination of the micrographs showed that the curvature is accomplished by bending the lattice planes. A possible explanation for the curvature in multilayer graphite sheets is discussed based on the conformation of graphite symmetry axes and the formation of sp 3 -like line defects in the sp 2 graphitic network

  5. High-speed X-ray phase tomography with Talbot interferometer and fringe scanning method

    International Nuclear Information System (INIS)

    Kibayashi, Shunsuke; Harasse, Sébastien; Yashiro, Wataru; Momose, Atsushi

    2012-01-01

    High-speed X-ray phase tomography based on the Fourier-transform method has been demonstrated with an X-ray Talbot interferometer using white synchrotron radiation. We report the experimental results of high-speed X-ray phase tomography with fringe-scanning method instead of Fourier-transform method to improve spatial resolution without a considerable increase of scan time. To apply fringe-scanning method to high speed tomography, we tested a scan that is a synchronous combination of one-way continuous movements of the sample rotation and the grating displacement. When this scanning method was combined with X-ray phase tomography, we were able to obtain a scan time of 5 s. A comparison of the image quality derived with the conventional approach and with the proposed approach using the fringe-scanning method showed that the latter had better spatial resolution.

  6. Response function and optimum configuration of semiconductor backscattered-electron detectors for scanning electron microscopes

    International Nuclear Information System (INIS)

    Rau, E. I.; Orlikovskiy, N. A.; Ivanova, E. S.

    2012-01-01

    A new highly efficient design for semiconductor detectors of intermediate-energy electrons (1–50 keV) for application in scanning electron microscopes is proposed. Calculations of the response function of advanced detectors and control experiments show that the efficiency of the developed devices increases on average twofold, which is a significant positive factor in the operation of modern electron microscopes in the mode of low currents and at low primary electron energies.

  7. CryoSat/SIRAL Cal1 Calibration Orbits

    Science.gov (United States)

    Scagliola, Michele; Fornari, Marco; Bouffard, Jerome; Parrinello, Tommaso

    2017-04-01

    The main payload of CryoSat is a Ku band pulsewidth limited radar altimeter, called SIRAL (Synthetic interferometric radar altimeter), that transmits pulses at a high pulse repetition frequency thus making the received echoes phase coherent and suitable for SAR processing. This allows to reach an along track resolution that is significantly improved with respect to traditional pulse-width limited altimeters. Due to the fact that SIRAL is a phase coherent pulse-width limited radar altimeter, a proper calibration approach has been developed. In fact, not only corrections for transfer function, gain and instrument path delay have to be computed (as in previous altimeters), but also corrections for phase (SAR/SARIn) and phase difference between the two receiving chains (SARIN only). Recalling that the CryoSat's orbit has a high inclination of 92° and it is non-sun-synchronous, the temperature of the SIRAL changes continuously along the orbit with a period of about 480 days and it is also function of the ascending/descending passes. By analysis of the CAL1 calibration corrections, it has been verified that the internal path delay and the instrument gain variation measured on the SIRAL are affected by the thermal status of the instrument and as a consequence they are expected to vary along the orbit. In order to gain knowledge on the calibration corrections (i.e. the instrument behavior) as function of latitude and temperature, it has been planned to command a few number of orbits where only CAL1 calibration acquisitions are continuously performed. The analysis of the CAL1 calibration corrections produced along the Calibration orbits can be also useful to verify whether the current calibration plan is able to provide sufficiently accurate corrections for the instrument acquisitions at any latitude. In 2016, the CryoSat/SIRAL Cal1 Calibration Orbits have been commanded two times, a first time the 20th of July 2016 and a second time the 24th of November 2016, and they

  8. Zeolites - a high resolution electron microscopy study

    International Nuclear Information System (INIS)

    Alfredsson, V.

    1994-10-01

    High resolution transmission electron microscopy (HRTEM) has been used to investigate a number of zeolites (EMT, FAU, LTL, MFI and MOR) and a member of the mesoporous M41S family. The electron optical artefact, manifested as a dark spot in the projected centre of the large zeolite channels, caused by insufficient transfer of certain reflections in the objective lens has been explained. The artefact severely hinders observation of materials confined in the zeolite channels and cavities. It is shown how to circumvent the artefact problem and how to image confined materials in spite of disturbance caused by the artefact. Image processing by means of a Wiener filter has been applied for removal of the artefact. The detailed surface structure of FAU has been investigated. Comparison of experimental micrographs with images simulated using different surface models indicates that the surface can be terminated in different ways depending on synthesis methods. The dealuminated form of FAU (USY) is covered by an amorphous region. Platinum incorporated in FAU has a preponderance to aggregate in the (111) twin planes, probably due to a local difference in cage structure with more spacious cages. It is shown that platinum is intra-zeolitic as opposed to being located on the external surface of the zeolite crystal. This could be deduced from tomography of ultra-thin sections among observations. HRTEM studies of the mesoporous MCM-41 show that the pores have a hexagonal shape and also supports the mechanistic model proposed which involves a cooperative formation of a mesophase including the silicate species as well as the surfactant. 66 refs, 24 figs

  9. Structural dissection of Ebola virus and its assembly determinants using cryo-electron tomography.

    Science.gov (United States)

    Bharat, Tanmay A M; Noda, Takeshi; Riches, James D; Kraehling, Verena; Kolesnikova, Larissa; Becker, Stephan; Kawaoka, Yoshihiro; Briggs, John A G

    2012-03-13

    Ebola virus is a highly pathogenic filovirus causing severe hemorrhagic fever with high mortality rates. It assembles heterogenous, filamentous, enveloped virus particles containing a negative-sense, single-stranded RNA genome packaged within a helical nucleocapsid (NC). We have used cryo-electron microscopy and tomography to visualize Ebola virus particles, as well as Ebola virus-like particles, in three dimensions in a near-native state. The NC within the virion forms a left-handed helix with an inner nucleoprotein layer decorated with protruding arms composed of VP24 and VP35. A comparison with the closely related Marburg virus shows that the N-terminal region of nucleoprotein defines the inner diameter of the Ebola virus NC, whereas the RNA genome defines its length. Binding of the nucleoprotein to RNA can assemble a loosely coiled NC-like structure; the loose coil can be condensed by binding of the viral matrix protein VP40 to the C terminus of the nucleoprotein, and rigidified by binding of VP24 and VP35 to alternate copies of the nucleoprotein. Four proteins (NP, VP24, VP35, and VP40) are necessary and sufficient to mediate assembly of an NC with structure, symmetry, variability, and flexibility indistinguishable from that in Ebola virus particles released from infected cells. Together these data provide a structural and architectural description of Ebola virus and define the roles of viral proteins in its structure and assembly.

  10. Electron diffraction and high-resolution transmission electron microscopy of the high temperature crystal structures of GexSb2Te3+x (x=1,2,3) phase change material

    NARCIS (Netherlands)

    Kooi, B.J.; de Hosson, J.T.M.

    2002-01-01

    The crystal structures of GeSb2Te4, Ge2Sb2Te5, and Ge3Sb2Te6 were determined using electron diffraction and high-resolution transmission electron microscopy. The structure determined for the former two crystals deviates from the ones proposed in the literature. These crystal structures were

  11. Near-Atomic Resolution Structure of a Plant Geminivirus Determined by Electron Cryomicroscopy.

    Science.gov (United States)

    Hipp, Katharina; Grimm, Clemens; Jeske, Holger; Böttcher, Bettina

    2017-08-01

    African cassava mosaic virus is a whitefly-transmitted geminivirus which forms unique twin particles of incomplete icosahedra that are joined at five-fold vertices, building an unusual waist. How its 22 capsomers interact within a half-capsid or across the waist is unknown thus far. Using electron cryo-microscopy and image processing, we determined the virion structure with a resolution of 4.2 Å and built an atomic model for its capsid protein. The inter-capsomer contacts mediated by the flexible N termini and loop regions differed within the half-capsids and at the waist, explaining partly the unusual twin structure. The tip of the pentameric capsomer is sealed by a plug formed by a turn region harboring the evolutionary conserved residue Y193. Basic amino acid residues inside the capsid form a positively charged pocket next to the five-fold axis of the capsomer suitable for binding DNA. Within this pocket, density most likely corresponding to DNA was resolved. Copyright © 2017 Elsevier Ltd. All rights reserved.

  12. High-resolution x-ray scattering studies of charge ordering in highly correlated electron systems

    International Nuclear Information System (INIS)

    Ghazi, M.E.

    2002-01-01

    Many important properties of transition metal oxides such as, copper oxide high-temperature superconductivity and colossal magnetoresistance (CMR) in manganites are due to strong electron-electron interactions, and hence these systems are called highly correlated systems. These materials are characterised by the coexistence of different kinds of order, including charge, orbital, and magnetic moment. This thesis contains high-resolution X-ray scattering studies of charge ordering in such systems namely the high-T C copper oxides isostructural system, La 2-x Sr x NiO 4 with various Sr concentrations (x = 0.33 - 0.2), and the CMR manganite system, Nd 1/2 Sr 1/2 MnO 3 . It also includes a review of charge ordering in a large variety of transition metal oxides, such as ferrates, vanadates, cobaltates, nickelates, manganites, and cuprates systems, which have been reported to date in the scientific literature. Using high-resolution synchrotron X-ray scattering, it has been demonstrated that the charge stripes exist in a series of single crystals of La 2-x Sr x NiO 4 with Sr concentrations (x = 0.33 - 0.2) at low temperatures. Satellite reflections due to the charge ordering were found with the wavevector (2ε, 0, 1) below the charge ordering transition temperature, T CO , where 2ε is the amount of separation from the corresponding Bragg peak. The charge stripes are shown to be two-dimensional in nature both by measurements of their correlation lengths and by measurement of the critical exponents of the charge stripe melting transition with an anomaly at x = 0.25. The results show by decreasing the hole concentration from the x = 0.33 to 0.2, the well-correlated charge stripes change to a glassy state at x = 0.25. The electronic transition into the charge stripe phase is second-order without any corresponding structural transition. Above the second-order transition critical scattering was observed due to fluctuations into the charge stripe phase. In a single-crystal of Nd

  13. Direct Visualization of Local Electromagnetic Field Structures by Scanning Transmission Electron Microscopy.

    Science.gov (United States)

    Shibata, Naoya; Findlay, Scott D; Matsumoto, Takao; Kohno, Yuji; Seki, Takehito; Sánchez-Santolino, Gabriel; Ikuhara, Yuichi

    2017-07-18

    The functional properties of materials and devices are critically determined by the electromagnetic field structures formed inside them, especially at nanointerface and surface regions, because such structures are strongly associated with the dynamics of electrons, holes and ions. To understand the fundamental origin of many exotic properties in modern materials and devices, it is essential to directly characterize local electromagnetic field structures at such defect regions, even down to atomic dimensions. In recent years, rapid progress in the development of high-speed area detectors for aberration-corrected scanning transmission electron microscopy (STEM) with sub-angstrom spatial resolution has opened new possibilities to directly image such electromagnetic field structures at very high-resolution. In this Account, we give an overview of our recent development of differential phase contrast (DPC) microscopy for aberration-corrected STEM and its application to many materials problems. In recent years, we have developed segmented-type STEM detectors which divide the detector plane into 16 segments and enable simultaneous imaging of 16 STEM images which are sensitive to the positions and angles of transmitted/scattered electrons on the detector plane. These detectors also have atomic-resolution imaging capability. Using these segmented-type STEM detectors, we show DPC STEM imaging to be a very powerful tool for directly imaging local electromagnetic field structures in materials and devices in real space. For example, DPC STEM can clearly visualize the local electric field variation due to the abrupt potential change across a p-n junction in a GaAs semiconductor, which cannot be observed by normal in-focus bright-field or annular type dark-field STEM imaging modes. DPC STEM is also very effective for imaging magnetic field structures in magnetic materials, such as magnetic domains and skyrmions. Moreover, real-time imaging of electromagnetic field structures can

  14. Processing a Biological Tissue from Cryo-SEM to Replica

    Czech Academy of Sciences Publication Activity Database

    Vaškovicová, Naděžda; Hrubanová, Kamila; Krzyžánek, Vladislav

    2016-01-01

    Roč. 22, S3 (2016), s. 236-237 ISSN 1431-9276 R&D Projects: GA MŠk(CZ) LO1212; GA MŠk ED0017/01/01 Institutional support: RVO:68081731 Keywords : Cryo-SEM * ACE600 Subject RIV: JA - Electronics ; Optoelectronics, Electrical Engineering Impact factor: 1.891, year: 2016

  15. Electron optical characteristics of a concave electrostatic electron mirror for a scanning electron microscope

    International Nuclear Information System (INIS)

    Hamarat, R.T.; Witzani, J.; Hoerl, E.M.

    1984-08-01

    Numerical computer calculations are used to explore the design characteristics of a concave electrostatic electron mirror for a mirror attachment for a conventional scanning electron microscope or an instrument designed totally as a scanning electron mirror microscope. The electron paths of a number of set-ups are calculated and drawn graphically in order to find the optimum shape and dimensions of the mirror geometry. This optimum configuration turns out to be the transition configuration between two cases of electron path deflection, towards the optical axis of the system and away from it. (Author)

  16. High-resolution imaging of magnetic fields using scanning superconducting quantum interference device (SQUID) microscopy

    Science.gov (United States)

    Fong de Los Santos, Luis E.

    Development of a scanning superconducting quantum interference device (SQUID) microscope system with interchangeable sensor configurations for imaging magnetic fields of room-temperature (RT) samples with sub-millimeter resolution. The low-critical-temperature (Tc) niobium-based monolithic SQUID sensor is mounted in the tip of a sapphire rod and thermally anchored to the cryostat helium reservoir. A 25 mum sapphire window separates the vacuum space from the RT sample. A positioning mechanism allows adjusting the sample-to-sensor spacing from the top of the Dewar. I have achieved a sensor-to-sample spacing of 100 mum, which could be maintained for periods of up to 4 weeks. Different SQUID sensor configurations are necessary to achieve the best combination of spatial resolution and field sensitivity for a given magnetic source. For imaging thin sections of geological samples, I used a custom-designed monolithic low-Tc niobium bare SQUID sensor, with an effective diameter of 80 mum, and achieved a field sensitivity of 1.5 pT/Hz1/2 and a magnetic moment sensitivity of 5.4 x 10-18 Am2/Hz1/2 at a sensor-to-sample spacing of 100 mum in the white noise region for frequencies above 100 Hz. Imaging action currents in cardiac tissue requires higher field sensitivity, which can only be achieved by compromising spatial resolution. I developed a monolithic low-Tc niobium multiloop SQUID sensor, with sensor sizes ranging from 250 mum to 1 mm, and achieved sensitivities of 480 - 180 fT/Hz1/2 in the white noise region for frequencies above 100 Hz, respectively. For all sensor configurations, the spatial resolution was comparable to the effective diameter and limited by the sensor-to-sample spacing. Spatial registration allowed us to compare high-resolution images of magnetic fields associated with action currents and optical recordings of transmembrane potentials to study the bidomain nature of cardiac tissue or to match petrography to magnetic field maps in thin sections of

  17. High-resolution electron microscopy study of electron-irradiation-induced crystalline-to-amorphous transition in α-SiC single crystals

    International Nuclear Information System (INIS)

    Inui, H.; Mori, H.; Sakata, T.

    1992-01-01

    An electron-irradiation-induced crystalline-to-amorphous (CA) transition in α-SiC has been studied by high-resolution electron microscopy (HREM). The irradiation-produced damage structure was examined as a function of dose of electrons by taking high-resolution maps extending from the unirradiated crystalline region to the completely amorphized region. In the intermediate region between those two regions, that is in the CA transition region, the damage structure was essentially a mixture of crystalline and amorphous phases. The volume fraction of the amorphous phase was found to increase with increasing dose of electrons and no discrete crystalline-amorphous interface was observed in the CA transition region. These facts indicate the heterogeneous and gradual nature of the CA transition. In the transition region close to the unirradiated crystalline region, a sort of fragmentation of the crystal lattice was observed to occur, crystallites with slightly different orientations with respect to the parent crystal were formed owing to the strain around the dispersed local amorphous regions. In the transition region close to the amorphized region, these crystallites were reduced in size and were embedded in an amorphous matrix. This damage structure is the result of the increased volume fraction of the amorphous phase. In the completely amorphized region, no lattice fringes were recognized in the HREM images. The atomistic process of the CA transition is discussed on the basis of the present results and those from previous studies. (Author)

  18. CryoSat Processing Prototype, how to generate LRM like echoes with SAR data and a Comparison to DUACS SLA over high latitudes

    Science.gov (United States)

    Picot, N.; Boy, F.; Desjonqueres, J.

    2012-12-01

    Like CryoSat, Sentinel3 embarks a doppler altimeter. While there is a long experience of LRM processing, SAR nadir looking data are new and will need in depth validation. Thanks to CryoSat data, the processing of SAR data can be experienced in orbit. The continuity to current altimeter data set (based on LRM acquisitions) has also to be analysed with details. A Cryosat Processing Prototype (C2P) has been developed on CNES side to prepare the CNES SAR ocean retracking study. this prototype allows to process SAR data in order to generate LRM like echoes on ground. Those CryoSat ocean products are routinely processed on CNES side and ingested in the SALP/DUACS system. CryoSat data have proved to be very accurate and very valuable for the ocean user community in the past monthes. For example, it has allowed to largely reduce the impact of the lost of the ESA ENVISAT mission as well as the long non availability of Jason-1 data. This paper will describe the system set up in place early 2012 to feed CryoSat data in the SALP/DUACS products and will present the routine data analysis . C2P CryoSat products will be compared with DUACS SLA estimates and a specific focus will be given over high latitudes knowing that CryoSat is the oinly mission providing sea surface estimates over latitudes above 66 degrees since the lost of the ESA ENVISAT mission.

  19. Performance of ultrahigh resolution electron microscope JEM-4000EX and some applications of high Tc superconductors

    International Nuclear Information System (INIS)

    Honda, T.; Ibe, K.; Ishida, Y.; Kersker, M.M.

    1989-01-01

    The high resolution electron microscope is powerful for modern materials science because of its direct observation capability for the atomic structure of materials. the JEM-4000EX, a 400 kV accelerating voltage electron microscope whose objective lens has a 1 mm spherical aberration coefficient, has a 0.168 nm theoretical resolving power. Using this microscope, atomic structure images of high Tc superconductor such as Y-Ba-Cu-O, Bi-Ca-Sr-Cu-O and Tl-Ca-Ba-Cu-O are reported

  20. Soft x-ray scanning microtomography with submicron resolution

    International Nuclear Information System (INIS)

    McNulty, I.; Haddad, W.S.; Trebes, J.E.; Anderson, E.H.

    1994-01-01

    Scanning soft x-ray microtomography was used to obtain high-resolution three-dimensional images of a microfabricated test object. Using a special rotation stage mounted on the scanning transmission x-ray microscope at the XIA Beamline at the National Synchrotron Light Source, we recorded nine two-dimensional projections of the 3D test object over an angular range of -50 degrees to +55 degrees. The x-ray wavelength was 3.6 nm and the radiation dose to the object per projection was approximately 2 x 10 6 Gy. The object consisted of two gold patterns supported on transparent silicon nitride membranes, separated by 4.75 Jim, with 100 to 300-nm wide and 65-nm thick features. We reconstructed a volumetric data set of the test object from the two-dimensional projections using an algebraic reconstruction technique algorithm. Features of the test object were resolved to ∼100 nm in transverse and longitudinal extent in three-dimensional images rendered from the volumetric set

  1. Cryo-electron Microscopy Structures of Expanded Poliovirus with VHHs Sample the Conformational Repertoire of the Expanded State.

    Science.gov (United States)

    Strauss, Mike; Schotte, Lise; Karunatilaka, Krishanthi S; Filman, David J; Hogle, James M

    2017-02-01

    By using cryo-electron microscopy, expanded 80S-like poliovirus virions (poliovirions) were visualized in complexes with four 80S-specific camelid VHHs (Nanobodies). In all four complexes, the VHHs bind to a site on the top surface of the capsid protein VP3, which is hidden in the native virus. Interestingly, although the four VHHs bind to the same site, the structures of the expanded virus differ in detail in each complex, suggesting that each of the Nanobodies has sampled a range of low-energy structures available to the expanded virion. By stabilizing unique structures of expanded virions, VHH binding permitted a more detailed view of the virus structure than was previously possible, leading to a better understanding of the expansion process that is a critical step in infection. It is now clear which polypeptide chains become disordered and which become rearranged. The higher resolution of these structures also revealed well-ordered conformations for the EF loop of VP2, the GH loop of VP3, and the N-terminal extensions of VP1 and VP2, which, in retrospect, were present in lower-resolution structures but not recognized. These structural observations help to explain preexisting mutational data and provide insights into several other stages of the poliovirus life cycle, including the mechanism of receptor-triggered virus expansion. When poliovirus infects a cell, it undergoes a change in its structure in order to pass RNA through its protein coat, but this altered state is short-lived and thus poorly understood. The structures of poliovirus bound to single-domain antibodies presented here capture the altered virus in what appear to be intermediate states. A careful analysis of these structures lets us better understand the molecular mechanism of infection and how these changes in the virus lead to productive-infection events. Copyright © 2017 American Society for Microbiology.

  2. High mass-resolution electron-ion-ion coincidence measurements on core-excited organic molecules

    CERN Document Server

    Tokushima, T; Senba, Y; Yoshida, H; Hiraya, A

    2001-01-01

    Total electron-ion-ion coincidence measurements on core excited organic molecules have been carried out with high mass resolution by using multimode (reflectron/linear) time-of-flight mass analyzer. From the ion correlation spectra of core excited CH sub 3 OH and CD sub 3 OH, the reaction pathway to form H sub 3 sup + (D sub 3 sup +) is identified as the elimination of three H (D) atoms from the methyl group, not as the inter-group (-CH sub 3 and -OH) interactions. In a PEPIPICO spectrum of acetylacetone (CH sub 3 COCH sub 2 COCH sub 3) measured by using a reflectron TOF, correlations between ions up to mass number 70 with one-mass resolution was recorded.

  3. Stereoscopic and photometric surface reconstruction in scanning electron microscopy

    International Nuclear Information System (INIS)

    Scherer, S.

    2000-01-01

    The scanning electron microscope (SEM) is one of the most important devices to examine microscopic structures as it offers images of a high contrast range with a large depth of focus. Nevertheless, three-dimensional measurements, as desired in fracture mechanics, have previously not been accomplished. This work presents a system for automatic, robust and dense surface reconstruction in scanning electron microscopy combining new approaches in shape from stereo and shape from photometric stereo. The basic theoretical assumption for a known adaptive window algorithm is shown not to hold in scanning electron microscopy. A constraint derived from this observation yields a new, simplified, hence faster calculation of the adaptive window. The correlation measure itself is obtained by a new ordinal measure coefficient. Shape from photometric stereo in the SEM is formulated by relating the image formation process with conventional photography. An iterative photometric ratio reconstruction is invented based on photometric ratios of backscatter electron images. The performance of the proposed system is evaluated using ground truth data obtained by three alternative shape recovery devices. Most experiments showed relative height accuracy within the tolerances of the alternative devices. (author)

  4. Zernike phase contrast cryo-electron tomography of sodium-driven flagellar hook-basal bodies from Vibrio alginolyticus.

    Science.gov (United States)

    Hosogi, Naoki; Shigematsu, Hideki; Terashima, Hiroyuki; Homma, Michio; Nagayama, Kuniaki

    2011-01-01

    Vibrio alginolyticus use flagella to swim. A flagellum consists of a filament, hook and basal body. The basal body is made up of a rod and several ring structures. This study investigates the structure of the T ring which is a unique component of the V. alginolyticus sodium ion-driven flagellar basal body. Using Zernike phase contrast (ZPC) cryo-electron tomography, we compared the 3D structures of purified hook-basal bodies (HBB) from a wild-type strain (KK148) and a deletion mutant lacking MotX and MotY (TH3), which are thought to form the T ring. ZPC images of HBBs had highly improved signal-to-noise ratio compared to conventional phase contrast images. We observed the outline of the HBBs from strains KK148 and TH3, and the TH3 mutant was missing its T ring. In the wild-type strain, the T ring was beneath the LP ring and seemed to form a ring shape with diameter of 32 nm. Copyright © 2010 Elsevier Inc. All rights reserved.

  5. Amyloid Structure and Assembly: Insights from Scanning Transmission Electron Microscopy

    Energy Technology Data Exchange (ETDEWEB)

    Goldsbury, C.; Wall, J.; Baxa, U.; Simon, M. N.; Steven, A. C.; Engel, A.; Aebi, U.; Muller, S. A.

    2011-01-01

    Amyloid fibrils are filamentous protein aggregates implicated in several common diseases such as Alzheimer's disease and type II diabetes. Similar structures are also the molecular principle of the infectious spongiform encephalopathies such as Creutzfeldt-Jakob disease in humans, scrapie in sheep, and of the so-called yeast prions, inherited non-chromosomal elements found in yeast and fungi. Scanning transmission electron microscopy (STEM) is often used to delineate the assembly mechanism and structural properties of amyloid aggregates. In this review we consider specifically contributions and limitations of STEM for the investigation of amyloid assembly pathways, fibril polymorphisms and structural models of amyloid fibrils. This type of microscopy provides the only method to directly measure the mass-per-length (MPL) of individual filaments. Made on both in vitro assembled and ex vivo samples, STEM mass measurements have illuminated the hierarchical relationships between amyloid fibrils and revealed that polymorphic fibrils and various globular oligomers can assemble simultaneously from a single polypeptide. The MPLs also impose strong constraints on possible packing schemes, assisting in molecular model building when combined with high-resolution methods like solid-state nuclear magnetic resonance (NMR) and electron paramagnetic resonance (EPR).

  6. Pathogen–host reorganization during Chlamydia invasion revealed by cryo-electron tomography

    Science.gov (United States)

    Nans, Andrea; Saibil, Helen R; Hayward, Richard D

    2014-01-01

    Invasion of host cells is a key early event during bacterial infection, but the underlying pathogen–host interactions are yet to be fully visualized in three-dimensional detail. We have captured snapshots of the early stages of bacterial-mediated endocytosis in situ by exploiting the small size of chlamydial elementary bodies (EBs) for whole-cell cryo-electron tomography. Chlamydiae are obligate intracellular bacteria that infect eukaryotic cells and cause sexually transmitted infections and trachoma, the leading cause of preventable blindness. We demonstrate that Chlamydia trachomatis LGV2 EBs are intrinsically polarized. One pole is characterized by a tubular inner membrane invagination, while the other exhibits asymmetric periplasmic expansion to accommodate an array of type III secretion systems (T3SSs). Strikingly, EBs orient with their T3SS-containing pole facing target cells, enabling the T3SSs to directly contact the cellular plasma membrane. This contact induces enveloping macropinosomes, actin-rich filopodia and phagocytic cups to zipper tightly around the internalizing bacteria. Once encapsulated into tight early vacuoles, EB polarity and the T3SSs are lost. Our findings reveal previously undescribed structural transitions in both pathogen and host during the initial steps of chlamydial invasion. PMID:24809274

  7. Resolução lamelar num novo microscópio eletrônico de varredura Lamellar resolution in a new scanning electron microscope

    Directory of Open Access Journals (Sweden)

    Hans-Jürgen Kestenbach

    1997-03-01

    Full Text Available RESUMO: Trabalhando com elétrons de baixa energia (na faixa de 1keV, o novo microscópio eletrônico de varredura dispensa a etapa de metalização e permite a observação direta da estrutura lamelar de polímeros semicristalinos, sem a necessidade de preparação de amostras. São apresentados exemplos da morfologia lamelar do PVDF em função das condições de processamento e da temperatura de cristalização, em filmes contendo as fases a, b e g. Um outro exemplo revela o crescimento inicial da camada transcristalina que se formou ao longo de uma fibra de polietileno de ultra-alto peso molecular embutida em matriz de polietileno de alta densidade.ABSTRACT: Working with low energy electrons (in the range of 1keV, the new scanning electron microscope permits the lamellar (supermolecular structure of semicrystalline polymers to be observed directly without the need of specimen coating or of any other sample preparation technique. Microscope performance is demonstrated by several examples of high resolution micrographs which show spherulitic, lamellar and fibrilar morphologies developed by the a, b and g phases of PVDF as a function of processing conditions and crystallization temperature. Another example reveals the early stages of transcrystalline layer formation in HDPE reinforced by UHMWPE fibers.

  8. Electron beam fabrication and characterization of high- resolution magnetic force microscopy tips

    NARCIS (Netherlands)

    Ruhrig, M.; Rührig, M.; Porthun, S.; Porthun, S.; Lodder, J.C.; Mc vitie, S.; Heyderman, L.J.; Johnston, A.B.; Chapman, J.N.

    1996-01-01

    The stray field, magnetic microstructure, and switching behavior of high‐resolution electron beam fabricated thin film tips for magnetic force microscopy (MFM) are investigated with different imaging modes in a transmission electron microscope (TEM). As the tiny smooth carbon needles covered with a

  9. Localization of fluorescently labeled structures in frozen-hydrated samples using integrated light electron microscopy.

    Science.gov (United States)

    Faas, F G A; Bárcena, M; Agronskaia, A V; Gerritsen, H C; Moscicka, K B; Diebolder, C A; van Driel, L F; Limpens, R W A L; Bos, E; Ravelli, R B G; Koning, R I; Koster, A J

    2013-03-01

    Correlative light and electron microscopy is an increasingly popular technique to study complex biological systems at various levels of resolution. Fluorescence microscopy can be employed to scan large areas to localize regions of interest which are then analyzed by electron microscopy to obtain morphological and structural information from a selected field of view at nm-scale resolution. Previously, an integrated approach to room temperature correlative microscopy was described. Combined use of light and electron microscopy within one instrument greatly simplifies sample handling, avoids cumbersome experimental overheads, simplifies navigation between the two modalities, and improves the success rate of image correlation. Here, an integrated approach for correlative microscopy under cryogenic conditions is presented. Its advantages over the room temperature approach include safeguarding the native hydrated state of the biological specimen, preservation of the fluorescence signal without risk of quenching due to heavy atom stains, and reduced photo bleaching. The potential of cryo integrated light and electron microscopy is demonstrated for the detection of viable bacteria, the study of in vitro polymerized microtubules, the localization of mitochondria in mouse embryonic fibroblasts, and for a search into virus-induced intracellular membrane modifications within mammalian cells. Copyright © 2012 Elsevier Inc. All rights reserved.

  10. CryoSat-2 Validation using CryoVEX 2011-12 Airborne Campaigns

    DEFF Research Database (Denmark)

    Skourup, Henriette; Forsberg, René; Kildegaard Rose, Stine

    Sat-2 by comparison to airborne and ground measurements. This is possible only through a major effort involving a large group of international partners. DTU Space has been involved in the CryoVEx campaigns with airborne activities since 2003. To validate the performance of the CryoSat-2 radar altimeter...... (SIRAL), the aircraft is equipped with an airborne version of the SIRAL altimeter (ASIRAS) together with a laser scanner. The campaigns are focused on five main validation sites: Devon ice cap (Canada), Austfonna ice cap (Svalbard), the EGIG line crossing the Greenland Ice Sheet, as well as the sea ice...... north of Alert and sea ice around Svalbard in the Fram Strait. Selected tracks were planned to match CryoSat-2 passes and a few of them were flown in formation flight with the Alfred Wegener Institute (AWI) Polar-5 carrying an EM-bird. This presentation summarizes the 2011-12 airborne campaigns...

  11. A graphene oxide-carbon nanotube grid for high-resolution transmission electron microscopy of nanomaterials

    International Nuclear Information System (INIS)

    Zhang Lina; Zhang Haoxu; Zhou Ruifeng; Chen Zhuo; Li Qunqing; Fan Shoushan; Jiang Kaili; Ge Guanglu; Liu Renxiao

    2011-01-01

    A novel grid for use in transmission electron microscopy is developed. The supporting film of the grid is composed of thin graphene oxide films overlying a super-aligned carbon nanotube network. The composite film combines the advantages of graphene oxide and carbon nanotube networks and has the following properties: it is ultra-thin, it has a large flat and smooth effective supporting area with a homogeneous amorphous appearance, high stability, and good conductivity. The graphene oxide-carbon nanotube grid has a distinct advantage when characterizing the fine structure of a mass of nanomaterials over conventional amorphous carbon grids. Clear high-resolution transmission electron microscopy images of various nanomaterials are obtained easily using the new grids.

  12. Diversity of extracellular vesicles in human ejaculates revealed by cryo-electron microscopy

    Directory of Open Access Journals (Sweden)

    Johanna L. Höög

    2015-11-01

    Full Text Available Human ejaculates contain extracellular vesicles (EVs, that to a large extent are considered to originate from the prostate gland, and are often denominated “prostasomes.” These EVs are important for human fertility, for example by promoting sperm motility and by inducing immune tolerance of the female immune system to the spermatozoa. So far, the EVs present in human ejaculate have not been studied in their native state, inside the seminal fluid without prior purification and isolation procedures. Using cryo-electron microscopy and tomography, we performed a comprehensive inventory of human ejaculate EVs. The sample was neither centrifuged, fixed, filtered or sectioned, nor were heavy metals added. Approximately 1,500 extracellular structures were imaged and categorized. The extracellular environment of human ejaculate was found to be diverse, with 5 major subcategories of EVs and 6 subcategories of extracellular membrane compartments, including lamellar bodies. Furthermore, 3 morphological features, including electron density, double membrane bilayers and coated surface, are described in all subcategories. This study reveals that the extracellular environment in human ejaculate is multifaceted. Several novel morphological EV subcategories are identified and clues to their cellular origin may be found in their morphology. This inventory is therefore important for developing future experimental approaches, and to interpret previously published data to understand the role of EVs for human male fertility.

  13. Mechanical design of a high-resolution x-ray powder diffractometer at the Advanced Photon Source.

    Energy Technology Data Exchange (ETDEWEB)

    Shu, D.; Lee, P.; Preissner, C.; Ramanathan, M.; Beno, M.; VonDreele, R.; Ranay, R.; Ribaud, L.; Kurtz, C.; Jiao, X.; Kline, D.; Jemian, P.; Toby, B.

    2007-01-01

    A novel high-resolution x-ray powder diffractometer has been designed and commissioned at the bending magnet beamline 11-BM at the Advanced Photon Source (APS), Argonne National Laboratory (ANL). This state-of-the-art instrument is designed to meet challenging mechanical and optical specifications for producing high-quality powder diffraction data with high throughput. The 2600 mm (H) X 2100 mm (L) X 1700 mm (W) diffractometer consists of five subassemblies: a customized two-circle goniometer with a 3-D adjustable supporting base; a twelve-channel high-resolution crystal analyzer system with an array of precision x-ray slits; a manipulator system for a twelve scintillator x-ray detectors; a 4-D sample manipulator with cryo-cooling capability; and a robot-based sample exchange automation system. The mechanical design of the diffractometer as well as the test results of its positioning performance are presented in this paper.

  14. Mechanical design of a high-resolution x-ray powder diffractometer at the Advanced Photon Source

    International Nuclear Information System (INIS)

    Shu, D.; Lee, P.; Preissner, C.; Ramanathan, M.; Beno, M.; VonDreele, R.; Ranay, R.; Ribaud, L.; Kurtz, C.; Jiao, X.; Kline, D.; Jemian, P.; Toby, B.

    2007-01-01

    A novel high-resolution x-ray powder diffractometer has been designed and commissioned at the bending magnet beamline 11-BM at the Advanced Photon Source (APS), Argonne National Laboratory (ANL). This state-of-the-art instrument is designed to meet challenging mechanical and optical specifications for producing high-quality powder diffraction data with high throughput. The 2600 mm (H) X 2100 mm (L) X 1700 mm (W) diffractometer consists of five subassemblies: a customized two-circle goniometer with a 3-D adjustable supporting base; a twelve-channel high-resolution crystal analyzer system with an array of precision x-ray slits; a manipulator system for a twelve scintillator x-ray detectors; a 4-D sample manipulator with cryo-cooling capability; and a robot-based sample exchange automation system. The mechanical design of the diffractometer as well as the test results of its positioning performance are presented in this paper.

  15. High-resolution spatiotemporal strain mapping reveals non-uniform deformation in micropatterned elastomers

    Science.gov (United States)

    Aksoy, B.; Rehman, A.; Bayraktar, H.; Alaca, B. E.

    2017-04-01

    Micropatterns are generated on a vast selection of polymeric substrates for various applications ranging from stretchable electronics to cellular mechanobiological systems. When these patterned substrates are exposed to external loading, strain field is primarily affected by the presence of microfabricated structures and similarly by fabrication-related defects. The capturing of such nonhomogeneous strain fields is of utmost importance in cases where study of the mechanical behavior with a high spatial resolution is necessary. Image-based non-contact strain measurement techniques are favorable and have recently been extended to scanning tunneling microscope and scanning electron microscope images for the characterization of mechanical properties of metallic materials, e.g. steel and aluminum, at the microscale. A similar real-time analysis of strain heterogeneity in elastomers is yet to be achieved during the entire loading sequence. The available measurement methods for polymeric materials mostly depend on cross-head displacement or precalibrated strain values. Thus, they suffer either from the lack of any real-time analysis, spatiotemporal distribution or high resolution in addition to a combination of these factors. In this work, these challenges are addressed by integrating a tensile stretcher with an inverted optical microscope and developing a subpixel particle tracking algorithm. As a proof of concept, the patterns with a critical dimension of 200 µm are generated on polydimethylsiloxane substrates and strain distribution in the vicinity of the patterns is captured with a high spatiotemporal resolution. In the field of strain measurement, there is always a tradeoff between minimum measurable strain value and spatial resolution. Current noncontact techniques on elastomers can deliver a strain resolution of 0.001% over a minimum length of 5 cm. More importantly, inhomogeneities within this quite large region cannot be captured. The proposed technique can

  16. High-resolution spatiotemporal strain mapping reveals non-uniform deformation in micropatterned elastomers

    International Nuclear Information System (INIS)

    Aksoy, B; Alaca, B E; Rehman, A; Bayraktar, H

    2017-01-01

    Micropatterns are generated on a vast selection of polymeric substrates for various applications ranging from stretchable electronics to cellular mechanobiological systems. When these patterned substrates are exposed to external loading, strain field is primarily affected by the presence of microfabricated structures and similarly by fabrication-related defects. The capturing of such nonhomogeneous strain fields is of utmost importance in cases where study of the mechanical behavior with a high spatial resolution is necessary. Image-based non-contact strain measurement techniques are favorable and have recently been extended to scanning tunneling microscope and scanning electron microscope images for the characterization of mechanical properties of metallic materials, e.g. steel and aluminum, at the microscale. A similar real-time analysis of strain heterogeneity in elastomers is yet to be achieved during the entire loading sequence. The available measurement methods for polymeric materials mostly depend on cross-head displacement or precalibrated strain values. Thus, they suffer either from the lack of any real-time analysis, spatiotemporal distribution or high resolution in addition to a combination of these factors. In this work, these challenges are addressed by integrating a tensile stretcher with an inverted optical microscope and developing a subpixel particle tracking algorithm. As a proof of concept, the patterns with a critical dimension of 200 µ m are generated on polydimethylsiloxane substrates and strain distribution in the vicinity of the patterns is captured with a high spatiotemporal resolution. In the field of strain measurement, there is always a tradeoff between minimum measurable strain value and spatial resolution. Current noncontact techniques on elastomers can deliver a strain resolution of 0.001% over a minimum length of 5 cm. More importantly, inhomogeneities within this quite large region cannot be captured. The proposed technique can

  17. Benchmarking NaI(Tl) Electron Energy Resolution Measurements

    International Nuclear Information System (INIS)

    Mengesha, Wondwosen; Valentine, J D.

    2002-01-01

    A technique for validating electron energy resolution results measured using the modified Compton coincidence technique (MCCT) has been developed. This technique relies on comparing measured gamma-ray energy resolution with calculated values that were determined using the measured electron energy resolution results. These gamma-ray energy resolution calculations were based on Monte Carlo photon transport simulations, the measured NaI(Tl) electron response, a simplified cascade sequence, and the measured electron energy resolution results. To demonstrate this technique, MCCT-measured NaI(Tl) electron energy resolution results were used along with measured gamma-ray energy resolution results from the same NaI(Tl) crystal. Agreement to within 5% was observed for all energies considered between the calculated and measured gamma-ray energy resolution results for the NaI(Tl) crystal characterized. The calculated gamma-ray energy resolution results were also compared with previously published gamma-ray energy resolution measurements with good agreement (<10%). In addition to describing the validation technique that was developed in this study and the results, a brief review of the electron energy resolution measurements made using the MCCT is provided. Based on the results of this study, it is believed that the MCCT-measured electron energy resolution results are reliable. Thus, the MCCT and this validation technique can be used in the future to characterize the electron energy resolution of other scintillators and to determine NaI(Tl) intrinsic energy resolution

  18. Pseudo LRM waveforms from CryoSat SARin acquisition

    Science.gov (United States)

    Scagliola, Michele; Fornari, Marco; Bouffard, Jerome; Parrinello, Tommaso; Féménias, Pierre

    2016-04-01

    CryoSat was launched on the 8th April 2010 and is the first European ice mission dedicated to the monitoring of precise changes in the thickness of polar ice sheets and floating sea ice. The main payload of CryoSat is a Ku-band pulsewidth limited radar altimeter, called SIRAL (Synthetic interferometric radar altimeter). When commanded in SARIn (synthetic aperture radar interferometry) mode, through coherent along-track processing of the returns received from two antennas, the interferometric phase related to the first arrival of the echo is used to retrieve the angle of arrival of the scattering in the across-track direction. When SIRAL operates in SAR or SARin mode, the obtained waveforms have an along-track resolution and a speckle reduction which is increased with respect to the pulse-limited waveforms. Anyway, in order to analyze the continuity of the geophysical retrieved parameters among different acquisition modes, techniques to transform SARin mode data to pseudo-LRM mode data are welcome. The transformation process is known as SAR reduction and it is worth recalling here that only approximate pseudo-LRM waveforms can be obtained in case of closed burst acquisitions, as SIRAL operates. A SAR reduction processing scheme has been developed to obtain pseudo-LRM waveforms from CryoSat SARin acquisition. As a trade-off between the along-track length on Earth surface contributing to one SARin pseudo-LRM waveform and the noisiness of the waveform itself, it has been chosen a SAR reduction approach based on the averaging of all the SARin echoes received each 20Hz, resulting in one pseudo-LRM waveform for each SARin burst given the SARin burst repetition period. SARin pseudo-LRM waveforms have been produced for CryoSat acquisition both on ice and sea surfaces, aiming at verifying the continuity of the retracked surface height over the ellipsoid between genuine LRM products and pseudo-LRM products. Moreover, the retracked height from the SARin pseudo-LRM has been

  19. Electron microscopy at atomic resolution

    Energy Technology Data Exchange (ETDEWEB)

    Gronsky, R.

    1983-11-01

    The direct imaging of atomic structure in solids has become increasingly easier to accomplish with modern transmission electron microscopes, many of which have an information retrieval limit near 0.2 nm point resolution. Achieving better resolution, particularly with any useful range of specimen tilting, requires a major design effort. This presentation describes the new Atomic Resolution Microscope (ARM), recently put into operation at the Lawrence Berkeley Laboratory. Capable of 0.18 nm or better interpretable resolution over a voltage range of 400 kV to 1000 kV with +- 40/sup 0/ biaxial specimen tilting, the ARM features a number of new electron-optical and microprocessor-control designs. These are highlighted, and its atomic resolution performance demonstrated for a selection of inorganic crystals.

  20. Electron microscopy at atomic resolution

    International Nuclear Information System (INIS)

    Gronsky, R.

    1983-11-01

    The direct imaging of atomic structure in solids has become increasingly easier to accomplish with modern transmission electron microscopes, many of which have an information retrieval limit near 0.2 nm point resolution. Achieving better resolution, particularly with any useful range of specimen tilting, requires a major design effort. This presentation describes the new Atomic Resolution Microscope (ARM), recently put into operation at the Lawrence Berkeley Laboratory. Capable of 0.18 nm or better interpretable resolution over a voltage range of 400 kV to 1000 kV with +- 40 0 biaxial specimen tilting, the ARM features a number of new electron-optical and microprocessor-control designs. These are highlighted, and its atomic resolution performance demonstrated for a selection of inorganic crystals

  1. Electron Beam Scanning in Industrial Applications

    Science.gov (United States)

    Jongen, Yves; Herer, Arnold

    1996-05-01

    Scanned electron beams are used within many industries for applications such as sterilization of medical disposables, crosslinking of wire and cables insulating jackets, polymerization and degradation of resins and biomaterials, modification of semiconductors, coloration of gemstones and glasses, removal of oxides from coal plant flue gasses, and the curing of advanced composites and other molded forms. X-rays generated from scanned electron beams make yet other applications, such as food irradiation, viable. Typical accelerators for these applications range in beam energy from 0.5MeV to 10 MeV, with beam powers between 5 to 500kW and scanning widths between 20 and 300 cm. Since precise control of dose delivery is required in many of these applications, the integration of beam characteristics, product conveyance, and beam scanning mechanisms must be well understood and optimized. Fundamental issues and some case examples are presented.

  2. Improving the Coastal Marine Gravity from CryoSat-2 Altimetry

    Science.gov (United States)

    Abulaitijiang, A.; Andersen, O. B.; Knudsen, P.

    2017-12-01

    The 7 years of CryoSat-2 satellite altimetry data can be potentially used to extract the high frequency components of the Earth gravity field beyond the Global Geopotential Models (GGMs) which corresponds to a resolution of 9.2 Km at the degree 2160. The Synthetic Aperture Radar (SAR) mode of the CryoSat-2 produced high precision along track observations and pushed the limits (qualified observations) to even closer to the coast by several kilometers. The conventional FFT method in deriving the marine gravity anomalies requires the input height anomalies to be gridded, and gridding in the irregular coastal zones (land-ocean transition zone) could introduce high frequency noise to the inversion. Therefore, Least Square Collocation (LSC) is preferred for the inversion. As part of a phd project, in this work, we will make use of the covariance function of the height anomalies in the coastal zones to derive the marine gravity anomalies. Using the conventional remove-compute-restore (only considering the GGMs) technique, the theoretical assumption of homogeneity and isotropy in the LSC algorithm is not always satisfied in the coastal regions and mountainous regions. Hence, the Topographic Correction (TC) using high resolution topographic grids is a critical step in the reduction of the gravity functionals (e.g., height anomaly and gravity anomaly), to comply with the theoretical assumption of LSC. In this work, the TC computation (both w.r.t. the height anomalies and gravity) will be conducted to test the performance of the signal reduction in several regions (patches) around Mediterranean, Chile, islands of Indonesia and Australian coast where the true gravity data is available. The derived marine gravity will be cross-validated against the ship-borne gravity observations. Earlier studies show that due to the presence of additional altimetry observations, 4 mGal accuracy can be achieved in the Greenland fjords, and equally good accuracy is expected from this work.

  3. Resolution Enhancement of Scanning Laser Acoustic Microscope Using Transverse Wave

    International Nuclear Information System (INIS)

    Ko, D. S.; Park, J. S.; Kim, Y. H.

    1997-01-01

    We studied the resolution enhancement of a novel scanning laser acoustic microscope (SLAM) using transverse waves. Mode conversion of the ultrasonic wave takes place at the liquid-solid interface and some energy of the insonifying longitudinal waves in the water will convert to transverse wave energy within the solid specimen. The resolution of SLAM depends on the size of detecting laser spot and the wavelength of the insonifying ultrasonic waves. Science the wavelength of the transverse wave is shorter than that of the longitudinal wave, we are able to achieve the high resolution by using transverse waves. In order to operate SLAM in the transverse wave mode, we made wedge for changing the incident angle. Our experimental results with model 2140 SLAM and an aluminum specimen showed higher contrast of the SLAM image in the transverse wave mode than that in the longitudinal wave mode

  4. State of the art in atomic resolution off-axis electron holography

    International Nuclear Information System (INIS)

    Linck, Martin; Freitag, Bert; Kujawa, Stephan; Lehmann, Michael; Niermann, Tore

    2012-01-01

    As proposed by Hannes Lichte, to resolve structure–property relations not only the question “Which atom is where?” but also the question “Which fields are around?” has to be answered. High-resolution off-axis electron holography opens up an access to these key questions in that it allows accessing the complete exit-wave of the object provided within the information limit of the microscope, i.e. amplitude and phase including atomic details such as position and species, and moreover, information about large area electric potentials and magnetic fields, which a conventional transmission electron microscope is blind for—also when using a Cs-corrector. For an excellent object exit-wave reconstruction, special care has to be taken on the hologram quality, i.e. interference fringe contrast and electron dose. Severe restrictions are given to signal resolution by the limited brightness of the electron source. Utilizing a new high-brightness Schottky field electron emitter in a state-of-the-art transmission electron microscope operated at 300 kV, the phase signal resolution at atomic resolution can significantly be enhanced. An improvement by at least a factor of 2.88 compared to the most recently reported single hologram at atomic resolution is found. To proof the applicability of this setup to real materials science problems, a grain boundary of gold has been investigated holographically. -- Highlights: ► Impact of the brightness on the reconstructed signal in electron holography. ► Factor 2.8 gain in signal quality by setup with a high brightness electron gun. ► Investigation of a grain boundary in gold with a state-of-the-art holography setup. ► A-posteriori aberration fine-tuning for true one Angstrom resolution in the object wave. ► Mistilt analysis on the atomic scale by numerical wave optics.

  5. Nanometer-resolution electron microscopy through micrometers-thick water layers

    Energy Technology Data Exchange (ETDEWEB)

    Jonge, Niels de, E-mail: niels.de.jonge@vanderbilt.edu [Vanderbilt University Medical Center, Department of Molecular Physiology and Biophysics, Nashville, TN 37232-0615 (United States); Oak Ridge National Laboratory, Materials Science and Technology Division, Oak Ridge, TN 37831-6064 (United States); Poirier-Demers, Nicolas; Demers, Hendrix [Universite de Sherbrooke, Electrical and Computer Engineering, Sherbrooke, Quebec J1K 2R1 (Canada); Peckys, Diana B. [Oak Ridge National Laboratory, Materials Science and Technology Division, Oak Ridge, TN 37831-6064 (United States); University of Tennessee, Center for Environmental Biotechnology, Knoxville, TN 37996-1605 (United States); Drouin, Dominique [Universite de Sherbrooke, Electrical and Computer Engineering, Sherbrooke, Quebec J1K 2R1 (Canada)

    2010-08-15

    Scanning transmission electron microscopy (STEM) was used to image gold nanoparticles on top of and below saline water layers of several micrometers thickness. The smallest gold nanoparticles studied had diameters of 1.4 nm and were visible for a liquid thickness of up to 3.3 {mu}m. The imaging of gold nanoparticles below several micrometers of liquid was limited by broadening of the electron probe caused by scattering of the electron beam in the liquid. The experimental data corresponded to analytical models of the resolution and of the electron probe broadening as function of the liquid thickness. The results were also compared with Monte Carlo simulations of the STEM imaging on modeled specimens of similar geometry and composition as used for the experiments. Applications of STEM imaging in liquid can be found in cell biology, e.g., to study tagged proteins in whole eukaryotic cells in liquid and in materials science to study the interaction of solid:liquid interfaces at the nanoscale.

  6. Special raster scanning for reduction of charging effects in scanning electron microscopy.

    Science.gov (United States)

    Suzuki, Kazuhiko; Oho, Eisaku

    2014-01-01

    A special raster scanning (SRS) method for reduction of charging effects is developed for the field of SEM. Both a conventional fast scan (horizontal direction) and an unusual scan (vertical direction) are adopted for acquiring raw data consisting of many sub-images. These data are converted to a proper SEM image using digital image processing techniques. About sharpness of the image and reduction of charging effects, the SRS is compared with the conventional fast scan (with frame-averaging) and the conventional slow scan. Experimental results show the effectiveness of SRS images. By a successful combination of the proposed scanning method and low accelerating voltage (LV)-SEMs, it is expected that higher-quality SEM images can be more easily acquired by the considerable reduction of charging effects, while maintaining the resolution. © 2013 Wiley Periodicals, Inc.

  7. High resolution electron microscopy of a small crack at the superficial layer of enamel.

    Science.gov (United States)

    Hayashi, Y

    1994-12-01

    A small enamel crack was investigated using a high resolution electron microscope. The inside of the crack was filled with aggregates of irregularly oriented plate-like crystals. Amorphous mineral deposits were observed among these aggregates at a low magnification. Selected area electron diffractions indicated that the plate-like crystals consisted of hydroxyapatite (OH-AP), and that the amorphous mineral deposits were a mixture of OH-AP and whitlockite. These findings indicate that this crack may have been formed by occlusal and/or masticatory stress, and that a natural occlusion might occur through mineral deposition at the small crack such as in this case.

  8. Electron Energy Resolution of the ATLAS TILECAL Modules with Flat Filter Method (July 2002 test beam)

    CERN Document Server

    Kulchitskii, Yu A; Vinogradov, V B

    2005-01-01

    The constructed ATLAS detector at the LHC will have the great physics discovery potential, in particular in the detection of a heavy Higgs boson. Calorimeters will play a crucial role in it. It is necessary to have confidence that the calorimeters will perform as expected. With the aim of understanding of performance of the ATLAS Tile hadronic calorimeter to electrons 12\\% of modules have been exposed in electron beams with various energies by three possible ways: cell-scan at $\\theta =20^o$ at the centers of the front face cells, $\\eta$-scan and tilerow scan at $\\theta = 90^o$ for the module side cells. We have extracted the electron energy resolutions of the $EBM-$ (ANL-44), $EBM+$ (IFA-42) and $BM$ (JINR-55) Modules of the ATLAS Tile Calorimeter at energies E = 10, 20, 50, 100 and 180 GeV and $\\theta = 20^o$ and $90^o $ and $\\eta$ scan from the July 2002 testbeam run data using the flat filter method of the PMT signal reconstruction. We have determined the statistical and constant terms for the electron en...

  9. Electron Energy Resolution of the ATLAS TILECAL Modules with Fit Filter Method (July 2002 test beam)

    CERN Document Server

    Kulchitskii, Yu A; Vinogradov, V B

    2006-01-01

    The constructed ATLAS detector at the LHC will have the great physics discovery potential, in particular in the detection of a heavy Higgs boson. Calorimeters will play a crucial role in it. It is necessary to have confidence that the calorimeters will perform as expected. With the aim of understanding of performance of the ATLAS Tile hadronic calorimeter to electrons 12\\% of modules have been exposed in electron beams with various energies by three possible ways: cell-scan at $\\theta =20^o$ at the centers of the front face cells, $\\eta$-scan and tilerow scan at $\\theta = 90^o$ for the module side cells. We have extracted the electron energy resolutions of the $EBM-$ (ANL-44), $EBM+$ (IFA-42) and $BM$ (JINR-55) Modules of the ATLAS Tile Calorimeter at energies E = 10, 20, 50, 100 and 180 GeV and $\\theta = 20^o$ and $90^o $ and $\\eta$ scan from the July 2002 testbeam run data using the fit filter method of the PMT signal reconstruction. We have determined the statistical and constant terms for the electron ene...

  10. The mechanism of PTFE and PE friction deposition: a combined scanning electron and scanning force microscopy study on highly oriented polymeric sliders

    NARCIS (Netherlands)

    Schönherr, Holger; Schaeben, H.; Vancso, Gyula J.

    1998-01-01

    The mechanism of friction deposition of polytetrafluoroethylene (PTFE) and polyethylene (PE) was studied by scanning electron (SEM) and scanning force microscopy (SFM) on the worn surfaces of PTFE and PE sliders that were used in friction deposition on glass substrates. These surfaces exhibited a

  11. Ultra-high resolution protein crystallography

    International Nuclear Information System (INIS)

    Takeda, Kazuki; Hirano, Yu; Miki, Kunio

    2010-01-01

    Many protein structures have been determined by X-ray crystallography and deposited with the Protein Data Bank. However, these structures at usual resolution (1.5< d<3.0 A) are insufficient in their precision and quantity for elucidating the molecular mechanism of protein functions directly from structural information. Several studies at ultra-high resolution (d<0.8 A) have been performed with synchrotron radiation in the last decade. The highest resolution of the protein crystals was achieved at 0.54 A resolution for a small protein, crambin. In such high resolution crystals, almost all of hydrogen atoms of proteins and some hydrogen atoms of bound water molecules are experimentally observed. In addition, outer-shell electrons of proteins can be analyzed by the multipole refinement procedure. However, the influence of X-rays should be precisely estimated in order to derive meaningful information from the crystallographic results. In this review, we summarize refinement procedures, current status and perspectives for ultra high resolution protein crystallography. (author)

  12. CryoSat-2 SIRAL Calibration and Performance

    Science.gov (United States)

    Fornari, M.; Scagliola, M.; Tagliani, N.; Parrinello, T.

    2012-12-01

    The main payload of CryoSat-2 is a Ku band pulse-width limited radar altimeter, called SIRAL (Synthetic interferometric radar altimeter), that transmits pulses at a high pulse repetition frequency thus making the received echoes phase coherent and suitable for azimuth processing. This allows to reach an along track resolution of about 250 meters which is a significant improvement over traditional pulse-width limited altimeters. Due to the fact that SIRAL is a phase coherent pulse-width limited radar altimeter, a proper calibration approach has been developed, including both an internal and external calibration. The internal calibration monitors the instrument impulse response and the transfer function, like traditional altimeters. In addition to that, the interferometer requires a special calibration developed ad hoc for SIRAL. The external calibration is performed with the use of a ground transponder, located in Svalbard, which receives SIRAL signal and sends the echo back to the satellite. Internal calibration data are processed on ground by the CryoSat-2 Instrument Processing Facility (IPF1) and then applied to the science data. In December 2012, two and a half years of calibration data will be available, which will be shown in this poster. The external calibration (transponder) data are processed and analyzed independently from the operational chain. The use of an external transponder has been very useful to determine instrument performance and for the tuning of the on-ground processor. This poster presents the transponder results in terms of range noise and datation error.

  13. The Development of a Scanning Soft X-Ray Microscope.

    Science.gov (United States)

    Rarback, Harvey Miles

    We have developed a scanning soft X-ray microscope, which can be used to image natural biological specimens at high resolution and with less damage than electron microscopy. The microscope focuses a monochromatic beam of synchrotron radiation to a nearly diffraction limited spot with the aid of a high resolution Fresnel zone plate, specially fabricated for us at the IBM Watson Research Center. The specimen at one atmosphere is mechanically scanned through the spot and the transmitted radiation is efficiently detected with a flow proportional counter. A computer forms a realtime transmission image of the specimen which is displayed on a color monitor. Our first generation optics have produced images of natural wet specimens at a resolution of 300 nm.

  14. All-inkjet-printed flexible electronics fabrication on a polymer substrate by low-temperature high-resolution selective laser sintering of metal nanoparticles

    International Nuclear Information System (INIS)

    Ko, Seung H; Pan Heng; Grigoropoulos, Costas P; Luscombe, Christine K; Frechet, Jean M J; Poulikakos, Dimos

    2007-01-01

    All-printed electronics is the key technology to ultra-low-cost, large-area electronics. As a critical step in this direction, we demonstrate that laser sintering of inkjet-printed metal nanoparticles enables low-temperature metal deposition as well as high-resolution patterning to overcome the resolution limitation of the current inkjet direct writing processes. To demonstrate this process combined with the implementation of air-stable carboxylate-functionalized polythiophenes, high-resolution organic transistors were fabricated in ambient pressure and room temperature without utilizing any photolithographic steps or requiring a vacuum deposition process. Local thermal control of the laser sintering process could minimize the heat-affected zone and the thermal damage to the substrate and further enhance the resolution of the process. This local nanoparticle deposition and energy coupling enable an environmentally friendly and cost-effective process as well as a low-temperature manufacturing sequence to realize large-area, flexible electronics on polymer substrates

  15. Real-Space Imaging of Carrier Dynamics of Materials Surfaces by Second-Generation Four-Dimensional Scanning Ultrafast Electron Microscopy

    KAUST Repository

    Sun, Jingya; Melnikov, Vasily; Khan, Jafar Iqbal; Mohammed, Omar F.

    2015-01-01

    , we establish a second generation of four-dimensional scanning ultrafast electron microscopy (4D S-UEM) and demonstrate the ability to record time-resolved images (snapshots) of material surfaces with 650 fs and ∼5 nm temporal and spatial resolutions

  16. CryoVex 2008

    DEFF Research Database (Denmark)

    Hvidegaard, Sine Munk; Forsberg, René; Skourup, Henriette

    2009-01-01

    Air Greenland. The main purpose was to collect coincident ASIRAS and laser elevation data form validation sites on land and sea ice and in addition offer logistical support to ground teams. Overflights of corner reflectors were done at main validation sites in order to calibrate the ASIRAS data....... The datasets from this campaign will be important for understanding the CryoSat-2 radar signals. The airborne part of CryoVEx 2008 was successfully carried out between April 15 and May 8 and the datasets have been stored and secured at DTU Space and Alfred Wegener Institute (AWI). Afterwards extensive data...

  17. Theoretical and experimental study of image formation in scanning transmission electron microscopy

    International Nuclear Information System (INIS)

    Prunier epouse Mory, Claudie

    1985-01-01

    This thesis contains a theoretical and experimental study of image formation in a dedicated scanning transmission electron microscope (STEM). Using a detailed description of the different optical elements between the field emission source and the specimen, one calculates the shape and size of the primary probe of electrons impinging on the sample. This modelization enables to estimate the spatial resolution in the different imaging or microanalytical modes. The influence of the specimen and the role of the various detectors are taken into account to calculate the point spread function of the instrument in STEM imaging modes. An experimental study of the characteristic properties of phase contrast bright field micrographs and incoherent dark field ones is performed by comparison of digitally recorded images in similar conditions. Spatial resolution, contrast and signal/noise ratio are assessed by correlation methods, Fourier analysis and statistical considerations; one can deduce the optimum focusing conditions. Limits such as the point resolution on quasi-atomic metallic clusters are determined and an analysis of the capabilities of signal mixing concludes this work. Applications are offered in various domains such as the visualisation of small metallic particles, biomolecules and unstained biological sections. (author) [fr

  18. Fluorescence photooxidation with eosin: a method for high resolution immunolocalization and in situ hybridization detection for light and electron microscopy

    Science.gov (United States)

    1994-01-01

    A simple method is described for high-resolution light and electron microscopic immunolocalization of proteins in cells and tissues by immunofluorescence and subsequent photooxidation of diaminobenzidine tetrahydrochloride into an insoluble osmiophilic polymer. By using eosin as the fluorescent marker, a substantial improvement in sensitivity is achieved in the photooxidation process over other conventional fluorescent compounds. The technique allows for precise correlative immunolocalization studies on the same sample using fluorescence, transmitted light and electron microscopy. Furthermore, because eosin is smaller in size than other conventional markers, this method results in improved penetration of labeling reagents compared to gold or enzyme based procedures. The improved penetration allows for three-dimensional immunolocalization using high voltage electron microscopy. Fluorescence photooxidation can also be used for high resolution light and electron microscopic localization of specific nucleic acid sequences by in situ hybridization utilizing biotinylated probes followed by an eosin-streptavidin conjugate. PMID:7519623

  19. SEM-microphotogrammetry, a new take on an old method for generating high-resolution 3D models from SEM images.

    Science.gov (United States)

    Ball, A D; Job, P A; Walker, A E L

    2017-08-01

    The method we present here uses a scanning electron microscope programmed via macros to automatically capture dozens of images at suitable angles to generate accurate, detailed three-dimensional (3D) surface models with micron-scale resolution. We demonstrate that it is possible to use these Scanning Electron Microscope (SEM) images in conjunction with commercially available software originally developed for photogrammetry reconstructions from Digital Single Lens Reflex (DSLR) cameras and to reconstruct 3D models of the specimen. These 3D models can then be exported as polygon meshes and eventually 3D printed. This technique offers the potential to obtain data suitable to reconstruct very tiny features (e.g. diatoms, butterfly scales and mineral fabrics) at nanometre resolution. Ultimately, we foresee this as being a useful tool for better understanding spatial relationships at very high resolution. However, our motivation is also to use it to produce 3D models to be used in public outreach events and exhibitions, especially for the blind or partially sighted. © 2017 The Authors Journal of Microscopy © 2017 Royal Microscopical Society.

  20. Low-cost, high-resolution scanning laser ophthalmoscope for the clinical environment

    Science.gov (United States)

    Soliz, P.; Larichev, A.; Zamora, G.; Murillo, S.; Barriga, E. S.

    2010-02-01

    Researchers have sought to gain greater insight into the mechanisms of the retina and the optic disc at high spatial resolutions that would enable the visualization of small structures such as photoreceptors and nerve fiber bundles. The sources of retinal image quality degradation are aberrations within the human eye, which limit the achievable resolution and the contrast of small image details. To overcome these fundamental limitations, researchers have been applying adaptive optics (AO) techniques to correct for the aberrations. Today, deformable mirror based adaptive optics devices have been developed to overcome the limitations of standard fundus cameras, but at prices that are typically unaffordable for most clinics. In this paper we demonstrate a clinically viable fundus camera with auto-focus and astigmatism correction that is easy to use and has improved resolution. We have shown that removal of low-order aberrations results in significantly better resolution and quality images. Additionally, through the application of image restoration and super-resolution techniques, the images present considerably improved quality. The improvements lead to enhanced visualization of retinal structures associated with pathology.