WorldWideScience

Sample records for replication electron microscopy

  1. Electron Microscopy.

    Science.gov (United States)

    Beer, Michael

    1980-01-01

    Reviews technical aspects of structure determination in biological electron microscopy (EM). Discusses low dose EM, low temperature microscopy, electron energy loss spectra, determination of mass or molecular weight, and EM of labeled systems. Cites 34 references. (CS)

  2. Electron Microscopy.

    Science.gov (United States)

    Beer, Michael

    1980-01-01

    Reviews technical aspects of structure determination in biological electron microscopy (EM). Discusses low dose EM, low temperature microscopy, electron energy loss spectra, determination of mass or molecular weight, and EM of labeled systems. Cites 34 references. (CS)

  3. The replication of Rocio virus in brain tissue of suckling mice. Study by electron microscopy.

    Science.gov (United States)

    Tanaka, H; Weigl, D R; de Souza Lopes, O

    1983-01-01

    By electron microscopy studies, Rocio virus particles were about 43 nm and spherically shaped. They were found within the cisternae of the endoplasmic reticulum and Golgi complex of infected neurons. No precursor particles were detected nor virus budding was evident.

  4. Electron Microscopy Center (EMC)

    Data.gov (United States)

    Federal Laboratory Consortium — The Electron Microscopy Center (EMC) at Argonne National Laboratory develops and maintains unique capabilities for electron beam characterization and applies those...

  5. Scanning ultrafast electron microscopy.

    Science.gov (United States)

    Yang, Ding-Shyue; Mohammed, Omar F; Zewail, Ahmed H

    2010-08-24

    Progress has been made in the development of four-dimensional ultrafast electron microscopy, which enables space-time imaging of structural dynamics in the condensed phase. In ultrafast electron microscopy, the electrons are accelerated, typically to 200 keV, and the microscope operates in the transmission mode. Here, we report the development of scanning ultrafast electron microscopy using a field-emission-source configuration. Scanning of pulses is made in the single-electron mode, for which the pulse contains at most one or a few electrons, thus achieving imaging without the space-charge effect between electrons, and still in ten(s) of seconds. For imaging, the secondary electrons from surface structures are detected, as demonstrated here for material surfaces and biological specimens. By recording backscattered electrons, diffraction patterns from single crystals were also obtained. Scanning pulsed-electron microscopy with the acquired spatiotemporal resolutions, and its efficient heat-dissipation feature, is now poised to provide in situ 4D imaging and with environmental capability.

  6. Scanning ultrafast electron microscopy

    Science.gov (United States)

    Yang, Ding-Shyue; Mohammed, Omar F.; Zewail, Ahmed H.

    2010-01-01

    Progress has been made in the development of four-dimensional ultrafast electron microscopy, which enables space-time imaging of structural dynamics in the condensed phase. In ultrafast electron microscopy, the electrons are accelerated, typically to 200 keV, and the microscope operates in the transmission mode. Here, we report the development of scanning ultrafast electron microscopy using a field-emission-source configuration. Scanning of pulses is made in the single-electron mode, for which the pulse contains at most one or a few electrons, thus achieving imaging without the space-charge effect between electrons, and still in ten(s) of seconds. For imaging, the secondary electrons from surface structures are detected, as demonstrated here for material surfaces and biological specimens. By recording backscattered electrons, diffraction patterns from single crystals were also obtained. Scanning pulsed-electron microscopy with the acquired spatiotemporal resolutions, and its efficient heat-dissipation feature, is now poised to provide in situ 4D imaging and with environmental capability. PMID:20696933

  7. Diagnostic electron microscopy

    Energy Technology Data Exchange (ETDEWEB)

    Dickersin, G.R.

    1988-01-01

    In this book the author presents a comprehensive reference text on diagnostic electron microscopy. Throughout the book he illustrates how ultrastructural identification can be helpful for the recognition of cell type and the identification of mechanisms of pathogenesis in various diseases. In addition to electron microscopy photographs, there are also numerous light microscopy photographs for comparison. This text presents the classification of neoplasms in the order and arrangement most familiar to the pathologist. Contents: Introduction; Diagram of a Normal Cell; Normal Cell Function; Embryology; Neoplasms; Infectious Agents; Metabolic Diseases; Renal Diseases; Skeletal Muscle and Peripheral Nerve Diseases; Index.

  8. Single particle electron microscopy

    NARCIS (Netherlands)

    Boekema, Egbert J.; Folea, Mihaela; Kouril, Roman; Kouřil, Roman

    2009-01-01

    Electron microscopy (EM) in combination with image analysis is a powerful technique to study protein structures at low, medium, and high resolution. Since electron micrographs of biological objects are very noisy, improvement of the signal-to-noise ratio by image processing is an integral part of EM

  9. Conventional transmission electron microscopy.

    Science.gov (United States)

    Winey, Mark; Meehl, Janet B; O'Toole, Eileen T; Giddings, Thomas H

    2014-02-01

    Researchers have used transmission electron microscopy (TEM) to make contributions to cell biology for well over 50 years, and TEM continues to be an important technology in our field. We briefly present for the neophyte the components of a TEM-based study, beginning with sample preparation through imaging of the samples. We point out the limitations of TEM and issues to be considered during experimental design. Advanced electron microscopy techniques are listed as well. Finally, we point potential new users of TEM to resources to help launch their project.

  10. Electronic detectors for electron microscopy.

    Science.gov (United States)

    Faruqi, A R; McMullan, G

    2011-08-01

    Electron microscopy (EM) is an important tool for high-resolution structure determination in applications ranging from condensed matter to biology. Electronic detectors are now used in most applications in EM as they offer convenience and immediate feedback that is not possible with film or image plates. The earliest forms of electronic detector used routinely in transmission electron microscopy (TEM) were charge coupled devices (CCDs) and for many applications these remain perfectly adequate. There are however applications, such as the study of radiation-sensitive biological samples, where film is still used and improved detectors would be of great value. The emphasis in this review is therefore on detectors for use in such applications. Two of the most promising candidates for improved detection are: monolithic active pixel sensors (MAPS) and hybrid pixel detectors (of which Medipix2 was chosen for this study). From the studies described in this review, a back-thinned MAPS detector appears well suited to replace film in for the study of radiation-sensitive samples at 300 keV, while Medipix2 is suited to use at lower energies and especially in situations with very low count rates. The performance of a detector depends on the energy of electrons to be recorded, which in turn is dependent on the application it is being used for; results are described for a wide range of electron energies ranging from 40 to 300 keV. The basic properties of detectors are discussed in terms of their modulation transfer function (MTF) and detective quantum efficiency (DQE) as a function of spatial frequency.

  11. Visualizing Single-molecule DNA Replication with Fluorescence Microscopy

    NARCIS (Netherlands)

    Tanner, Nathan A.; Loparo, Joseph J.; Oijen, Antoine M. van

    2009-01-01

    We describe a simple fluorescence microscopy-based real-time method for observing DNA replication at the single-molecule level. A circular, forked DNA template is attached to a functionalized glass coverslip and replicated extensively after introduction of replication proteins and nucleotides. The

  12. Visualizing Single-molecule DNA Replication with Fluorescence Microscopy

    NARCIS (Netherlands)

    Tanner, Nathan A.; Loparo, Joseph J.; Oijen, Antoine M. van

    2009-01-01

    We describe a simple fluorescence microscopy-based real-time method for observing DNA replication at the single-molecule level. A circular, forked DNA template is attached to a functionalized glass coverslip and replicated extensively after introduction of replication proteins and nucleotides. The g

  13. Advanced computing in electron microscopy

    CERN Document Server

    Kirkland, Earl J

    2010-01-01

    This book features numerical computation of electron microscopy images as well as multislice methods High resolution CTEM and STEM image interpretation are included in the text This newly updated second edition will bring the reader up to date on new developments in the field since the 1990's The only book that specifically addresses computer simulation methods in electron microscopy

  14. Soil microstructure and electron microscopy

    Science.gov (United States)

    Smart, P.; Fryer, J. R.

    1988-01-01

    As part of the process of comparing Martian soils with terrestial soils, high resolution electron microscopy and associated techniques should be used to examine the finer soil particles, and various techniques of electron and optical microscopy should be used to examine the undisturbed structure of Martian soils. To examine the structure of fine grained portions of the soil, transmission electron microscopy may be required. A striking feature of many Martian soils is their red color. Although the present-day Martian climate appears to be cold, this color is reminiscent of terrestial tropical red clays. Their chemical contents are broadly similar.

  15. Electronic Blending in Virtual Microscopy

    Science.gov (United States)

    Maybury, Terrence S.; Farah, Camile S.

    2010-01-01

    Virtual microscopy (VM) is a relatively new technology that transforms the computer into a microscope. In essence, VM allows for the scanning and transfer of glass slides from light microscopy technology to the digital environment of the computer. This transition is also a function of the change from print knowledge to electronic knowledge, or as…

  16. Four-dimensional electron microscopy.

    Science.gov (United States)

    Zewail, Ahmed H

    2010-04-09

    The discovery of the electron over a century ago and the realization of its dual character have given birth to one of the two most powerful imaging instruments: the electron microscope. The electron microscope's ability to resolve three-dimensional (3D) structures on the atomic scale is continuing to affect different fields, including materials science and biology. In this Review, we highlight recent developments and inventions made by introducing the fourth dimension of time in electron microscopy. Today, ultrafast electron microscopy (4D UEM) enables a resolution that is 10 orders of magnitude better than that of conventional microscopes, which are limited by the video-camera rate of recording. After presenting the central concept involved, that of single-electron stroboscopic imaging, we discuss prototypical applications, which include the visualization of complex structures when unfolding on different length and time scales. The developed UEM variant techniques are several, and here we illucidate convergent-beam and near-field imaging, as well as tomography and scanning-pulse microscopy. We conclude with current explorations in imaging of nanomaterials and biostructures and an outlook on possible future directions in space-time, 4D electron microscopy.

  17. 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...

  18. Correlative fluorescence and electron microscopy.

    Science.gov (United States)

    Schirra, Randall T; Zhang, Peijun

    2014-10-01

    Correlative fluorescence and electron microscopy (CFEM) is a multimodal technique that combines dynamic and localization information from fluorescence methods with ultrastructural data from electron microscopy, to give new information about how cellular components change relative to the spatiotemporal dynamics within their environment. In this review, we will discuss some of the basic techniques and tools of the trade for utilizing this attractive research method, which is becoming a very powerful tool for biology labs. The information obtained from correlative methods has proven to be invaluable in creating consensus between the two types of microscopy, extending the capability of each, and cutting the time and expense associated with using each method separately for comparative analysis. The realization of the advantages of these methods in cell biology has led to rapid improvement in the protocols and has ushered in a new generation of instruments to reach the next level of correlation--integration.

  19. Electron Microscopy of Intracellular Protozoa.

    Science.gov (United States)

    1983-08-01

    described by Casero et al.(1). For electron microscopy, 50 x 106 organisms in 5 ml of incubation medium (1) were treated with 10 0 DMSO(control cultures... Casero , R.A., Klayman, D.L., Childs, G.E., Scoville, J.P., and Desjardins, R.E. 1980. Activity of 2-acetylpyridine thiosemicarbazones against

  20. Electron microscopic analysis of rotavirus assembly-replication intermediates

    Energy Technology Data Exchange (ETDEWEB)

    Boudreaux, Crystal E.; Kelly, Deborah F. [Virginia Tech Carilion School of Medicine and Research Institute, Roanoke, VA (United States); McDonald, Sarah M., E-mail: mcdonaldsa@vtc.vt.edu [Virginia Tech Carilion School of Medicine and Research Institute, Roanoke, VA (United States); Department of Biomedical Sciences and Pathobiology, Virginia—Maryland Regional College of Veterinary Medicine, Blacksburg, VA (United States)

    2015-03-15

    Rotaviruses (RVs) replicate their segmented, double-stranded RNA genomes in tandem with early virion assembly. In this study, we sought to gain insight into the ultrastructure of RV assembly-replication intermediates (RIs) using transmission electron microscopy (EM). Specifically, we examined a replicase-competent, subcellular fraction that contains all known RV RIs. Three never-before-seen complexes were visualized in this fraction. Using in vitro reconstitution, we showed that ~15-nm doughnut-shaped proteins in strings were nonstructural protein 2 (NSP2) bound to viral RNA transcripts. Moreover, using immunoaffinity-capture EM, we revealed that ~20-nm pebble-shaped complexes contain the viral RNA polymerase (VP1) and RNA capping enzyme (VP3). Finally, using a gel purification method, we demonstrated that ~30–70-nm electron-dense, particle-shaped complexes represent replicase-competent core RIs, containing VP1, VP3, and NSP2 as well as capsid proteins VP2 and VP6. The results of this study raise new questions about the interactions among viral proteins and RNA during the concerted assembly–replicase process. - Highlights: • Rotaviruses replicate their genomes in tandem with early virion assembly. • Little is known about rotavirus assembly-replication intermediates. • Assembly-replication intermediates were imaged using electron microscopy.

  1. Correlated Light Microscopy and Electron Microscopy

    NARCIS (Netherlands)

    Sjollema, Klaas A.; Schnell, Ulrike; Kuipers, Jeroen; Kalicharan, Ruby; Giepmans, Ben N. G.; MullerReichert, T; Verkade, P

    2012-01-01

    Understanding where, when, and how biomolecules (inter)act is crucial to uncover fundamental mechanisms in cell biology. Recent developments in fluorescence light microscopy (FLM) allow protein imaging in living cells and at the near molecular level. However, fluorescence microscopy only reveals

  2. Correlated Light Microscopy and Electron Microscopy

    NARCIS (Netherlands)

    Sjollema, Klaas A.; Schnell, Ulrike; Kuipers, Jeroen; Kalicharan, Ruby; Giepmans, Ben N. G.; MullerReichert, T; Verkade, P

    2012-01-01

    Understanding where, when, and how biomolecules (inter)act is crucial to uncover fundamental mechanisms in cell biology. Recent developments in fluorescence light microscopy (FLM) allow protein imaging in living cells and at the near molecular level. However, fluorescence microscopy only reveals sel

  3. Correlative Stochastic Optical Reconstruction Microscopy and Electron Microscopy

    OpenAIRE

    Doory Kim; Deerinck, Thomas J.; Sigal, Yaron M.; Babcock, Hazen P.; Ellisman, Mark H.; Xiaowei Zhuang

    2015-01-01

    Correlative fluorescence light microscopy and electron microscopy allows the imaging of spatial distributions of specific biomolecules in the context of cellular ultrastructure. Recent development of super-resolution fluorescence microscopy allows the location of molecules to be determined with nanometer-scale spatial resolution. However, correlative super-resolution fluorescence microscopy and electron microscopy (EM) still remains challenging because the optimal specimen preparation and ima...

  4. Bridging fluorescence microscopy and electron microscopy

    NARCIS (Netherlands)

    Giepmans, Ben N. G.

    Development of new fluorescent probes and fluorescence microscopes has led to new ways to study cell biology. With the emergence of specialized microscopy units at most universities and research centers, the use of these techniques is well within reach for a broad research community. A major

  5. Bridging fluorescence microscopy and electron microscopy

    NARCIS (Netherlands)

    Giepmans, Ben N. G.

    2008-01-01

    Development of new fluorescent probes and fluorescence microscopes has led to new ways to study cell biology. With the emergence of specialized microscopy units at most universities and research centers, the use of these techniques is well within reach for a broad research community. A major breakth

  6. Polyethyleneimine as tracer for electron microscopy

    NARCIS (Netherlands)

    Schurer, Jacob Willem

    1980-01-01

    In this thesis the development of a tracer particle for use in electron microscopy is described. Attempts were made to use this tracer particle in immuno-electron microscopy and to trace negatively charged tissue components. ... Zie: Summary

  7. Synthetic incoherence for electron microscopy.

    Science.gov (United States)

    Levine, Zachary H; Dunstan, Robyn M

    2007-08-01

    Tomographic studies of submicrometer samples in materials science using electron microscopy have been inhibited by diffraction effects. In the present work, we describe a practical method for ameliorating these effects. First, we find an analytic expression for the mutual coherence function for hollow-cone illumination. Then, we use this analytic expression to propose a Gaussian weighting of hollow-cone illumination, which we name tapered solid-cone illumination, and present an analytic expression for its mutual coherence function. Finally, we investigate numerically an n-ring approximation to tapered solid-cone illumination. The results suggest a method for removing diffraction effects and hence enabling tomography.

  8. Liquid Cell Transmission Electron Microscopy

    Science.gov (United States)

    Liao, Hong-Gang; Zheng, Haimei

    2016-05-01

    Liquid cell transmission electron microscopy (TEM) has attracted significant interest in recent years. With nanofabricated liquid cells, it has been possible to image through liquids using TEM with subnanometer resolution, and many previously unseen materials dynamics have been revealed. Liquid cell TEM has been applied to many areas of research, ranging from chemistry to physics, materials science, and biology. So far, topics of study include nanoparticle growth and assembly, electrochemical deposition and lithiation for batteries, tracking and manipulation of nanoparticles, catalysis, and imaging of biological materials. In this article, we first review the development of liquid cell TEM and then highlight progress in various areas of research. In the study of nanoparticle growth, the electron beam can serve both as the illumination source for imaging and as the input energy for reactions. However, many other research topics require the control of electron beam effects to minimize electron beam damage. We discuss efforts to understand electron beam-liquid matter interactions. Finally, we provide a perspective on future challenges and opportunities in liquid cell TEM.

  9. Electron microscopy and forensic practice

    Science.gov (United States)

    Kotrlý, Marek; Turková, Ivana

    2013-05-01

    Electron microanalysis in forensic practice ranks among basic applications used in investigation of traces (latents, stains, etc.) from crime scenes. Applying electron microscope allows for rapid screening and receiving initial information for a wide range of traces. SEM with EDS/WDS makes it possible to observe topography surface and morphology samples and examination of chemical components. Physical laboratory of the Institute of Criminalistics Prague use SEM especially for examination of inorganic samples, rarely for biology and other material. Recently, possibilities of electron microscopy have been extended considerably using dual systems with focused ion beam. These systems are applied mainly in study of inner micro and nanoparticles , thin layers (intersecting lines in graphical forensic examinations, analysis of layers of functional glass, etc.), study of alloys microdefects, creating 3D particles and aggregates models, etc. Automated mineralogical analyses are a great asset to analysis of mineral phases, particularly soils, similarly it holds for cathode luminescence, predominantly colour one and precise quantitative measurement of their spectral characteristics. Among latest innovations that are becoming to appear also at ordinary laboratories are TOF - SIMS systems and micro Raman spectroscopy with a resolution comparable to EDS/WDS analysis (capable of achieving similar level as through EDS/WDS analysis).

  10. Correlative stochastic optical reconstruction microscopy and electron microscopy.

    Directory of Open Access Journals (Sweden)

    Doory Kim

    Full Text Available Correlative fluorescence light microscopy and electron microscopy allows the imaging of spatial distributions of specific biomolecules in the context of cellular ultrastructure. Recent development of super-resolution fluorescence microscopy allows the location of molecules to be determined with nanometer-scale spatial resolution. However, correlative super-resolution fluorescence microscopy and electron microscopy (EM still remains challenging because the optimal specimen preparation and imaging conditions for super-resolution fluorescence microscopy and EM are often not compatible. Here, we have developed several experiment protocols for correlative stochastic optical reconstruction microscopy (STORM and EM methods, both for un-embedded samples by applying EM-specific sample preparations after STORM imaging and for embedded and sectioned samples by optimizing the fluorescence under EM fixation, staining and embedding conditions. We demonstrated these methods using a variety of cellular targets.

  11. Measurement of replication structures at the nanometer scale using super-resolution light microscopy.

    Science.gov (United States)

    Baddeley, D; Chagin, V O; Schermelleh, L; Martin, S; Pombo, A; Carlton, P M; Gahl, A; Domaing, P; Birk, U; Leonhardt, H; Cremer, C; Cardoso, M C

    2010-01-01

    DNA replication, similar to other cellular processes, occurs within dynamic macromolecular structures. Any comprehensive understanding ultimately requires quantitative data to establish and test models of genome duplication. We used two different super-resolution light microscopy techniques to directly measure and compare the size and numbers of replication foci in mammalian cells. This analysis showed that replication foci vary in size from 210 nm down to 40 nm. Remarkably, spatially modulated illumination (SMI) and 3D-structured illumination microscopy (3D-SIM) both showed an average size of 125 nm that was conserved throughout S-phase and independent of the labeling method, suggesting a basic unit of genome duplication. Interestingly, the improved optical 3D resolution identified 3- to 5-fold more distinct replication foci than previously reported. These results show that optical nanoscopy techniques enable accurate measurements of cellular structures at a level previously achieved only by electron microscopy and highlight the possibility of high-throughput, multispectral 3D analyses.

  12. Four-dimensional ultrafast electron microscopy.

    Science.gov (United States)

    Lobastov, Vladimir A; Srinivasan, Ramesh; Zewail, Ahmed H

    2005-05-17

    Electron microscopy is arguably the most powerful tool for spatial imaging of structures. As such, 2D and 3D microscopies provide static structures with subnanometer and increasingly with angstrom-scale spatial resolution. Here we report the development of 4D ultrafast electron microscopy, whose capability imparts another dimension to imaging in general and to dynamics in particular. We demonstrate its versatility by recording images and diffraction patterns of crystalline and amorphous materials and images of biological cells. The electron packets, which were generated with femtosecond laser pulses, have a de Broglie wavelength of 0.0335 angstroms at 120 keV and have as low as one electron per pulse. With such few particles, doses of few electrons per square ångstrom, and ultrafast temporal duration, the long sought after but hitherto unrealized quest for ultrafast electron microscopy has been realized. Ultrafast electron microscopy should have an impact on all areas of microscopy, including biological imaging.

  13. 3D correlative light and electron microscopy of cultured cells using serial blockface scanning electron microscopy

    Science.gov (United States)

    Lerner, Thomas R.; Burden, Jemima J.; Nkwe, David O.; Pelchen-Matthews, Annegret; Domart, Marie-Charlotte; Durgan, Joanne; Weston, Anne; Jones, Martin L.; Peddie, Christopher J.; Carzaniga, Raffaella; Florey, Oliver; Marsh, Mark; Gutierrez, Maximiliano G.

    2017-01-01

    ABSTRACT The processes of life take place in multiple dimensions, but imaging these processes in even three dimensions is challenging. Here, we describe a workflow for 3D correlative light and electron microscopy (CLEM) of cell monolayers using fluorescence microscopy to identify and follow biological events, combined with serial blockface scanning electron microscopy to analyse the underlying ultrastructure. The workflow encompasses all steps from cell culture to sample processing, imaging strategy, and 3D image processing and analysis. We demonstrate successful application of the workflow to three studies, each aiming to better understand complex and dynamic biological processes, including bacterial and viral infections of cultured cells and formation of entotic cell-in-cell structures commonly observed in tumours. Our workflow revealed new insight into the replicative niche of Mycobacterium tuberculosis in primary human lymphatic endothelial cells, HIV-1 in human monocyte-derived macrophages, and the composition of the entotic vacuole. The broad application of this 3D CLEM technique will make it a useful addition to the correlative imaging toolbox for biomedical research. PMID:27445312

  14. 3D correlative light and electron microscopy of cultured cells using serial blockface scanning electron microscopy.

    Science.gov (United States)

    Russell, Matthew R G; Lerner, Thomas R; Burden, Jemima J; Nkwe, David O; Pelchen-Matthews, Annegret; Domart, Marie-Charlotte; Durgan, Joanne; Weston, Anne; Jones, Martin L; Peddie, Christopher J; Carzaniga, Raffaella; Florey, Oliver; Marsh, Mark; Gutierrez, Maximiliano G; Collinson, Lucy M

    2017-01-01

    The processes of life take place in multiple dimensions, but imaging these processes in even three dimensions is challenging. Here, we describe a workflow for 3D correlative light and electron microscopy (CLEM) of cell monolayers using fluorescence microscopy to identify and follow biological events, combined with serial blockface scanning electron microscopy to analyse the underlying ultrastructure. The workflow encompasses all steps from cell culture to sample processing, imaging strategy, and 3D image processing and analysis. We demonstrate successful application of the workflow to three studies, each aiming to better understand complex and dynamic biological processes, including bacterial and viral infections of cultured cells and formation of entotic cell-in-cell structures commonly observed in tumours. Our workflow revealed new insight into the replicative niche of Mycobacterium tuberculosis in primary human lymphatic endothelial cells, HIV-1 in human monocyte-derived macrophages, and the composition of the entotic vacuole. The broad application of this 3D CLEM technique will make it a useful addition to the correlative imaging toolbox for biomedical research. © 2017. Published by The Company of Biologists Ltd.

  15. Proximity Scanning Transmission Electron Microscopy/Spectroscopy

    CERN Document Server

    Hwang, Ing-Shouh

    2016-01-01

    Here a new microscopic method is proposed to image and characterize very thin samples like few-layer materials, organic molecules, and nanostructures with nanometer or sub-nanometer resolution using electron beams of energies lower than 20 eV. The microscopic technique achieves high resolution through the proximity (or near-field) effect, as in scanning tunneling microscopy (STM), while it also allows detection of transmitted electrons for imaging and spectroscopy, as in scanning transmission electron microscopy (STEM). This proximity transmission electron microscopy (PSTEM) does not require any lens to focus the electron beam. It also allows detailed characterization of the interaction of low-energy electron with materials. PSTEM can operate in a way very similar to scanning tunneling microscopy, which provides high-resolution imaging of geometric and electronic structures of the sample surface. In addition, it allows imaging and characterization of the interior structures of the sample based on the detected...

  16. Fast electron microscopy via compressive sensing

    Science.gov (United States)

    Larson, Kurt W; Anderson, Hyrum S; Wheeler, Jason W

    2014-12-09

    Various technologies described herein pertain to compressive sensing electron microscopy. A compressive sensing electron microscope includes a multi-beam generator and a detector. The multi-beam generator emits a sequence of electron patterns over time. Each of the electron patterns can include a plurality of electron beams, where the plurality of electron beams is configured to impart a spatially varying electron density on a sample. Further, the spatially varying electron density varies between each of the electron patterns in the sequence. Moreover, the detector collects signals respectively corresponding to interactions between the sample and each of the electron patterns in the sequence.

  17. Atomic resolution 3D electron diffraction microscopy

    Energy Technology Data Exchange (ETDEWEB)

    Miao, Jianwei; Ohsuna, Tetsu; Terasaki, Osamu; O' Keefe, Michael A.

    2002-03-01

    Electron lens aberration is the major barrier limiting the resolution of electron microscopy. Here we describe a novel form of electron microscopy to overcome electron lens aberration. By combining coherent electron diffraction with the oversampling phasing method, we show that the 3D structure of a 2 x 2 x 2 unit cell nano-crystal (framework of LTA [Al12Si12O48]8) can be ab initio determined at the resolution of 1 Angstrom from a series of simulated noisy diffraction pattern projections with rotation angles ranging from -70 degrees to +70 degrees in 5 degrees increments along a single rotation axis. This form of microscopy (which we call 3D electron diffraction microscopy) does not require any reference waves, and can image the 3D structure of nanocrystals, as well as non-crystalline biological and materials science samples, with the resolution limited only by the quality of sample diffraction.

  18. Ultrafast Science Opportunities with Electron Microscopy

    Energy Technology Data Exchange (ETDEWEB)

    DURR, HERMANN; Wang, X.J., ed.

    2016-04-28

    X-rays and electrons are two of the most fundamental probes of matter. When the Linac Coherent Light Source (LCLS), the world’s first x-ray free electron laser, began operation in 2009, it transformed ultrafast science with the ability to generate laser-like x-ray pulses from the manipulation of relativistic electron beams. This document describes a similar future transformation. In Transmission Electron Microscopy, ultrafast relativistic (MeV energy) electron pulses can achieve unsurpassed spatial and temporal resolution. Ultrafast temporal resolution will be the next frontier in electron microscopy and can ideally complement ultrafast x-ray science done with free electron lasers. This document describes the Grand Challenge science opportunities in chemistry, material science, physics and biology that arise from an MeV ultrafast electron diffraction & microscopy facility, especially when coupled with linac-based intense THz and X-ray pump capabilities.

  19. Transmission Electron Microscopy Physics of Image Formation

    CERN Document Server

    Kohl, Helmut

    2008-01-01

    Transmission Electron Microscopy: Physics of Image Formation presents the theory of image and contrast formation, and the analytical modes in transmission electron microscopy. The principles of particle and wave optics of electrons are described. Electron-specimen interactions are discussed for evaluating the theory of scattering and phase contrast. Also discussed are the kinematical and dynamical theories of electron diffraction and their applications for crystal-structure analysis and imaging of lattices and their defects. X-ray microanalysis and electron energy-loss spectroscopy are treated as analytical methods. Specimen damage and contamination by electron irradiation limits the resolution for biological and some inorganic specimens. This fifth edition includes discussion of recent progress, especially in the area of aberration correction and energy filtering; moreover, the topics introduced in the fourth edition have been updated. Transmission Electron Microscopy: Physics of Image Formation is written f...

  20. Fundamental Technical Elements of Freeze-fracture/Freeze-etch in Biological Electron Microscopy

    Science.gov (United States)

    Freeze-fracture/freeze-etch describes a process whereby specimens, typically biological or nanomaterial in nature, are frozen, fractured, and replicated to generate a carbon/platinum "cast" intended for examination by transmission electron microscopy. Specimens are subjected to u...

  1. Fundamental Technical Elements of Freeze-fracture/Freeze-etch in Biological Electron Microscopy

    Science.gov (United States)

    Freeze-fracture/freeze-etch describes a process whereby specimens, typically biological or nanomaterial in nature, are frozen, fractured, and replicated to generate a carbon/platinum "cast" intended for examination by transmission electron microscopy. Specimens are subjected to u...

  2. Advanced electron microscopy for advanced materials.

    Science.gov (United States)

    Van Tendeloo, Gustaaf; Bals, Sara; Van Aert, Sandra; Verbeeck, Jo; Van Dyck, Dirk

    2012-11-08

    The idea of this Review is to introduce newly developed possibilities of advanced electron microscopy to the materials science community. Over the last decade, electron microscopy has evolved into a full analytical tool, able to provide atomic scale information on the position, nature, and even the valency atoms. This information is classically obtained in two dimensions (2D), but can now also be obtained in 3D. We show examples of applications in the field of nanoparticles and interfaces.

  3. Aberration Correction in Electron Microscopy

    CERN Document Server

    Rose, Harald H

    2005-01-01

    The resolution of conventional electron microscopes is limited by spherical and chromatic aberrations. Both defects are unavoidable in the case of static rotationally symmetric electromagnetic fields (Scherzer theorem). Multipole correctors and electron mirrros have been designed and built, which compensate for these aberrations. The principles of correction will be demonstrated for the tetrode mirror, the quadrupole-octopole corrector and the hexapole corrector. Electron mirrors require a magnetic beam separator free of second-order aberrations. The multipole correctors are highly symmetric telescopic systems compensating for the defects of the objective lens. The hexapole corrector has the most simple structure yet eliminates only the spherical aberration, whereas the mirror and the quadrupole-octopole corrector are able to correct for both aberrations. Chromatic correction is achieved in the latter corrector by cossed electric and magnetic quadrupoles acting as first-order Wien filters. Micrographs obtaine...

  4. Digital detectors for electron microscopy

    CERN Document Server

    Faruqi, A R

    2002-01-01

    Film has traditionally been used for recording images in transmission electron microscopes but there is an essential need for computer-interfaced electronic detectors. Cooled-CCD detectors, developed over the past few years, though not ideal, are increasingly used as the preferred detection system in a number of applications. We describe briefly the design of CCD-based detectors, along with their main properties, which have been used in electron crystallography. A newer detector design with a much bigger sensitive area, incorporating a 2x2 tiled array of CCDs with tapered fibre optics will overcome some of the limitations of existing CCD detectors. We also describe some preliminary results for 8 keV imaging, from (direct detection) silicon hybrid pixel detectors, which offer advantages over CCDs in terms of better spatial resolution, faster readout with minimal noise.

  5. High-energy electron diffraction and microscopy

    CERN Document Server

    Peng, L M; Whelan, M J

    2011-01-01

    This book provides a comprehensive introduction to high energy electron diffraction and elastic and inelastic scattering of high energy electrons, with particular emphasis on applications to modern electron microscopy. Starting from a survey of fundamental phenomena, the authors introduce the most important concepts underlying modern understanding of high energy electron diffraction. Dynamical diffraction in transmission (THEED) and reflection (RHEED) geometries is treated using ageneral matrix theory, where computer programs and worked examples are provided to illustrate the concepts and to f

  6. Low voltage transmission electron microscopy of graphene.

    Science.gov (United States)

    Bachmatiuk, Alicja; Zhao, Jiong; Gorantla, Sandeep Madhukar; Martinez, Ignacio Guillermo Gonzalez; Wiedermann, Jerzy; Lee, Changgu; Eckert, Juergen; Rummeli, Mark Hermann

    2015-02-04

    The initial isolation of graphene in 2004 spawned massive interest in this two-dimensional pure sp(2) carbon structure due to its incredible electrical, optical, mechanical, and thermal effects. This in turn led to the rapid development of various characterization tools for graphene. Examples include Raman spectroscopy and scanning tunneling microscopy. However, the one tool with the greatest prowess for characterizing and studying graphene is the transmission electron microscope. State-of-the-art (scanning) transmission electron microscopes enable one to image graphene with atomic resolution, and also to conduct various other characterizations simultaneously. The advent of aberration correctors was timely in that it allowed transmission electron microscopes to operate with reduced acceleration voltages, so that damage to graphene is avoided while still providing atomic resolution. In this comprehensive review, a brief introduction is provided to the technical aspects of transmission electron microscopes relevant to graphene. The reader is then introduced to different specimen preparation techniques for graphene. The different characterization approaches in both transmission electron microscopy and scanning transmission electron microscopy are then discussed, along with the different aspects of electron diffraction and electron energy loss spectroscopy. The use of graphene for other electron microscopy approaches such as in-situ investigations is also presented. © 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  7. Electron Microscopy of Intracellular Protozoa.

    Science.gov (United States)

    1980-08-01

    often with disrupted plasma membranes and a matrix which was vacuolated and less electron- dense than normal (figure 7). The merozoites were covered...Plasmodium brasilianum. J. Infect. Dis., 75: 1-32. -~ ~.Clak, .A., Allison, A.C., Cox, F.E., 1976. Protection of mice against Babesia and Plasmodium with BCG ...binding trypanosome were observed in each case (Fig 6). Lack of enhanced uptake by cells of BCG -treated mice. BCG (Mycobacterium bovis) treatment of mice

  8. Compressed Sensing and Electron Microscopy

    Science.gov (United States)

    2010-01-01

    rescanning the material many times as long as the beam intensity is low enough. Strontium Titanite and M1 catalysts are examples of materials of this type...The electron beam width is smaller than the atomic spacing. A typical setting in STEM is that the beam width is a fraction of an angstrom while the... atomic spacing is at least 3 angstroms. When the beam is centered near a particular atom or atom column, the beam produces an intensity distribution

  9. Transmission electron microscopy of composites

    Science.gov (United States)

    Pirouz, P.; Farmer, S. C.; Ernst, F.; Chung, J.

    1988-01-01

    Since interphase-interfaces are often both the structurally weakest and chemically least stable regions of a composite material, they are critical determinants of such macrostructural characteristics as tensile strength and fracture toughness. Attention is presently given to the use of TEM for the study of interfaces between dissimilar materials; electron-diffraction, analytical, and high-resolution forms of TEM are employed, for the cases of both structural and semiconductor composites. The materials studied are SiC/Si, GaP/Si, and SiC fiber- and whisker-reinforced Si3N4.

  10. Magnetic contrast in threshold photoemission electron microscopy

    NARCIS (Netherlands)

    Veghel, Marinus Godefridus Adrianus van

    2004-01-01

    In threshold photoemission electron microscopy (threshold PEEM), photoelectrons are excited by UV photons with an energy just above the photoemission threshold. The lateral intensity distribution of these electrons is then imaged by an electrostatic lens system. In this thesis, the possibilities o

  11. Electron Microscopy of Living Cells During in Situ Fluorescence Microscopy.

    Science.gov (United States)

    Liv, Nalan; van Oosten Slingeland, Daan S B; Baudoin, Jean-Pierre; Kruit, Pieter; Piston, David W; Hoogenboom, Jacob P

    2016-01-26

    We present an approach toward dynamic nanoimaging: live fluorescence of cells encapsulated in a bionanoreactor is complemented with in situ scanning electron microscopy (SEM) on an integrated microscope. This allows us to take SEM snapshots on-demand, that is, at a specific location in time, at a desired region of interest, guided by the dynamic fluorescence imaging. We show that this approach enables direct visualization, with EM resolution, of the distribution of bioconjugated quantum dots on cellular extensions during uptake and internalization.

  12. Analysis of cytokinesis by electron microscopy.

    Science.gov (United States)

    König, J; Borrego-Pinto, J; Streichert, D; Munzig, M; Lenart, P; Müller-Reichert, T

    2017-01-01

    Following up on a chapter on the Correlative Light and Electron Microscopy of Early Caenorhabditis elegans Embryos in Mitosis (MCB 79, 101-119), we present an adaptation of our established protocol for the ultrastructural analysis of either permeabilized or injected embryonic systems. We prepared both drug-treated early C. elegans embryos and fluorescently labeled sea urchin embryos of Lytechinus pictus for ultrastructural studies on animal cytokinesis. Here we focus on the initial preparation steps of postmitotic embryos for high-pressure freezing and subsequent electron microscopy with an emphasis on electron tomography. The advantages and limitations of our extended protocol will be discussed. Copyright © 2017 Elsevier Inc. All rights reserved.

  13. Electron Microscopy of Natural and Epitaxial Diamond

    Science.gov (United States)

    Posthill, J. B.; George, T.; Malta, D. P.; Humphreys, T. P.; Rudder, R. A.; Hudson, G. C.; Thomas, R. E.; Markunas, R. J.

    1993-01-01

    Semiconducting diamond films have the potential for use as a material in which to build active electronic devices capable of operating at high temperatures or in high radiation environments. Ultimately, it is preferable to use low-defect-density single crystal diamond for device fabrication. We have previously investigated polycrystalline diamond films with transmission electron microscopy (TEM) and scanning electron microscopy (SEM), and homoepitaxial films with SEM-based techniques. This contribution describes some of our most recent observations of the microstructure of natural diamond single crystals and homoepitaxial diamond thin films using TEM.

  14. Environmental scanning electron microscopy in cell biology.

    Science.gov (United States)

    McGregor, J E; Staniewicz, L T L; Guthrie Neé Kirk, S E; Donald, A M

    2013-01-01

    Environmental scanning electron microscopy (ESEM) (1) is an imaging technique which allows hydrated, insulating samples to be imaged under an electron beam. The resolution afforded by this technique is higher than conventional optical microscopy but lower than conventional scanning electron microscopy (CSEM). The major advantage of the technique is the minimal sample preparation needed, making ESEM quick to use and the images less susceptible to the artifacts that the extensive sample preparation usually required for CSEM may introduce. Careful manipulation of both the humidity in the microscope chamber and the beam energy are nevertheless essential to prevent dehydration and beam damage artifacts. In some circumstances it is possible to image live cells in the ESEM (2).In the following sections we introduce the fundamental principles of ESEM imaging before presenting imaging protocols for plant epidermis, mammalian cells, and bacteria. In the first two cases samples are imaged using the secondary electron (topographic) signal, whereas a transmission technique is employed to image bacteria.

  15. Illuminating Electron Microscopy of Photocatalysts

    DEFF Research Database (Denmark)

    Cavalca, Filippo

    rearrangement of the illuminated photocatalysts as well as localized charging effects and variations. We aim to investigate the reaction to gas and light exposure at the nanoscale. References [1] Herrmann, J. M., Top. Catal. 2005, 34, (1-4), 49-65. [2] Tsujimoto, M., S. Moriguchi, et al., J. Electron. Microsc......, composition and operation of photocatalysts and to provide information on the compounds inner arrangement and a fundamental contribution for their further optimization [2]. We want to construct a novel specimen holder capable of shining light onto samples inside the TEM allowing real time in situ experiments....... The holder is implemented with a laser diode and an optical system that guides the light onto the sample surface with maximum power transmission. The source can be changed and tuned according to the needs, in principle spanning the whole visible and UV light spectrum. It is possible to use the device inside...

  16. Scanning Electron Microscopy Sample Preparation and Imaging.

    Science.gov (United States)

    Nguyen, Jenny Ngoc Tran; Harbison, Amanda M

    2017-01-01

    Scanning electron microscopes allow us to reach magnifications of 20-130,000× and resolve compositional and topographical images with intense detail. These images are created by bombarding a sample with electrons in a focused manner to generate a black and white image from the electrons that bounce off of the sample. The electrons are detected using positively charged detectors. Scanning electron microscopy permits three-dimensional imaging of desiccated specimens or wet cells and tissues by using variable pressure chambers. SEM ultrastructural analysis and intracellular imaging supplement light microscopy for molecular profiling of prokaryotes, plants, and mammals. This chapter demonstrates how to prepare and image samples that are (a) desiccated and conductive, (b) desiccated and nonconductive but coated with an electron conductive film using a gold sputter coater, and (c) wet and maintained in a hydrated state using a Deben Coolstage.

  17. Comparative study of electron microscopy and scanning probe microscopy in photosynthetic research

    OpenAIRE

    MATĚNOVÁ, Martina

    2009-01-01

    The aim of this study is to compare the ability of transmission electron microscopy, scanning electron microscopy and atomic force microscopy to visualize individual protein complexes. The principle of electron microscopy and atomic force microscopy is explained. For comparision of these methods well characterized photosynthetic complexes LH1, LH2, PSI and PSII were selected.

  18. [Pili annulati. A scanning electron microscopy study].

    Science.gov (United States)

    Lalević-Vasić, B; Polić, D

    1988-01-01

    A case of ringed hair studied by light and electron microscopy is reported. The patient, a 20-year old girl, had been presenting with the hair abnormality since birth. At naked eye examination the hairs were dry, 6 to 7 cm long, and they showed dull and shining areas giving the scalp hair a scintillating appearance (fig. 1). Several samples of hair were taken and examined by light microscopy under white and polarized light. Hair shafts and cryo-fractured surfaces were examined by scanning electron microscopy. RESULTS. 1. Light microscopy. Lesions were found in every hair examined. There were abnormal, opaque and fusiform areas alternating with normal areas all along the hair shaft (fig. 2). The abnormal areas resulted from intracortical air-filled cavities. Fractures similar to those of trichorrhexis nodosa were found in the opaque areas of the distal parts of the hairs. 2. Scanning electron microscopy. A. Hair shaft surface. The abnormal areas showed a longitudinal, "curtain-like" folding of the cuticular cells which had punctiform depressions on their surface and worn free edges (fig. 4, 5, 6); trichorrhexis-type fractures were seen in the distal parts of the hair shafts (fig. 7, 8). Normal areas regularly presented with longitudinal, superficial, short and non-systematized depressions (fig. 9); the cuticular cells were worn, and there were places where the denuded cortex showed dissociated cortical fibres (fig. 10).(ABSTRACT TRUNCATED AT 250 WORDS)

  19. The future of high resolution electron microscopy

    Institute of Scientific and Technical Information of China (English)

    D Van Dyck

    2000-01-01

    The state of the art and the future in quantitative high resolution electron microscopy are discussed in the framework of parameter estimation. Reconstruction methods are then to be considered as direct methods to yield a starting structure for further refinement. With the increasing flexibility of the instruments, computer aided experimental strategy will become important.

  20. Transmission electron microscopy characterization of nanomaterials

    CERN Document Server

    2014-01-01

    Third volume of a 40volume series on nanoscience and nanotechnology, edited by the renowned scientist Challa S.S.R. Kumar. This handbook gives a comprehensive overview about Transmission electron microscopy characterization of nanomaterials. Modern applications and state-of-the-art techniques are covered and make this volume an essential reading for research scientists in academia and industry.

  1. Energetic materials research using scanning electron microscopy

    NARCIS (Netherlands)

    Elshout, J.J.M.H. van den; Duvalois, W.; Benedetto, G.L. Di; Bouma, R.H.B.; Heijden, A.E.D.M. van der

    2016-01-01

    A key-technique for the research of energetic materials is scanning electron microscopy. In this paper several examples are given of characterization studies on energetic materials, including a solid composite propellant formulation. Results of the characterization of energetic materials using scann

  2. Phosphogypsum surface characterisation using scanning electron microscopy

    Directory of Open Access Journals (Sweden)

    Rajković Miloš B.

    2003-01-01

    Full Text Available This paper presents the results of application of Scanning Electron Microscopy (SEM to examinations of the samples of natural gypsum and phosphogypsum. Phosphogypsum has a well developed crystalline structure, and appear in two polymorphous forms, of rombic and hexagonal shape crystals. Natural gypsum has a poorly crystalline structure. The differences in crystalline structure influence the chemical behavior of these row materials.

  3. Energetic materials research using scanning electron microscopy

    NARCIS (Netherlands)

    Elshout, J.J.M.H. van den; Duvalois, W.; Benedetto, G.L. Di; Bouma, R.H.B.; Heijden, A.E.D.M. van der

    2016-01-01

    A key-technique for the research of energetic materials is scanning electron microscopy. In this paper several examples are given of characterization studies on energetic materials, including a solid composite propellant formulation. Results of the characterization of energetic materials using

  4. Photoemission electron microscopy and scanning electron microscopy of Magnetospirillum magnetotacticum's magnetosome chains.

    Science.gov (United States)

    Keutner, Christoph; von Bohlen, Alex; Berges, Ulf; Espeter, Philipp; Schneider, Claus M; Westphal, Carsten

    2014-10-01

    Magnetotactic bacteria are of great interdisciplinary interest, since a vast field of applications from magnetic recording media to medical nanorobots is conceivable. A key feature for a further understanding is the detailed knowledge about the magnetosome chain within the bacteria. We report on two preparation procedures suitable for UHV experiments in reflective geometry. Further, we present the results of scanning electron microscopy, as well as the first photoemission electron microscopy experiments, both accessing the magnetosomes within intact magnetotactic bacteria and compare these to scanning electron microscopy data from the literature. From the images, we can clearly identify individual magnetosomes within their chains.

  5. Photoemission electron microscopy, a tool for plasmonics

    Energy Technology Data Exchange (ETDEWEB)

    Douillard, L., E-mail: ludovic.douillard@cea.fr; Charra, F.

    2013-08-15

    Highlights: •Brief review of photoemission electron microscopy PEEM as a tool for plasmonics, •PEEM gives access to (tip free) near field maps of subwavelength length resolution, •PEEM makes use of a true optical excitation scheme, exhibiting strong connections to ultrafast optical spectrometries. -- Abstract: A key challenge to plasmonics is the development of experimental tools allowing access to the spatial distribution of the optical near field at the nanometre scale. A recent approach for mapping the near optical field is the use of the photoemission electron microscopy PEEM. Indeed, photoemission can be strongly enhanced upon excitation of surface plasmons. By collecting the photoemitted electrons, two-dimensional intensity maps reflecting the actual distribution of the optical near-field are obtained. In the following a brief overview of the possibilities of the photoemission electron microscopy as a tool for plasmonics is given. Main focuses will be set on experimental results regarding the mapping of the near optical field at nanometer scale, the investigation of the spatio-temporal dynamics of plasmon-polariton waves and the manipulation at will of the near field.

  6. 4D electron microscopy: principles and applications.

    Science.gov (United States)

    Flannigan, David J; Zewail, Ahmed H

    2012-10-16

    The transmission electron microscope (TEM) is a powerful tool enabling the visualization of atoms with length scales smaller than the Bohr radius at a factor of only 20 larger than the relativistic electron wavelength of 2.5 pm at 200 keV. The ability to visualize matter at these scales in a TEM is largely due to the efforts made in correcting for the imperfections in the lens systems which introduce aberrations and ultimately limit the achievable spatial resolution. In addition to the progress made in increasing the spatial resolution, the TEM has become an all-in-one characterization tool. Indeed, most of the properties of a material can be directly mapped in the TEM, including the composition, structure, bonding, morphology, and defects. The scope of applications spans essentially all of the physical sciences and includes biology. Until recently, however, high resolution visualization of structural changes occurring on sub-millisecond time scales was not possible. In order to reach the ultrashort temporal domain within which fundamental atomic motions take place, while simultaneously retaining high spatial resolution, an entirely new approach from that of millisecond-limited TEM cameras had to be conceived. As shown below, the approach is also different from that of nanosecond-limited TEM, whose resolution cannot offer the ultrafast regimes of dynamics. For this reason "ultrafast electron microscopy" is reserved for the field which is concerned with femtosecond to picosecond resolution capability of structural dynamics. In conventional TEMs, electrons are produced by heating a source or by applying a strong extraction field. Both methods result in the stochastic emission of electrons, with no control over temporal spacing or relative arrival time at the specimen. The timing issue can be overcome by exploiting the photoelectric effect and using pulsed lasers to generate precisely timed electron packets of ultrashort duration. The spatial and temporal resolutions

  7. Correlative photoactivated localization and scanning electron microscopy.

    Directory of Open Access Journals (Sweden)

    Benjamin G Kopek

    Full Text Available The ability to localize proteins precisely within subcellular space is crucial to understanding the functioning of biological systems. Recently, we described a protocol that correlates a precise map of fluorescent fusion proteins localized using three-dimensional super-resolution optical microscopy with the fine ultrastructural context of three-dimensional electron micrographs. While it achieved the difficult simultaneous objectives of high photoactivated fluorophore preservation and ultrastructure preservation, it required a super-resolution optical and specialized electron microscope that is not available to many researchers. We present here a faster and more practical protocol with the advantage of a simpler two-dimensional optical (Photoactivated Localization Microscopy (PALM and scanning electron microscope (SEM system that retains the often mutually exclusive attributes of fluorophore preservation and ultrastructure preservation. As before, cryosections were prepared using the Tokuyasu protocol, but the staining protocol was modified to be amenable for use in a standard SEM without the need for focused ion beam ablation. We show the versatility of this technique by labeling different cellular compartments and structures including mitochondrial nucleoids, peroxisomes, and the nuclear lamina. We also demonstrate simultaneous two-color PALM imaging with correlated electron micrographs. Lastly, this technique can be used with small-molecule dyes as demonstrated with actin labeling using phalloidin conjugated to a caged dye. By retaining the dense protein labeling expected for super-resolution microscopy combined with ultrastructural preservation, simplifying the tools required for correlative microscopy, and expanding the number of useful labels we expect this method to be accessible and valuable to a wide variety of researchers.

  8. Scanning electron microscopy of superficial white onychomycosis*

    Science.gov (United States)

    de Almeida Jr., Hiram Larangeira; Boabaid, Roberta Oliveira; Timm, Vitor; Silva, Ricardo Marques e; de Castro, Luis Antonio Suita

    2015-01-01

    Superficial white onychomycosis is characterized by opaque, friable, whitish superficial spots on the nail plate. We examined an affected halux nail of a 20-year-old male patient with scanning electron microscopy. The mycological examination isolated Trichophyton mentagrophytes. Abundant hyphae with the formation of arthrospores were found on the nail's surface, forming small fungal colonies. These findings showed the great capacity for dissemination of this form of onychomycosis. PMID:26560225

  9. Correlative photoactivated localization and scanning electron microscopy.

    Science.gov (United States)

    Kopek, Benjamin G; Shtengel, Gleb; Grimm, Jonathan B; Clayton, David A; Hess, Harald F

    2013-01-01

    The ability to localize proteins precisely within subcellular space is crucial to understanding the functioning of biological systems. Recently, we described a protocol that correlates a precise map of fluorescent fusion proteins localized using three-dimensional super-resolution optical microscopy with the fine ultrastructural context of three-dimensional electron micrographs. While it achieved the difficult simultaneous objectives of high photoactivated fluorophore preservation and ultrastructure preservation, it required a super-resolution optical and specialized electron microscope that is not available to many researchers. We present here a faster and more practical protocol with the advantage of a simpler two-dimensional optical (Photoactivated Localization Microscopy (PALM)) and scanning electron microscope (SEM) system that retains the often mutually exclusive attributes of fluorophore preservation and ultrastructure preservation. As before, cryosections were prepared using the Tokuyasu protocol, but the staining protocol was modified to be amenable for use in a standard SEM without the need for focused ion beam ablation. We show the versatility of this technique by labeling different cellular compartments and structures including mitochondrial nucleoids, peroxisomes, and the nuclear lamina. We also demonstrate simultaneous two-color PALM imaging with correlated electron micrographs. Lastly, this technique can be used with small-molecule dyes as demonstrated with actin labeling using phalloidin conjugated to a caged dye. By retaining the dense protein labeling expected for super-resolution microscopy combined with ultrastructural preservation, simplifying the tools required for correlative microscopy, and expanding the number of useful labels we expect this method to be accessible and valuable to a wide variety of researchers.

  10. Transmission electron microscopy in micro-nanoelectronics

    CERN Document Server

    Claverie, Alain

    2013-01-01

    Today, the availability of bright and highly coherent electron sources and sensitive detectors has radically changed the type and quality of the information which can be obtained by transmission electron microscopy (TEM). TEMs are now present in large numbers not only in academia, but also in industrial research centers and fabs.This book presents in a simple and practical way the new quantitative techniques based on TEM which have recently been invented or developed to address most of the main challenging issues scientists and process engineers have to face to develop or optimize sem

  11. Electron Microscopy of Nanostructures in Cells

    DEFF Research Database (Denmark)

    Købler, Carsten

    with cells is therefore increasingly more relevant from both an engineering and a toxicological viewpoint. My work involves developing and exploring electron microscopy (EM) for imaging nanostructures in cells, for the purpose of understanding nanostructure-cell interactions in terms of their possibilities...... in science and concerns in toxicology. In the present work, EM methods for imaging nanostructure-cell interactions have been explored, and the complex interactions documented and ordered. In particular the usability of the focused ion beam scanning electron microscope (FIB-SEM) was explored. Using EM...

  12. Transmission Electron Microscopy and Diffractometry of Materials

    CERN Document Server

    Fultz, Brent

    2013-01-01

    This book explains concepts of transmission electron microscopy (TEM) and x-ray diffractometry (XRD) that are important for the characterization of materials. The fourth edition adds important new techniques of TEM such as electron tomography, nanobeam diffraction, and geometric phase analysis. A new chapter on neutron scattering completes the trio of x-ray, electron and neutron diffraction. All chapters were updated and revised for clarity. The book explains the fundamentals of how waves and wavefunctions interact with atoms in solids, and the similarities and differences of using x-rays, electrons, or neutrons for diffraction measurements. Diffraction effects of crystalline order, defects, and disorder in materials are explained in detail. Both practical and theoretical issues are covered. The book can be used in an introductory-level or advanced-level course, since sections are identified by difficulty. Each chapter includes a set of problems to illustrate principles, and the extensive Appendix includes la...

  13. Metallothioneins for correlative light and electron microscopy.

    Science.gov (United States)

    Fernández de Castro, Isabel; Sanz-Sánchez, Laura; Risco, Cristina

    2014-01-01

    Structural biologists have been working for decades on new strategies to identify proteins in cells unambiguously. We recently explored the possibilities of using the small metal-binding protein, metallothionein (MT), as a tag to detect proteins in transmission electron microscopy. It had been reported that, when fused with a protein of interest and treated in vitro with gold salts, a single MT tag will build an electron-dense gold cluster ~1 nm in diameter; we provided proof of this principle by demonstrating that MT can be used to detect intracellular proteins in bacteria and eukaryotic cells. The method, which is compatible with a variety of sample processing techniques, allows specific detection of proteins in cells with exceptional sensitivity. We illustrated the applicability of the technique in a series of studies to visualize the intracellular distribution of bacterial and viral proteins. Immunogold labeling was fundamental to confirm the specificity of the MT-gold method. When proteins were double-tagged with green fluorescent protein and MT, direct correlative light and electron microscopy allowed visualization of the same macromolecular complexes with different spatial resolutions. MT-gold tagging might also become a useful tool for mapping proteins into the 3D-density maps produced by (cryo)-electron tomography. New protocols will be needed for double or multiple labeling of proteins, using different versions of MT with fluorophores of different colors. Further research is also necessary to render the MT-gold labeling procedure compatible with immunogold labeling on Tokuyasu cryosections and with cryo-electron microscopy of vitreous sections.

  14. Mechanisms of decoherence in electron microscopy.

    Science.gov (United States)

    Howie, A

    2011-06-01

    The understanding and where possible the minimisation of decoherence mechanisms in electron microscopy were first studied in plasmon loss, diffraction contrast images but are of even more acute relevance in high resolution TEM phase contrast imaging and electron holography. With the development of phase retrieval techniques they merit further attention particularly when their effect cannot be eliminated by currently available energy filters. The roles of electronic excitation, thermal diffuse scattering, transition radiation and bremsstrahlung are examined here not only in the specimen but also in the electron optical column. Terahertz-range aloof beam electronic excitation appears to account satisfactorily for recent observations of decoherence in electron holography. An apparent low frequency divergence can emerge for the calculated classical bremsstrahlung event probability but can be ignored for photon wavelengths exceeding the required coherence distance or path lengths in the equipment. Most bremsstrahlung event probabilities are negligibly important except possibly in large-angle bending magnets or mandolin systems. A more reliable procedure for subtracting thermal diffuse scattering from diffraction pattern intensities is proposed. Copyright © 2010 Elsevier B.V. All rights reserved.

  15. Microfluidic system for transmission electron microscopy

    Energy Technology Data Exchange (ETDEWEB)

    Ring, Elisabeth A [ORNL; De Jonge, Niels [ORNL

    2010-01-01

    We present a microfluidic system that maintains liquid flow in a specimen chamber for (scanning) transmission electron microscope ((S)TEM) imaging. The specimen chamber consists of two ultra-thin silicon nitride windows supported by silicon microchips. They are placed in a specimen holder that seals the sample from the vacuum in the electron microscope, and incorporates tubing to and from the sample connected to a syringe pump outside the microscope. Using results obtained from fluorescence microscopy of microspheres flowing through the system, an equation to characterize the liquid flow through the system was calibrated. Gold nanoparticles of diameters of 30 and 100 nm moving in liquid were imaged with a 200 kV STEM. It was concluded that despite strong influences from Brownian motion, and sensitivity to small changes in the depth of the bypass channel, the electron microscopy flow data matched the calculated flow speed within an order of magnitude. The system allows for rapid (within a minute) liquid exchange, which can potentially be used, for example, to investigate the response of specimens, e.g., eukaryotic-, or bacterial cells, to certain stimuli.

  16. Scanning Electron Microscopy of the Presbylarynx.

    Science.gov (United States)

    Gonçalves, Tatiana Maria; Dos Santos, Daniela Carvalho; Pessin, Adriana Bueno Benito; Martins, Regina Helena Garcia

    2016-06-01

    To describe the findings on the presbylarynx under scanning electron microscopy. Cadaver study. Universidade Estadual Paulista (Botucatu, São Paulo, Brazil). Sixteen vocal folds were removed during necropsies and distributed into 2 age groups: control (n = 8; aged 30-50 years) and elderly (n = 8; aged 75-92 years). The right vocal fold was dissected, fixed in glutaraldehyde 2.5%, and prepared for scanning electron microscopy. The thickness of the epithelium was measured using a scandium morphometric digital program. In the control group, the epithelium had 5 to 7 overlapped cell layers, rare desquamation cells, and little undulation with protruding intercellular junctions. The lamina propria showed a uniform network of collagen and elastic fibers in the superficial layer. A dense network of collagen was identified in the deeper layer. In the elderly group, the epithelium was atrophic (2-3 cells), with more desquamation cells and intercellular junctions delimited by deep sulci. The epithelial thickness was lower in elderly than in controls (mean [SD], 221.64 [145.90] µm vs 41.79 [21.40] µm, respectively). The lamina propria had a dense and irregular distribution of collagen and elastic fibers in the superficial layer. In the deep layers, the collagen fibers formed a true fibrotic and rigid skeleton. Scanning electron microscopy identified several changes in the elderly larynx, differentiating it from the controls. These alterations are probably related to the aging process of the vocal folds. However, the exact interpretation of these findings requires additional studies, even to the molecular level, having the fibroblasts as targets. © American Academy of Otolaryngology—Head and Neck Surgery Foundation 2016.

  17. Metrological investigation of nanostructured polymer surfaces replication using atomic force microscopy

    DEFF Research Database (Denmark)

    Quagliotti, D.; Tosello, G.; Hansen, H. N.

    2015-01-01

    Polymer specimens have been manufactured by injection moulding and measured by atomic force microscopy (AFM) with the aim to investigate the possibility of replicating their surfaces with good fidelity at the sub-μm dimensional scale. Three different cases with surface features in the 100 nm...

  18. Applications of orientation mapping by scanning and transmission electron microscopy

    DEFF Research Database (Denmark)

    Juul Jensen, D.

    1997-01-01

    The potentials of orientation mapping techniques (in the following referred to as OIM) for studies of thermomechanical processes are analysed. Both transmission electron microscopy (TEM) and scanning electron microscopy (SEM) based OIM techniques are considered. Among the thermomechanical processes...

  19. Spatial Resolution in Scanning Electron Microscopy and Scanning Transmission Electron Microscopy Without a Specimen Vacuum Chamber.

    Science.gov (United States)

    Nguyen, Kayla X; Holtz, Megan E; Richmond-Decker, Justin; Muller, David A

    2016-08-01

    A long-standing goal of electron microscopy has been the high-resolution characterization of specimens in their native environment. However, electron optics require high vacuum to maintain an unscattered and focused probe, a challenge for specimens requiring atmospheric or liquid environments. Here, we use an electron-transparent window at the base of a scanning electron microscope's objective lens to separate column vacuum from the specimen, enabling imaging under ambient conditions, without a specimen vacuum chamber. We demonstrate in-air imaging of specimens at nanoscale resolution using backscattered scanning electron microscopy (airSEM) and scanning transmission electron microscopy. We explore resolution and contrast using Monte Carlo simulations and analytical models. We find that nanometer-scale resolution can be obtained at gas path lengths up to 400 μm, although contrast drops with increasing gas path length. As the electron-transparent window scatters considerably more than gas at our operating conditions, we observe that the densities and thicknesses of the electron-transparent window are the dominant limiting factors for image contrast at lower operating voltages. By enabling a variety of detector configurations, the airSEM is applicable to a wide range of environmental experiments including the imaging of hydrated biological specimens and in situ chemical and electrochemical processes.

  20. An electron microscopy appraisal of tensile fracture in metallic glasses

    NARCIS (Netherlands)

    Matthews, D. T. A.; Ocelik, V.; Bronsveld, P. M.; De Hosson, J. Th. M.

    2008-01-01

    Three glass-forming alloy compositions were chosen for ribbon production and subsequent electron microscopy studies. In situ tensile testing with transmission electron microscopy (TEM), followed by ex situ TEM and ex situ scanning electron microscopy (SEM), allowed the deformation processes in tensi

  1. An electron microscopy appraisal of tensile fracture in metallic glasses

    NARCIS (Netherlands)

    Matthews, D. T. A.; Ocelik, V.; Bronsveld, P. M.; De Hosson, J. Th. M.

    Three glass-forming alloy compositions were chosen for ribbon production and subsequent electron microscopy studies. In situ tensile testing with transmission electron microscopy (TEM), followed by ex situ TEM and ex situ scanning electron microscopy (SEM), allowed the deformation processes in

  2. Electron microscopy study of refractory ceramic fibers.

    Science.gov (United States)

    MacKinnon, P A; Lentz, T J; Rice, C H; Lockey, J E; Lemasters, G K; Gartside, P S

    2001-10-01

    In epidemiological studies designed to identify potential health risks of exposures to synthetic vitreous fibers, the characterization of airborne fiber dimensions may be essential for assessing mechanisms of fiber toxicity. Toward this end, air sampling was conducted as part of an industry-wide study of workers potentially exposed to airborne fibrous dusts during the manufacture of refractory ceramic fibers (RCF) and RCF products. Analyses of a subset of samples obtained on the sample filter as well as on the conductive sampling cowl were performed using both scanning electron microscopy (SEM) and transmission electron microscopy (TEM) to characterize dimensions of airborne fibers. Comparison was made of bivariate fiber size distributions (length and diameter) from air samples analyzed by SEM and by TEM techniques. Results of the analyses indicate that RCF size distributions include fibers small enough in diameter ( 60 microm) may go undetected by TEM, as evidenced by the proportion of fibers in this category for TEM and SEM analyses (1% and 5%, respectively). Limitations of the microscopic techniques and differences in fiber-sizing rules for each method are believed to have contributed to the variation among fiber-sizing results. It was concluded from these data that further attempts to characterize RCF exposure in manufacturing and related operations should include analysis by TEM and SEM, since the smallest diameter fibers are not resolved with SEM and the fibers of longer length are not sized by TEM.

  3. Characterization of nanomaterials with transmission electron microscopy

    KAUST Repository

    Anjum, Dalaver H.

    2016-08-01

    The field of nanotechnology is about research and development on materials whose at least one dimension is in the range of 1 to 100 nanometers. In recent years, the research activity for developing nano-materials has grown exponentially owing to the fact that they offer better solutions to the challenges faced by various fields such as energy, food, and environment. In this paper, the importance of transmission electron microscopy (TEM) based techniques is demonstrated for investigating the properties of nano-materials. Specifically the nano-materials that are investigated in this report include gold nano-particles (Au-NPs), silver atom-clusters (Ag-ACs), tantalum single-atoms (Ta-SAs), carbon materials functionalized with iron cobalt (Fe-Co) NPs and titania (TiO2) NPs, and platinum loaded Ceria (Pt-CeO2) Nano composite. TEM techniques that are employed to investigate nano-materials include aberration corrected bright-field TEM (BF-TEM), high-angle dark-field scanning TEM (HAADF-STEM), electron energy-loss spectroscopy (EELS), and BF-TEM electron tomography (ET). With the help presented of results in this report, it is proved herein that as many TEM techniques as available in a given instrument are essential for a comprehensive nano-scale analysis of nanomaterials.

  4. 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.

  5. Scanning electron microscopy of molluscum contagiosum*

    Science.gov (United States)

    de Almeida Jr, Hiram Larangeira; Abuchaim, Martha Oliveira; Schneider, Maiko Abel; Marques, Leandra; de Castro, Luis Antônio Suíta

    2013-01-01

    Molluscum contagiosum is a disease caused by a poxvirus. It is more prevalent in children up to 5 years of age. There is a second peak of incidence in young adults. In order to examine its ultrastructure, three lesions were curetted without disruption, cut transversely with a scalpel, and routinely processed for scanning electron microscopy (SEM). The oval structure of molluscum contagiosum could be easily identified. In its core, there was a central umbilication and just below this depression, there was a keratinized tunnel. Under higher magnification, a proliferation similar to the epidermis was seen. Moreover, there were areas of cells disposed like a mosaic. Under higher magnification, rounded structures measuring 0.4 micron could be observed at the end of the keratinized tunnel and on the surface of the lesion. PMID:23539009

  6. Transmission electron microscopy and diffractometry of materials

    CERN Document Server

    Fultz, Brent

    2001-01-01

    This book teaches graduate students the concepts of trans- mission electron microscopy (TEM) and x-ray diffractometry (XRD) that are important for the characterization of materi- als. It emphasizes themes common to both techniques, such as scattering from atoms and the formation and analysis of dif- fraction patterns. It also describes unique aspects of each technique, especially imaging and spectroscopy in the TEM. The textbook thoroughly develops both introductory and ad- vanced-level material, using over 400 accompanying illustra- tions. Problems are provided at the end of each chapter to reinforce key concepts. Simple citatioins of rules are avoi- ded as much as possible, and both practical and theoretical issues are explained in detail. The book can be used as both an introductory and advanced-level graduate text since sec- tions/chapters are sorted according to difficulty and grou- ped for use in quarter and semester courses on TEM and XRD.

  7. Scanning electron microscopy physics of image formation and microanalysis

    CERN Document Server

    Reimer, Ludwig

    1998-01-01

    Scanning Electron Microscopy provides a description of the physics of electron-probe formation and of electron-specimen interations The different imaging and analytical modes using secondary and backscattered electrons, electron-beam-induced currents, X-ray and Auger electrons, electron channelling effects, and cathodoluminescence are discussed to evaluate specific contrasts and to obtain quantitative information

  8. Medipix 2 detector applied to low energy electron microscopy

    NARCIS (Netherlands)

    Gastel, van R.; Sikharulidze, I.; Schramm, S.; Abrahams, J.P.; Poelsema, B.; Tromp, R.M.; Molen, van der S.J.

    2009-01-01

    Low energy electron microscopy (LEEM) and photo-emission electron microscopy (PEEM) traditionally use microchannel plates (MCPs), a phosphor screen and a CCD-camera to record images and diffraction patterns. In recent years, however, MCPs have become a limiting factor for these types of microscopy.

  9. Correlative light and electron microscopy : strategies and applications

    NARCIS (Netherlands)

    Driel, Linda Francina van

    2011-01-01

    Correlative light and electron microscopy (CLEM) refers to the observation of the same structures or ultrastructures with both light microscopy (LM) and electron microscopy (EM). LM provides an overview of the studied material, and enables the quick localization of structures that are fluorescently

  10. Analyzing Lysosome-Related Organelles by Electron Microscopy

    KAUST Repository

    Hurbain, Ilse

    2017-04-29

    Intracellular organelles have a particular morphological signature that can only be appreciated by ultrastructural analysis at the electron microscopy level. Optical imaging and associated methodologies allow to explore organelle localization and their dynamics at the cellular level. Deciphering the biogenesis and functions of lysosomes and lysosome-related organelles (LROs) and their dysfunctions requires their visualization and detailed characterization at high resolution by electron microscopy. Here, we provide detailed protocols for studying LROs by transmission electron microscopy. While conventional electron microscopy and its recent improvements is the method of choice to investigate organelle morphology, immunoelectron microscopy allows to localize organelle components and description of their molecular make up qualitatively and quantitatively.

  11. Electronic cameras for low-light microscopy.

    Science.gov (United States)

    Rasnik, Ivan; French, Todd; Jacobson, Ken; Berland, Keith

    2013-01-01

    This chapter introduces to electronic cameras, discusses the various parameters considered for evaluating their performance, and describes some of the key features of different camera formats. The chapter also presents the basic understanding of functioning of the electronic cameras and how these properties can be exploited to optimize image quality under low-light conditions. Although there are many types of cameras available for microscopy, the most reliable type is the charge-coupled device (CCD) camera, which remains preferred for high-performance systems. If time resolution and frame rate are of no concern, slow-scan CCDs certainly offer the best available performance, both in terms of the signal-to-noise ratio and their spatial resolution. Slow-scan cameras are thus the first choice for experiments using fixed specimens such as measurements using immune fluorescence and fluorescence in situ hybridization. However, if video rate imaging is required, one need not evaluate slow-scan CCD cameras. A very basic video CCD may suffice if samples are heavily labeled or are not perturbed by high intensity illumination. When video rate imaging is required for very dim specimens, the electron multiplying CCD camera is probably the most appropriate at this technological stage. Intensified CCDs provide a unique tool for applications in which high-speed gating is required. The variable integration time video cameras are very attractive options if one needs to acquire images at video rate acquisition, as well as with longer integration times for less bright samples. This flexibility can facilitate many diverse applications with highly varied light levels.

  12. Life cycle of phytoreoviruses visualized by electron microscopy and tomography

    Directory of Open Access Journals (Sweden)

    Naoyuki eMiyazaki

    2013-10-01

    Full Text Available Rice dwarf virus (RDV and Rice gall dwarf virus (RGDV, members of the genus Phytoreovirus in the family Reoviridae, are known as agents of rice disease, because their spread results in substantial economic damage in many Asian countries. These viruses are transmitted via insect vectors, and they multiply both in the plants and in the insect vectors. Structural information about the viruses and their interactions with cellular components in the life cycle are essential for understanding viral infection and replication mechanisms. The life cycle of the viruses involves various cellular events such as cell entry, synthesis of viral genome and proteins, assembly of viral components, viral egress from infected cells, and intra- and inter-cellular transports. This review focuses on the major events underlying the life cycle of phytoreoviruses, which has been visualized by various EM imaging techniques, including cryo-electron microscopy and tomography, and demonstrates the advantage of the advanced EM imaging techniques to investigate the viral infection and replication mechanisms.

  13. Advanced electron microscopy characterization of multimetallic nanoparticles

    Science.gov (United States)

    Khanal, Subarna Raj

    Research in noble metal nanoparticles has led to exciting progress in a versatile array of applications. For the purpose of better tailoring of nanoparticles activities and understanding the correlation between their structures and properties, control over the composition, shape, size and architecture of bimetallic and multimetallic nanomaterials plays an important role on revealing their new or enhanced functions for potentials application. Advance electron microscopy techniques were used to provide atomic scale insights into the structure-properties of different materials: PtPd, Au-Au3Cu, Cu-Pt, AgPd/Pt and AuCu/Pt nanoparticles. The objective of this work is to understand the physical and chemical properties of nanomaterials and describe synthesis, characterization, surface properties and growth mechanism of various bimetallic and multimetallic nanoparticles. The findings have provided us with novel and significant insights into the physical and chemical properties of noble metal nanoparticles. Different synthesis routes allowed us to synthesize bimetallic: Pt-Pd, Au-Au3Cu, Cu-Pt and trimetallic: AgPd/Pt, AuCu/Pt, core-shell and alloyed nanoparticles with monodispersed sizes, controlled shapes and tunable surface properties. For example, we have synthesized the polyhedral PtPd core-shell nanoparticles with octahedral, decahedral, and triangular plates. Decahedral PtPd core-shell structures are novel morphologies for this system. For the first time we fabricated that the Au core and Au3Cu alloyed shell nanoparticles passivated with CuS2 surface layers and characterized by Cs-corrected scanning transmission electron microscopy. The analysis of the high-resolution micrographs reveals that these nanoparticles have decahedral structure with shell periodicity, and that each of the particles is composed by Au core and Au3Cu ordered superlattice alloyed shell surrounded by CuS 2 surface layer. Additionally, we have described both experimental and theoretical methods of

  14. The EIGER detector for low-energy electron microscopy and photoemission electron microscopy.

    Science.gov (United States)

    Tinti, G; Marchetto, H; Vaz, C A F; Kleibert, A; Andrä, M; Barten, R; Bergamaschi, A; Brückner, M; Cartier, S; Dinapoli, R; Franz, T; Fröjdh, E; Greiffenberg, D; Lopez-Cuenca, C; Mezza, D; Mozzanica, A; Nolting, F; Ramilli, M; Redford, S; Ruat, M; Ruder, Ch; Schädler, L; Schmidt, Th; Schmitt, B; Schütz, F; Shi, X; Thattil, D; Vetter, S; Zhang, J

    2017-09-01

    EIGER is a single-photon-counting hybrid pixel detector developed at the Paul Scherrer Institut, Switzerland. It is designed for applications at synchrotron light sources with photon energies above 5 keV. Features of EIGER include a small pixel size (75 µm × 75 µm), a high frame rate (up to 23 kHz), a small dead-time between frames (down to 3 µs) and a dynamic range up to 32-bit. In this article, the use of EIGER as a detector for electrons in low-energy electron microscopy (LEEM) and photoemission electron microscopy (PEEM) is reported. It is demonstrated that, with only a minimal modification to the sensitive part of the detector, EIGER is able to detect electrons emitted or reflected by the sample and accelerated to 8-20 keV. The imaging capabilities are shown to be superior to the standard microchannel plate detector for these types of applications. This is due to the much higher signal-to-noise ratio, better homogeneity and improved dynamic range. In addition, the operation of the EIGER detector is not affected by radiation damage from electrons in the present energy range and guarantees more stable performance over time. To benchmark the detector capabilities, LEEM experiments are performed on selected surfaces and the magnetic and electronic properties of individual iron nanoparticles with sizes ranging from 8 to 22 nm are detected using the PEEM endstation at the Surface/Interface Microscopy (SIM) beamline of the Swiss Light Source.

  15. Quantitative characterization of electron detectors for transmission electron microscopy.

    Science.gov (United States)

    Ruskin, Rachel S; Yu, Zhiheng; Grigorieff, Nikolaus

    2013-12-01

    A new generation of direct electron detectors for transmission electron microscopy (TEM) promises significant improvement over previous detectors in terms of their modulation transfer function (MTF) and detective quantum efficiency (DQE). However, the performance of these new detectors needs to be carefully monitored in order to optimize imaging conditions and check for degradation over time. We have developed an easy-to-use software tool, FindDQE, to measure MTF and DQE of electron detectors using images of a microscope's built-in beam stop. Using this software, we have determined the DQE curves of four direct electron detectors currently available: the Gatan K2 Summit, the FEI Falcon I and II, and the Direct Electron DE-12, under a variety of total dose and dose rate conditions. We have additionally measured the curves for the Gatan US4000 and TVIPS TemCam-F416 scintillator-based cameras. We compare the results from our new method with published curves. Copyright © 2013 Elsevier Inc. All rights reserved.

  16. Low energy electron microscopy imaging using Medipix2 detector

    NARCIS (Netherlands)

    Sikharulidze, I.; Gastel, van R.; Schramm, S.; Abrahams, J.P.; Poelsema, B.; Tromp, R.M.; Molen, van der S.J.

    2011-01-01

    Low Energy Electron Microscopy (LEEM) and Photo-Emission Electron Microscopy (PEEM) predominantly use a combination of microchannel plate (MCP), phosphor screen and optical camera to record images formed by 10–20 keV electrons. We have tested the performance of a LEEM/PEEM instrument with a Medipix2

  17. Low-temperature electron microscopy: techniques and protocols.

    Science.gov (United States)

    Fleck, Roland A

    2015-01-01

    Low-temperature electron microscopy endeavors to provide "solidification of a biological specimen by cooling with the aim of minimal displacement of its components through the use of low temperature as a physical fixation strategy" (Steinbrecht and Zierold, Cryotechniques in biological electron microscopy. Springer-Verlag, Berlin, p 293, 1987). The intention is to maintain confidence that the tissue observed retains the morphology and dimensions of the living material while also ensuring soluble cellular components are not displaced. As applied to both scanning and transmission electron microscopy, cryo-electron microscopy is a strategy whereby the application of low-temperature techniques are used to reduce or remove processing artifacts which are commonly encountered in more conventional room temperature electron microscopy techniques which rely heavily on chemical fixation and heavy metal staining. Often, cryo-electron microscopy allows direct observation of specimens, which have not been stained or chemically fixed.

  18. Electron microscopy in the investigation of asthenozoospermia.

    Science.gov (United States)

    Mobberley, M A

    2010-01-01

    Asthenozoospermia, defined as low sperm motility, is a significant cause of subfertility in men. Its origins are diverse and in some instances cannot be ascertained. However, severely reduced motility can often be associated with abnormalities in the structure of the sperm tails, which can only be detected by transmission electron microscopy (TEM). In this respect, TEM is an important adjunct to the traditional methods of semen analysis. This review examines the development of the current state of knowledge of sperm tail abnormalities. These may be genetic in origin, or they may be acquired as a result of extrinsic factors. At present, consistent molecular markers are not available to characterise many of the genetic defects. However, TEM can distinguish specific defects of genetic origin and the non-specific structural anomalies that are typical of an acquired condition. It can also differentiate sperm structural anomalies from necrospermia, or sperm death, which is another significant cause of asthenozoospermia. In this modern era of assisted reproduction, it is possible in some instances to circumvent the problems of sperm immotility and to achieve fertilisation and pregnancy using intracytoplasmic sperm injection (ICSI). However, because of the possible genetic origin of asthenozoospermia, many scientists working in the field of infertility believe that it is of the utmost importance to investigate the causes of asthenozoospermia. This review considers the continuing relevance of TEM to the evaluation of sperm tail abnormalities in the context of current reproductive techniques.

  19. Electron microscopy and theoretical modeling of cochleates.

    Science.gov (United States)

    Nagarsekar, Kalpa; Ashtikar, Mukul; Thamm, Jana; Steiniger, Frank; Schacher, Felix; Fahr, Alfred; May, Sylvio

    2014-11-11

    Cochleates are self-assembled cylindrical condensates that consist of large rolled-up lipid bilayer sheets and represent a novel platform for oral and systemic delivery of therapeutically active medicinal agents. With few preceding investigations, the physical basis of cochleate formation has remained largely unexplored. We address the structure and stability of cochleates in a combined experimental/theoretical approach. Employing different electron microscopy methods, we provide evidence for cochleates consisting of phosphatidylserine and calcium to be hollow tubelike structures with a well-defined constant lamellar repeat distance and statistically varying inner and outer radii. To rationalize the relation between inner and outer radii, we propose a theoretical model. Based on the minimization of a phenomenological free energy expression containing a bending, adhesion, and frustration contribution, we predict the optimal tube dimensions of a cochleate and estimate ratios of material constants for cochleates consisting of phosphatidylserines with varied hydrocarbon chain structures. Knowing and understanding these ratios will ultimately benefit the successful formulation of cochleates for drug delivery applications.

  20. Imaging Cytoskeleton Components by Electron Microscopy

    Science.gov (United States)

    Svitkina, Tatyana

    2016-01-01

    The cytoskeleton is a complex of detergent-insoluble components of the cytoplasm playing critical roles in cell motility, shape generation, and mechanical properties of a cell. Fibrillar polymers—actin filaments, microtubules, and intermediate filaments—are major constituents of the cytoskeleton, which constantly change their organization during cellular activities. The actin cytoskeleton is especially polymorphic, as actin filaments can form multiple higher order assemblies performing different functions. Structural information about cytoskeleton organization is critical for understanding its functions and mechanisms underlying various forms of cellular activity. Because of the nanometer-scale thickness of cytoskeletal fibers, electron microscopy (EM) is a key tool to determine the structure of the cytoskeleton. This article describes application of rotary shadowing (or metal replica) EM for visualization of the cytoskeleton. The procedure is applicable to thin cultured cells growing on glass coverslips and consists of detergent extraction of cells to expose their cytoskeleton, chemical fixation to provide stability, ethanol dehydration and critical point drying to preserve three-dimensionality, rotary shadowing with platinum to create contrast, and carbon coating to stabilize replicas. This technique provides easily interpretable three-dimensional images, in which individual cytoskeletal fibers are clearly resolved, and individual proteins can be identified by immunogold labeling. More importantly, replica EM is easily compatible with live cell imaging, so that one can correlate the dynamics of a cell or its components, e.g., expressed fluorescent proteins, with high resolution structural organization of the cytoskeleton in the same cell. PMID:26498781

  1. X-ray and electron microscopy of actinide materials.

    Science.gov (United States)

    Moore, Kevin T

    2010-06-01

    Actinide materials demonstrate a wide variety of interesting physical properties in both bulk and nanoscale form. To better understand these materials, a broad array of microscopy techniques have been employed, including transmission electron microscopy (TEM), electron energy-loss spectroscopy (EELS), energy dispersive X-ray spectroscopy (EDXS), high-angle annular dark-field imaging (HAADF), scanning electron microscopy (SEM), wavelength dispersive X-ray spectroscopy (WDXS), electron back scattered diffraction (EBSD), scanning tunneling microscopy (STM), atomic force microscopy (AFM), and scanning transmission X-ray microscopy (STXM). Here these techniques will be reviewed, highlighting advances made in the physics, materials science, chemistry, and biology of actinide materials through microscopy. Construction of a spin-polarized TEM will be discussed, considering its potential for examining the nanoscale magnetic structure of actinides as well as broader materials and devices, such as those for computational magnetic memory. Copyright 2009 Elsevier Ltd. All rights reserved.

  2. Ballistic-electron-emission Microscopy of Semiconductor Heterostructures

    Science.gov (United States)

    Bell, L. Douglas; Narayanamurti, Venkatesh

    1997-01-01

    Balistic-electron-emission microscopy has developed from its beginning as a probe of Schottky barriers into a powerful nanometer-scale method for characterizing semiconductor interfaces and hot-electron transport.

  3. Scanning transmission electron microscopy imaging and analysis

    CERN Document Server

    Pennycook, Stephen J

    2011-01-01

    Provides the first comprehensive treatment of the physics and applications of this mainstream technique for imaging and analysis at the atomic level Presents applications of STEM in condensed matter physics, materials science, catalysis, and nanoscience Suitable for graduate students learning microscopy, researchers wishing to utilize STEM, as well as for specialists in other areas of microscopy Edited and written by leading researchers and practitioners

  4. Quantitative annular dark field electron microscopy using single electron signals.

    Science.gov (United States)

    Ishikawa, Ryo; Lupini, Andrew R; Findlay, Scott D; Pennycook, Stephen J

    2014-02-01

    One of the difficulties in analyzing atomic resolution electron microscope images is that the sample thickness is usually unknown or has to be fitted from parameters that are not precisely known. An accurate measure of thickness, ideally on a column-by-column basis, parameter free, and with single atom accuracy, would be of great value for many applications, such as matching to simulations. Here we propose such a quantification method for annular dark field scanning transmission electron microscopy by using the single electron intensity level of the detector. This method has the advantage that we can routinely quantify annular dark field images operating at both low and high beam currents, and under high dynamic range conditions, which is useful for the quantification of ultra-thin or light-element materials. To facilitate atom counting at the atomic scale we use the mean intensity in an annular dark field image averaged over a primitive cell, with no free parameters to be fitted. To illustrate the potential of our method, we demonstrate counting the number of Al (or N) atoms in a wurtzite-type aluminum nitride single crystal at each primitive cell over the range of 3-99 atoms.

  5. EdU Incorporation for FACS and Microscopy Analysis of DNA Replication in Budding Yeast.

    Science.gov (United States)

    Talarek, Nicolas; Petit, Julie; Gueydon, Elisabeth; Schwob, Etienne

    2015-01-01

    DNA replication is a key determinant of chromosome segregation and stability in eukaryotes. The yeast Saccharomyces cerevisiae has been extensively used for cell cycle studies, yet simple but key parameters such as the fraction of cells in S phase in a population or the subnuclear localization of DNA synthesis have been difficult to gather for this organism. 5-ethynyl-2'-deoxyuridine (EdU) is a thymidine analogue that can be incorporated in vivo and later detected using copper-catalyzed azide alkyne cycloaddition (Click reaction) without prior DNA denaturation. This chapter describes a budding yeast strain and conditions that allow rapid EdU incorporation at moderate extracellular concentrations, followed by its efficient detection for the analysis of DNA replication in single cells by flow cytometry and fluorescence microscopy.

  6. Automated Scanning Electron Microscopy Analysis of Sampled Aerosol

    DEFF Research Database (Denmark)

    Bluhme, Anders Brostrøm; Kling, Kirsten; Mølhave, Kristian

    development of an automated software-based analysis of aerosols using Scanning Electron Microscopy (SEM) and Scanning Transmission Electron Microscopy (STEM) coupled with Energy-Dispersive X-ray Spectroscopy (EDS). The automated analysis will be capable of providing both detailed physical and chemical single...

  7. A MONTE CARLO SIMULATION OF SECONDARY ELECTRON AND BACKSCATTERED ELECTRON IMAGES IN SCANNING ELECTRON MICROSCOPY

    Institute of Scientific and Technical Information of China (English)

    H.M. Li; Z.J. Ding

    2005-01-01

    A new parallel Monte Carlo simulation method of secondary electron (SE) and backscattered electron images (BSE) of scanning electron microscopy (SEM) for a complex geometric structure has been developed. This paper describes briefly the simulation method and the modification to the conventional sampling method for the step length. Example simulation results have been obtained for several artificial structures.

  8. Particles and waves in electron optics and microscopy

    CERN Document Server

    Pozzi, Giulio

    2016-01-01

    Advances in Imaging and Electron Physics merges two long-running serials, Advances in Electronics and Electron Physics and Advances in Optical and Electron Microscopy. The series features extended articles on the physics of electron devices (especially semiconductor devices), particle optics at high and low energies, microlithography, image science, digital image processing, electromagnetic wave propagation, electron microscopy, and the computing methods used in all these domains. * Contains contributions from leading authorities on the subject matter* Informs and updates all the latest developments in the field of imaging and electron physics* Provides practitioners interested in microscopy, optics, image processing, mathematical morphology, electromagnetic fields, electron, and ion emission with a valuable resource* 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 pro...

  9. Quantitative Scanning Transmission Electron Microscopy of Electronic and Nanostructured Materials

    Science.gov (United States)

    Yankovich, Andrew B.

    Electronic and nanostructured materials have been investigated using advanced scanning transmission electron microscopy (STEM) techniques. The first topic is the microstructure of Ga and Sb-doped ZnO. Ga-doped ZnO is a candidate transparent conducting oxide material. The microstructure of GZO thin films grown by MBE under different growth conditions and different substrates were examined using various electron microscopy (EM) techniques. The microstructure, prevalent defects, and polarity in these films strongly depend on the growth conditions and substrate. Sb-doped ZnO nanowires have been shown to be the first route to stable p-type ZnO. Using Z-contrast STEM, I have showed that an unusual microstructure of Sb-decorated head-to-head inversion domain boundaries and internal voids contain all the Sb in the nanowires and cause the p-type conduction. InGaN thin films and InGaN / GaN quantum wells (QW) for light emitting diodes are the second topic. Low-dose Z-contrast STEM, PACBED, and EDS on InGaN QW LED structures grown by MOCVD show no evidence for nanoscale composition variations, contradicting previous reports. In addition, a new extended defect in GaN and InGaN was discovered. The defect consists of a faceted pyramid-shaped void that produces a threading dislocation along the [0001] growth direction, and is likely caused by carbon contamination during growth. Non-rigid registration (NRR) and high-precision STEM of nanoparticles is the final topic. NRR is a new image processing technique that corrects distortions arising from the serial nature of STEM acquisition that previously limited the precision of locating atomic columns and counting the number of atoms in images. NRR was used to demonstrate sub-picometer precision in STEM images of single crystal Si and GaN, the best achieved in EM. NRR was used to measure the atomic surface structure of Pt nanoacatalysts and Au nanoparticles, which revealed new bond length variation phenomenon of surface atoms. In

  10. Value of electron microscopy in the diagnosis of glomerular diseases.

    Science.gov (United States)

    Darouich, Sihem; Goucha, Rym Louzir; Jaafoura, Mohamed Habib; Moussa, Fatma Ben; Zekri, Semy; Maiz, Hédi Ben

    2010-04-01

    To evaluate the contribution of electron microscopy to the final diagnosis of glomerulopathies, the authors established a prospective study during the first semester of 2006. A total of 52 kidney biopsies were performed with 3 samples for light microscopy, immunofluorescence, and electron microscopy. Among these renal biopsies, only 20 were examined with electron microscopy because the diagnosis made on the basis of conventional methods had remained unclear or doubtful. In 18 cases, electron microscopy was undertaken for the investigation of primary kidney disease. The 2 remaining cases were transplant biopsies. In this series of 20 patients, there were 3 children with an average age of 9 years and 17 adults with an average age of 35.5 years. Fifteen patients (75%) were nephrotic. The study revealed that electron microscopy was essential for diagnosis in 8 cases (40%) and was helpful in 12 cases (60%). In conclusion, the results showed that the ultrastructural study provides essential or helpful information in many cases of glomerular diseases, and therefore electron microscopy should be considered an important tool of diagnostic renal pathology. As was recommended, it is important to reserve renal tissue for ultrastructural study unless electron microscopy can be routinely used in all biopsies. Thus, this technique could be performed wherever a renal biopsy has to be ultrastructurally evaluated.

  11. 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.

  12. Transmission electron microscopy physics of image formation and microanalysis

    CERN Document Server

    Reimer, Ludwig

    1993-01-01

    "Transmission Electron Microscopy" presents the theory of image and contrastformation, and the analytical modes in transmission electron microscopy Theprinciples of particle and wave optics of electrons are described Electron-specimen interactions are discussed for evaluating the theory of scattering and phase contrast Also analysed are the kinetical and dynamical theories of electron diffraction and their applications for crystal-structure determination and imaging of lattices and their defects X-ray microanalysis and electron energy-loss spectroscopy are treated as analytical methods The third edition includes a brief discussionof Schottky emission guns, some clarification of minor details, and references to the recent literature

  13. Electron-beam-assisted Scanning Tunneling Microscopy Of Insulating Surfaces

    CERN Document Server

    Bullock, E T

    2000-01-01

    Insulating materials are widely used in electronic devices. Bulk insulators and insulating films pose unique challenges for high resolution study since most commonly used charged particle surface analysis techniques are incompatible with insulating surfaces and materials. A, method of performing scanning tunneling microscopy (STM) on insulating surfaces has been investigated. The method is referred to as electron-beam assisted scanning tunneling microscopy (e-BASTM). It is proposed that by coupling the STM and the scanning electron microscopy (SEM) as one integrated device, that insulating materials may be studied, obtaining both high spatial resolution, and topographic and electronic resolution. The premise of the technique is based on two physical consequences of the interaction of an energetic electron beam (PE) with a material. First, when an electron beam is incident upon a material, low level material electrons are excited into conduction band states. For insulators, with very high secondary electron yi...

  14. Near-infrared branding efficiently correlates light and electron microscopy.

    Science.gov (United States)

    Bishop, Derron; Nikić, Ivana; Brinkoetter, Mary; Knecht, Sharmon; Potz, Stephanie; Kerschensteiner, Martin; Misgeld, Thomas

    2011-06-05

    The correlation of light and electron microscopy of complex tissues remains a major challenge. Here we report near-infrared branding (NIRB), which facilitates such correlation by using a pulsed, near-infrared laser to create defined fiducial marks in three dimensions in fixed tissue. As these marks are fluorescent and can be photo-oxidized to generate electron contrast, they can guide re-identification of previously imaged structures as small as dendritic spines by electron microscopy.

  15. Resolution Versus Error for Computational Electron Microscopy

    Energy Technology Data Exchange (ETDEWEB)

    Luzi, Lorenzo; Stevens, Andrew; Yang, Hao; Browning, Nigel D.

    2017-07-01

    Images that are collected via scanning transmission electron microscopy (STEM) can be undersampled to avoid damage to the specimen while maintaining resolution [1, 2]. We have used BPFA to impute missing data and reduce noise [3]. The reconstruction is typically evaluated using the peak signal-to-noise ratio (PSNR). This measure is too conservative for STEM images and we propose that the Fourier ring correlation (FRC) is used instead to evaluate the reconstruction. We are not concerned with exact reconstruction of the truth image, and therefore PSNR is a conservative estimation of the quality of the reconstruction. Instead, we are concerned with the visual resolution of the image and whether atoms can be distinguished. We have evaluated the reconstruction of a simulated STEM image using the FRC and compared the results with the PSNR measurements. The FRC captures the resolution of the image and is not affected by a large MSE if the atom peaks are still distinguishable. The noisy and reconstructed images are shown in Figure 1. The simulated STEM image was sampled at 100%, 80%, 40%, and 20% of the original pixels to simulate an undersampled scan. The reconstruction was done using BPFA with a patch size of 10 x 10 and no overlapping patches. Not having overlapping patches produces inferior results but they are still acceptable. The dictionary size is 64 and 30 iterations were completed during each reconstruction. The 100% image was denoised instead of reconstructed. Poisson noise was applied to the simulated image with λ values of 500, 50, and 5 to simulate lower imaging dose. The original simulated STEM image was also included in our calculations and was generated using a dose of 1000. The simulated STEM image is 100 by 100 pixels and has essentially no high frequency components. The image reconstruction tends to smooth the data, also resulting in no high frequency components. This causes the FRC of the two images to be large at higher resolutions and may be

  16. Image Resolution in Scanning Transmission Electron Microscopy

    Energy Technology Data Exchange (ETDEWEB)

    Pennycook, S. J.; Lupini, A.R.

    2008-06-26

    Digital images captured with electron microscopes are corrupted by two fundamental effects: shot noise resulting from electron counting statistics and blur resulting from the nonzero width of the focused electron beam. The generic problem of computationally undoing these effects is called image reconstruction and for decades has proved to be one of the most challenging and important problems in imaging science. This proposal concerned the application of the Pixon method, the highest-performance image-reconstruction algorithm yet devised, to the enhancement of images obtained from the highest-resolution electron microscopes in the world, now in operation at Oak Ridge National Laboratory.

  17. Transmission electron microscopy physics of image formation and microanalysis

    CERN Document Server

    Reimer, Ludwig

    1989-01-01

    The aim of this book is to present the theory of image and contrast formation and the analytical modes in transmission electron microscopy The principles of particle and wave optics of electrons are described Electron-specimen interactions are discussed for evaluating the theory of scattering and phase contrast Also discussed are the kinematical and dynamical theories of electron diffraction and their applications for crystal structure determination and imaging of lattice defects X-ray microanalysis and energy-loss spectroscopy are treated as analytical methods The second edition includes discussion of recent progress, especially in the areas of energy-loss spectroscopy, crystal-lattice imaging and reflection electron microscopy

  18. Transmission electron microscopy physics of image formation and microanalysis

    CERN Document Server

    Reimer, Ludwig

    1997-01-01

    Transmission Electron Microscopy presents the theory of image and contrast formation, and the analytical modes in transmission electron microscopy. The principles of particle and wave optics of electrons are described. Electron-specimen interactions are discussed for evaluating the theory of scattering and phase contrast. Also discussed are the kinematical and dynamical theories of electron diffraction and their applications for crystal-structure analysis and imaging of lattices and their defects. X-ray micronanalysis and electron energy-loss spectroscopy are treated as analytical methods. Specimen damage and contamination by electron irradiation limits the resolution for biological and some inorganic specimens. This fourth edition includes discussion of recent progress, especially in the area of Schottky emission guns, convergent-beam electron diffraction, electron tomography, holography and the high resolution of crystal lattices.

  19. Structure of Wet Specimens in Electron Microscopy

    Science.gov (United States)

    Parsons, D. F.

    1974-01-01

    Discussed are past work and recent advances in the use of electron microscopes for viewing structures immersed in gas and liquid. Improved environmental chambers make it possible to examine wet specimens easily. (Author/RH)

  20. Structure of Wet Specimens in Electron Microscopy

    Science.gov (United States)

    Parsons, D. F.

    1974-01-01

    Discussed are past work and recent advances in the use of electron microscopes for viewing structures immersed in gas and liquid. Improved environmental chambers make it possible to examine wet specimens easily. (Author/RH)

  1. Writing silica structures in liquid with scanning transmission electron microscopy.

    Science.gov (United States)

    van de Put, Marcel W P; Carcouët, Camille C M C; Bomans, Paul H H; Friedrich, Heiner; de Jonge, Niels; Sommerdijk, Nico A J M

    2015-02-04

    Silica nanoparticles are imaged in solution with scanning transmission electron microscopy (STEM) using a liquid cell with silicon nitride (SiN) membrane windows. The STEM images reveal that silica structures are deposited in well-defined patches on the upper SiN membranes upon electron beam irradiation. The thickness of the deposits is linear with the applied electron dose. Scanning electron microscopy (SEM) and atomic force microscopy (AFM) demonstrate that the deposited patches are a result of the merging of the original 20 nm-diameter nanoparticles, and that the related surface roughness depends on the electron dose rate used. Using this approach, sub-micrometer scale structures are written on the SiN in liquid by controlling the electron exposure as function of the lateral position.

  2. Stimulated excitation electron microscopy and spectroscopy

    Energy Technology Data Exchange (ETDEWEB)

    Howie, A.

    2015-04-15

    Recent advances in instrumentation for electron optics and spectroscopy have prompted exploration of ultra-low excitations such as phonons, bond vibrations and Johnson noise. These can be excited not just with fast electrons but also thermally or by other external sources of radiation. The near-field theory of electron energy loss and gain provides a convenient platform for analysing these processes. Possibilities for selected phonon mapping and imaging are discussed. Effects should certainly be observable in atomic resolution structure imaging but diffraction contrast imaging could perhaps be more informative. Additional exciting prospects to be explored include the transition from phonon excitation to single atom recoil and the boosting of energy loss and gain signals with tuned laser illumination. - Highlights: • Electron energy gains and losses measure thermal or laser boosting of excitations. • Electron energy gains and losses are conveniently analysed by near field theory. • Diffraction contrast theory is relevant for phonon imaging by electrons. • The transition from phonon excitation to single atom recoil deserves study.

  3. Atomic-level Electron Microscopy of Metal and Alloy Electrocatalysts

    DEFF Research Database (Denmark)

    Deiana, Davide

    by means of ex situ Scanning Transmission Electron Microscopy (STEM) in combination with in situ indirect nanoplasmonic sensing. Secondly, electron microscopy imaging and spectroscopy have been used for the characterisation of novel metal alloy nanoparticle electrocatalysts for the Oxygen Reduction......This thesis presents the application of transmission electron microscopy techniques towards the characterisation of novel metal nanoparticle catalysts. Two main subjects have been covered: first, the sintering-resistance behaviour of monomodal mass-selected Pt cluster catalysts have been studied...... peroxide H2O2. The active surface is predicted to be formed by reactive Pt or Pd atoms surrounded by more inert Hg atoms. Electrochemical measurements on the two catalysts have shown performance exceeding the current state-of-the-art in both forms of extended surface and nanoparticles. Electron microscopy...

  4. Photoemission electron microscopy and scanning electron microscopy of Magnetospirillum magnetotacticum’s magnetosome chains

    Energy Technology Data Exchange (ETDEWEB)

    Keutner, Christoph [Technische Univ. Dortmund, Dortmung (Germany); von Bohlen, Alex [Leibniz-Institut fur Analytische Wissenschaften, Dortmund (Germany); Berges, Ulf [Technische Univ. Dortmund, Dortmung (Germany); Espeter, Philipp [Technische Univ. Dortmund, Dortmung (Germany); Schneider, Claus M. [Peter Grunberg Institut, Julich (Germany); Westphal, Carsten [Technische Univ. Dortmund, Dortmung (Germany)

    2014-10-07

    Magnetotactic bacteria are of great interdisciplinary interest, since a vast field of applications from magnetic recording media to medical nanorobots is conceivable. A key feature for a further understanding is the detailed knowledge about the magnetosome chain within the bacteria. We report on two preparation procedures suitable for UHV experiments in reflective geometry. Further, we present the results of scanning electron microscopy, as well as the first photoemission electron microscopy experiments, both accessing the magnetosomes within intact magnetotactic bacteria and compare these to scanning electron microscopy data from the literature. From the images, we can clearly identify individual magnetosomes within their chains.

  5. In situ transmission electron microscopy for magnetic nanostructures

    DEFF Research Database (Denmark)

    Ngo, Duc-The; Kuhn, Luise Theil

    2016-01-01

    Nanomagnetism is a subject of great interest because of both application and fundamental aspects in which understanding of the physical and electromagnetic structure of magnetic nanostructures is essential to explore the magnetic properties. Transmission electron microscopy (TEM) is a powerful tool......-structure correlation. This paper aims at reviewing and discussing in situ TEM magnetic imaging studies, including Lorentz microscopy and electron holography in TEM, applied to the research of magnetic nanostructures....

  6. Thermal Balloon Endometrial Ablation: Safety Aspects Evaluated by Serosal Temperature, Light Microscopy and Electron Microscopy

    DEFF Research Database (Denmark)

    Andersen, L F; Meinert, L; Junge, Jette

    1998-01-01

    subsequent hysterectomy the extent of thermal damage into the myometrium was assessed by light and electron microscopy. RESULTS: The highest temperature measured on the uterine serosa was 39.1 degrees C. Coagulation of the myometrium adjacent to the endometrium could be demonstrated by light microscopy...... in all patients, with a maximum depth of 11.5 mm. By electron microscopy no influence of heat could be demonstrated beyond 15 mm from the endometrial surface. CONCLUSION: Up to 16 min of thermal balloon endometrial ablation therapy can destroy the endometrium and the submucosal layers. The myometrium...

  7. Thermal Balloon Endometrial Ablation: Safety Aspects Evaluated by Serosal Temperature, Light Microscopy and Electron Microscopy

    DEFF Research Database (Denmark)

    Andersen, L F; Meinert, L; Junge, Jette

    1998-01-01

    subsequent hysterectomy the extent of thermal damage into the myometrium was assessed by light and electron microscopy. RESULTS: The highest temperature measured on the uterine serosa was 39.1 degrees C. Coagulation of the myometrium adjacent to the endometrium could be demonstrated by light microscopy...... in all patients, with a maximum depth of 11.5 mm. By electron microscopy no influence of heat could be demonstrated beyond 15 mm from the endometrial surface. CONCLUSION: Up to 16 min of thermal balloon endometrial ablation therapy can destroy the endometrium and the submucosal layers. The myometrium...

  8. Thermal balloon endometrial ablation: safety aspects evaluated by serosal temperature, light microscopy and electron microscopy

    DEFF Research Database (Denmark)

    Andersen, L F; Meinert, L; Rygaard, Carsten

    1998-01-01

    subsequent hysterectomy the extent of thermal damage into the myometrium was assessed by light and electron microscopy. RESULTS: The highest temperature measured on the uterine serosa was 39.1 degrees C. Coagulation of the myometrium adjacent to the endometrium could be demonstrated by light microscopy...... in all patients, with a maximum depth of 11.5 mm. By electron microscopy no influence of heat could be demonstrated beyond 15 mm from the endometrial surface. CONCLUSION: Up to 16 min of thermal balloon endometrial ablation therapy can destroy the endometrium and the submucosal layers. The myometrium...

  9. High-resolution imaging by scanning electron microscopy of semithin sections in correlation with light microscopy.

    Science.gov (United States)

    Koga, Daisuke; Kusumi, Satoshi; Shodo, Ryusuke; Dan, Yukari; Ushiki, Tatsuo

    2015-12-01

    In this study, we introduce scanning electron microscopy (SEM) of semithin resin sections. In this technique, semithin sections were adhered on glass slides, stained with both uranyl acetate and lead citrate, and observed with a backscattered electron detector at a low accelerating voltage. As the specimens are stained in the same manner as conventional transmission electron microscopy (TEM), the contrast of SEM images of semithin sections was similar to TEM images of ultrathin sections. Using this technique, wide areas of semithin sections were also observed by SEM, without the obstruction of grids, which was inevitable for traditional TEM. This study also applied semithin section SEM to correlative light and electron microscopy. Correlative immunofluorescence microscopy and immune-SEM were performed in semithin sections of LR white resin-embedded specimens using a FluoroNanogold-labeled secondary antibody. Because LR white resin is hydrophilic and electron stable, this resin is suitable for immunostaining and SEM observation. Using correlative microscopy, the precise localization of the primary antibody was demonstrated by fluorescence microscopy and SEM. This method has great potential for studies examining the precise localization of molecules, including Golgi- and ER-associated proteins, in correlation with LM and SEM.

  10. Electron Microscopy of Biological Materials at the Nanometer Scale

    Science.gov (United States)

    Kourkoutis, Lena Fitting; Plitzko, Jürgen M.; Baumeister, Wolfgang

    2012-08-01

    Electron microscopy of biological matter uses three different imaging modalities: (a) electron crystallography, (b) single-particle analysis, and (c) electron tomography. Ideally, these imaging modalities are applied to frozen-hydrated samples to ensure an optimal preservation of the structures under scrutiny. Cryo-electron microscopy of biological matter has made important advances in the past decades. It has become a research tool that further expands the scope of structural research into unique areas of cell and molecular biology, and it could augment the materials research portfolio in the study of soft and hybrid materials. This review addresses how researchers using transmission electron microscopy can derive structural information at high spatial resolution from fully hydrated specimens, despite their sensitivity to ionizing radiation, despite the adverse conditions of high vacuum for samples that have to be kept in aqueous environments, and despite their low contrast resulting from weakly scattering building blocks.

  11. Exploring the third dimension: volume electron microscopy comes of age.

    Science.gov (United States)

    Peddie, Christopher J; Collinson, Lucy M

    2014-06-01

    Groundbreaking advances in volume electron microscopy and specimen preparation are enabling the 3-dimensional visualisation of specimens with unprecedented detail, and driving a gratifying resurgence of interest in the ultrastructural examination of cellular systems. Serial section techniques, previously the domain of specialists, are becoming increasingly automated with the development of systems such as the automatic tape-collecting ultramicrotome, and serial blockface and focused ion beam scanning electron microscopes. These changes are rapidly broadening the scope of biomedical studies to which volume electron microscopy techniques can be applied beyond the brain. Further innovations in microscope design are also in the pipeline, which have the potential to enhance the speed and quality of data collection. The recent introduction of integrated light and electron microscopy systems will revolutionise correlative light and volume electron microscopy studies, by enabling the sequential collection of data from light and electron imaging modalities without intermediate specimen manipulation. In doing so, the acquisition of comprehensive functional information and direct correlation with ultrastructural details within a 3-dimensional reference space will become routine. The prospects for volume electron microscopy are therefore bright, and the stage is set for a challenging and exciting future.

  12. Cryo scanning electron microscopy of Plasmodium falciparum-infected erythrocytes.

    Science.gov (United States)

    Hempel, Casper

    2017-07-01

    Plasmodium falciparum invades erythrocytes as an essential part of their life cycle. While living inside erythrocytes, the parasite remodels the cell's intracellular organization as well as its outer surface. Late trophozoite-stage parasites and schizonts introduce numerous small protrusions on the erythrocyte surface, called knobs. Current methods for studying these knobs include atomic force microscopy and electron microscopy. Standard electron microscopy methods rely on chemical fixation and dehydration modifying cell size. Here, a novel method is presented using rapid freezing and scanning electron microscopy under cryogenic conditions allowing for high resolution and magnification of erythrocytes. This novel technique can be used for precise estimates of knob density and for studies on cytoadhesion. © 2017 APMIS. Published by John Wiley & Sons Ltd.

  13. Telocytes and putative stem cells in the lungs: electron microscopy, electron tomography and laser scanning microscopy.

    Science.gov (United States)

    Popescu, Laurentiu M; Gherghiceanu, Mihaela; Suciu, Laura C; Manole, Catalin G; Hinescu, Mihail E

    2011-09-01

    This study describes a novel type of interstitial (stromal) cell - telocytes (TCs) - in the human and mouse respiratory tree (terminal and respiratory bronchioles, as well as alveolar ducts). TCs have recently been described in pleura, epicardium, myocardium, endocardium, intestine, uterus, pancreas, mammary gland, etc. (see www.telocytes.com ). TCs are cells with specific prolongations called telopodes (Tp), frequently two to three per cell. Tp are very long prolongations (tens up to hundreds of μm) built of alternating thin segments known as podomers (≤ 200 nm, below the resolving power of light microscope) and dilated segments called podoms, which accommodate mitochondria, rough endoplasmic reticulum and caveolae. Tp ramify dichotomously, making a 3-dimensional network with complex homo- and heterocellular junctions. Confocal microscopy reveals that TCs are c-kit- and CD34-positive. Tp release shed vesicles or exosomes, sending macromolecular signals to neighboring cells and eventually modifying their transcriptional activity. At bronchoalveolar junctions, TCs have been observed in close association with putative stem cells (SCs) in the subepithelial stroma. SCs are recognized by their ultrastructure and Sca-1 positivity. Tp surround SCs, forming complex TC-SC niches (TC-SCNs). Electron tomography allows the identification of bridging nanostructures, which connect Tp with SCs. In conclusion, this study shows the presence of TCs in lungs and identifies a TC-SC tandem in subepithelial niches of the bronchiolar tree. In TC-SCNs, the synergy of TCs and SCs may be based on nanocontacts and shed vesicles.

  14. Transmission electron microscopy of mercury metal

    KAUST Repository

    Anjum, Dalaver H.

    2016-03-28

    Summary: Transmission electron microcopy (TEM) analysis of liquid metals, especially mercury (Hg), is difficult to carry out because their specimen preparation poses a daunting task due to the unique surface properties of these metals. This paper reports a cryoTEM study on Hg using a novel specimen preparation technique. Hg metal is mixed with water using sonication and quenched in liquid ethane cryogen. This technique permits research into the morphological, phase and structural properties of Hg at nanoscale dimensions. © 2016 Royal Microscopical Society.

  15. Magnetic Force Microscopy Using Electron-Beam Fabricated Tips

    NARCIS (Netherlands)

    Rührig, M.; Porthun, S.; Lodder, J.C.

    1994-01-01

    We used a new concept of tip preparation for magnetic force microscopy (MFM) proposed recently based on coating electron beam deposited carbon needles with appropriate magnetic thin film materials. In combining the advantages of electron beam fabricated needles with those of already widely used thin

  16. Environmental Transmission Electron Microscopy in an Aberration-Corrected Environment

    DEFF Research Database (Denmark)

    Hansen, Thomas W.; Wagner, Jakob B.

    2012-01-01

    The increasing use of environmental transmission electron microscopy (ETEM) in materials science provides exciting new possibilities for investigating chemical reactions and understanding both the interaction of fast electrons with gas molecules and the effect of the presence of gas on high-resol...

  17. Photon-induced near-field electron microscopy.

    Science.gov (United States)

    Barwick, Brett; Flannigan, David J; Zewail, Ahmed H

    2009-12-17

    In materials science and biology, optical near-field microscopies enable spatial resolutions beyond the diffraction limit, but they cannot provide the atomic-scale imaging capabilities of electron microscopy. Given the nature of interactions between electrons and photons, and considering their connections through nanostructures, it should be possible to achieve imaging of evanescent electromagnetic fields with electron pulses when such fields are resolved in both space (nanometre and below) and time (femtosecond). Here we report the development of photon-induced near-field electron microscopy (PINEM), and the associated phenomena. We show that the precise spatiotemporal overlap of femtosecond single-electron packets with intense optical pulses at a nanostructure (individual carbon nanotube or silver nanowire in this instance) results in the direct absorption of integer multiples of photon quanta (nhomega) by the relativistic electrons accelerated to 200 keV. By energy-filtering only those electrons resulting from this absorption, it is possible to image directly in space the near-field electric field distribution, obtain the temporal behaviour of the field on the femtosecond timescale, and map its spatial polarization dependence. We believe that the observation of the photon-induced near-field effect in ultrafast electron microscopy demonstrates the potential for many applications, including those of direct space-time imaging of localized fields at interfaces and visualization of phenomena related to photonics, plasmonics and nanostructures.

  18. In situ Transmission Electron Microscopy of catalyst sintering

    DEFF Research Database (Denmark)

    DeLaRiva, Andrew T.; Hansen, Thomas Willum; Challa, Sivakumar R.

    2013-01-01

    Recent advancements in the field of electron microscopy, such as aberration correctors, have now been integrated into Environmental Transmission Electron Microscopes (TEMs), making it possible to study the behavior of supported metal catalysts under operating conditions at atomic resolution. Here......, we focus on in situ electron microscopy studies of catalysts that shed light on the mechanistic aspects of catalyst sintering. Catalyst sintering is an important mechanism for activity loss, especially for catalysts that operate at elevated temperatures. Literature from the past decade is reviewed...

  19. Electron microscopy methods in studies of cultural heritage sites

    Science.gov (United States)

    Vasiliev, A. L.; Kovalchuk, M. V.; Yatsishina, E. B.

    2016-11-01

    The history of the development and application of scanning electron microscopy (SEM), transmission electron microscopy (TEM), and energy-dispersive X-ray microanalysis (EDXMA) in studies of cultural heritage sites is considered. In fact, investigations based on these methods began when electron microscopes became a commercial product. Currently, these methods, being developed and improved, help solve many historical enigmas. To date, electron microscopy combined with microanalysis makes it possible to investigate any object, from parchment and wooden articles to pigments, tools, and objects of art. Studies by these methods have revealed that some articles were made by ancient masters using ancient "nanotechnologies"; hence, their comprehensive analysis calls for the latest achievements in the corresponding instrumental methods and sample preparation techniques.

  20. A correlative optical microscopy and scanning electron microscopy approach to locating nanoparticles in brain tumors.

    Science.gov (United States)

    Kempen, Paul J; Kircher, Moritz F; de la Zerda, Adam; Zavaleta, Cristina L; Jokerst, Jesse V; Mellinghoff, Ingo K; Gambhir, Sanjiv S; Sinclair, Robert

    2015-01-01

    The growing use of nanoparticles in biomedical applications, including cancer diagnosis and treatment, demands the capability to exactly locate them within complex biological systems. In this work a correlative optical and scanning electron microscopy technique was developed to locate and observe multi-modal gold core nanoparticle accumulation in brain tumor models. Entire brain sections from mice containing orthotopic brain tumors injected intravenously with nanoparticles were imaged using both optical microscopy to identify the brain tumor, and scanning electron microscopy to identify the individual nanoparticles. Gold-based nanoparticles were readily identified in the scanning electron microscope using backscattered electron imaging as bright spots against a darker background. This information was then correlated to determine the exact location of the nanoparticles within the brain tissue. The nanoparticles were located only in areas that contained tumor cells, and not in the surrounding healthy brain tissue. This correlative technique provides a powerful method to relate the macro- and micro-scale features visible in light microscopy with the nanoscale features resolvable in scanning electron microscopy.

  1. Ion sputtered deposit analysis by electron microscopy

    Energy Technology Data Exchange (ETDEWEB)

    Lundquist, T.R.; Kraus, B.; Swann, P.R. (GATAN, Inc., Warrendale, PA (USA))

    1983-12-15

    The collected deposit formed by sputtering a specimen provides a permanent record of the elemental surface composition. For analysis by X-rays or energy loss in a transmission electron microscope, all the sputtered particles (both ions and neutrals) are collected on a carbon or SiO thin film. Surface analysis can be obtained by exposing different areas of the specimen to the ion beam. Information available in the angular distributions of sputtered particles is retained on the thin film substrate. Depth profiling can be performed by the sequential exposure of different areas of the thin film substrate to the sputtered specimen particles. Examples from stainless steels and silicon compounds are given. The advantage of this ion sputtered deposit analysis (ISDA) technique, apart from its collection efficiency, is its ability to store permanently all the elemental information obtained from a particular experiment. This information can then be processed in a parallel or serial fashion at any time after the sputtering experiment.

  2. Scanning electron microscopy of bacteria Tetrasphaera duodecadis.

    Science.gov (United States)

    Arroyo, E; Enríquez, L; Sánchez, A; Ovalle, M; Olivas, A

    2014-01-01

    This study reports the characterization of the Tetrasphaera duodecadis bacteria and the techniques used therein. In order to evaluate the morphological characteristics of the T. duodecadis bacteria scanning electron microscope (SEM) was used throughout its different growth stages. These microorganisms were grown in vitamin B12 broths with 1% tryptone, 0.2% yeast extract, and 0.1% glucose. The turbidimetric method was employed for the determination of bacterial concentration and growth curve. The SEM results show small agglomerates of 0.8 ± 0.05 µm during the lag phase, and rod-like shapes during the exponential phase with similar shapes in the stationary phase.

  3. Scanning electron microscopy of Dalkon Shield tails.

    Science.gov (United States)

    Bank, H L; Williamson, H O

    1983-09-01

    Scanning electron micrographs of Dalkon Shield tails removed from asymptomatic patients show a variety of microbes and debris throughout their entire length. Apparently, even in undamaged tails, bacterial flora thrive in the protein-rich environment within the multifilament tail. The presence of microbes in the portion of the tail beyond the double knot indicates that an alternative mechanism of microbial transport can occur. Since transient endometritis often occurs immediately after insertion of intrauterine devices, microbes may come in contact with both exposed ends of the multifilament tail and be drawn into the tail by capillary action from the uterine environment down the tail toward the double knot as well as upward from the vagina. Such microorganisms could serve as an inoculum for infection.

  4. Contributed review: Review of integrated correlative light and electron microscopy.

    Science.gov (United States)

    Timmermans, F J; Otto, C

    2015-01-01

    New developments in the field of microscopy enable to acquire increasing amounts of information from large sample areas and at an increased resolution. Depending on the nature of the technique, the information may reveal morphological, structural, chemical, and still other sample characteristics. In research fields, such as cell biology and materials science, there is an increasing demand to correlate these individual levels of information and in this way to obtain a better understanding of sample preparation and specific sample properties. To address this need, integrated systems were developed that combine nanometer resolution electron microscopes with optical microscopes, which produce chemically or label specific information through spectroscopy. The complementary information from electron microscopy and light microscopy presents an opportunity to investigate a broad range of sample properties in a correlated fashion. An important part of correlating the differences in information lies in bridging the different resolution and image contrast features. The trend to analyse samples using multiple correlated microscopes has resulted in a new research field. Current research is focused, for instance, on (a) the investigation of samples with nanometer scale distribution of inorganic and organic materials, (b) live cell analysis combined with electron microscopy, and (c) in situ spectroscopic and electron microscopy analysis of catalytic materials, but more areas will benefit from integrated correlative microscopy.

  5. Contributed Review: Review of integrated correlative light and electron microscopy

    Energy Technology Data Exchange (ETDEWEB)

    Timmermans, F. J.; Otto, C. [Medical Cell Biophysics Group, MIRA Institute, University of Twente, P.O. Box 217, 7500 AE Enschede (Netherlands)

    2015-01-15

    New developments in the field of microscopy enable to acquire increasing amounts of information from large sample areas and at an increased resolution. Depending on the nature of the technique, the information may reveal morphological, structural, chemical, and still other sample characteristics. In research fields, such as cell biology and materials science, there is an increasing demand to correlate these individual levels of information and in this way to obtain a better understanding of sample preparation and specific sample properties. To address this need, integrated systems were developed that combine nanometer resolution electron microscopes with optical microscopes, which produce chemically or label specific information through spectroscopy. The complementary information from electron microscopy and light microscopy presents an opportunity to investigate a broad range of sample properties in a correlated fashion. An important part of correlating the differences in information lies in bridging the different resolution and image contrast features. The trend to analyse samples using multiple correlated microscopes has resulted in a new research field. Current research is focused, for instance, on (a) the investigation of samples with nanometer scale distribution of inorganic and organic materials, (b) live cell analysis combined with electron microscopy, and (c) in situ spectroscopic and electron microscopy analysis of catalytic materials, but more areas will benefit from integrated correlative microscopy.

  6. Ultra-high resolution electron microscopy

    Science.gov (United States)

    Oxley, Mark P.; Lupini, Andrew R.; Pennycook, Stephen J.

    2017-02-01

    The last two decades have seen dramatic advances in the resolution of the electron microscope brought about by the successful correction of lens aberrations that previously limited resolution for most of its history. We briefly review these advances, the achievement of sub-Ångstrom resolution and the ability to identify individual atoms, their bonding configurations and even their dynamics and diffusion pathways. We then present a review of the basic physics of electron scattering, lens aberrations and their correction, and an approximate imaging theory for thin crystals which provides physical insight into the various different imaging modes. Then we proceed to describe a more exact imaging theory starting from Yoshioka’s formulation and covering full image simulation methods using Bloch waves, the multislice formulation and the frozen phonon/quantum excitation of phonons models. Delocalization of inelastic scattering has become an important limiting factor at atomic resolution. We therefore discuss this issue extensively, showing how the full-width-half-maximum is the appropriate measure for predicting image contrast, but the diameter containing 50% of the excitation is an important measure of the range of the interaction. These two measures can differ by a factor of 5, are not a simple function of binding energy, and full image simulations are required to match to experiment. The Z-dependence of annular dark field images is also discussed extensively, both for single atoms and for crystals, and we show that temporal incoherence must be included accurately if atomic species are to be identified through matching experimental intensities to simulations. Finally we mention a few promising directions for future investigation.

  7. Collective electronic effects in scanning probe microscopy

    Science.gov (United States)

    Passian, Ali

    The surface plasmon dispersion relations are calculated for a metal coated dielectric probe above a dielectric half space with and without metal coating. Employing prolate spheroidal coordinate system this configuration was modeled as confocal single-sheeted hyperboloids of revolution superimposed on planar domains. The involved media are characterized by frequency dependent, spatially local dielectric functions. Due to subwavelength dimensions of the region of interest, nonretarded electrodynamics is utilized to derive exact analytical expressions describing the resonant surface modes. The dispersion relations are studied as functions of the parameter that defines the hyperboloidal boundaries of the tip and the corresponding coating, and as functions of the involved coating thicknesses. Both parallel and perpendicular polarizations are considered. The results are simulated numerically and limiting cases are discussed with comparison to the Cartesian thin foil case. Using this new type of probe-substrate configuration, the surface plasmon coupling mechanism is investigated experimentally utilizing a scanning probe microscope, and the signal strength acquired by the probe is measured as a function of the distance between the probe and the sample. This is repeated at three different wavelengths of the incident p-polarized photons used to stimulate surface plasmons in the thin metal foil. The results are compared with the theory. Utilizing the prolate spheroidal coordinate system, the related and relevant problem of the Coulomb interaction of a dielectric probe tip with a uniform field existing above a semiinfinite, homogeneous dielectric substrate was studied. This is of interest in atomic force microscopy when the sample surface is electrically charged. The induced polarization surface charge density and the field distribution at the bounding surface of the dielectric medium with the geometry of a single-sheeted hyperboloid of revolution located above the dielectric

  8. Photon gating in four-dimensional ultrafast electron microscopy.

    Science.gov (United States)

    Hassan, Mohammed T; Liu, Haihua; Baskin, John Spencer; Zewail, Ahmed H

    2015-10-20

    Ultrafast electron microscopy (UEM) is a pivotal tool for imaging of nanoscale structural dynamics with subparticle resolution on the time scale of atomic motion. Photon-induced near-field electron microscopy (PINEM), a key UEM technique, involves the detection of electrons that have gained energy from a femtosecond optical pulse via photon-electron coupling on nanostructures. PINEM has been applied in various fields of study, from materials science to biological imaging, exploiting the unique spatial, energy, and temporal characteristics of the PINEM electrons gained by interaction with a "single" light pulse. The further potential of photon-gated PINEM electrons in probing ultrafast dynamics of matter and the optical gating of electrons by invoking a "second" optical pulse has previously been proposed and examined theoretically in our group. Here, we experimentally demonstrate this photon-gating technique, and, through diffraction, visualize the phase transition dynamics in vanadium dioxide nanoparticles. With optical gating of PINEM electrons, imaging temporal resolution was improved by a factor of 3 or better, being limited only by the optical pulse widths. This work enables the combination of the high spatial resolution of electron microscopy and the ultrafast temporal response of the optical pulses, which provides a promising approach to attain the resolution of few femtoseconds and attoseconds in UEM.

  9. Transmission Electron Microscopy of Itokawa Regolith Grains

    Science.gov (United States)

    Keller, Lindsay P.; Berger, E. L.

    2013-01-01

    Introduction: In a remarkable engineering achievement, the JAXA space agency successfully recovered the Hayabusa space-craft in June 2010, following a non-optimal encounter and sur-face sampling mission to asteroid 25143 Itokawa. These are the first direct samples ever obtained and returned from the surface of an asteroid. The Hayabusa samples thus present a special op-portunity to directly investigate the evolution of asteroidal sur-faces, from the development of the regolith to the study of the effects of space weathering. Here we report on our preliminary TEM measurements on two Itokawa samples. Methods: We were allocated particles RA-QD02-0125 and RA-QD02-0211. Both particles were embedded in low viscosity epoxy and thin sections were prepared using ultramicrotomy. High resolution images and electron diffraction data were ob-tained using a JEOL 2500SE 200 kV field-emission scanning-transmission electron microscope. Quantitative maps and anal-yses were obtained using a Thermo thin-window energy-dispersive x-ray (EDX) spectrometer. Results: Both particles are olivine-rich (Fo70) with µm-sized inclusions of FeS and have microstructurally complex rims. Par-ticle RA-QD02-0125 is rounded and has numerous sub-µm grains attached to its surface including FeS, albite, olivine, and rare melt droplets. Solar flare tracks have not been observed, but the particle is surrounded by a continuous 50 nm thick, stuctur-ally disordered rim that is compositionally similar to the core of the grain. One of the surface adhering grains is pyrrhotite show-ing a S-depleted rim (8-10 nm thick) with nanophase Fe metal grains (<5 nm) decorating the outermost surface. The pyrrhotite displays a complex superstructure in its core that is absent in the S-depleted rim. Particle RA-QD02-0211 contains solar flare particle tracks (2x109 cm-2) and shows a structurally disordered rim 100 nm thick. The track density corresponds to a surface exposure of 103-104 years based on the track production rate

  10. 78 FR 16707 - Certain Electronic Devices Having Placeshifting or Display Replication Functionality and Products...

    Science.gov (United States)

    2013-03-18

    .... International Trade Commission has received a complaint entitled Certain Electronic Devices Having Placeshifting... From the Federal Register Online via the Government Publishing Office ] INTERNATIONAL TRADE COMMISSION Certain Electronic Devices Having Placeshifting or Display Replication Functionality and...

  11. Focused ion beam scanning electron microscopy in biology.

    Science.gov (United States)

    Kizilyaprak, C; Daraspe, J; Humbel, B M

    2014-06-01

    Since the end of the last millennium, the focused ion beam scanning electron microscopy (FIB-SEM) has progressively found use in biological research. This instrument is a scanning electron microscope (SEM) with an attached gallium ion column and the 2 beams, electrons and ions (FIB) are focused on one coincident point. The main application is the acquisition of three-dimensional data, FIB-SEM tomography. With the ion beam, some nanometres of the surface are removed and the remaining block-face is imaged with the electron beam in a repetitive manner. The instrument can also be used to cut open biological structures to get access to internal structures or to prepare thin lamella for imaging by (cryo-) transmission electron microscopy. Here, we will present an overview of the development of FIB-SEM and discuss a few points about sample preparation and imaging.

  12. 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.

  13. Evaluations of carbon nanotube field emitters for electron microscopy

    Science.gov (United States)

    Nakahara, Hitoshi; Kusano, Yoshikazu; Kono, Takumi; Saito, Yahachi

    2009-11-01

    Brightness of carbon nanotube (CNT) emitters was already reported elsewhere. However, brightness of electron emitter is affected by a virtual source size of the emitter, which strongly depends on electron optical configuration around the emitter. In this work, I- V characteristics and brightness of a CNT emitter are measured under a practical field emission electron gun (e-gun) configuration to investigate availability of CNT for electron microscopy. As a result, it is obtained that an emission area of MWNT is smaller than its tip surface area, and the emission area corresponds to a five-membered-ring with 2nd nearest six-membered-rings on the MWNT cap surface. Reduced brightness of MWNT is measured as at least 2.6×109 A/m 2 sr V. It is concluded that even a thick MWNT has enough brightness under a practical e-gun electrode configuration and suitable for electron microscopy.

  14. Evaluations of carbon nanotube field emitters for electron microscopy

    Energy Technology Data Exchange (ETDEWEB)

    Nakahara, Hitoshi, E-mail: nakahara@nagoya-u.jp [Department of Quantum Engineering, Graduate School of Engineering, Nagoya University, Furo-cho, Chikusa-ku, Nagoya 464-8603 (Japan); Kusano, Yoshikazu; Kono, Takumi; Saito, Yahachi [Department of Quantum Engineering, Graduate School of Engineering, Nagoya University, Furo-cho, Chikusa-ku, Nagoya 464-8603 (Japan)

    2009-11-30

    Brightness of carbon nanotube (CNT) emitters was already reported elsewhere. However, brightness of electron emitter is affected by a virtual source size of the emitter, which strongly depends on electron optical configuration around the emitter. In this work, I-V characteristics and brightness of a CNT emitter are measured under a practical field emission electron gun (e-gun) configuration to investigate availability of CNT for electron microscopy. As a result, it is obtained that an emission area of MWNT is smaller than its tip surface area, and the emission area corresponds to a five-membered-ring with 2nd nearest six-membered-rings on the MWNT cap surface. Reduced brightness of MWNT is measured as at least 2.6x10{sup 9} A/m{sup 2} sr V. It is concluded that even a thick MWNT has enough brightness under a practical e-gun electrode configuration and suitable for electron microscopy.

  15. Imaging hydrated microbial extracellular polymers: Comparative analysis by electron microscopy

    Energy Technology Data Exchange (ETDEWEB)

    Dohnalkova, A.C.; Marshall, M. J.; Arey, B. W.; Williams, K. H.; Buck, E. C.; Fredrickson, J. K.

    2011-01-01

    Microbe-mineral and -metal interactions represent a major intersection between the biosphere and geosphere but require high-resolution imaging and analytical tools for investigating microscale associations. Electron microscopy has been used extensively for geomicrobial investigations and although used bona fide, the traditional methods of sample preparation do not preserve the native morphology of microbiological components, especially extracellular polymers. Herein, we present a direct comparative analysis of microbial interactions using conventional electron microscopy approaches of imaging at room temperature and a suite of cryogenic electron microscopy methods providing imaging in the close-to-natural hydrated state. In situ, we observed an irreversible transformation of the hydrated bacterial extracellular polymers during the traditional dehydration-based sample preparation that resulted in their collapse into filamentous structures. Dehydration-induced polymer collapse can lead to inaccurate spatial relationships and hence could subsequently affect conclusions regarding nature of interactions between microbial extracellular polymers and their environment.

  16. Transmission Electron Microscopy Characterization of Helium Bubbles in Aged Plutonium

    Energy Technology Data Exchange (ETDEWEB)

    Schwartz, A J; Wall, M A; Zocco, T G; Blobaum, K M

    2004-11-02

    The self-irradiation damage generated by alpha decay of plutonium results in the formation of lattice defects, helium, and uranium atoms. Over time, microstructural evolution resulting from the self-irradiation may influence the physical and mechanical properties of the material. In order to assess microstructural changes, we have developed and applied procedures for the specimen preparation, handling, and transmission electron microscopy characterization of Pu alloys. These transmission electron microscopy investigations of Pu-Ga alloys ranging in age up to 42-years old reveal the presence of nanometer-sized helium bubbles. The number density of bubbles and the average size have been determined for eight different aged materials.

  17. Electron microscopy of microwave-synthesized rare-earth chromites

    OpenAIRE

    Schmidt, Rainer; Prado-Gonjal, Jesus; Avila, David; Amador, Ulises; Moran, Emilio

    2014-01-01

    The perovskite rare-earth (RE) chromite series (RE)CrO3 (RE = La, Ce, Pr, Nd, Sm, Eu, Gd, Tb, Dy, Y, Ho, Er, Tm, Yb, Lu) has been synthesized in our laboratory using microwave techniques. In this work we will demonstrate how X-ray diffraction (XRD), Rietveld refinement of XRD pattern and complementary High Resolution Transmission Electron Microscopy (HRTEM) were used to confirm that the desired crystal structure had been formed. Field-emission scanning electron microscopy (FE-SEM) gave clear ...

  18. Self-labelling enzymes as universal tags for fluorescence microscopy, super-resolution microscopy and electron microscopy.

    Science.gov (United States)

    Liss, Viktoria; Barlag, Britta; Nietschke, Monika; Hensel, Michael

    2015-12-08

    Research in cell biology demands advanced microscopy techniques such as confocal fluorescence microscopy (FM), super-resolution microscopy (SRM) and transmission electron microscopy (TEM). Correlative light and electron microscopy (CLEM) is an approach to combine data on the dynamics of proteins or protein complexes in living cells with the ultrastructural details in the low nanometre scale. To correlate both data sets, markers functional in FM, SRM and TEM are required. Genetically encoded markers such as fluorescent proteins or self-labelling enzyme tags allow observations in living cells. Various genetically encoded tags are available for FM and SRM, but only few tags are suitable for CLEM. Here, we describe the red fluorescent dye tetramethylrhodamine (TMR) as a multimodal marker for CLEM. TMR is used as fluorochrome coupled to ligands of genetically encoded self-labelling enzyme tags HaloTag, SNAP-tag and CLIP-tag in FM and SRM. We demonstrate that TMR can additionally photooxidize diaminobenzidine (DAB) to an osmiophilic polymer visible on TEM sections, thus being a marker suitable for FM, SRM and TEM. We evaluated various organelle markers with enzymatic tags in mammalian cells labelled with TMR-coupled ligands and demonstrate the use as efficient and versatile DAB photooxidizer for CLEM approaches.

  19. Scanning electron microscopy and transmission electron microscopy study of hot-deformed gamma-TiAl-based alloy microstructure.

    Science.gov (United States)

    Chrapoński, J; Rodak, K

    2006-09-01

    The aim of this work was to assess the changes in the microstructure of hot-deformed specimens made of alloys containing 46-50 at.% Al, 2 at.% Cr and 2 at.% Nb (and alloying additions such as carbon and boron) with the aid of scanning electron microscopy and transmission electron microscopy techniques. After homogenization and heat treatment performed in order to make diverse lamellae thickness, the specimens were compressed at 1000 degrees C. Transmission electron microscopy examinations of specimens after the compression test revealed the presence of heavily deformed areas with a high density of dislocation. Deformation twins were also observed. Dynamically recrystallized grains were revealed. For alloys no. 2 and no. 3, the recovery and recrystallization processes were more extensive than for alloy no. 1.

  20. Transmission electron microscopy of the preclinical phase of experimental phytophotodermatitis

    Directory of Open Access Journals (Sweden)

    Hiram Larangeira de Almeida Jr

    2008-01-01

    Full Text Available OBJECTIVE: To examine the epidermis in induced phytophotodermatitis using transmission electron microscopy in order to detect histologic changes even before lesions are visible by light microscopy. INTRODUCTION: In the first six hours after the experimental induction of phytophotodermatitis, no changes are detectable by light microscopy. Only after 24 hours can keratinocyte necrosis and epidermal vacuolization be detected histologically, and blisters form by 48 hours. METHODS: The dorsum of four adult rats (Rattus norvegicus was manually epilated. After painting the right half of the rat with the peel juice of Tahiti lemon, they were exposed to sunlight for eight minutes under general anesthesia. The left side was used as the control and exposed to sunlight only. Biopsies were performed immediately after photoinduction and one and two hours later, and the tissue was analyzed by transmission electron microscopy. RESULTS: No histological changes were seen on the control side. Immediately after induction, vacuolization in keratinocytes was observed. After one hour, desmosomal changes were also observed in addition to vacuolization. Keratin filaments were not attached to the desmosomal plaque. Free desmosomes and membrane ruptures were also seen. At two hours after induction, similar changes were found, and granular degeneration of keratin was also observed. DISCUSSION: The interaction of sunlight and psoralens generates a photoproduct that damages keratinocyte proteins, leading to keratinocyte necrosis and blister formation. CONCLUSIONS: Transmission electron microscopy can detect vacuolization, lesions of the membrane, and desmosomes in the first two hours after experimental induction of phytophotodermatitis.

  1. A direct electron detector for time-resolved MeV electron microscopy

    Energy Technology Data Exchange (ETDEWEB)

    Vecchione, T.; Denes, P.; Jobe, R. K.; Johnson, I. J.; Joseph, J. M.; Li, R. K.; Perazzo, A.; Shen, X.; Wang, X. J.; Weathersby, S. P.; Yang, J.; Zhang, D.

    2017-03-01

    The introduction of direct electron detectors enabled the structural biology revolution of cryogenic electron microscopy. Direct electron detectors are now expected to have a similarly dramatic impact on time-resolved MeV electron microscopy, particularly by enabling both spatial and temporal jitter correction. Here we report on the commissioning of a direct electron detector for time-resolved MeV electron microscopy. The direct electron detector demonstrated MeV single electron sensitivity and is capable of recording megapixel images at 180 Hz. The detector has a 15-bit dynamic range, better than 30-μmμm spatial resolution and less than 20 analogue-to-digital converter count RMS pixel noise. The unique capabilities of the direct electron detector and the data analysis required to take advantage of these capabilities are presented. The technical challenges associated with generating and processing large amounts of data are also discussed.

  2. Progress & perspectives for atomic-resolution electron microscopy

    OpenAIRE

    Smith, David J.

    2010-01-01

    The transmission electron microscope (TEM) has evolved into a highly sophisticated instrument that is ideally suited to the characterization of advanced materials. Atomic-level information is routinely accessible using both fixed-beam and scanning TEMs. This report briefly considers developments in the field of atomic-resolution electron microscopy. Recent activities include renewed attention to on-line microscope control (‘autotuning’), and assessment and correction of aberrations. Aberratio...

  3. Detective quantum efficiency of electron area detectors in electron microscopy

    Energy Technology Data Exchange (ETDEWEB)

    McMullan, G., E-mail: gm2@mrc-lmb.cam.ac.uk [MRC Laboratory of Molecular Biology, Hills Road, Cambridge CB2 0QH (United Kingdom); Chen, S.; Henderson, R.; Faruqi, A.R. [MRC Laboratory of Molecular Biology, Hills Road, Cambridge CB2 0QH (United Kingdom)

    2009-08-15

    Recent progress in detector design has created the need for a careful side-by-side comparison of the modulation transfer function (MTF) and resolution-dependent detective quantum efficiency (DQE) of existing electron detectors with those of detectors based on new technology. We present MTF and DQE measurements for four types of detector: Kodak SO-163 film, TVIPS 224 charge coupled device (CCD) detector, the Medipix2 hybrid pixel detector, and an experimental direct electron monolithic active pixel sensor (MAPS) detector. Film and CCD performance was measured at 120 and 300 keV, while results are presented for the Medipix2 at 120 keV and for the MAPS detector at 300 keV. In the case of film, the effects of electron backscattering from both the holder and the plastic support have been investigated. We also show that part of the response of the emulsion in film comes from light generated in the plastic support. Computer simulations of film and the MAPS detector have been carried out and show good agreement with experiment. The agreement enables us to conclude that the DQE of a backthinned direct electron MAPS detector is likely to be equal to, or better than, that of film at 300 keV.

  4. Detective quantum efficiency of electron area detectors in electron microscopy.

    Science.gov (United States)

    McMullan, G; Chen, S; Henderson, R; Faruqi, A R

    2009-08-01

    Recent progress in detector design has created the need for a careful side-by-side comparison of the modulation transfer function (MTF) and resolution-dependent detective quantum efficiency (DQE) of existing electron detectors with those of detectors based on new technology. We present MTF and DQE measurements for four types of detector: Kodak SO-163 film, TVIPS 224 charge coupled device (CCD) detector, the Medipix2 hybrid pixel detector, and an experimental direct electron monolithic active pixel sensor (MAPS) detector. Film and CCD performance was measured at 120 and 300 keV, while results are presented for the Medipix2 at 120 keV and for the MAPS detector at 300 keV. In the case of film, the effects of electron backscattering from both the holder and the plastic support have been investigated. We also show that part of the response of the emulsion in film comes from light generated in the plastic support. Computer simulations of film and the MAPS detector have been carried out and show good agreement with experiment. The agreement enables us to conclude that the DQE of a backthinned direct electron MAPS detector is likely to be equal to, or better than, that of film at 300 keV.

  5. Microfluidic chip for high resolution transmission electron microscopy

    DEFF Research Database (Denmark)

    2013-01-01

    A Microfluidic chip (100) for transmission electron microscopy has a monolithic body (101) with a front side (102) and a back side (103). The monolithic body (101) comprises an opening (104) on the back side (103) extending in a vertical direction from the back side (103) to a membrane (107...

  6. Quantifying Nanoscale Order in Amorphous Materials via Fluctuation Electron Microscopy

    Science.gov (United States)

    Bogle, Stephanie Nicole

    2009-01-01

    Fluctuation electron microscopy (FEM) has been used to study the nanoscale order in various amorphous materials. The method is explicitly sensitive to 3- and 4-body atomic correlation functions in amorphous materials; this is sufficient to establish the existence of structural order on the nanoscale, even when the radial distribution function…

  7. Collaboration at the Nanoscale: Exploring Viral Genetics with Electron Microscopy

    Science.gov (United States)

    Duboise, S. Monroe; Moulton, Karen D.; Jamison, Jennifer L.

    2009-01-01

    The Maine Science Corps is a project sponsored by the National Science Foundation's (NSF) Graduate Teaching Fellows in K-12 Education (GK-12 ) program. Through this program, the University of Southern Maine's (USM) virology and transmission electron microscopy (TEM) research group provides high school teachers and students in rural areas with…

  8. Detection of parvoviruses in wolf feces by electron microscopy

    Science.gov (United States)

    Muneer, M.A.; Farah, I.O.; Pomeroy, K.A.; Goyal, S.M.; Mech, L.D.

    1988-01-01

    One hundred fifteen wolf (Canis lupus) feces were collected between 1980 and 1984 from northeastern Minnesota and were examined for canine parvovirus by negative contrast electron microscopy. Of these, seven (6%) samples revealed the presence of parvovirus. Some of these viruses were able to grow in cell cultures forming intranuclear inclusion bodies and giant cells.

  9. Ultrastructure of Proechinophthirus zumpti (Anoplura, Echinophthiriidae by scanning electron microscopy

    Directory of Open Access Journals (Sweden)

    Dolores del Carmen Castro

    2002-09-01

    Full Text Available The ultrastructure of Proechinophthirus zumpti Werneck, 1955, mainly the external chorionic features of the egg, is described through electronic microscopy techniques. This species was first cited in Argentina, infesting Arctocephalus australis (Zimmermann, 1873. The morphological adaptations of adults and nymphs are described in both species of Proechinophthirus parasitic on Otariidae: P. fluctus (Ferris, 1916 and P. zumpti.

  10. Metals on BN Studied by High Resolution Transmission Electron Microscopy

    Science.gov (United States)

    Bangert, U.; Zan, R.; Ramasse, Q.; Jalil, Rashid; Riaz, Ibstam; Novoselov, K. S.

    2012-07-01

    Metal impurities, gold and nickel, have been deliberately introduced into boron-nitride (BN) sheets. The structural and topographic properties of doped BN have been studied by aberration corrected scanning transmission electron microscopy (STEM). Analysis revealed that metal atoms cluster preferentially in/on contaminated areas. The metal coverage on BN is almost the same for the same evaporated amount of 1 Å.

  11. Electron microscopy studies on MoS2 nanocrystals

    DEFF Research Database (Denmark)

    Hansen, Lars Pilsgaard

    Industrial-style MoS2-based hydrotreating catalysts are studied using electron microscopy. The MoS2 nanostructures are imaged with single-atom sensitivity to reveal the catalytically important edge structures. Furthermore, the in-situ formation of MoS2 crystals is imaged for the first time....

  12. Fc-mediated immune precipitation. III. Visualization by electron microscopy

    DEFF Research Database (Denmark)

    Møller, NPH; Christiansen, Gunna

    1983-01-01

    Fc-mediated interactions between immune complexes are of major importance for the precipitin reaction. In the present study these interactions were investigated by means of electron microscopy. Keyhole limpet haemocyanin (KLH) was adsorbed to a thin glow charged carbon supporting film and reacted...

  13. Collaboration at the Nanoscale: Exploring Viral Genetics with Electron Microscopy

    Science.gov (United States)

    Duboise, S. Monroe; Moulton, Karen D.; Jamison, Jennifer L.

    2009-01-01

    The Maine Science Corps is a project sponsored by the National Science Foundation's (NSF) Graduate Teaching Fellows in K-12 Education (GK-12 ) program. Through this program, the University of Southern Maine's (USM) virology and transmission electron microscopy (TEM) research group provides high school teachers and students in rural areas with…

  14. Quantifying Nanoscale Order in Amorphous Materials via Fluctuation Electron Microscopy

    Science.gov (United States)

    Bogle, Stephanie Nicole

    2009-01-01

    Fluctuation electron microscopy (FEM) has been used to study the nanoscale order in various amorphous materials. The method is explicitly sensitive to 3- and 4-body atomic correlation functions in amorphous materials; this is sufficient to establish the existence of structural order on the nanoscale, even when the radial distribution function…

  15. Automated data collection in single particle electron microscopy

    Science.gov (United States)

    Tan, Yong Zi; Cheng, Anchi; Potter, Clinton S.; Carragher, Bridget

    2016-01-01

    Automated data collection is an integral part of modern workflows in single particle electron microscopy (EM) research. This review surveys the software packages available for automated single particle EM data collection. The degree of automation at each stage of data collection is evaluated, and the capabilities of the software packages are described. Finally, future trends in automation are discussed. PMID:26671944

  16. Transmission electron microscopy investigation of Bi-2223/Ag tapes

    DEFF Research Database (Denmark)

    Andersen, L.G.; Bals, S.; Tendeloo, G. Van

    2001-01-01

    The microstructure of (Bi,Pb)(2)Sr2Ca2CuOx (Bi-2223) tapes has been investigated by means of transmission electron microscopy (TEM) and high-resolution TEM. The emphasis has been placed on: (1) an examination of the grain morphology and size, (2) grain and colony boundary angles, which are formed...

  17. Microstress contrast in scanning electron acoustic microscopy of ceramics

    Science.gov (United States)

    Cantrell, John H.; Qian, Menglu

    1991-01-01

    A mathematical model of image contrast in scanning electron acoustic microscopy (SEAM) due to the effect of residual stresses in materials is presented. It is found that in regions near the ends of the radial cracks induced by Vickers indentation the SEAM micrographs reveal a rather large variation of the acoustic output signal.

  18. Preparation of Articular Cartilage Specimens for Scanning Electron Microscopy.

    Science.gov (United States)

    Stupina, T A

    2016-08-01

    We developed and adapted a technology for preparation of articular cartilage specimens for scanning electron microscopy. The method includes prefixation processing, fixation, washing, and dehydration of articular cartilage specimens with subsequent treatment in camphene and air-drying. The technological result consists in prevention of deformation of the articular cartilage structures. The method is simpler and cheaper than the known technologies.

  19. The Electron Microscopy eXchange (EMX) initiative

    Science.gov (United States)

    Marabini, Roberto; Ludtke, Steven J.; Murray, Stephen C.; Chiu, Wah; de la Rosa-Trevín, Jose M.; Patwardhan, Ardan; Heymann, J. Bernard; Carazo, Jose M.

    2016-01-01

    Three-dimensional electron microscopy (3DEM) of ice-embedded samples allows the structural analysis of large biological macromolecules close to their native state. Different techniques have been developed during the last forty years to process cryo-electron microscopy (cryo-EM) data. Not surprisingly, success in analysis and interpretation is highly correlated with the continuous development of image processing packages. The field has matured to the point where further progress in data and methods sharing depends on an agreement between the packages on how to describe common image processing tasks. Such standardization will facilitate the use of software as well as seamless collaboration, allowing the sharing of rich information between different platforms. Our aim here is to describe the Electron Microscopy eXchange (EMX) initiative, launched at the 2012 Instruct Image Processing Center Developer Workshop, with the intention of developing a first set of standard conventions for the interchange of information for single-particle analysis (EMX version 1.0). These conventions cover the specification of the metadata for micrograph and particle images, including contrast transfer function (CTF) parameters and particle orientations. EMX v1.0 has already been implemented in the Bsoft, EMAN, Xmipp and Scipion image processing packages. It has been and will be used in the CTF and EMDataBank Validation Challenges respectively. It is also being used in EMPIAR, the Electron Microscopy Pilot Image Archive, which stores raw image data related to the 3DEM reconstructions in EMDB. PMID:26873784

  20. The Electron Microscopy eXchange (EMX) initiative.

    Science.gov (United States)

    Marabini, Roberto; Ludtke, Steven J; Murray, Stephen C; Chiu, Wah; de la Rosa-Trevín, Jose M; Patwardhan, Ardan; Heymann, J Bernard; Carazo, Jose M

    2016-05-01

    Three-dimensional electron microscopy (3DEM) of ice-embedded samples allows the structural analysis of large biological macromolecules close to their native state. Different techniques have been developed during the last forty years to process cryo-electron microscopy (cryo-EM) data. Not surprisingly, success in analysis and interpretation is highly correlated with the continuous development of image processing packages. The field has matured to the point where further progress in data and methods sharing depends on an agreement between the packages on how to describe common image processing tasks. Such standardization will facilitate the use of software as well as seamless collaboration, allowing the sharing of rich information between different platforms. Our aim here is to describe the Electron Microscopy eXchange (EMX) initiative, launched at the 2012 Instruct Image Processing Center Developer Workshop, with the intention of developing a first set of standard conventions for the interchange of information for single-particle analysis (EMX version 1.0). These conventions cover the specification of the metadata for micrograph and particle images, including contrast transfer function (CTF) parameters and particle orientations. EMX v1.0 has already been implemented in the Bsoft, EMAN, Xmipp and Scipion image processing packages. It has been and will be used in the CTF and EMDataBank Validation Challenges respectively. It is also being used in EMPIAR, the Electron Microscopy Pilot Image Archive, which stores raw image data related to the 3DEM reconstructions in EMDB.

  1. Electron microscopy in cell biology: integrating structure and function

    NARCIS (Netherlands)

    Koster, A.J.; Klumperman, J.

    2003-01-01

    Electron microscopy (EM) is at the highest-resolution limit of a spectrum of complementary morphological techniques. When combined with molecular detection methods, EM is the only technique with sufficient resolution to localize proteins to small membrane subdomains in the context of the cell. Recen

  2. Multicolor Electron Microscopy for Simultaneous Visualization of Multiple Molecular Species

    NARCIS (Netherlands)

    Adams, Stephen R; Mackey, Mason R; Ramachandra, Ranjan; Palida Lemieux, Sakina F; Steinbach, Paul; Bushong, Eric A; Butko, Margaret T; Giepmans, Ben N G; Ellisman, Mark H; Tsien, Roger Y

    2016-01-01

    Electron microscopy (EM) remains the primary method for imaging cellular and tissue ultrastructure, although simultaneous localization of multiple specific molecules continues to be a challenge for EM. We present a method for obtaining multicolor EM views of multiple subcellular components. The meth

  3. Electron microscopy studies on MoS2 nanocrystals

    DEFF Research Database (Denmark)

    Hansen, Lars Pilsgaard

    Industrial-style MoS2-based hydrotreating catalysts are studied using electron microscopy. The MoS2 nanostructures are imaged with single-atom sensitivity to reveal the catalytically important edge structures. Furthermore, the in-situ formation of MoS2 crystals is imaged for the first time....

  4. A national facility for biological cryo-electron microscopy

    Energy Technology Data Exchange (ETDEWEB)

    Saibil, Helen R., E-mail: h.saibil@mail.cryst.bbk.ac.uk [Birkbeck College, Malet Street, London WC1E 7HX (United Kingdom); Grünewald, Kay [University of Oxford, Oxford OX3 7BN (United Kingdom); Stuart, David I. [University of Oxford, Oxford OX3 7BN (United Kingdom); Diamond Light Source, Didcot OX11 0DE (United Kingdom); Birkbeck College, Malet Street, London WC1E 7HX (United Kingdom)

    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. Statistiscal Experimental Design for Quantitative Atomic Resolution Transmission Electron Microscopy

    NARCIS (Netherlands)

    Van Aert, S.

    2003-01-01

    Statistical experimental design is applied to set up quantitative atomic resolution transmission electron microscopy experiments. In such experiments, observations of the atomic structure of the object under study are always subject to spontaneous fluctuations. As a result of these fluctuations, the

  6. 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 partner

  7. Scanning electron microscopy physics of image formation and microanalysis

    CERN Document Server

    Reimer, Ludwig

    1985-01-01

    The aim of this book is to outline the physics of image formation, electron­ specimen interactions, imaging modes, the interpretation of micrographs and the use of quantitative modes "in scanning electron microscopy (SEM). lt forms a counterpart to Transmission Electron Microscopy (Vol. 36 of this Springer Series in Optical Sciences) . The book evolved from lectures delivered at the University of Münster and from a German text entitled Raster-Elektronenmikroskopie (Springer-Verlag), published in collaboration with my colleague Gerhard Pfefferkorn. In the introductory chapter, the principles of the SEM and of electron­ specimen interactions are described, the most important imaging modes and their associated contrast are summarized, and general aspects of eiemental analysis by x-ray and Auger electron emission are discussed. The electron gun and electron optics are discussed in Chap. 2 in order to show how an electron probe of small diameter can be formed, how the elec­ tron beam can be blanked at high fre...

  8. High-resolution low-dose scanning transmission electron microscopy.

    Science.gov (United States)

    Buban, James P; Ramasse, Quentin; Gipson, Bryant; Browning, Nigel D; Stahlberg, Henning

    2010-01-01

    During the past two decades instrumentation in scanning transmission electron microscopy (STEM) has pushed toward higher intensity electron probes to increase the signal-to-noise ratio of recorded images. While this is suitable for robust specimens, biological specimens require a much reduced electron dose for high-resolution imaging. We describe here protocols for low-dose STEM image recording with a conventional field-emission gun STEM, while maintaining the high-resolution capability of the instrument. Our findings show that a combination of reduced pixel dwell time and reduced gun current can achieve radiation doses comparable to low-dose TEM.

  9. Three-dimensional volume imaging with electron microscopy toward connectome.

    Science.gov (United States)

    Ohno, Nobuhiko; Katoh, Mitsuhiko; Saitoh, Yurika; Saitoh, Sei; Ohno, Shinichi

    2015-02-01

    Ultrastructural analyses with electron microscopy have provided indispensable information to understand physiology and pathology of the nervous system. Recent advancement in imaging methodology paved the way for complete reconstruction of the neuronal connection map in the central nervous system, which is termed 'connectome' and would provide key insights to understand the functions of the brain. The critical advancement includes serial ultrastructural observation with scanning electron microscopy (SEM) instead of conventional serial sectioning transmission electron microscopy along with specific tissue preparation methods to increase heavy metal deposition for efficient SEM imaging. The advanced imaging methods using SEM have distinct advantages and disadvantages in multiple aspects, such as resolution and imaging speed, and should be selected depending on the observation conditions, such as target tissue sizes, required spatial resolution and necessity for re-observation. Dealing with the huge dataset remained to be a major obstacle, and automation in segmentation and 3D reconstruction would be critical to understand neuronal circuits in a larger volume of the brain. Future improvement in acquisition and analyses of the morphological data obtained with the advanced SEM imaging is awaited to elucidate the significance of whole connectome as the structural basis of the consciousness, intelligence and memory of a subject. © The Author 2014. 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.

  10. Transmission electron microscopy reveals distinct macrophage- and tick cell-specific morphological stages of Ehrlichia chaffeensis.

    Directory of Open Access Journals (Sweden)

    Sarah E Dedonder

    Full Text Available BACKGROUND: Ehrlichia chaffeensis is an emerging tick-borne rickettsial pathogen responsible for human monocytic ehrlichiosis. Despite the induction of an active host immune response, the pathogen has evolved to persist in its vertebrate and tick hosts. Understanding how the organism progresses in tick and vertebrate host cells is critical in identifying effective strategies to block the pathogen transmission. Our recent molecular and proteomic studies revealed differences in numerous expressed proteins of the organism during its growth in different host environments. METHODOLOGY/PRINCIPAL FINDINGS: Transmission electron microscopy analysis was performed to assess morphological changes in the bacterium within macrophages and tick cells. The stages of pathogen progression observed included the attachment of the organism to the host cells, its engulfment and replication within a morulae by binary fission and release of the organisms from infected host cells by complete host cell lysis or by exocytosis. E. chaffeensis grown in tick cells was highly pleomorphic and appears to replicate by both binary fission and filamentous type cell divisions. The presence of Ehrlichia-like inclusions was also observed within the nucleus of both macrophages and tick cells. This observation was confirmed by confocal microscopy and immunoblot analysis. CONCLUSIONS/SIGNIFICANCE: Morphological differences in the pathogen's progression, replication, and processing within macrophages and tick cells provide further evidence that E. chaffeensis employs unique host-cell specific strategies in support of adaptation to vertebrate and tick cell environments.

  11. Electron transparent graphene windows for environmental scanning electron microscopy in liquids and dense gases.

    Science.gov (United States)

    Stoll, Joshua D; Kolmakov, Andrei

    2012-12-21

    Due to its ultrahigh electron transmissivity in a wide electron energy range, molecular impermeability, high electrical conductivity and excellent mechanical stiffness, suspended graphene membranes appear to be a nearly ideal window material for in situ (in vivo) environmental electron microscopy of nano- and mesoscopic objects (including bio-medical samples) immersed in liquids and/or in dense gaseous media. In this paper, taking advantage of a small modification of the graphene transfer protocol onto metallic and SiN supporting orifices, reusable environmental cells with exchangeable graphene windows have been designed. Using colloidal gold nanoparticles (50 nm) dispersed in water as model objects for scanning electron microscopy in liquids as proof of concept, different conditions for imaging through the graphene membrane were tested. Limiting factors for electron microscopy in liquids, such as electron beam induced water radiolysis and damage of the graphene membrane at high electron doses, are discussed.

  12. Engineering Electrochemical Setups for Electron Microscopy of Liquid Processes

    DEFF Research Database (Denmark)

    Jensen, Eric; Burrows, Andrew

    systems; the electrochemical scanning-electron-microscopy cell (EC-SEM cell) and the TEM chip (version 1 and 2), and verifying that they can be used, for liquid phase experiments, in SEM and TEM. These systems allow imaging of liquids and of objects in the liquid. They also include electrical connection......This work focuses on creating tools for imaging liquid samples at atmospheric pressure and room temperature in two different electron microscopes; the scanning electron microscope (SEM) and the transmission electron microscope (TEM). The main focus of the project was the fabrication of the two...... to the liquid allowing measurement of electrical signals or the application of a voltage to the liquid during imaging. In a SEM, where there is a comparatively large chamber to work with, a polycarbonate holder with micro-fluidic channels was fabricated to contain the liquid. The EC-SEM chip with an electron...

  13. Vibrational and optical spectroscopies integrated with environmental transmission electron microscopy

    Energy Technology Data Exchange (ETDEWEB)

    Picher, Matthieu; Mazzucco, Stefano [Center for Nanoscale Science and Technology, National Institute of Standards and Technology, Gaithersburg, MD 20899-6203 (United States); Institute for Research in Electronics and Applied Physics, University of Maryland, College Park, MD 20740 (United States); Blankenship, Steve [Center for Nanoscale Science and Technology, National Institute of Standards and Technology, Gaithersburg, MD 20899-6203 (United States); Sharma, Renu, E-mail: renu.sharma@nist.gov [Center for Nanoscale Science and Technology, National Institute of Standards and Technology, Gaithersburg, MD 20899-6203 (United States)

    2015-03-15

    Here, we present a measurement platform for collecting multiple types of spectroscopy data during high-resolution environmental transmission electron microscopy observations of dynamic processes. Such coupled measurements are made possible by a broadband, high-efficiency, free-space optical system. The critical element of the system is a parabolic mirror, inserted using an independent hollow rod and placed below the sample holder which can focus a light on the sample and/or collect the optical response. We demonstrate the versatility of this optical setup by using it to combine in situ atomic-scale electron microscopy observations with Raman spectroscopy. The Raman data is also used to measure the local temperature of the observed sample area. Other applications include, but are not limited to: cathodo- and photoluminescence spectroscopy, and use of the laser as a local, high-rate heating source. - Highlights: • Broadband, high-efficiency design adaptable to other electron microscopes. • Raman spectroscopy integrated with environmental transmission electron microscopy. • Raman spectra peak frequency shifts enable measurement of local sample temperature. • Multiple types of optical spectroscopy enabled, e.g. cathodoluminescence.

  14. Low-energy electron beams through ultra-thin foils, applications for electron microscopy

    NARCIS (Netherlands)

    Van Aken, R.H.

    2005-01-01

    This thesis has discussed two electron microscopy applications that make use of ultra-thin foils: the tunnel junction emitter and the low-energy foil corrector. Both applications have in common that the electron beam is sent through the thin foil at low energy. Part of the electrons will scatter in

  15. An adjustable electron achromat for cathode lens microscopy

    Energy Technology Data Exchange (ETDEWEB)

    Tromp, R.M., E-mail: rtromp@us.ibm.com [IBM T.J. Watson Research Center, 1101 Kitchawan Road, Yorktown Heights, NY 10598 (United States); Leiden Institute of Physics, Kamerlingh Onnes Laboratory, Niels Bohrweg 2, 2333 CA Leiden (Netherlands)

    2015-12-15

    Chromatic aberration correction in light optics began with the invention of a two-color-corrected achromatic crown/flint lens doublet by Chester Moore Hall in 1730. Such color correction is necessary because any single glass shows dispersion (i.e. its index of refraction changes with wavelength), which can be counteracted by combining different glasses with different dispersions. In cathode lens microscopes (such as Photo Electron Emission Microscopy – PEEM) we encounter a similar situation, where the chromatic aberration coefficient of the cathode lens shows strong dispersion, i.e. depends (non-linearly) on the energy with which the electrons leave the sample. Here I show how a cathode lens in combination with an electron mirror can be configured as an adjustable electron achromat. The lens/mirror combination can be corrected at two electron energies by balancing the settings of the electron mirror against the settings of the cathode lens. The achromat can be adjusted to deliver optimum performance, depending on the requirements of a specific experiment. Going beyond the achromat, an apochromat would improve resolution and transmission by a very significant margin. I discuss the requirements and outlook for such a system, which for now remains a wish waiting for fulfilment. - Highlights: • The properties of cathode objective lens plus electron mirror are discussed. • In analogy with light-optical achromats, cathode lens plus mirror can be configured as an electron achromat. • Unlike light optics, the electron achromat can be adjusted to best fulfill experimental requirements.

  16. 78 FR 73563 - Certain Electronic Devices Having Placeshifting or Display Replication Functionality and Products...

    Science.gov (United States)

    2013-12-06

    ... From the Federal Register Online via the Government Publishing Office INTERNATIONAL TRADE COMMISSION Certain Electronic Devices Having Placeshifting or Display Replication Functionality and Products... in Default; Termination of Investigation AGENCY: U.S. International Trade Commission. ACTION:...

  17. Microfabricated high-bandpass foucault aperture for electron microscopy

    Science.gov (United States)

    Glaeser, Robert; Cambie, Rossana; Jin, Jian

    2014-08-26

    A variant of the Foucault (knife-edge) aperture is disclosed that is designed to provide single-sideband (SSB) contrast at low spatial frequencies but retain conventional double-sideband (DSB) contrast at high spatial frequencies in transmission electron microscopy. The aperture includes a plate with an inner open area, a support extending from the plate at an edge of the open area, a half-circle feature mounted on the support and located at the center of the aperture open area. The radius of the half-circle portion of reciprocal space that is blocked by the aperture can be varied to suit the needs of electron microscopy investigation. The aperture is fabricated from conductive material which is preferably non-oxidizing, such as gold, for example.

  18. Combined time-lapse cinematography and immuno-electron microscopy.

    Science.gov (United States)

    Balfour, B M; Goscicka, T; MacKenzie, J L; Gautam, A; Tate, M; Clark, J

    1990-04-01

    A method was developed to record interactions between mobile non-adherent immunocytes by time-lapse cinematography and then to study the same cells by immuno-electron microscopy, using monoclonal antibodies against surface components. For this purpose a modified stage was designed to fit an inverted microscope. The attachment included a device to cool the culture chamber with N2 gas, a micro-injector for monoclonal antibody and immuno-gold treatment, and two pairs of washing needles to change the medium without disturbance. The technique was first employed to study the formation of aggregates around the antigen-presenting cells in cultures containing cells from hyper-immunized animals. Recently peripheral blood cells from normal subjects and patients with immune deficiency syndromes were stimulated with pokeweed mitogen, cluster formation was recorded, and the cells were processed for immuno-electron microscopy.

  19. Scanning electron microscopy: preparation and imaging for SEM.

    Science.gov (United States)

    Jones, Chris G

    2012-01-01

    Scanning electron microscopy (SEM) has been almost universally applied for the surface examination and characterization of both natural and man-made objects. Although an invasive technique, developments in electron microscopy over the years has given the microscopist a much clearer choice in how invasive the technique will be. With the advent of low vacuum SEM in the 1970s (The environmental cold stage, 1970) and environmental SEM in the late 1980s (J Microsc 160(pt. 1):9-19, 1989), it is now possible in some circumstances to examine samples without preparation. However, for the examination of biological tissue and cells it is still advisable to chemically fix, dehydrate, and coat samples for SEM imaging and analysis. This chapter aims to provide an overview of SEM as an imaging tool, and a general introduction to some of the methods applied for the preparation of samples.

  20. Microfabricated high-bandpass foucault aperture for electron microscopy

    Energy Technology Data Exchange (ETDEWEB)

    Glaeser, Robert; Cambie, Rossana; Jin, Jian

    2014-08-26

    A variant of the Foucault (knife-edge) aperture is disclosed that is designed to provide single-sideband (SSB) contrast at low spatial frequencies but retain conventional double-sideband (DSB) contrast at high spatial frequencies in transmission electron microscopy. The aperture includes a plate with an inner open area, a support extending from the plate at an edge of the open area, a half-circle feature mounted on the support and located at the center of the aperture open area. The radius of the half-circle portion of reciprocal space that is blocked by the aperture can be varied to suit the needs of electron microscopy investigation. The aperture is fabricated from conductive material which is preferably non-oxidizing, such as gold, for example.

  1. Scanning electron microscopy of primate chorionic villi following ultrasonic microdissection.

    Science.gov (United States)

    King, B F

    1991-01-01

    Villi from human, macaque and baboon placentae were subjected to ultrasonication after prolonged osmication, and examined by scanning electron microscopy. The technique was often successful in removing the overlying trophoblast and revealing expanses of the trophoblastic basal lamina, a conclusion corroborated by transmission electron microscopy. These preparations bore a remarkable similarity in appearance to microvascular cast preparations of the fetal vasculature. Relatively straight parallel tubules appeared to correspond in position to the location of fetal vessels in intermediate villi, whereas portions of the basal laminae of terminal villi were in the form of convoluted, branched cylinders similar to SEM images of fetal capillaries of terminal villi. The basal lamina did not have evidence of pores as has been described in some basal laminae.

  2. Diagnostic electron microscopy and the influence of Dr. Juan Rosai.

    Science.gov (United States)

    Wick, Mark R

    2016-09-01

    Transmission electron microscopy (TEM) was introduced by Ruska and Knoll as a laboratory technique in 1933. Thereafter, several decades passed before the methods required for its optimal implementation were fully developed. Early uses of TEM were in Botany, rather than in Medicine; however, isolated publications did catalog the ultrastructural characteristics of several individual human tumor types. Finally, in 1968, Rosai and Rodriguez authored an important article, introducing the concept that TEM could be used for the differential diagnosis of histologically similar neoplasms. This publication heralded the steadily increasing application of TEM in anatomic pathology over the following decade, including continuing contributions by Dr. Juan Rosai. This brief review summarizes his influence on clinical electron microscopy, and lists some of the lesions for which that procedure is still a useful means of analysis.

  3. Using electron microscopy to calculate optical properties of biological samples

    OpenAIRE

    Wu, Wenli; Radosevich, Andrew J.; Eshein, Adam; Nguyen, The-Quyen; Yi, Ji; Cherkezyan, Lusik; Roy, Hemant K.; Szleifer, Igal; Backman, Vadim

    2016-01-01

    The microscopic structural origins of optical properties in biological media are still not fully understood. Better understanding these origins can serve to improve the utility of existing techniques and facilitate the discovery of other novel techniques. We propose a novel analysis technique using electron microscopy (EM) to calculate optical properties of specific biological structures. This method is demonstrated with images of human epithelial colon cell nuclei. The spectrum of anisotropy...

  4. Blotting protein complexes from native gels to electron microscopy grids.

    Science.gov (United States)

    Knispel, Roland Wilhelm; Kofler, Christine; Boicu, Marius; Baumeister, Wolfgang; Nickell, Stephan

    2012-01-08

    We report a simple and generic method for the direct transfer of protein complexes separated by native gel electrophoresis to electron microscopy grids. After transfer, sufficient material remains in the gel for identification and characterization by mass spectrometry. The method should facilitate higher-throughput single-particle analysis by substantially reducing the time needed for protein purification, as demonstrated for three complexes from Thermoplasma acidophilum.

  5. Practical aspects of monochromators developed for transmission electron microscopy

    Science.gov (United States)

    Kimoto, Koji

    2014-01-01

    A few practical aspects of monochromators recently developed for transmission electron microscopy are briefly reviewed. The basic structures and properties of four monochromators, a single Wien filter monochromator, a double Wien filter monochromator, an omega-shaped electrostatic monochromator and an alpha-shaped magnetic monochromator, are outlined. The advantages and side effects of these monochromators in spectroscopy and imaging are pointed out. A few properties of the monochromators in imaging, such as spatial or angular chromaticity, are also discussed. PMID:25125333

  6. SCANNING ELECTRON MICROSCOPY STUDY OF FILLED SILICONE RUBBER

    Institute of Scientific and Technical Information of China (English)

    LI Yufu; YANG Qiyun; LI Guangliang

    1988-01-01

    The fracture surfaces of a number of silicone vulcanizates were investigated by the use of scanning electron microscopy (SEM). It was found that the difference in the presence and absence of filler, the variation of its surface modification as well as the history of thermal aging of the vulcanizates, all of these factors made difference in surface morphology of the fractured surface. This was correlated with the strength of the vulcanizates. The reinforcing effect of filler and the process of fracture were discussed.

  7. Single-particle cryo-electron microscopy of macromolecular assemblies

    OpenAIRE

    Cheng, Kimberley

    2009-01-01

    In this thesis, single-particle cryo-electron microscopy (cryo-EM) was used to study the structure of three macromolecular assemblies: the two hemocyanin isoforms from Rapana thomasiana, the Pyrococcus furiosus chaperonin, and the ribosome from Escherichia coli. Hemocyanins are large respiratory proteins in arthropods and molluscs. Most molluscan hemocyanins exist as two distinct isoforms composed of related polypeptides. In most species the two isoforms differ in terms of their oligomeric st...

  8. In Situ Electron Microscopy of Lactomicroselenium Particles in Probiotic Bacteria

    Directory of Open Access Journals (Sweden)

    Gabor Nagy

    2016-06-01

    Full Text Available Electron microscopy was used to test whether or not (a in statu nascendi synthesized, and in situ measured, nanoparticle size does not differ significantly from the size of nanoparticles after their purification; and (b the generation of selenium is detrimental to the bacterial strains that produce them. Elemental nano-sized selenium produced by probiotic latic acid bacteria was used as a lactomicroselenium (lactomicroSel inhibitor of cell growth in the presence of lactomicroSel, and was followed by time-lapse microscopy. The size of lactomicroSel produced by probiotic bacteria was measured in situ and after isolation and purification. For these measurements the TESLA BS 540 transmission electron microscope was converted from analog (aTEM to digital processing (dTEM, and further to remote-access internet electron microscopy (iTEM. Lactobacillus acidophilus produced fewer, but larger, lactomicroSel nanoparticles (200–350 nm than Lactobacillus casei (L. casei, which generated many, smaller lactomicroSel particles (85–200 nm and grains as a cloudy, less electrodense material. Streptococcus thermophilus cells generated selenoparticles (60–280 nm in a suicidic manner. The size determined in situ in lactic acid bacteria was significantly lower than those measured by scanning electron microscopy after the isolation of lactomicroSel particles obtained from lactobacilli (100–500 nm, but higher relative to those isolated from Streptococcus thermopilus (50–100 nm. These differences indicate that smaller lactomicroSel particles could be more toxic to the producing bacteria themselves and discrepancies in size could have implications with respect to the applications of selenium nanoparticles as prebiotics.

  9. Elimination of Ferromagnetic Particles Aggregation for Investigation by Electron Microscopy

    Directory of Open Access Journals (Sweden)

    О.S. Kuzema

    2011-01-01

    Full Text Available It has been described the device for sample preparation of highly dispersed ferromagnetic powders including micropowders for permanent magnets, magnetic carriers, machine and mechanism components’ wear products contained in lubricants for investigation of these materials by light and electron microscopy. The device eliminates the coalescence of ferromagnetic particles and improves reliability of the results of such objects investigation. The technique of such device application has been described and exemplified for various materials investigation.

  10. Quantitative high resolution electron microscopy of grain boundaries

    Energy Technology Data Exchange (ETDEWEB)

    Campbell, G.H., King, W.E., Cohen, D., Carter, C.B.

    1996-12-12

    The {Sigma}11 (113)/[1{bar 1}0] symmetric tilt grain boundary has been characterized by high resolution transmission electron microscopy. The method by which the images are prepared for analysis is described. The statistics of the image data have been found to follow a normal distribution. The electron-optical imaging parameters used to acquire the image have been determined by nonlinear least-square image simulation optimization within the perfect crystal region of the micrograph. A similar image simulation optimization procedure is used to determine the atom positions which provide the best match between the experimental image and the image simulation.

  11. Electron microscopy of some rock phosphate dissolving bacteria and fungi.

    Science.gov (United States)

    Gaur, A C; Arora, D; Prakash, N

    1979-01-01

    Bacteria Pseudomonas striata, Bacillus polymyxa, B. megaterium and B. pulvifaciens, and fungi Aspergillus awamori, A. niger and Penicillium digitatum dissolve tricalcium phosphate and, much less, Mussorie and Udaipur rock phosphate. The solubilizing power of fungi was higher than that of bacteria, the highest being with A. awamori and A. niger, and with P. striata. Electron microscopy of the various cultures showed an electron-dense layer on the bacterial surface after negative staining. The size of phosphate particles decreased by the microbial action, with tricalcium phosphate from 140--250 to 30--90 nm after three weeks of incubation.

  12. Fixation methods for electron microscopy of human and other liver

    Institute of Scientific and Technical Information of China (English)

    Eddie; Wisse; Filip; Braet; Hans; Duimel; Celien; Vreuls; Ger; Koek; Steven; WM; Olde; Damink; Maartje; AJ; van; den; Broek; Bart; De; Geest; Cees; HC; Dejong; Chise; Tateno; Peter; Frederik

    2010-01-01

    For an electron microscopic study of the liver,expertise and complicated,time-consuming processing of hepatic tissues and cells is needed.The interpretation of electron microscopy(EM) images requires knowledge of the liver fine structure and experience with the numerous artifacts in fixation,embedding,sectioning,contrast staining and microscopic imaging.Hence,the aim of this paper is to present a detailed summary of different methods for the preparation of hepatic cells and tissue,for the purpose of preserv...

  13. 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.

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

    Science.gov (United States)

    Müller, E; Gerthsen, D

    2017-02-01

    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 InxGa1-xAs 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.

  15. Transmission electron microscopy physics of image formation and microanalysis

    CERN Document Server

    Reimer, Ludwig

    1984-01-01

    The aim of this book is to outline the physics of image formation, electron­ specimen interactions and image interpretation in transmission electron mic­ roscopy. The book evolved from lectures delivered at the University of Munster and is a revised version of the first part of my earlier book Elek­ tronenmikroskopische Untersuchungs- und Priiparationsmethoden, omitting the part which describes specimen-preparation methods. In the introductory chapter, the different types of electron microscope are compared, the various electron-specimen interactions and their applications are summarized and the most important aspects of high-resolution, analytical and high-voltage electron microscopy are discussed. The optics of electron lenses is discussed in Chapter 2 in order to bring out electron-lens properties that are important for an understanding of the function of an electron microscope. In Chapter 3, the wave optics of elec­ trons and the phase shifts by electrostatic and magnetic fields are introduced; Fresne...

  16. The Role of Electron Microscopy in Studying the Continuum of Changes in Membranous Structures during Poliovirus Infection.

    Science.gov (United States)

    Rossignol, Evan D; Yang, Jie E; Bullitt, Esther

    2015-10-12

    Replication of the poliovirus genome is localized to cytoplasmic replication factories that are fashioned out of a mixture of viral proteins, scavenged cellular components, and new components that are synthesized within the cell due to viral manipulation/up-regulation of protein and phospholipid synthesis. These membranous replication factories are quite complex, and include markers from multiple cytoplasmic cellular organelles. This review focuses on the role of electron microscopy in advancing our understanding of poliovirus RNA replication factories. Structural data from the literature provide the basis for interpreting a wide range of biochemical studies that have been published on virus-induced lipid biosynthesis. In combination, structural and biochemical experiments elucidate the dramatic membrane remodeling that is a hallmark of poliovirus infection. Temporal and spatial membrane modifications throughout the infection cycle are discussed. Early electron microscopy studies of morphological changes following viral infection are re-considered in light of more recent data on viral manipulation of lipid and protein biosynthesis. These data suggest the existence of distinct subcellular vesicle populations, each of which serves specialized roles in poliovirus replication processes.

  17. Transmission electron microscopy and atomic force microscopy characterization of nickel deposition on bacterial cells

    Institute of Scientific and Technical Information of China (English)

    2007-01-01

    Recently bacterial cells have become attractive biological templates for the fabrication of metal nano- structures or nanomaterials due to their inherent small size, various standard geometrical shapes and abundant source. In this paper, nickel-coated bacterial cells (gram-negative bacteria of Escherichia coli) were fabricated via electroless chemical plating. Atomic force microscopy (AFM) and transmission electron microscopy (TEM) characterization results reveal evident morphological difference between bacterial cells before and after deposition with nickel. The bare cells with smooth surface presented transverse outspreading effect at mica surface. Great changes took place in surface roughness for those bacterial cells after metallization. A large number of nickel nanoparticles were observed to be equably distributed at bacterial surface after activation and subsequent metallization. Furthermore, ultra thin section analytic results validated the presence and uniformity of thin nickel coating at bacterial surface after metallization.

  18. Investigation of Nematode Diversity using Scanning Electron Microscopy and Fluorescent Microscopy

    Science.gov (United States)

    Seacor, Taylor; Howell, Carina

    2013-03-01

    Nematode worms account for the vast majority of the animals in the biosphere. They are colossally important to global public health as parasites, and to agriculture both as pests and as beneficial inhabitants of healthy soil. Amphid neurons are the anterior chemosensory neurons in nematodes, mediating critical behaviors including chemotaxis and mating. We are examining the cellular morphology and external anatomy of amphid neurons, using fluorescence microscopy and scanning electron microscopy, respectively, of a wide range of soil nematodes isolated in the wild. We use both classical systematics (e.g. diagnostic keys) and molecular markers (e.g. ribosomal RNA) to classify these wild isolates. Our ultimate aim is to build a detailed anatomical database in order to dissect genetic pathways of neuronal development and function across phylogeny and ecology. Research supported by NSF grants 092304, 0806660, 1058829 and Lock Haven University FPDC grants

  19. Probing cytotoxicity of nanoparticles and organic compounds using scanning proton microscopy, scanning electron microscopy and fluorescence microscopy

    Energy Technology Data Exchange (ETDEWEB)

    Tong Yongpeng [Institute of Nuclear Techniques, Shenzhen University, Nanhai Avenue 3688, Shenzhen 518060 (China)], E-mail: yongpengt@yahoo.com.cn; Li Changming [School of Chemical and Biomedical Engineering, Nanyang Technological University, Singapore 637457 (Singapore); Liang Feng [Institute Pasteur of Shanghai, Chinese Academy of Sciences, Shanghai 200025 (China); Chen Jianmin [Shenzhen Municipal Hospital for Chronic Disease Control and Prevention, Guangdong 518020 (China); Zhang Hong; Liu Guoqing; Sun Huibin [Institute of Nuclear Techniques, Shenzhen University, Nanhai Avenue 3688, Shenzhen 518060 (China); Luong, John H.T. [Biotechnology Research Institute, National Research Council Canada, Montreal, Quebec, H4P 2R2 (Canada)

    2008-12-15

    Scanning proton microscopy, scanning electron microscopy (SEM) and fluorescence microscopy have been used to probe the cytotoxicity effect of benzo[a]pyrene (BaP), ethidium bromide (EB) and nanoparticles (ZnO, Al{sub 2}O{sub 3} and TiO{sub 2}) on a T lymphoblastic leukemia Jurkat cell line. The increased calcium ion (from CaCl{sub 2}) in the culture medium stimulated the accumulation of BaP and EB inside the cell, leading to cell death. ZnO, Al{sub 2}O{sub 3} and TiO{sub 2} nanoparticles, however, showed a protective effect against these two organic compounds. Such inorganic nanoparticles complexed with BaP or EB which became less toxic to the cell. Fe{sub 2}O{sub 3} nanoparticles as an insoluble particle model scavenged by macrophage were investigated in rats. They were scavenged out of the lung tissue about 48 h after infection. This result suggest that some insoluble inorganic nanoparticles of PM (particulate matters) showed protective effects on organic toxins induced acute toxic effects as they can be scavenged by macrophage cells. Whereas, some inorganic ions such as calcium ion in PM may help environmental organic toxins to penetrate cell membrane and induce higher toxic effect.

  20. Low energy electron point source microscopy: beyond imaging.

    Science.gov (United States)

    Beyer, André; Gölzhäuser, Armin

    2010-09-01

    Low energy electron point source (LEEPS) microscopy has the capability to record in-line holograms at very high magnifications with a fairly simple set-up. After the holograms are numerically reconstructed, structural features with the size of about 2 nm can be resolved. The achievement of an even higher resolution has been predicted. However, a number of obstacles are known to impede the realization of this goal, for example the presence of electric fields around the imaged object, electrostatic charging or radiation induced processes. This topical review gives an overview of the achievements as well as the difficulties in the efforts to shift the resolution limit of LEEPS microscopy towards the atomic level. A special emphasis is laid on the high sensitivity of low energy electrons to electrical fields, which limits the structural determination of the imaged objects. On the other hand, the investigation of the electrical field around objects of known structure is very useful for other tasks and LEEPS microscopy can be extended beyond the task of imaging. The determination of the electrical resistance of individual nanowires can be achieved by a proper analysis of the corresponding LEEPS micrographs. This conductivity imaging may be a very useful application for LEEPS microscopes.

  1. Big Data Analytics for Scanning Transmission Electron Microscopy Ptychography

    Science.gov (United States)

    Jesse, S.; Chi, M.; Belianinov, A.; Beekman, C.; Kalinin, S. V.; Borisevich, A. Y.; Lupini, A. R.

    2016-05-01

    Electron microscopy is undergoing a transition; from the model of producing only a few micrographs, through the current state where many images and spectra can be digitally recorded, to a new mode where very large volumes of data (movies, ptychographic and multi-dimensional series) can be rapidly obtained. Here, we discuss the application of so-called “big-data” methods to high dimensional microscopy data, using unsupervised multivariate statistical techniques, in order to explore salient image features in a specific example of BiFeO3 domains. Remarkably, k-means clustering reveals domain differentiation despite the fact that the algorithm is purely statistical in nature and does not require any prior information regarding the material, any coexisting phases, or any differentiating structures. While this is a somewhat trivial case, this example signifies the extraction of useful physical and structural information without any prior bias regarding the sample or the instrumental modality. Further interpretation of these types of results may still require human intervention. However, the open nature of this algorithm and its wide availability, enable broad collaborations and exploratory work necessary to enable efficient data analysis in electron microscopy.

  2. Big Data Analytics for Scanning Transmission Electron Microscopy Ptychography.

    Science.gov (United States)

    Jesse, S; Chi, M; Belianinov, A; Beekman, C; Kalinin, S V; Borisevich, A Y; Lupini, A R

    2016-05-23

    Electron microscopy is undergoing a transition; from the model of producing only a few micrographs, through the current state where many images and spectra can be digitally recorded, to a new mode where very large volumes of data (movies, ptychographic and multi-dimensional series) can be rapidly obtained. Here, we discuss the application of so-called "big-data" methods to high dimensional microscopy data, using unsupervised multivariate statistical techniques, in order to explore salient image features in a specific example of BiFeO3 domains. Remarkably, k-means clustering reveals domain differentiation despite the fact that the algorithm is purely statistical in nature and does not require any prior information regarding the material, any coexisting phases, or any differentiating structures. While this is a somewhat trivial case, this example signifies the extraction of useful physical and structural information without any prior bias regarding the sample or the instrumental modality. Further interpretation of these types of results may still require human intervention. However, the open nature of this algorithm and its wide availability, enable broad collaborations and exploratory work necessary to enable efficient data analysis in electron microscopy.

  3. Scanning transmission electron microscopy imaging dynamics at low accelerating voltages

    Energy Technology Data Exchange (ETDEWEB)

    Lugg, N.R. [School of Physics, University of Melbourne, Parkville, Victoria 3010 (Australia); Findlay, S.D. [Institute of Engineering Innovation, The University of Tokyo, Tokyo 116-0013 (Japan); Shibata, N. [Institute of Engineering Innovation, The University of Tokyo, Tokyo 116-0013 (Japan); PRESTO, Japan Science and Technology Agency, Saitama 332-0012 (Japan); Mizoguchi, T. [Institute of Industrial Science, The University of Tokyo, Tokyo 153-8505 (Japan); D' Alfonso, A.J. [School of Physics, University of Melbourne, Parkville, Victoria 3010 (Australia); Allen, L.J., E-mail: lja@unimelb.edu.au [School of Physics, University of Melbourne, Parkville, Victoria 3010 (Australia); Ikuhara, Y. [Institute of Engineering Innovation, The University of Tokyo, Tokyo 116-0013 (Japan); Nanostructures Research Laboratory, Japan Fine Ceramic Center, Nagoya 456-8587 (Japan); WPI Advanced Institute for Materials Research, Tohoku University, Sendai 980-8577 (Japan)

    2011-07-15

    Motivated by the desire to minimize specimen damage in beam sensitive specimens, there has been a recent push toward using relatively low accelerating voltages (<100kV) in scanning transmission electron microscopy. To complement experimental efforts on this front, this paper seeks to explore the variations with accelerating voltage of the imaging dynamics, both of the channelling of the fast electron and of the inelastic interactions. High-angle annular-dark field, electron energy loss spectroscopic imaging and annular bright field imaging are all considered. -- Highlights: {yields} Both elastic and inelastic scattering in STEM are acceleration voltage dependent. {yields} HAADF, EELS and ABF imaging are assessed with a view to optimum imaging. {yields} Lower accelerating voltages improve STEM EELS contrast in very thin crystals. {yields} Higher accelerating voltages give better STEM EELS contrast in thicker crystals. {yields} At fixed resolution, higher accelerating voltage aids ABF imaging of light elements.

  4. Electron microscopy of primary cell cultures in solution and correlative optical microscopy using ASEM

    Energy Technology Data Exchange (ETDEWEB)

    Hirano, Kazumi; Kinoshita, Takaaki [Laboratory of Cell Biology, Department of Bioinformatics, Faculty of Engineering, Soka University, 1-236 Tangi-machi, Hachioji, Tokyo 192-8577 (Japan); Uemura, Takeshi [Department of Molecular Neurobiology and Pharmacology, Graduate School of Medicine, University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-0033 (Japan); Department of Molecular and Cellular Physiology, Shinshu University School of Medicine, 3-1-1 Asahi, Matsumoto, Nagano 390-8621 (Japan); Motohashi, Hozumi [Department of Gene Expression Regulation, Institute of Development, Aging and Cancer, Tohoku University, 4-1 Seiryo-cho, Aoba-ku, Sendai 980-8575 (Japan); Watanabe, Yohei; Ebihara, Tatsuhiko [Biomedical Research Institute, National Institute of Industrial Science and Technology (AIST), 1-1-1 Higashi, Tsukuba 305-8566 (Japan); Nishiyama, Hidetoshi [JEOL Ltd., 1-2 Musashino 3-chome, Akishima, Tokyo 196-8558 (Japan); Sato, Mari [Biomedical Research Institute, National Institute of Industrial Science and Technology (AIST), 1-1-1 Higashi, Tsukuba 305-8566 (Japan); Suga, Mitsuo [JEOL Ltd., 1-2 Musashino 3-chome, Akishima, Tokyo 196-8558 (Japan); Maruyama, Yuusuke; Tsuji, Noriko M. [Biomedical Research Institute, National Institute of Industrial Science and Technology (AIST), 1-1-1 Higashi, Tsukuba 305-8566 (Japan); Yamamoto, Masayuki [Department of Medical Biochemistry, Tohoku University Graduate School of Medicine, 2-1 Seiryo-cho, Aoba-ku, Sendai 980-8575 (Japan); Nishihara, Shoko, E-mail: shoko@soka.ac.jp [Laboratory of Cell Biology, Department of Bioinformatics, Faculty of Engineering, Soka University, 1-236 Tangi-machi, Hachioji, Tokyo 192-8577 (Japan); Sato, Chikara, E-mail: ti-sato@aist.go.jp [Biomedical Research Institute, National Institute of Industrial Science and Technology (AIST), 1-1-1 Higashi, Tsukuba 305-8566 (Japan)

    2014-08-01

    Correlative light-electron microscopy of cells in a natural environment of aqueous liquid facilitates high-throughput observation of protein complex formation. ASEM allows the inverted SEM to observe the wet sample from below, while an optical microscope observes it from above quasi-simultaneously. The disposable ASEM dish with a silicon nitride (SiN) film window can be coated variously to realize the primary-culture of substrate-sensitive cells in a few milliliters of culture medium in a stable incubator environment. Neuron differentiation, neural networking, proplatelet-formation and phagocytosis were captured by optical or fluorescence microscopy, and imaged at high resolution by gold-labeled immuno-ASEM with/without metal staining. Fas expression on the cell surface was visualized, correlated to the spatial distribution of F-actin. Axonal partitioning was studied using primary-culture neurons, and presynaptic induction by GluRδ2-N-terminus-linked fluorescent magnetic beads was correlated to the presynaptic-marker Bassoon. Further, megakaryocytes secreting proplatelets were captured, and P-selectins with adherence activity were localized to some of the granules present by immuno-ASEM. The phagocytosis of lactic acid bacteria by dendritic cells was also imaged. Based on these studies, ASEM correlative microscopy promises to allow the study of various mesoscopic-scale dynamics in the near future. - Highlights: • In situ correlative light electron microscopy of samples in open solution by ASEM. • Primary cultures for in-solution CLEM by developing SiN-film coating methods • First visualization of fluorescent magnetic beads in aqueous solution by CLEM. • Presynaptic induction of neurons by GluRδ2-N-terminus-coated beads studied by CLEM. • Axonal partitioning, bacterial phagocytosis, platelet formation imaged by CLEM.

  5. Electron microscopy of flatworms standard and cryo-preparation methods.

    Science.gov (United States)

    Salvenmoser, Willi; Egger, Bernhard; Achatz, Johannes G; Ladurner, Peter; Hess, Michael W

    2010-01-01

    Electron microscopy (EM) has long been indispensable for flatworm research, as most of these worms are microscopic in dimension and provide only a handful of characters recognizable by eye or light microscopy. Therefore, major progress in understanding the histology, systematics, and evolution of this animal group relied on methods capable of visualizing ultrastructure. The rise of molecular and cellular biology renewed interest in such ultrastructural research. In the light of recent developments, we offer a best-practice guide for users of transmission EM and provide a comparison of well-established chemical fixation protocols with cryo-processing methods (high-pressure freezing/freeze-substitution, HPF/FS). The organisms used in this study include the rhabditophorans Macrostomum lignano, Polycelis nigra and Dugesia gonocephala, as well as the acoel species Isodiametra pulchra.

  6. Ultrahigh Voltage Electron Microscopy Links Neuroanatomy and Neuroscience/Neuroendocrinology

    Directory of Open Access Journals (Sweden)

    Hirotaka Sakamoto

    2012-01-01

    Full Text Available The three-dimensional (3D analysis of anatomical ultrastructures is extremely important in most fields of biological research. Although it is very difficult to perform 3D image analysis on exact serial sets of ultrathin sections, 3D reconstruction from serial ultrathin sections can generally be used to obtain 3D information. However, this technique can only be applied to small areas of a specimen because of technical and physical difficulties. We used ultrahigh voltage electron microscopy (UHVEM to overcome these difficulties and to study the chemical neuroanatomy of 3D ultrastructures. This methodology, which links UHVEM and light microscopy, is a useful and powerful tool for studying molecular and/or chemical neuroanatomy at the ultrastructural level.

  7. Generation and application of bessel beams in electron microscopy

    Energy Technology Data Exchange (ETDEWEB)

    Grillo, Vincenzo, E-mail: vincenzo.grillo@cnr.it [CNR-Istituto Nanoscienze, Centro S3, Via G Campi 213/a, I-41125 Modena (Italy); CNR-IMEM, Parco Area delle Scienze 37/A, I-43124 Parma (Italy); Harris, Jérémie [Department of Physics, University of Ottawa, 25 Templeton St., Ottawa, Ontario, Canada K1N 6N5 (Canada); Gazzadi, Gian Carlo [CNR-Istituto Nanoscienze, Centro S3, Via G Campi 213/a, I-41125 Modena (Italy); Balboni, Roberto [CNR-IMM Bologna, Via P. Gobetti 101, 40129 Bologna (Italy); Mafakheri, Erfan [Dipartimento di Fisica Informatica e Matematica, Università di Modena e Reggio Emilia, via G Campi 213/a, I-41125 Modena (Italy); Dennis, Mark R. [H.H. Wills Physics Laboratory, University of Bristol, Bristol BS8 1TL (United Kingdom); Frabboni, Stefano [CNR-Istituto Nanoscienze, Centro S3, Via G Campi 213/a, I-41125 Modena (Italy); Dipartimento di Fisica Informatica e Matematica, Università di Modena e Reggio Emilia, via G Campi 213/a, I-41125 Modena (Italy); Boyd, Robert W.; Karimi, Ebrahim [Department of Physics, University of Ottawa, 25 Templeton St., Ottawa, Ontario, Canada K1N 6N5 (Canada)

    2016-07-15

    We report a systematic treatment of the holographic generation of electron Bessel beams, with a view to applications in electron microscopy. We describe in detail the theory underlying hologram patterning, as well as the actual electron-optical configuration used experimentally. We show that by optimizing our nanofabrication recipe, electron Bessel beams can be generated with relative efficiencies reaching 37±3%. We also demonstrate by tuning various hologram parameters that electron Bessel beams can be produced with many visible rings, making them ideal for interferometric applications, or in more highly localized forms with fewer rings, more suitable for imaging. We describe the settings required to tune beam localization in this way, and explore beam and hologram configurations that allow the convergences and topological charges of electron Bessel beams to be controlled. We also characterize the phase structure of the Bessel beams generated with our technique, using a simulation procedure that accounts for imperfections in the hologram manufacturing process. - Highlights: • Bessel beams with different convergence, topological charge, visible fringes are demonstrated. • The relation between the Fresnel hologram and the probe shape is explained by detailed calculations and experiments. • Among the holograms here presented the highest relative efficiency is 37%, the best result ever reached for blazed holograms.

  8. Generation and application of bessel beams in electron microscopy.

    Science.gov (United States)

    Grillo, Vincenzo; Harris, Jérémie; Gazzadi, Gian Carlo; Balboni, Roberto; Mafakheri, Erfan; Dennis, Mark R; Frabboni, Stefano; Boyd, Robert W; Karimi, Ebrahim

    2016-07-01

    We report a systematic treatment of the holographic generation of electron Bessel beams, with a view to applications in electron microscopy. We describe in detail the theory underlying hologram patterning, as well as the actual electron-optical configuration used experimentally. We show that by optimizing our nanofabrication recipe, electron Bessel beams can be generated with relative efficiencies reaching 37±3%. We also demonstrate by tuning various hologram parameters that electron Bessel beams can be produced with many visible rings, making them ideal for interferometric applications, or in more highly localized forms with fewer rings, more suitable for imaging. We describe the settings required to tune beam localization in this way, and explore beam and hologram configurations that allow the convergences and topological charges of electron Bessel beams to be controlled. We also characterize the phase structure of the Bessel beams generated with our technique, using a simulation procedure that accounts for imperfections in the hologram manufacturing process. Copyright © 2016 Elsevier B.V. All rights reserved.

  9. 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.

  10. Electron and Light Microscopy Techniques Suitable for Studying Fatigue Damage in a Crystallized Glass Ceramic

    Science.gov (United States)

    Harrell, Shelley; Zaretsky, Erwin V.

    1961-01-01

    The crystals of Pyroceram are randomly oriented and highly reflective so that standard microscopy techniques are not satisfactory for studying this material. Standard replicating procedures proved difficult to use. New microscopy techniques and procedures have therefore been developed. A method for locating, orienting, and identifying specific areas to be viewed with an electron microscope is described. This method not require any special equipment. Plastic replicas were found to be unsatisfactory because of their tendency to adhere to Pryoceram. This caused them to tear when released or resulted in artifacts. Preshadowed silicon monoxide replicas were satisfactory but required a releasing agent. A method of depositing the releasing agent is described. To polish specimens without evidence of fire-polishing, it was found necessary to use a vibratory polishing technique. Chrome oxide was used as the abrasive and either water or kerosene as the lubricant. Vibratory polishing is extremely slow, but surfaces so polished show no evidence of fire polishing, even when examined by electron microscopy. The most satisfactory etching process used for Pyroceram 9608 consisted of a primary etch of 5 milliliters of hydrochloric acid (concentrated), 5 milliliters of hydrogen fluoride (45 percent), and 45 milliliters of water, and a secondary etch with methyl alcohol replacing the water. Best results were obtained with total etching times from 25 to 30 seconds. Staining of the Pyroceram surface with a Sanford's marker was found to be an expedient way to reduce the glare of reflected light.

  11. Rapid detection of intracellular nanoparticles by electron microscopy

    Directory of Open Access Journals (Sweden)

    Kyoung Hwan Lee

    2010-03-01

    Full Text Available Recently, a number of nanoparticle carriers have provided new platforms for research in biotechnology and biomedicine. A particularly interest in these fields is the monitoring of nanoparticle delivery to target cells. Since the structures involved are on a nanometer scale, high resolution imaging, such as electron microscopy, is required. Aside from assessing the structural characteristics of the target sites localized with the nanoparticles, an electron microscope can also be used to observe the biological effects of the nanoparticles on the cells. It can also be used to test and detect a wide range of fluorescent nanoparticles and nanoassemblies. Although this approach has many advantages, most researchers are unwilling to try electron microscopy due to the complicated specimen preparation procedures and time-consuming process. Here, we developed a method to simplify the sample preparation and shorten the total processing time. In particular, double staining was removed, and cryo-preparation was included. Using this simple and rapid sample preparation, we were able to observe nanoparticles with high-contrast images of the cellular organelles. This efficient detection method can be applied to studies on nanoparticle drug delivery systems and nanoparticle-cell interactions.

  12. 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.

  13. Electron microscopy study of direct laser deposited IN718

    Energy Technology Data Exchange (ETDEWEB)

    Ding, R.G., E-mail: r.ding@bham.ac.uk [School of Metallurgy and Materials, The University of Birmingham, Edgbaston, Birmingham B15 2TT (United Kingdom); Huang, Z.W.; Li, H.Y. [School of Metallurgy and Materials, The University of Birmingham, Edgbaston, Birmingham B15 2TT (United Kingdom); Mitchell, I.; Baxter, G. [Rolls-Royce plc., Derby DE24 8BJ (United Kingdom); Bowen, P. [School of Metallurgy and Materials, The University of Birmingham, Edgbaston, Birmingham B15 2TT (United Kingdom)

    2015-08-15

    The microstructure of direct laser deposited (DLD) IN718 has been investigated in detail using scanning electron microscopy (SEM) and transmission electron microscopy (TEM). The results confirm that the dendrite core microstructure can be linked to the cooling rate experienced during the deposition. A ~ 100 μm wide δ partially dissolved region in the IN718 substrate was observed close to the substrate/deposit boundary. In the deposited IN718, γ/Laves eutectic constituent is the predominant minor microconstituent. Irregular and regular (small) (Nb,Ti)C carbides and a mixture of the carbides and Laves were observed. Most M{sub 3}B{sub 2} borides were nucleated around a (Nb,Ti)C carbide. Needles of δ phase precipitated from the Laves phase were also observed. A complex constituent (of Laves, δ, α-Cr, γ″, and γ matrix) is reported in IN718 for the first time. The formation of α-Cr particles could be related to Cr rejection during the formation and growth of Cr-depleted δ phase. - Highlights: • Secondary phases in IN718 deposits were identified using electron diffraction and EDS. • MC, M{sub 3}B{sub 2}, γ/Laves eutectic and γ/NbC/Laves eutectic were observed. • Needle-like δ phases were precipitated from the Laves phase. • A complex constituent (Laves, δ, α-Cr, γ″ and γ) was reported for the first time.

  14. Aberration-Coreected Electron Microscopy at Brookhaven National Laboratory

    Energy Technology Data Exchange (ETDEWEB)

    Zhu,Y.; Wall, J.

    2008-04-01

    The last decade witnessed the rapid development and implementation of aberration correction in electron optics, realizing a more-than-70-year-old dream of aberration-free electron microscopy with a spatial resolution below one angstrom [1-9]. With sophisticated aberration correctors, modern electron microscopes now can reveal local structural information unavailable with neutrons and x-rays, such as the local arrangement of atoms, order/disorder, electronic inhomogeneity, bonding states, spin configuration, quantum confinement, and symmetry breaking [10-17]. Aberration correction through multipole-based correctors, as well as the associated improved stability in accelerating voltage, lens supplies, and goniometers in electron microscopes now enables medium-voltage (200-300kV) microscopes to achieve image resolution at or below 0.1nm. Aberration correction not only improves the instrument's spatial resolution but, equally importantly, allows larger objective lens pole-piece gaps to be employed thus realizing the potential of the instrument as a nanoscale property-measurement tool. That is, while retaining high spatial resolution, we can use various sample stages to observe the materials response under various temperature, electric- and magnetic- fields, and atmospheric environments. Such capabilities afford tremendous opportunities to tackle challenging science and technology issues in physics, chemistry, materials science, and biology. The research goal of the electron microscopy group at the Dept. of Condensed Matter Physics and Materials Science and the Center for Functional Nanomaterials, as well as the Institute for Advanced Electron Microscopy, Brookhaven National Laboratory (BNL), is to elucidate the microscopic origin of the physical- and chemical-behavior of materials, and the role of individual, or groups of atoms, especially in their native functional environments. We plan to accomplish this by developing and implementing various quantitative

  15. Copper Decoration of Carbon Nanotubes and High Resolution Electron Microscopy

    Science.gov (United States)

    Probst, Camille

    A new process of decorating carbon nanotubes with copper was developed for the fabrication of nanocomposite aluminum-nanotubes. The process consists of three stages: oxidation, activation and electroless copper plating on the nanotubes. The oxidation step was required to create chemical function on the nanotubes, essential for the activation step. Then, catalytic nanoparticles of tin-palladium were deposited on the tubes. Finally, during the electroless copper plating, copper particles with a size between 20 and 60 nm were uniformly deposited on the nanotubes surface. The reproducibility of the process was shown by using another type of carbon nanotube. The fabrication of nanocomposites aluminum-nanotubes was tested by aluminum vacuum infiltration. Although the infiltration of carbon nanotubes did not produce the expected results, an interesting electron microscopy sample was discovered during the process development: the activated carbon nanotubes. Secondly, scanning transmitted electron microscopy (STEM) imaging in SEM was analysed. The images were obtained with a new detector on the field emission scanning electron microscope (Hitachi S-4700). Various parameters were analysed with the use of two different samples: the activated carbon nanotubes (previously obtained) and gold-palladium nanodeposits. Influences of working distance, accelerating voltage or sample used on the spatial resolution of images obtained with SMART (Scanning Microscope Assessment and Resolution Testing) were analysed. An optimum working distance for the best spatial resolution related to the sample analysed was found for the imaging in STEM mode. Finally, relation between probe size and spatial resolution of backscattered electrons (BSE) images was studied. An image synthesis method was developed to generate the BSE images from backscattered electrons coefficients obtained with CASINO software. Spatial resolution of images was determined using SMART. The analysis shown that using a probe

  16. In situ and operando transmission electron microscopy of catalytic materials

    DEFF Research Database (Denmark)

    Crozier, Peter A.; Hansen, Thomas Willum

    2015-01-01

    Catalytic nanomaterials play a major role in chemical conversions and energy transformations. Understanding how materials control and regulate surface reactions is a major objective for fundamental research on heterogeneous catalysts. In situ environmental transmission electron microscopy (ETEM......) is a powerful technique for revealing the atomic structures of materials at elevated temperatures in the presence of reactive gases. This approach can allow the structure-reactivity relations underlying catalyst functionality to be investigated. Thus far, ETEM has been limited by the absence of in situ...

  17. Simultaneous orientation and thickness mapping in transmission electron microscopy

    Energy Technology Data Exchange (ETDEWEB)

    Tyutyunnikov, Dmitry, E-mail: dmitry.tyutyunnikov@uni-ulm.de [Institute for Experimental Physics, University of Ulm, Albert-Einstein-Allee 11, 89081 Ulm (Germany); Burak Özdöl, V. [National Center for Electron Microscopy, MS 72-150 Lawrence Berkeley National Laboratory, Berkeley, CA 94720 (United States); Koch, Christoph T. [Institute for Experimental Physics, University of Ulm, Albert-Einstein-Allee 11, 89081 Ulm (Germany)

    2015-03-15

    In this paper we introduce an approach for simultaneous thickness and orientation mapping of crystalline samples by means of transmission electron microscopy. We show that local thickness and orientation values can be extracted from experimental dark-field (DF) image data acquired at different specimen tilts. The method has been implemented to automatically acquire the necessary data and then map thickness and crystal orientation for a given region of interest. We have applied this technique to a specimen prepared from a commercial semiconductor device, containing multiple 22 nm technology transistor structures. The performance and limitations of our method are discussed and compared to those of other techniques available.

  18. Metallocarbohedrenes: Transmission Electron Microscopy of Mass Gated Deposits

    Science.gov (United States)

    Castleman, M. E. Lyn, Jr.

    2002-03-01

    Titanium and zirconium Met-Car cluster ions have been detected from the direct laser vaporization of metal-graphite mixtures using time-of-flight mass spectrometry. Optimization of the production conditions enabled sufficient intensities to mass select and deposit Met-Cars on surfaces. High-resolution transmission electron microscopy images of mass gated Met-Car species reveals deposited nanocrystals 2 nm in diameter. Diffraction patterns indicate the presence of multiple species and shows that the deposits have spatial orientation. Lattice parameters have been extracted. The implication of the findings will be discussed. Support for the work has been from the AFOSR F49620-01-1-0122.

  19. Electron microscopy of a Gd-Ba-Cu-O superconductor

    Science.gov (United States)

    Ramesh, R.; Thomas, G.; Meng, R. L.; Hor, P. H.; Chu, C. W.

    1989-01-01

    An electron microscopy study has been carried out to characterize the microstructure of a sintered Gd-Ba-Cu-O superconductor alloy. The GdBa2Cu3O(7-x) phase in the oxygen annealed sample is orthorhombic, while in the vacuum annealed sample it is tetragonal. It is shown that the details of the fine structure in the 001-line zone axis convergent beam patterns can be used to distinguish between the orthorhombic form and the tetragonal form. In addition to this matrix phase, an amorphous phase is frequently observed at the triple grain junctions. Gd-rich inclusions have been observed inside the matrix phase.

  20. Measurement of dihedral angles by scanning electron microscopy.

    Science.gov (United States)

    Achutaramayya, G.; Scott, W. D.

    1973-01-01

    The extension of Hoover's (1971) technique to the case of dihedral-angle measurement is described. Dihedral angles are often determined by interferometry on thermally grooved grain boundaries to obtain information on relative interfacial energies. In the technique considered the measured angles approach the true angles as the tilt angle approaches 90 deg. It is pointed out that the scanning electron microscopy method provides a means of seeing the real root of a groove at a lateral magnification which is higher than that obtainable with interferometry.

  1. Correction of bubble size distributions from transmission electron microscopy observations

    Energy Technology Data Exchange (ETDEWEB)

    Kirkegaard, P.; Eldrup, M.; Horsewell, A.; Skov Pedersen, J.

    1996-01-01

    Observations by transmission electron microscopy of a high density of gas bubbles in a metal matrix yield a distorted size distribution due to bubble overlap and bubble escape from the surface. A model is described that reconstructs 3-dimensional bubble size distributions from 2-dimensional projections on taking these effects into account. Mathematically, the reconstruction is an ill-posed inverse problem, which is solved by regularization technique. Extensive Monte Carlo simulations support the validity of our model. (au) 1 tab., 32 ills., 32 refs.

  2. Neuron Segmentation in Electron Microscopy Images Using Partial Differential Equations.

    Science.gov (United States)

    Jones, Cory; Sayedhosseini, Mojtaba; Ellisman, Mark; Tasdizen, Tolga

    2013-01-01

    In connectomics, neuroscientists seek to identify the synaptic connections between neurons. Segmentation of cell membranes using supervised learning algorithms on electron microscopy images of brain tissue is often done to assist in this effort. Here we present a partial differential equation with a novel growth term to improve the results of a supervised learning algorithm. We also introduce a new method for representing the resulting image that allows for a more dynamic thresholding to further improve the result. Using these two processes we are able to close small to medium sized gaps in the cell membrane detection and improve the Rand error by as much as 9% over the initial supervised segmentation.

  3. Microstructural studies of dental amalgams using analytical transmission electron microscopy

    Science.gov (United States)

    Hooghan, Tejpal Kaur

    Dental amalgams have been used for centuries as major restorative materials for decaying teeth. Amalgams are prepared by mixing alloy particles which contain Ag, Sn, and Cu as the major constituent elements with liquid Hg. The study of microstructure is essential in understanding the setting reactions and improving the properties of amalgams. Until the work reported in this dissertation, optical microscopy (OM), scanning electron microscopy (SEM), and x-ray diffractometry (XRD) were used commonly to analyze amalgam microstructures. No previous systematic transmission electron microscopy (TEM) study has been performed due to sample preparation difficulties and composite structure of dental amalgams. The goal of this research was to carry out detailed microstructural and compositional studies of dental amalgams. This was accomplished using the enhanced spatial resolution of the TEM and its associated microanalytical techniques, namely, scanning transmission electron microscopy (STEM), x-ray energy dispersive spectroscopy (XEDS) and micro-microdiffraction (mumuD). A new method was developed for thinning amalgam samples to electron transparency using the "wedge technique." Velvalloy, a low-Cu amalgam, and Tytin, a high-Cu amalgam, were the two amalgams characterized. Velvalloy is composed of a Agsb2Hgsb3\\ (gammasb1)/HgSnsb{7-9}\\ (gammasb2) matrix surrounding unreacted Agsb3Sn (gamma) particles. In addition, hitherto uncharacterized reaction layers between Agsb3Sn(gamma)/Agsb2Hgsb3\\ (gammasb2)\\ and\\ Agsb2Hgsb3\\ (gammasb1)/HgSnsb{7-9}\\ (gammasb2) were observed and analyzed. An Ag-Hg-Sn (betasb1) phase was clearly identified for the first time. In Tytin, the matrix consists of Agsb2Hgsb3\\ (gammasb1) grains. Fine precipitates of Cusb6Snsb5\\ (etasp') are embedded inside the gammasb1 and at the grain boundaries. These precipitates are responsible for the improved creep resistance of Tytin compared to Velvalloy. The additional Cu has completely eliminated the gammasb

  4. Measurement of dihedral angles by scanning electron microscopy.

    Science.gov (United States)

    Achutaramayya, G.; Scott, W. D.

    1973-01-01

    The extension of Hoover's (1971) technique to the case of dihedral-angle measurement is described. Dihedral angles are often determined by interferometry on thermally grooved grain boundaries to obtain information on relative interfacial energies. In the technique considered the measured angles approach the true angles as the tilt angle approaches 90 deg. It is pointed out that the scanning electron microscopy method provides a means of seeing the real root of a groove at a lateral magnification which is higher than that obtainable with interferometry.

  5. Microstrain in Nanocrystalline Copper by High Resolution Electron Microscopy

    Institute of Scientific and Technical Information of China (English)

    MIN Changping; RUAN Xuefeng; ZOU Huamin

    2009-01-01

    The elastic microstrains in a crystallite of electrodeposited nanocrystalline copper were investigated by analyzing the high resolution electron microscopy(HRTEM)image.The mi-crostrain was considered as consisting of two parts,in which the uniform part was determined with fast Fourier transformation of the HRTEM image,while the non-uniform part of the microstrain in the crystallite was measured by means of peak finding.Atomic column spacing measurements show that the crystal lattice is contracted in the longitudinal direction,while expanded in the transverse direction of the elliptical crystallite,indicating that the variation of microstrain exists mainly near the grain boundary.

  6. Confocal Microscopy for Modeling Electron Microbeam Irradiation of Skin

    Energy Technology Data Exchange (ETDEWEB)

    Miller, John H.; Chrisler, William B.; Wang, Xihai; Sowa, Marianne B.

    2011-08-01

    For radiation exposures employing targeted sources such as particle microbeams, the deposition of energy and dose will depend on the spatial heterogeneity of the spample. Although cell structural variations are relatively minor for two-dimensional cell cultures, they can vary significantly for fully differential tissues. Employing high-resolution confocal microscopy, we have determined the spatial distribution, size, and shape of epidermal kerantinocyte nuclei for the full-thickness EpiDerm skin model (MatTek, Ashland, VA). Application of these data to claculate the microdosimetry and microdistribution of energy deposition by an electron microbeam is discussed.

  7. GENRES OF ELECTRONIC LITERATURE IN THE CONTEXT OF GENRE REPLICATION

    Directory of Open Access Journals (Sweden)

    Basova, M.V.

    2016-06-01

    Full Text Available The article reviews some genres of electronic literary discourse that are prevalent in modern Russian literary Internet (“Rulinet”. Analyzing net literature works as secondary genres we reveal some features of primary genres. It is found that as the genre system of Internet communication develops, the models of discourse transformation become more complicated.

  8. GENRES OF ELECTRONIC LITERATURE IN THE CONTEXT OF GENRE REPLICATION

    OpenAIRE

    BASOVA M.V.; Rozhdestvenskaya, O.Ju.; Usacheva, O.Ju.

    2016-01-01

    The article reviews some genres of electronic literary discourse that are prevalent in modern Russian literary Internet (“Rulinet”). Analyzing net literature works as secondary genres we reveal some features of primary genres. It is found that as the genre system of Internet communication develops, the models of discourse transformation become more complicated.

  9. Nanofabrication by advanced electron microscopy using intense and focused beam∗

    Science.gov (United States)

    Furuya, Kazuo

    2008-01-01

    The nanogrowth and nanofabrication of solid substances using an intense and focused electron beam are reviewed in terms of the application of scanning and transmission electron microscopy (SEM, TEM and STEM) to control the size, position and structure of nanomaterials. The first example discussed is the growth of freestanding nanotrees on insulator substrates by TEM. The growth process of the nanotrees was observed in situ and analyzed by high-resolution TEM (HRTEM) and was mainly controlled by the intensity of the electron beam. The second example is position- and size-controlled nanofabrication by STEM using a focused electron beam. The diameters of the nanostructures grown ranged from 4 to 20 nm depending on the size of the electron beam. Magnetic nanostructures were also obtained using an iron-containing precursor gas, Fe(CO)5. The freestanding iron nanoantennas were examined by electron holography. The magnetic field was observed to leak from the nanostructure body which appeared to act as a ‘nanomagnet’. The third example described is the effect of a vacuum on the size and growth process of fabricated nanodots containing W in an ultrahigh-vacuum field-emission TEM (UHV-FE-TEM). The size of the dots can be controlled by changing the dose of electrons and the partial pressure of the precursor. The smallest particle size obtained was about 1.5 nm in diameter, which is the smallest size reported using this method. Finally, the importance of a smaller probe and a higher electron-beam current with atomic resolution is emphasized and an attempt to develop an ultrahigh-vacuum spherical aberration corrected STEM (Cs-corrected STEM) at NIMS is reported. PMID:27877936

  10. Sample heating system for spin-polarized scanning electron microscopy.

    Science.gov (United States)

    Kohashi, Teruo; Motai, Kumi

    2013-08-01

    A sample-heating system for spin-polarized scanning electron microscopy (spin SEM) has been developed and used for microscopic magnetization analysis at temperatures up to 500°C. In this system, a compact ceramic heater and a preheating operation keep the ultra-high vacuum conditions while the sample is heated during spin SEM measurement. Moreover, the secondary-electron collector, which is arranged close to the sample, was modified so that it is not damaged at high temperatures. The system was used to heat a Co(1000) single-crystal sample from room temperature up to 500°C, and the magnetic-domain structures were observed. Changes of the domain structures were observed around 220 and 400°C, and these changes are considered to be due to phase transitions of this sample.

  11. Theory and application of scanning electron acoustic microscopy

    Science.gov (United States)

    Cantrell, John H.; Qian, Menglu; Chen, Ruiyi; Yost, William T.

    1992-01-01

    A three-dimensional theoretical model based on the application of the thermal conduction and Navier equations to a chopped electron beam incident on a disk specimen is used to obtain the particle displacement field in the specimen. The results lead to a consideration of the signal generation, spatial resolution, and contrast mechanisms in scanning electron acoustic microscopy (SEAM). The model suggests that the time-variant heat source produced by the beam chopping generates driving source, thermal wave, and acoustic wave displacements simultaneously in the specimen. Evidence of the correctness of the prediction is obtained from the mathematically similar problem of pulsed laser light injection into a tank of water. High speed Schlieren photographs taken following laser injection show the simultaneous evolution of thermal and acoustic waveforms. Examples of contrast reversal, stress-induced contrast, and acoustic zone contrast and resolution with SEAM are presented and explained in terms of the model features.

  12. Some applications of microanalytical electron microscopy in materials research

    Energy Technology Data Exchange (ETDEWEB)

    Thomas, G.

    1985-10-01

    Electron microscopy has made extraordinary progress over the past 30 years and has become an indispensible tool for research in materials science. In this paper a review is given of some applications of microdiffraction and microanalysis in our current materials science research projects at the University of California, Berkeley. The topics discussed include: (1) The problem of solute atom partitioning in steels; this includes the difficulties of measuring carbon contents and methods of utilizing diffraction, lattice imaging, energy dispersive x-ray (EDXS) and electron energy loss (EELS) spectroscopies and atom probe analysis will be illustrated. (2) Utilization of CBED and EDXS techniques in zirconia ceramics research. (3) Applications of CBED to the study of el-Fe2O3 particles used in magnetic recording systems. (4) Applications of CBED and EDXS to rare earth permanent magnets. (5) Channelling enhanced microanalysis. 50 refs., 21 figs.

  13. Electron microscopy of gold nanoparticles at atomic resolution

    Science.gov (United States)

    Azubel, Maia; Koivisto, Jaakko; Malola, Sami; Bushnell, David; Hura, Greg L.; Koh, Ai Leen; Tsunoyama, Hironori; Tsukuda, Tatsuya; Pettersson, Mika; Häkkinen, Hannu; Kornberg, Roger D.

    2014-01-01

    Structure determination of gold nanoparticles (AuNPs) is necessary for understanding their physical and chemical properties, and only one AuNP larger than 1 nm in diameter, an Au102NP, has been solved to atomic resolution. Whereas the Au102NP structure was determined by X-ray crystallography, other large AuNPs have proved refractory to this approach. Here we report the structure determination of an Au68NP at atomic resolution by aberration-corrected transmission electron microscopy (AC-TEM), performed with the use of a minimal electron dose, an approach that should prove applicable to metal NPs in general. The structure of the Au68NP was supported by small angle X-ray scattering (SAXS) and by comparison of observed infrared (IR) absorption spectra with calculations by density functional theory (DFT). PMID:25146285

  14. Visualizing aquatic bacteria by light and transmission electron microscopy.

    Science.gov (United States)

    Silva, Thiago P; Noyma, Natália P; Duque, Thabata L A; Gamalier, Juliana P; Vidal, Luciana O; Lobão, Lúcia M; Chiarini-Garcia, Hélio; Roland, Fábio; Melo, Rossana C N

    2014-01-01

    The understanding of the functional role of aquatic bacteria in microbial food webs is largely dependent on methods applied to the direct visualization and enumeration of these organisms. While the ultrastructure of aquatic bacteria is still poorly known, routine observation of aquatic bacteria by light microscopy requires staining with fluorochromes, followed by filtration and direct counting on filter surfaces. Here, we used a new strategy to visualize and enumerate aquatic bacteria by light microscopy. By spinning water samples from varied tropical ecosystems in a cytocentrifuge, we found that bacteria firmly adhere to regular slides, can be stained by fluorochoromes with no background formation and fast enumerated. Significant correlations were found between the cytocentrifugation and filter-based methods. Moreover, preparations through cytocentrifugation were more adequate for bacterial viability evaluation than filter-based preparations. Transmission electron microscopic analyses revealed a morphological diversity of bacteria with different internal and external structures, such as large variation in the cell envelope and capsule thickness, and presence or not of thylakoid membranes. Our results demonstrate that aquatic bacteria represent an ultrastructurally diverse population and open avenues for easy handling/quantification and better visualization of bacteria by light microscopy without the need of filter membranes.

  15. 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.

  16. Transmission electron microscopy a textbook for materials science

    CERN Document Server

    Williams, David B

    1996-01-01

    Electron microscopy has revolutionized our understanding the extraordinary intellectual demands required of the mi­ of materials by completing the processing-structure-prop­ croscopist in order to do the job properly: crystallography, erties links down to atomistic levels. It now is even possible diffraction, image contrast, inelastic scattering events, and to tailor the microstructure (and meso structure ) of materials spectroscopy. Remember, these used to be fields in them­ to achieve specific sets of properties; the extraordinary abili­ selves. Today, one has to understand the fundamentals ties of modem transmission electron microscopy-TEM­ of all of these areas before one can hope to tackle signifi­ instruments to provide almost all of the structural, phase, cant problems in materials science. TEM is a technique of and crystallographic data allow us to accomplish this feat. characterizing materials down to the atomic limits. It must Therefore, it is obvious that any curriculum in modem mate­ be use...

  17. 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).

  18. Volume scanning electron microscopy for imaging biological ultrastructure.

    Science.gov (United States)

    Titze, Benjamin; Genoud, Christel

    2016-11-01

    Electron microscopy (EM) has been a key imaging method to investigate biological ultrastructure for over six decades. In recent years, novel volume EM techniques have significantly advanced nanometre-scale imaging of cells and tissues in three dimensions. Previously, this had depended on the slow and error-prone manual tasks of cutting and handling large numbers of sections, and imaging them one-by-one with transmission EM. Now, automated volume imaging methods mostly based on scanning EM (SEM) allow faster and more reliable acquisition of serial images through tissue volumes and achieve higher z-resolution. Various software tools have been developed to manipulate the acquired image stacks and facilitate quantitative analysis. Here, we introduce three volume SEM methods: serial block-face electron microscopy (SBEM), focused ion beam SEM (FIB-SEM) and automated tape-collecting ultramicrotome SEM (ATUM-SEM). We discuss and compare their capabilities, provide an overview of the full volume SEM workflow for obtaining 3D datasets and showcase different applications for biological research.

  19. Golgi apparatus analyzed by cryo-electron microscopy.

    Science.gov (United States)

    Han, Hong-Mei; Bouchet-Marquis, Cedric; Huebinger, Jan; Grabenbauer, Markus

    2013-10-01

    In 1898, the Golgi apparatus was discovered by light microscopy, and since the 1950s, the ultrastructure composition is known by electron microscopic investigation. The complex three-dimensional morphology fascinated researchers and was sometimes even the driving force to develop novel visualization techniques. However, the highly dynamic membrane systems of Golgi apparatus are delicate and prone to fixation artifacts. Therefore, the understanding of Golgi morphology and its function has been improved significantly with the development of better preparation methods. Nowadays, cryo-fixation is the method of choice to arrest instantly all dynamic and physiological processes inside cells, tissues, and small organisms. Embedded in amorphous ice, such samples can be further processed by freeze substitution or directly analyzed in their fully hydrated state by cryo-electron microscopy and tomography. Even though the overall morphology of vitrified Golgi stacks is comparable to well-prepared and resin-embedded samples, previously unknown structural details can be observed solely based on their native density. At this point, any further improvement of sample preparation would gain novel insights, perhaps not in terms of general morphology, but on fine structural details of this dynamic organelle.

  20. A national facility for biological cryo-electron microscopy.

    Science.gov (United States)

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

    2015-01-01

    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.

  1. Hybrid fluorescence and electron cryo-microscopy for simultaneous electron and photon imaging.

    Science.gov (United States)

    Iijima, Hirofumi; Fukuda, Yoshiyuki; Arai, Yoshihiro; Terakawa, Susumu; Yamamoto, Naoki; Nagayama, Kuniaki

    2014-01-01

    Integration of fluorescence light and transmission electron microscopy into the same device would represent an important advance in correlative microscopy, which traditionally involves two separate microscopes for imaging. To achieve such integration, the primary technical challenge that must be solved regards how to arrange two objective lenses used for light and electron microscopy in such a manner that they can properly focus on a single specimen. To address this issue, both lateral displacement of the specimen between two lenses and specimen rotation have been proposed. Such movement of the specimen allows sequential collection of two kinds of microscopic images of a single target, but prevents simultaneous imaging. This shortcoming has been made up by using a simple optical device, a reflection mirror. Here, we present an approach toward the versatile integration of fluorescence and electron microscopy for simultaneous imaging. The potential of simultaneous hybrid microscopy was demonstrated by fluorescence and electron sequential imaging of a fluorescent protein expressed in cells and cathodoluminescence imaging of fluorescent beads. Copyright © 2013 Elsevier Inc. All rights reserved.

  2. Electron microscopy investigations of nanoparticles for cancer diagnostic applications

    Science.gov (United States)

    Koh, Ai Leen

    This dissertation concerns electron microscopy characterization of magnetic (MNP) and surface enhanced Raman scattering (SERS) nanoparticles for in-vitro cancer diagnostic applications. Electron microscopy is an essential characterization tool owing to its (sub) nanometer spatial resolution. Structural information about the nanoparticles can be obtained using transmission electron microscopy (TEM), which can in turn be correlated to their physical characteristics. The scanning electron microscope (SEM) has excellent depth of field and can be effectively utilized to obtain high resolution information about nanoparticles binding onto cell surfaces. Part One of this thesis focuses on MNPs for bio-sensing and detection applications. As a preliminary study, chemically-synthesized, commercially-available iron oxide nanoparticles were compared against their laboratory-synthesized counterparts to assess their suitability for this application. The motivation for this initial study came about due to the lack of published data on commercially available iron oxide nanoparticles. TEM studies show that the latter are "beads" composed of multiple iron oxide cores encapsulated by a polymer shell, with large standard deviations in core diameter. Laboratory-synthesized iron oxide nanoparticles, on the other hand, are single core particles with small variations in diameter and therefore are expected to be better candidates for the required application. A key limitation in iron oxide nanoparticles is their relatively weak magnetic signals. The development of high moment Synthetic Anti-Ferromagnetic (SAF) nanoparticles aims to overcome this issue. SAFs are a novel class of MNPs fabricated using nanoimprint lithography, direct deposition of multilayer structure and final suspension into liquid medium (water). TEM analyses of cross-section specimens reveal that the SAFs possess characteristics similar to those of sputtered magnetic multilayer thin films. Their layered structure is

  3. Imaging domains in transmission electron microscopy (invited) (abstract)

    Science.gov (United States)

    Mishra, R. K.

    1987-04-01

    Magnetic domain walls and domains inside thin electron transparent specimens of ferromagnetic materials can be imaged using the Fresnel and Focault techniques in a transmission electron microscope. Combined with the diffraction, microstructural and microchemical capabilities of modern microscopes, Lorentz microscopy offers one of the most powerful tools to study structure-property relationships in magnetic materials. In addition, using this technique, it is possible to deduce the local magnetization distribution around inhomogeneities and complex Bloch and Néel walls. Lorentz images can be used to quantitatively measure domain wall thickness and estimate domain wall energy. With modified sample holders and pole pieces, one can study in situ domain wall motion and the interaction of domains with microstructural features such as second phases, grain boundaries, structural defects, etc. All these will be illustrated with examples of Lorentz images from soft and hard magnets with special emphasis on the Nd-Fe-B hard magnets. Finally, the limitations of the Lorentz imaging technique utilizing the deflected electron intensities will be outlined and a new technique which utilizes the phase changes in the electron beam as it passes through the material in a scanning transmission microscope will be reviewed.

  4. Characterization of strained semiconductor structures using transmission electron microscopy

    Energy Technology Data Exchange (ETDEWEB)

    Oezdoel, Vasfi Burak

    2011-08-15

    Today's state-of-the-art semiconductor electronic devices utilize the charge transport within very small volumes of the active device regions. The structural, chemical and optical material properties in these small dimensions can critically affect the performance of these devices. The present thesis is focused on the nanometer scale characterization of the strain state in semiconductor structures using transmission electron microscopy (TEM). Although high-resolution TEM has shown to provide the required accuracy at the nanometer scale, optimization of imaging conditions is necessary for accurate strain measurements. An alternative HRTEM method based on strain mapping on complex-valued exit face wave functions is developed to reduce the artifacts arising from objective lens aberrations. However, a much larger field of view is crucial for mapping strain in the active regions of complex structures like latest generation metal-oxide-semiconductor field-effect transistors (MOSFETs). To overcome this, a complementary approach based on electron holography is proposed. The technique relies on the reconstruction of the phase shifts in the diffracted electron beams from a focal series of dark-field images using recently developed exit-face wave function reconstruction algorithm. Combining high spatial resolution, better than 1 nm, with a field of view of about 1 {mu}m in each dimension, simultaneous strain measurements on the array of MOSFETs are possible. Owing to the much lower electron doses used in holography experiments when compared to conventional quantitative methods, the proposed approach allows to map compositional distribution in electron beam sensitive materials such as InGaN heterostructures without alteration of the original morphology and chemical composition. Moreover, dark-field holography experiments can be performed on thicker specimens than the ones required for high-resolution TEM, which in turn reduces the thin foil relaxation. (orig.)

  5. Bright-field scanning confocal electron microscopy using a double aberration-corrected transmission electron microscope

    Energy Technology Data Exchange (ETDEWEB)

    Wang, Peng; Behan, Gavin; Kirkland, Angus I. [Department of Materials, University of Oxford, Parks Road, Oxford OX1 3PH (United Kingdom); Nellist, Peter D., E-mail: peter.nellist@materials.ox.ac.uk [Department of Materials, University of Oxford, Parks Road, Oxford OX1 3PH (United Kingdom); Cosgriff, Eireann C.; D' Alfonso, Adrian J.; Morgan, Andrew J.; Allen, Leslie J. [School of Physics, University of Melbourne, Parkville, Victoria 3010 (Australia); Hashimoto, Ayako [Advanced Nano-characterization Center, National Institute for Materials Science (NIMS), 3-13 Sakura, Tsukuba 305-0003 (Japan); Takeguchi, Masaki [Advanced Nano-characterization Center, National Institute for Materials Science (NIMS), 3-13 Sakura, Tsukuba 305-0003 (Japan); High Voltage Electron Microscopy Station, NIMS, 3-13 Sakura, Tsukuba 305-0003 (Japan); Mitsuishi, Kazutaka [Advanced Nano-characterization Center, National Institute for Materials Science (NIMS), 3-13 Sakura, Tsukuba 305-0003 (Japan); Quantum Dot Research Center, NIMS, 3-13 Sakura, Tsukuba 305-0003 (Japan); Shimojo, Masayuki [High Voltage Electron Microscopy Station, NIMS, 3-13 Sakura, Tsukuba 305-0003 (Japan); Advanced Science Research Laboratory, Saitama Institute of Technology, 1690 Fusaiji, Fukaya 369-0293 (Japan)

    2011-06-15

    Scanning confocal electron microscopy (SCEM) offers a mechanism for three-dimensional imaging of materials, which makes use of the reduced depth of field in an aberration-corrected transmission electron microscope. The simplest configuration of SCEM is the bright-field mode. In this paper we present experimental data and simulations showing the form of bright-field SCEM images. We show that the depth dependence of the three-dimensional image can be explained in terms of two-dimensional images formed in the detector plane. For a crystalline sample, this so-called probe image is shown to be similar to a conventional diffraction pattern. Experimental results and simulations show how the diffracted probes in this image are elongated in thicker crystals and the use of this elongation to estimate sample thickness is explored. -- Research Highlights: {yields} The confocal probe image in a scanning confocal electron microscopy image reveals information about the thickness and height of the crystalline layer. {yields} The form of the contrast in a three-dimensional bright-field scanning confocal electron microscopy image can be explained in terms of the confocal probe image. {yields} Despite the complicated form of the contrast in bright-field scanning confocal electron microscopy, we see that depth information is transferred on a 10 nm scale.

  6. Optimized Negative-Staining Electron Microscopy for Lipoprotein Studies

    Science.gov (United States)

    Zhang, Lei; Tong, Huimin; Garewal, Mark; Ren, Gang

    2012-01-01

    Background Negative-staining (NS), a rapid, simple and conventional technique of electron microscopy (EM), has been commonly used to initially study the morphology and structure of proteins for half a century. Certain NS protocols however can cause artifacts, especially for structurally flexible or lipid-related proteins, such as lipoproteins. Lipoproteins were often observed in the form of rouleau as lipoprotein particles appeared to be stacked together by conventional NS protocols. The flexible components of lipoproteins, i.e. lipids and amphipathic apolipoproteins, resulted in the lipoprotein structure being sensitive to the NS sample preparation parameters, such as operational procedures, salt concentrations, and the staining reagents. Scope of review The most popular NS protocols that have been used to examine lipoprotein morphology and structure were reviewed. Major conclusions The comparisons show that an optimized NS (OpNS) protocol can eliminate the rouleau artifacts of lipoproteins, and that the lipoproteins are similar in size and shape as statistically measured from two EM methods, OpNS and cryo-electron microscopy (cryo-EM). OpNS is a high-throughput, high-contrast and high-resolution (near 1 nm, but rarely better than 1 nm) method which has been used to discover the mechanics of a small protein, 53 kDa cholesterol ester transfer protein (CETP), and the structure of an individual particle of a single protein by individual-particle electron tomography (IPET), i.e. a 14 Å-resolution IgG antibody three-dimensional map. General significance It is suggested that OpNS can be used as a general protocol to study the structure of proteins, especially highly dynamic proteins with equilibrium-fluctuating structures. PMID:23032862

  7. Electron Microscopy of Staphylococcus aureus Cell Wall Lysis

    Science.gov (United States)

    Virgilio, R.; González, C.; Muñoz, Nubia; Mendoza, Silvia

    1966-01-01

    Virgilio, Rafael (Escuela de Química y Farmacia, Universidad de Chile, Santiago, Chile), C. González, Nubia Muñoz, and Silvia Mendoza. Electron microscopy of Staphylococcus aureus cell wall lysis. J. Bacteriol. 91:2018–2024. 1966.—A crude suspension of Staphylococcus aureus cell walls (strain Cowan III) in buffer solution was shown by electron microscopy to lyse slightly after 16 hr, probably owing to the action of autolysin. The lysis was considerably faster and more intense after the addition of lysozyme. A remarkable reduction in thickness and rigidity of the cell walls, together with the appearance of many irregular protrusions in their outlines, was observed after 2 hr; after 16 hr, there remained only a few recognizable cell wall fragments but many residual particulate remnants. When autolysin was previously inactivated by trypsin, there was a complete inhibition of the lytic action of lysozyme; on the other hand, when autolysin was inactivated by heat and lysozyme was added, a distinct decrease in the thickness of the cell walls was observed, but there was no destruction of the walls. The lytic action of lysozyme, after treatment with hot 5% trichloroacetic acid, gave rise to a marked dissolution of the structure of the cell walls, which became lost against the background, without, however, showing ostensible alteration of wall outlines. From a morphological point of view, the lytic action of autolysin plus lysozyme was quite different from that of trichloroacetic acid plus lysozyme, as shown by electron micrographs, but in both cases it was very intense. This would suggest different mechanisms of action for these agents. Images PMID:5939482

  8. Preliminary Study of In Vivo Formed Dental Plaque Using Confocal Microscopy and Scanning Electron Microscopy

    Directory of Open Access Journals (Sweden)

    KA. Al-Salihi

    2009-12-01

    Full Text Available Objective: Confocal laser scanning microscopy (CLSM is relatively a new light microscopical imaging technique with a wide range of applications in biological sciences. The primary value of CLSM for the biologist is its ability to provide optical sections from athree-dimensional specimen. The present study was designed to assess the thickness and content of in vivo accumulated dental plaque using CLSM and scanning electron microscopy (SEM.Materials and Methods: Acroflat lower arch splints (acrylic appliance were worn by five participants for three days without any disturbance. The formed plaques were assessed using CLSM combined with vital fluorescence technique and SEM.Results: In this study accumulated dental plaque revealed varied plaque microflora vitality and thickness according to participant’s oral hygiene. The thickness of plaque smears ranged from 40.32 to 140.72 μm and 65.00 to 128.88 μm for live (vital and dead accumulated microorganisms, respectively. Meanwhile, the thickness of plaque on the appliance ranged from 101 μm to 653 μm. CLSM revealed both dead and vital bacteria on the surface of the dental plaque. In addition, SEM revealed layers of various bacterial aggregations in all dental plaques.Conclusion: This study offers a potent non-invasive tool to evaluate and assess the dental plaque biofilm, which is a very important factor in the development of dental caries.

  9. Engineering and Characterization of Collagen Networks Using Wet Atomic Force Microscopy and Environmental Scanning Electron Microscopy

    Science.gov (United States)

    Osborn, Jenna; Coffey, Tonya; Conrad, Brad; Burris, Jennifer; Hester, Brooke

    2014-03-01

    Collagen is an abundant protein and its monomers covalently crosslink to form fibrils which form fibers which contribute to forming macrostructures like tendon or bone. While the contribution is well understood at the macroscopic level, it is not well known at the fibril level. We wish to study the mechanical properties of collagen for networks of collagen fibers that vary in size and density. We present here a method to synthesize collagen networks from monomers and that allows us to vary the density of the networks. By using biotynilated collagen and a surface that is functionalized with avidin, we generate two-dimensional collagen networks across the surface of a silicon wafer. During network synthesis, the incubation time is varied from 30 minutes to 3 hours or temperature is varied from 25°C to 45°C. The two-dimensional collagen network created in the process is characterized using environmental atomic force microscopy (AFM) and scanning electron microscopy (SEM). The network density is measured by the number of strands in one frame using SPIP software. We expect that at body temperature (37°C) and with longer incubation times, the network density should increase.

  10. Fluctuation Electron Microscopy of Amorphous and Polycrystalline Materials

    Science.gov (United States)

    Rezikyan, Aram

    Fluctuation Electron Microscopy (FEM) has become an effective materials' structure characterization technique, capable of probing medium-range order (MRO) that may be present in amorphous materials. Although its sensitivity to MRO has been exercised in numerous studies, FEM is not yet a quantitative technique. The holdup has been the discrepancy between the computed kinematical variance and the experimental variance, which previously was attributed to source incoherence. Although high-brightness, high coherence, electron guns are now routinely available in modern electron microscopes, they have not eliminated this discrepancy between theory and experiment. The main objective of this thesis was to explore, and to reveal, the reasons behind this conundrum. The study was started with an analysis of the speckle statistics of tilted dark-field TEM images obtained from an amorphous carbon sample, which confirmed that the structural ordering is sensitively detected by FEM. This analysis also revealed the inconsistency between predictions of the source incoherence model and the experimentally observed variance. FEM of amorphous carbon, amorphous silicon and ultra nanocrystalline diamond samples was carried out in an attempt to explore the conundrum. Electron probe and sample parameters were varied to observe the scattering intensity variance behavior. Results were compared to models of probe incoherence, diffuse scattering, atom displacement damage, energy loss events and multiple scattering. Models of displacement decoherence matched the experimental results best. Decoherence was also explored by an interferometric diffraction method using bilayer amorphous samples, and results are consistent with strong displacement decoherence in addition to temporal decoherence arising from the electron source energy spread and energy loss events in thick samples. It is clear that decoherence plays an important role in the long-standing discrepancy between experimental FEM and its

  11. On mapping subangstrom electron clouds with force microscopy.

    Science.gov (United States)

    Wright, C Alan; Solares, Santiago D

    2011-11-09

    In 2004 Hembacher et al. (Science 2004, 305, 380-383) reported simultaneous higher-harmonics atomic force mocroscopy (AFM)/scanning tunneling microscopy (STM) images acquired while scanning a graphite surface with a tungsten tip. They interpreted the observed subatomic features in the AFM images as the signature of lobes of increased electron density at the tungsten tip apex. Although these intriguing images have stirred controversy, an in-depth theoretical feasibility study has not yet been produced. Here we report on the development of a method for simulating higher harmonics AFM images and its application to the same system. Our calculations suggest that four lobes of increased electron density are expected to be present at a W(001) tip apex atom and that the corresponding higher harmonics AFM images of graphite can exhibit 4-fold symmetry features. Despite these promising results, open questions remain since the calculated amplitudes of the higher harmonics generated by the short-range forces are on the order of hundredths of picometers, leading to very small corrugations in the theoretical images. Additionally, the complex, intermittent nature of the tip-sample interaction, which causes constant readjustment of the tip and sample orbitals as the tip approaches and retracts from the surface, prevents a direct quantitative connection between the electron density and the AFM image features.

  12. Robust image alignment for cryogenic transmission electron microscopy.

    Science.gov (United States)

    McLeod, Robert A; Kowal, Julia; Ringler, Philippe; Stahlberg, Henning

    2017-03-01

    Cryo-electron microscopy recently experienced great improvements in structure resolution due to direct electron detectors with improved contrast and fast read-out leading to single electron counting. High frames rates enabled dose fractionation, where a long exposure is broken into a movie, permitting specimen drift to be registered and corrected. The typical approach for image registration, with high shot noise and low contrast, is multi-reference (MR) cross-correlation. Here we present the software package Zorro, which provides robust drift correction for dose fractionation by use of an intensity-normalized cross-correlation and logistic noise model to weight each cross-correlation in the MR model and filter each cross-correlation optimally. Frames are reliably registered by Zorro with low dose and defocus. Methods to evaluate performance are presented, by use of independently-evaluated even- and odd-frame stacks by trajectory comparison and Fourier ring correlation. Alignment of tiled sub-frames is also introduced, and demonstrated on an example dataset. Zorro source code is available at github.com/CINA/zorro. Copyright © 2016 Elsevier Inc. All rights reserved.

  13. Improved Zernike-type phase contrast for transmission electron microscopy.

    Science.gov (United States)

    Koeck, P J B

    2015-07-01

    Zernike phase contrast has been recognized as a means of recording high-resolution images with high contrast using a transmission electron microscope. This imaging mode can be used to image typical phase objects such as unstained biological molecules or cryosections of biological tissue. According to the original proposal discussed in Danev and Nagayama (2001) and references therein, the Zernike phase plate applies a phase shift of π/2 to all scattered electron beams outside a given scattering angle and an image is recorded at Gaussian focus or slight underfocus (below Scherzer defocus). Alternatively, a phase shift of -π/2 is applied to the central beam using the Boersch phase plate. The resulting image will have an almost perfect contrast transfer function (close to 1) from a given lowest spatial frequency up to a maximum resolution determined by the wave length, the amount of defocus and the spherical aberration of the microscope. In this paper, I present theory and simulations showing that this maximum spatial frequency can be increased considerably without loss of contrast by using a Zernike or Boersch phase plate that leads to a phase shift between scattered and unscattered electrons of only π /4, and recording images at Scherzer defocus. The maximum resolution can be improved even more by imaging at extended Scherzer defocus, though at the cost of contrast loss at lower spatial frequencies. © 2015 The Authors Journal of Microscopy © 2015 Royal Microscopical Society.

  14. Transmission electron microscopy in molecular structural biology: A historical survey.

    Science.gov (United States)

    Harris, J Robin

    2015-09-01

    In this personal, historic account of macromolecular transmission electron microscopy (TEM), published data from the 1940s through to recent times is surveyed, within the context of the remarkable progress that has been achieved during this time period. The evolution of present day molecular structural biology is described in relation to the associated biological disciplines. The contribution of numerous electron microscope pioneers to the development of the subject is discussed. The principal techniques for TEM specimen preparation, thin sectioning, metal shadowing, negative staining and plunge-freezing (vitrification) of thin aqueous samples are described, with a selection of published images to emphasise the virtues of each method. The development of digital image analysis and 3D reconstruction is described in detail as applied to electron crystallography and reconstructions from helical structures, 2D membrane crystals as well as single particle 3D reconstruction of icosahedral viruses and macromolecules. The on-going development of new software, algorithms and approaches is highlighted before specific examples of the historical progress of the structural biology of proteins and viruses are presented.

  15. A transmission electron microscopy study of presolar hibonite

    CERN Document Server

    Zega, Thomas J; Nittler, Larry R; Stroud, Rhonda M

    2011-01-01

    We report isotopic and microstructural data on five presolar hibonite grains identified in an acid residue of the Krymka LL3.1 ordinary chondrite. Isotopic measurements by secondary ion mass spectrometry (SIMS) verified a presolar circumstellar origin for the grains. Transmission electron microscopy (TEM) examination of the crystal structure and chemistry of the grains was enabled by in situ sectioning and lift-out with a focused-ion-beam scanning-electron microscope. Comparisons of isotopic compositions with models indicate that four of the five grains formed in low-mass stars that evolved through the red-giant/asymptotic-giant branches, whereas one grain formed in the ejecta of a Type II supernova. Selected-area electron-diffraction patterns show that all grains are single crystals of hibonite. Some grains contain stacking faults and small spreads in orientation that can be attributed to a combination of growth defects and mechanical processing by grain-grain collisions. The similar structure of the superno...

  16. Scanning electron microscopy of the neuropathology of murine cerebral malaria

    Directory of Open Access Journals (Sweden)

    Brenneis Christian

    2006-11-01

    Full Text Available Abstract Background The mechanisms leading to death and functional impairments due to cerebral malaria (CM are yet not fully understood. Most of the knowledge about the pathomechanisms of CM originates from studies in animal models. Though extensive histopathological studies of the murine brain during CM are existing, alterations have not been visualized by scanning electron microscopy (SEM so far. The present study investigates the neuropathological features of murine CM by applying SEM. Methods C57BL/6J mice were infected with Plasmodium berghei ANKA blood stages. When typical symptoms of CM developed perfused brains were processed for SEM or light microscopy, respectively. Results Ultrastructural hallmarks were disruption of vessel walls, parenchymal haemorrhage, leukocyte sequestration to the endothelium, and diapedesis of macrophages and lymphocytes into the Virchow-Robin space. Villous appearance of observed lymphocytes were indicative of activated state. Cerebral oedema was evidenced by enlargement of perivascular spaces. Conclusion The results of the present study corroborate the current understanding of CM pathophysiology, further support the prominent role of the local immune system in the neuropathology of CM and might expose new perspectives for further interventional studies.

  17. Char porosity characterisation by scanning electron microscopy and image analysis

    Energy Technology Data Exchange (ETDEWEB)

    Soerensen, H.S.; Rosenberg, P.; Petersen, H.I.; Soerensen, L.H. [Danfoss A/S, Nordborg (Denmark)

    2000-09-01

    No significant change in either the morphotype composition or the macroporosity (pores {gt}5 {mu}m) in the 0-30 wt.% char burnout interval were revealed by reflected light microscopy or image analysis. Two high temperature char series from a Tertiary South American coal (C1) and a Permian Gondwana coal (C2) were therefore examined by scanning electron microscopy to provide information on the combustion process up to {approximately} 60 wt% char burnout. This study documents a significant mesopore ({approximately} 0.1-5 {mu}m) development on the fused chars in the burnout interval studied. A method to quantify the size and amount of the mesopores is described and both the parameters increased with increasing char burnout. Above a char burnout of {approximately} 30 wt% an increase in macroporosity was detected and ascribed to coalescence of mesopores to form large pores. Although the measurement of mesoporosity is restricted to fused chars, i.e. pores in fragments and the char morphotypes inertoid, fusinoid and solid could not be measured, the consideration of mesoporosity seems to be fundamental in understanding, evaluating and modelling combustion processes in the char burnout interval studied. 7 refs., 9 figs., 4 tabs.

  18. Using electron microscopy to calculate optical properties of biological samples.

    Science.gov (United States)

    Wu, Wenli; Radosevich, Andrew J; Eshein, Adam; Nguyen, The-Quyen; Yi, Ji; Cherkezyan, Lusik; Roy, Hemant K; Szleifer, Igal; Backman, Vadim

    2016-11-01

    The microscopic structural origins of optical properties in biological media are still not fully understood. Better understanding these origins can serve to improve the utility of existing techniques and facilitate the discovery of other novel techniques. We propose a novel analysis technique using electron microscopy (EM) to calculate optical properties of specific biological structures. This method is demonstrated with images of human epithelial colon cell nuclei. The spectrum of anisotropy factor g, the phase function and the shape factor D of the nuclei are calculated. The results show strong agreement with an independent study. This method provides a new way to extract the true phase function of biological samples and provides an independent validation for optical property measurement techniques.

  19. Photoemission Electron Microscopy as a Tool for Studying Steel Grains

    Science.gov (United States)

    Roese, Peter; Keutner, Christoph; Berges, Ulf; Espeter, Philipp; Westphal, Carsten

    2017-03-01

    Key properties of steel like stability, weldability, or ability for absorbing deformation energy are defined by their grain structure. The knowledge about their micrometer and submicrometer structure is of particular interest for tailor-cut macroscopic steel properties. We report on photoemission electron microscopy studies which in principle yield a higher magnification than comparable optical techniques. A flat surface without any topographic features was obtained by applying a non-etching preparation procedure. PEEM images showed very tiny phase islands embedded within a steel phase matrix. Furthermore, we developed an analysis procedure for PEEM images for dual-phase steels. As a result, it is possible to identify the individual work functions of different steel phases at the surface.

  20. Conditioning of mealybug (Hemiptera: Pseudococcidae by Scanning Electron Microscopy.

    Directory of Open Access Journals (Sweden)

    Melissa Palma-Jiménez

    2015-06-01

    Full Text Available The aim of this work was to determine the methodology for an adequate conditioning for the cleaning of mealybugs specimens and its correct observation. This work was done in the laboratory of the Research Center in Microscopic Structures (CIEMIC of the University of Costa Rica, in 2012. Four types of methodologies were implemented, which evidenced a gradual improvement of the observation of the ultrastructures through the Scanning Electron Microscopy. Every process was described in detail. The best results were showed with 10% xylene (in some cases it was feasible using 95-100% ethanol. It allowed to remove the wax from the body of the insect, avoiding its collapse, and observing the specific ultrastructures of the individual. This approach will reduce the time and cost of future taxonomic research of mealybugs.

  1. Electron Microscopy Structural Insights into CPAP Oligomeric Behavior

    DEFF Research Database (Denmark)

    Alvarez-Cabrera, Ana L; Delgado, Sandra; Gil-Carton, David

    2017-01-01

    that represent obtaining the protein in a soluble, homogeneous state for structural studies. Our work constitutes a systematic structural analysis on multiple oligomers of HsCPAP(897)(-1338), using single-particle electron microscopy (EM) of negatively stained (NS) samples. Based on image classification...... into clearly different regular 3D maps (putatively corresponding to dimers and tetramers) and direct observation of individual images representing other complexes of HsCPAP(897-1338) (i.e., putative flexible monomers and higher-order multimers), we report a dynamic oligomeric behavior of this protein, where...... of atomic models into the NS 3D maps at resolutions around 20 Å is performed only to complement our experimental data, allowing us to hypothesize on the oligomeric composition of the different complexes. In this way, the current EM work represents an initial step toward the structural characterization...

  2. Fluctuation electron microscopy studies of complex structured materials

    Science.gov (United States)

    Zhao, Gongpu; Rougée, Annick; Buseck, Peter; Treacy, Michael

    2008-03-01

    Fluctuation electron microscopy (FEM) is a hybrid imaging-diffraction technique. This technique is particularly sensitive to paracrystalline structures of dimension 0.5-2 nm, which are difficult to detect by either imaging or diffraction techniques alone. It has been successfully deployed to study paracrystalline structures in amorphous silicon, germanium thin film. This technique has also been used to study metallic glasses and oxide glasses. Until now, FEM has not been used to study disordered geological materials. In this talk we present our FEM studies of shungite, a naturally occurring disordered carbonaceous material, reveal that trace quantities of tightly curved graphene structures such as C60, or fragments of C60, is present in shungite. We also present results from our study of metamict zircon, whose crystal structure is destroyed by self-radiation during naturally occurring α decay events. Work is in progress to study the structural evolution during the metamictization process.

  3. Scanning electron microscopy of congenital corneal leukomas (Peters' anomaly).

    Science.gov (United States)

    Polack, F M; Graue, E L

    1979-08-01

    Specimens of three corneas in two patients with Peter's anomaly were obtained at the time of penetrating keratoplasty and studied by scanning and transmission electron microscopy. In one patient, the anomaly was monocular, and the endothelial surface showed a central defect in Descemet's layer with isolated rounded defects in the midperiphery. Fine collagenous material covered the posterior surface. The other two specimens were obtained from a patient with rubella syndrome without cataracts. The cornea showed malformation of Descemet's membrane with fibroblastic overgrowth on the endothelial layer. Epithelial-like cells and leukocytes were also found. The congenital central leukoma we believe was caused by adhesion of the pupillary membrane in our first patient, and possibly was inflammatory in our second patient.

  4. Electron Microscopy Analysis of the Nucleolus of Trypanosoma cruzi

    Science.gov (United States)

    López-Velázquez, Gabriel; Hernández, Roberto; López-Villaseñor, Imelda; Reyes-Vivas, Horacio; Segura-Valdez, María De L.; Jiménez-García, Luis F.

    2005-08-01

    The nucleolus is the main site for synthesis and processing of ribosomal RNA in eukaryotes. In mammals, plants, and yeast the nucleolus has been extensively characterized by electron microscopy, but in the majority of the unicellular eukaryotes no such studies have been performed. Here we used ultrastructural cytochemical and immunocytochemical techniques as well as three-dimensional reconstruction to analyze the nucleolus of Trypanosoma cruzi, which is an early divergent eukaryote of medical importance. In T. cruzi epimastigotes the nucleolus is a spherical intranuclear ribonucleoprotein organelle localized in a relatively central position within the nucleus. Dense fibrillar and granular components but not fibrillar centers were observed. In addition, nuclear bodies resembling Cajal bodies were observed associated to the nucleolus in the surrounding nucleoplasm. Our results provide additional morphological data to better understand the synthesis and processing of the ribosomal RNA in kinetoplastids.

  5. In Situ Transmission Electron Microscopy of Cadmium Selenide Nanorod Sublimation.

    Science.gov (United States)

    Hellebusch, Daniel J; Manthiram, Karthish; Beberwyck, Brandon J; Alivisatos, A Paul

    2015-02-19

    In situ electron microscopy is used to observe the morphological evolution of cadmium selenide nanorods as they sublime under vacuum at a series of elevated temperatures. Mass loss occurs anisotropically along the nanorod's long axis. At temperatures close to the sublimation threshold, the phase change occurs from both tips of the nanorods and proceeds unevenly with periods of rapid mass loss punctuated by periods of relative stability. At higher temperatures, the nanorods sublime at a faster, more uniform rate, but mass loss occurs from only a single end of the rod. We propose a mechanism that accounts for the observed sublimation behavior based on the terrace-ledge-kink (TLK) model and how the nanorod surface chemical environment influences the kinetic barrier of sublimation.

  6. Transmission Electron Microscopy (TEM) investigations of ancient Egyptian cosmetic powders

    Science.gov (United States)

    Deeb, C.; Walter, P.; Castaing, J.; Penhoud, P.; Veyssière, P.

    The processing technologies available during the time of ancient Egypt are of present concern to the field of Archaeology and Egyptology. Materials characterization is the best tool for establishing the processing history of archaeological objects. In this study, transmission electron microscopy (TEM) is used, in addition to other techniques, for phase identification and study of the microstructure and characteristic defect structures in ancient Egyptian cosmetic powders. These powders generally consist of a mix of Pb-containing mineral phases: galena (PbS), cerussite (PbCO3), and phosgenite (Pb2Cl2CO3), among others. Modern materials are fabricated according to recipes found in ancient texts to mimic the processing of ancient times and to compare with the archaeological specimens. In particular, a comparison between the dislocation structures of PbS crystals deformed in the laboratory and PbS from archaeological specimens from the collections of the Louvre Museum is presented .

  7. Scanning electron microscopy of ascospores of Debaryomyces and Saccharomyces.

    Science.gov (United States)

    Kurtzman, C P; Smiley, M J; Baker, F L

    1975-02-28

    Ascospores from species of Debaryomyces and the Torulaspora-group of Saccharomyces were examined by scanning electron microscopy. Ornamentation on ascospores of D. hansenii varied from short to long interconnected ridges or broad based, elongated conical protuberances. A spiral rigde system was detected on the ascospores of D. marama, but wart-like protuberances occurred on those of D. cantarelli, D. castellii, D. coudertii, D. formicarius, D. phaffii, D. vanriji and D. yarrowii. Ascospores of D. halotolerans did not have protuberances and the species appears to be identical with Pichia farinosa. Wart-like protuberances also were found on ascospores of S. delbrueckii, S. microellipsodes, S. rosei, S. inconspicuus, S. fermentati, S. montanus and S. vafer, but the ascospore surface of S. pretoriensis was covered by fine ridges. Short tapered ridges covered the ascospores of S. kloeckerianus.

  8. Variability of Protein Structure Models from Electron Microscopy.

    Science.gov (United States)

    Monroe, Lyman; Terashi, Genki; Kihara, Daisuke

    2017-03-02

    An increasing number of biomolecular structures are solved by electron microscopy (EM). However, the quality of structure models determined from EM maps vary substantially. To understand to what extent structure models are supported by information embedded in EM maps, we used two computational structure refinement methods to examine how much structures can be refined using a dataset of 49 maps with accompanying structure models. The extent of structure modification as well as the disagreement between refinement models produced by the two computational methods scaled inversely with the global and the local map resolutions. A general quantitative estimation of deviations of structures for particular map resolutions are provided. Our results indicate that the observed discrepancy between the deposited map and the refined models is due to the lack of structural information present in EM maps and thus these annotations must be used with caution for further applications.

  9. Photoemission Electron Microscopy as a Tool for Studying Steel Grains

    Science.gov (United States)

    Roese, Peter; Keutner, Christoph; Berges, Ulf; Espeter, Philipp; Westphal, Carsten

    2017-01-01

    Key properties of steel like stability, weldability, or ability for absorbing deformation energy are defined by their grain structure. The knowledge about their micrometer and submicrometer structure is of particular interest for tailor-cut macroscopic steel properties. We report on photoemission electron microscopy studies which in principle yield a higher magnification than comparable optical techniques. A flat surface without any topographic features was obtained by applying a non-etching preparation procedure. PEEM images showed very tiny phase islands embedded within a steel phase matrix. Furthermore, we developed an analysis procedure for PEEM images for dual-phase steels. As a result, it is possible to identify the individual work functions of different steel phases at the surface.

  10. Investigation of the Remineralization Effect Tnrough Scanning Electron Microscopy

    Directory of Open Access Journals (Sweden)

    Damyanova Dobrinka M

    2016-05-01

    Full Text Available Background: Local fluoride varnishes have been widely used as a method of non-operative treatment and for caries preventive interventions for more than three decades. Purpose: Evaluation of the remineralization effect by means of electron microscopy of mineralization varnish - Clinpro ™ White Varnish with TCP (Tri-Calcium phosphate (3M. Materials and Methods: The material used is from 20 temporary intact teeth, extracted due to physiological change with permanent teeth, with a completely preserved structure and anatomy of crowns and fully physiologically resorbed roots. For the purposes of the study a scanning electron microscope JEOL JSM 6390 is used with an attachment for element analysis (EDS INCA of Oxford. Prepared samples are pre-coated with gold (cathode sputtering with apparatus JEOL JFC – 1200 to obtain a better contrast of the SEM image of early carious lesions on the smooth surfaces of the temporary teeth, with predilection for development of caries with a d1 threshold. For this purpose the two processes were monitored occurring continuously on the enamel surfacede- and remineralization. Performed was computer processing of the digital images. Results: There is presence of certain minerals deposited in the embossed enamel prisms after of remineralization. The chemical analysis established the presence of calcium (Ca2 + , around the organic matrix. Demineralised surface has pores present of around 1%, which is visible through the enamel on the surface of the deciduous teeth looking like filled and pores looking like partially covered, filled with newly formed and growing crystals. The crystals, which are hydroxylapatite, fluorapatite or fluorhydroxiapatite gradually connect, growing and forming mineral structure filling the microscopi defects and the pores from the demineralisation in the surface enamel prismless layer

  11. In situ transmission electron microscopy experimentation of nanostructured materials

    Science.gov (United States)

    Alducin, Diego

    Due to the remarkable mechanical and electrical properties some nanostructured materials possess, it is important to be able to quantitatively characterize how these materials react under different types of stimulus. In situ transmission electron microscopy is a unique technique that allows the user to fully observe and record the crystallographic behavior of such materials undergoing a variety of tests. The crystallographic orientations silver nanowires were mapped in order to understand the structure and facets due to its geometry. Measuring the toughness and yield of the material led us to understand the anisotropic behavior of AgNWs. Depending on whether a load is applied to either a boundary between facets or on a facet will change the mechanical strength of the nanowire. By measuring the resistivity of the this material during deformation has also led us to understand that the intrinsic defects in the crystal structure of nanowires will change the way the material reacts to an electric potential. We have been also able to completely map the crystallographic orientations of very complex geometries of gold nanoparticles and characterize the weak forces involved in the manipulation if these nanoparticles. Finally, the elasticity of MoS2 was tested and found to be exponentially dependent upon the thickness of the nanosheets. However, the resistivity of this material does not seem to be affected by any type of deformation it is subjected to. The complete categorization of how materials interact with external stimulus while comparing the changes observed in its crystal structure is essential to understanding the underlying properties of nanostructured materials, which would not be possible without in situ transmisison electron microscopy experimentation.

  12. Monte Carlo simulation of secondary electron images for real sample structures in scanning electron microscopy.

    Science.gov (United States)

    Zhang, P; Wang, H Y; Li, Y G; Mao, S F; Ding, Z J

    2012-01-01

    Monte Carlo simulation methods for the study of electron beam interaction with solids have been mostly concerned with specimens of simple geometry. In this article, we propose a simulation algorithm for treating arbitrary complex structures in a real sample. The method is based on a finite element triangular mesh modeling of sample geometry and a space subdivision for accelerating simulation. Simulation of secondary electron image in scanning electron microscopy has been performed for gold particles on a carbon substrate. Comparison of the simulation result with an experiment image confirms that this method is effective to model complex morphology of a real sample.

  13. Scanning electron microscopy and electron probe X-ray microanalysis (SEM-EPMA) of pink teeth

    Energy Technology Data Exchange (ETDEWEB)

    Ikeda, N.; Watanabe, G.; Harada, A.; Suzuki, T.

    1988-11-01

    Samples of postmortem pink teeth were investigated by scanning electron microscopy and electron probe X-ray microanalysis. Fracture surfaces of the dentin in pink teeth were noticeably rough and revealed many more smaller dentinal tubules than those of the control white teeth. Electron probe X-ray microanalysis showed that the pink teeth contained iron which seemed to be derived from blood hemoglobin. The present study confirms that under the same circumstance red coloration of teeth may occur more easily in the teeth in which the dentin is less compact and contains more dentinal tubules.

  14. Morphological classification of bioaerosols from composting using scanning electron microscopy

    Energy Technology Data Exchange (ETDEWEB)

    Tamer Vestlund, A. [Institute for Energy and Resource Technology, Environmental Science and Technology Department, School of Applied Sciences, Cranfield University, Building 40, Bedfordshire MK43 0AL (United Kingdom); FIRA International Ltd., Maxwell Road, Stevenage, Herts SG1 2EW (United Kingdom); Al-Ashaab, R.; Tyrrel, S.F.; Longhurst, P.J.; Pollard, S.J.T. [Institute for Energy and Resource Technology, Environmental Science and Technology Department, School of Applied Sciences, Cranfield University, Building 40, Bedfordshire MK43 0AL (United Kingdom); Drew, G.H., E-mail: g.h.drew@cranfield.ac.uk [Institute for Energy and Resource Technology, Environmental Science and Technology Department, School of Applied Sciences, Cranfield University, Building 40, Bedfordshire MK43 0AL (United Kingdom)

    2014-07-15

    Highlights: • Bioaerosols were captured using the filter method. • Bioaerosols were analysed using scanning electron microscope. • Bioaerosols were classified on the basis of morphology. • Single small cells were found more frequently than aggregates and larger cells. • Smaller cells may disperse further than heavier aggregate structures. - Abstract: This research classifies the physical morphology (form and structure) of bioaerosols emitted from open windrow composting. Aggregation state, shape and size of the particles captured are reported alongside the implications for bioaerosol dispersal after release. Bioaerosol sampling took place at a composting facility using personal air filter samplers. Samples were analysed using scanning electron microscopy. Particles were released mainly as small (<1 μm) single, spherical cells, followed by larger (>1 μm) single cells, with aggregates occurring in smaller proportions. Most aggregates consisted of clusters of 2–3 particles as opposed to chains, and were <10 μm in size. No cells were attached to soil debris or wood particles. These small single cells or small aggregates are more likely to disperse further downwind from source, and cell viability may be reduced due to increased exposure to environmental factors.

  15. Materials characterisation by angle-resolved scanning transmission electron microscopy

    Science.gov (United States)

    Müller-Caspary, Knut; Oppermann, Oliver; Grieb, Tim; Krause, Florian F.; Rosenauer, Andreas; Schowalter, Marco; Mehrtens, Thorsten; Beyer, Andreas; Volz, Kerstin; Potapov, Pavel

    2016-11-01

    Solid-state properties such as strain or chemical composition often leave characteristic fingerprints in the angular dependence of electron scattering. Scanning transmission electron microscopy (STEM) is dedicated to probe scattered intensity with atomic resolution, but it drastically lacks angular resolution. Here we report both a setup to exploit the explicit angular dependence of scattered intensity and applications of angle-resolved STEM to semiconductor nanostructures. Our method is applied to measure nitrogen content and specimen thickness in a GaNxAs1‑x layer independently at atomic resolution by evaluating two dedicated angular intervals. We demonstrate contrast formation due to strain and composition in a Si- based metal-oxide semiconductor field effect transistor (MOSFET) with GexSi1‑x stressors as a function of the angles used for imaging. To shed light on the validity of current theoretical approaches this data is compared with theory, namely the Rutherford approach and contemporary multislice simulations. Inconsistency is found for the Rutherford model in the whole angular range of 16–255 mrad. Contrary, the multislice simulations are applicable for angles larger than 35 mrad whereas a significant mismatch is observed at lower angles. This limitation of established simulations is discussed particularly on the basis of inelastic scattering.

  16. From the physics of secondary electron emission to image contrasts in scanning electron microscopy.

    Science.gov (United States)

    Cazaux, Jacques

    2012-01-01

    Image formation in scanning electron microscopy (SEM) is a combination of physical processes, electron emissions from the sample, and of a technical process related to the detection of a fraction of these electrons. For the present survey of image contrasts in SEM, simplified considerations in the physics of the secondary electron emission yield, δ, are combined with the effects of a partial collection of the emitted secondary electrons. Although some consideration is initially given to the architecture of modern SEM, the main attention is devoted to the material contrasts with the respective roles of the sub-surface and surface compositions of the sample, as well as with the roles of the field effects in the vacuum gap. The recent trends of energy filtering in normal SEM and the reduction of the incident energy to a few electron volts in very low-energy electron microscopy are also considered. For an understanding by the SEM community, the mathematical expressions are explained with simple physical arguments.

  17. Infection Counter: Automated Quantification of in Vitro Virus Replication by Fluorescence Microscopy

    Directory of Open Access Journals (Sweden)

    Siân Culley

    2016-07-01

    Full Text Available The ability to accurately and reliably quantify viral infection is essential to basic and translational virology research. Here, we describe a simple and robust automated method for using fluorescence microscopy to estimate the proportion of virally infected cells in a monolayer. We provide details of the automated analysis workflow along with a freely available open-source ImageJ plugin, Infection Counter, for performing image quantification. Using hepatitis C virus (HCV as an example, we have experimentally verified our method, demonstrating that it is equivalent, if not better, than the established focus-forming assay. Finally, we used Infection Counter to assess the anti-HCV activity of SMBz-CsA, a non-immunosuppressive cyclosporine analogue.

  18. 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.

  19. Overview of electron crystallography of membrane proteins: crystallization and screening strategies using negative stain electron microscopy.

    Science.gov (United States)

    Nannenga, Brent L; Iadanza, Matthew G; Vollmar, Breanna S; Gonen, Tamir

    2013-01-01

    Electron cryomicroscopy, or cryoEM, is an emerging technique for studying the three-dimensional structures of proteins and large macromolecular machines. Electron crystallography is a branch of cryoEM in which structures of proteins can be studied at resolutions that rival those achieved by X-ray crystallography. Electron crystallography employs two-dimensional crystals of a membrane protein embedded within a lipid bilayer. The key to a successful electron crystallographic experiment is the crystallization, or reconstitution, of the protein of interest. This unit describes ways in which protein can be expressed, purified, and reconstituted into well-ordered two-dimensional crystals. A protocol is also provided for negative stain electron microscopy as a tool for screening crystallization trials. When large and well-ordered crystals are obtained, the structures of both protein and its surrounding membrane can be determined to atomic resolution.

  20. Probing Individual Ice Nucleation Events with Environmental Scanning Electron Microscopy

    Science.gov (United States)

    Wang, Bingbing; China, Swarup; Knopf, Daniel; Gilles, Mary; Laskin, Alexander

    2016-04-01

    Heterogeneous ice nucleation is one of the processes of critical relevance to a range of topics in the fundamental and the applied science and technologies. Heterogeneous ice nucleation initiated by particles proceeds where microscopic properties of particle surfaces essentially control nucleation mechanisms. Ice nucleation in the atmosphere on particles governs the formation of ice and mixed phase clouds, which in turn influence the Earth's radiative budget and climate. Heterogeneous ice nucleation is still insufficiently understood and poses significant challenges in predictive understanding of climate change. We present a novel microscopy platform allowing observation of individual ice nucleation events at temperature range of 193-273 K and relative humidity relevant for ice formation in the atmospheric clouds. The approach utilizes a home built novel ice nucleation cell interfaced with Environmental Scanning Electron Microscope (IN-ESEM system). The IN-ESEM system is applied for direct observation of individual ice formation events, determining ice nucleation mechanisms, freezing temperatures, and relative humidity onsets. Reported microanalysis of the ice nucleating particles (INP) include elemental composition detected by the energy dispersed analysis of X-rays (EDX), and advanced speciation of the organic content in particles using scanning transmission x-ray microscopy with near edge X-ray absorption fine structure spectroscopy (STXM/NEXAFS). The performance of the IN-ESEM system is validated through a set of experiments with kaolinite particles with known ice nucleation propensity. We demonstrate an application of the IN-ESEM system to identify and characterize individual INP within a complex mixture of ambient particles.

  1. From electron energy-loss spectroscopy to multi-dimensional and multi-signal electron microscopy.

    Science.gov (United States)

    Colliex, Christian

    2011-01-01

    This review intends to illustrate how electron energy-loss spectroscopy (EELS) techniques in the electron microscope column have evolved over the past 60 years. Beginning as a physicist tool to measure basic excitations in solid thin foils, EELS techniques have gradually become essential for analytical purposes, nowadays pushed to the identification of individual atoms and their bonding states. The intimate combination of highly performing techniques with quite efficient computational tools for data processing and ab initio modeling has opened the way to a broad range of novel imaging modes with potential impact on many different fields. The combination of Angström-level spatial resolution with an energy resolution down to a few tenths of an electron volt in the core-loss spectral domain has paved the way to atomic-resolved elemental and bonding maps across interfaces and nanostructures. In the low-energy range, improved energy resolution has been quite efficient in recording surface plasmon maps and from them electromagnetic maps across the visible electron microscopy (EM) domain, thus bringing a new view to nanophotonics studies. Recently, spectrum imaging of the emitted photons under the primary electron beam and the spectacular introduction of time-resolved techniques down to the femtosecond time domain, have become innovative keys for the development and use of a brand new multi-dimensional and multi-signal electron microscopy.

  2. Electron Microscopy Observation of Biomineralization within Wood Tissues of Kurogaki

    Directory of Open Access Journals (Sweden)

    Kazue Tazaki

    2017-07-01

    Full Text Available Interactions between minerals and microorganisms play a crucial role in living wood tissues. However, living wood tissues have never been studied in the field. Fortunately, we found several kurogaki (black persimmon; Diospyros kaki trees at Tawara in Kanazawa, Ishikawa, Japan. Here, we report the characterization of kurogaki based on scanning electron microscopy equipped with energy-dispersive spectroscopy (SEM-EDS and transmission electron microscopy (TEM, associated with inductively coupled plasma-mass spectrometry (ICP-MS analyses, X-ray fluorescence analyses (XRF and X-ray powder diffraction (XRD analyses. This study aims to illustrate the ability of various microorganisms associated with biominerals within wood tissues of kurogaki, as shown by SEM-EDS elemental content maps and TEM images. Kurogaki grows very slowly and has extremely hard wood, known for its striking black and beige coloration, referred to as a “peacock pattern”. However, the scientific data for kurogaki are very limited. The black “peacock pattern” of the wood mainly comprises cellulose and high levels of crystal cristobalite. As per the XRD results, the black taproot contains mineralized 7 Å clays (kaolinite, cellulose, apatite and cristobalite associated with many microorganisms. The chemical compositions of the black and beige portions of the black persimmon tree were obtained by ICP-MS analyses. Particular elements such as abundant Ca, Mg, K, P, Mn, Ba, S, Cl, Fe, Na, and Al were concentrated in the black region, associated with Pb and Sr elements. SEM-EDS semi-qualitative analyses of kurogaki indicated an abundance of P and Ca in microorganisms in the black region, associated with Pb, Sr, S, Mn, and Mg elements. On the other hand, XRF and XRD mineralogical data showed that fresh andesite, weathered andesite, and the soils around the roots of kurogaki correlate with biomineralization of the black region in kurogaki roots, showing clay minerals (kaolinite and

  3. Correlating intravital multi-photon microscopy to 3D electron microscopy of invading tumor cells using anatomical reference points.

    Directory of Open Access Journals (Sweden)

    Matthia A Karreman

    Full Text Available Correlative microscopy combines the advantages of both light and electron microscopy to enable imaging of rare and transient events at high resolution. Performing correlative microscopy in complex and bulky samples such as an entire living organism is a time-consuming and error-prone task. Here, we investigate correlative methods that rely on the use of artificial and endogenous structural features of the sample as reference points for correlating intravital fluorescence microscopy and electron microscopy. To investigate tumor cell behavior in vivo with ultrastructural accuracy, a reliable approach is needed to retrieve single tumor cells imaged deep within the tissue. For this purpose, fluorescently labeled tumor cells were subcutaneously injected into a mouse ear and imaged using two-photon-excitation microscopy. Using near-infrared branding, the position of the imaged area within the sample was labeled at the skin level, allowing for its precise recollection. Following sample preparation for electron microscopy, concerted usage of the artificial branding and anatomical landmarks enables targeting and approaching the cells of interest while serial sectioning through the specimen. We describe here three procedures showing how three-dimensional (3D mapping of structural features in the tissue can be exploited to accurately correlate between the two imaging modalities, without having to rely on the use of artificially introduced markers of the region of interest. The methods employed here facilitate the link between intravital and nanoscale imaging of invasive tumor cells, enabling correlating function to structure in the study of tumor invasion and metastasis.

  4. Using advanced electron microscopy for the characterization of catalytic materials

    Science.gov (United States)

    Pyrz, William D.

    Catalysis will continue to be vitally important to the advancement and sustainability of industrialized societies. Unfortunately, the petroleum-based resources that currently fuel the energy and consumer product needs of an advancing society are becoming increasingly difficult and expensive to extract as supplies diminish and the quality of sources degrade. Therefore, the development of sustainable energy sources and the improvement of the carbon efficiency of existing chemical processes are critical. Further challenges require that these initiatives are accomplished in an environmentally friendly fashion since the effects of carbon-based emissions are proving to be a serious threat to global climate stability. In this dissertation, materials being developed for sustainable energy and process improvement initiatives are studied. Our approach is to use materials characterization, namely advanced electron microscopy, to analyze the targeted systems at the nano- or Angstrom-scale with the goal of developing useful relationships between structure, composition, crystalline order, morphology, and catalytic performance. One area of interest is the complex Mo-V-M-O (M=Te, Sb, Ta, Nb) oxide system currently being developed for the selective oxidation/ammoxidation of propane to acrylic acid or acrylonitrile, respectively. Currently, the production of acrylic acid and acrylonitrile rely on propylene-based processes, yet significant cost savings could be realized if the olefin-based feeds could be replaced by paraffin-based ones. The major challenge preventing this feedstock replacement is the development of a suitable paraffin-activating catalyst. Currently, the best candidate is the Mo-V-Nb-Te-O complex oxide catalyst that is composed of two majority phases that are commonly referred to as M1 and M2. However, there is a limited understanding of the roles of each component with respect to how they contribute to catalyst stability and the reaction mechanism. Aberration

  5. Capturing enveloped viruses on affinity grids for downstream cryo-electron microscopy applications

    Science.gov (United States)

    Electron microscopy cryo-electron microscopy and cryo-electron tomography are essential techniques used for characterizing basic virus morphology and determining the three-dimensional structure of viruses. Enveloped viruses, which contain an outer lipoprotein coat, constitute the largest group of pa...

  6. Low-energy electron microscopy on two-dimensional systems : : growth, potentiometry and band structure mapping

    NARCIS (Netherlands)

    Kautz, Jaap

    2015-01-01

    Low Energy Electron Microscopy (LEEM) is a microscopy technique typically used to study surface processes. The sample is illuminated with a parallel beam of electrons under normal incidence and the reflected electrons are projected onto a pixelated detector, where an image is formed. In the first

  7. Automated Quantitative Rare Earth Elements Mineralogy by Scanning Electron Microscopy

    Science.gov (United States)

    Sindern, Sven; Meyer, F. Michael

    2016-09-01

    Increasing industrial demand of rare earth elements (REEs) stems from the central role they play for advanced technologies and the accelerating move away from carbon-based fuels. However, REE production is often hampered by the chemical, mineralogical as well as textural complexity of the ores with a need for better understanding of their salient properties. This is not only essential for in-depth genetic interpretations but also for a robust assessment of ore quality and economic viability. The design of energy and cost-efficient processing of REE ores depends heavily on information about REE element deportment that can be made available employing automated quantitative process mineralogy. Quantitative mineralogy assigns numeric values to compositional and textural properties of mineral matter. Scanning electron microscopy (SEM) combined with a suitable software package for acquisition of backscatter electron and X-ray signals, phase assignment and image analysis is one of the most efficient tools for quantitative mineralogy. The four different SEM-based automated quantitative mineralogy systems, i.e. FEI QEMSCAN and MLA, Tescan TIMA and Zeiss Mineralogic Mining, which are commercially available, are briefly characterized. Using examples of quantitative REE mineralogy, this chapter illustrates capabilities and limitations of automated SEM-based systems. Chemical variability of REE minerals and analytical uncertainty can reduce performance of phase assignment. This is shown for the REE phases parisite and synchysite. In another example from a monazite REE deposit, the quantitative mineralogical parameters surface roughness and mineral association derived from image analysis are applied for automated discrimination of apatite formed in a breakdown reaction of monazite and apatite formed by metamorphism prior to monazite breakdown. SEM-based automated mineralogy fulfils all requirements for characterization of complex unconventional REE ores that will become

  8. Cryogenic transmission electron microscopy nanostructural study of shed microparticles.

    Directory of Open Access Journals (Sweden)

    Liron Issman

    Full Text Available Microparticles (MPs are sub-micron membrane vesicles (100-1000 nm shed from normal and pathologic cells due to stimulation or apoptosis. MPs can be found in the peripheral blood circulation of healthy individuals, whereas elevated concentrations are found in pregnancy and in a variety of diseases. Also, MPs participate in physiological processes, e.g., coagulation, inflammation, and angiogenesis. Since their clinical properties are important, we have developed a new methodology based on nano-imaging that provides significant new data on MPs nanostructure, their composition and function. We are among the first to characterize by direct-imaging cryogenic transmitting electron microscopy (cryo-TEM the near-to-native nanostructure of MP systems isolated from different cell types and stimulation procedures. We found that there are no major differences between the MP systems we have studied, as most particles were spherical, with diameters from 200 to 400 nm. However, each MP population is very heterogeneous, showing diverse morphologies. We investigated by cryo-TEM the effects of standard techniques used to isolate and store MPs, and found that either high-g centrifugation of MPs for isolation purposes, or slow freezing to -80 °C for storage introduce morphological artifacts, which can influence MP nanostructure, and thus affect the efficiency of these particles as future diagnostic tools.

  9. Non-thermal plasma mills bacteria: Scanning electron microscopy observations

    Energy Technology Data Exchange (ETDEWEB)

    Lunov, O., E-mail: lunov@fzu.cz; Churpita, O.; Zablotskii, V.; Jäger, A.; Dejneka, A. [Institute of Physics AS CR, Prague 18221 (Czech Republic); Deyneka, I. G.; Meshkovskii, I. K. [St. Petersburg State University of Information Technologies, Mechanics and Optics, St. Petersburg 197101 (Russian Federation); Syková, E. [Institute of Experimental Medicine AS CR, Prague 14220 (Czech Republic); Kubinová, Š. [Institute of Physics AS CR, Prague 18221 (Czech Republic); Institute of Experimental Medicine AS CR, Prague 14220 (Czech Republic)

    2015-02-02

    Non-thermal plasmas hold great promise for a variety of biomedical applications. To ensure safe clinical application of plasma, a rigorous analysis of plasma-induced effects on cell functions is required. Yet mechanisms of bacteria deactivation by non-thermal plasma remain largely unknown. We therefore analyzed the influence of low-temperature atmospheric plasma on Gram-positive and Gram-negative bacteria. Using scanning electron microscopy, we demonstrate that both Gram-positive and Gram-negative bacteria strains in a minute were completely destroyed by helium plasma. In contrast, mesenchymal stem cells (MSCs) were not affected by the same treatment. Furthermore, histopathological analysis of hematoxylin and eosin–stained rat skin sections from plasma–treated animals did not reveal any abnormalities in comparison to control ones. We discuss possible physical mechanisms leading to the shred of bacteria under non-thermal plasma irradiation. Our findings disclose how helium plasma destroys bacteria and demonstrates the safe use of plasma treatment for MSCs and skin cells, highlighting the favorability of plasma applications for chronic wound therapy.

  10. Fossil micro-organisms evidenced by electronic microscopy

    Energy Technology Data Exchange (ETDEWEB)

    Prashnowsky, A.A.; Oberlies, F.; Burger, K.

    1983-04-01

    Fossil microorganisms in colonies and in the form of isolated cells (iron bacteria, fungi, actinomycetes etc.) were detected by electron microscopy of rocks containing remains of plant roots, carbonaceous substance, and strata of clay iron stone with ooids. These findings suggest an environment favourable to bacterial activity during sedimentation in the Upper Carboniferous and during the later processes of peat and coal formation. They also suggest that bacterial processes are an important factor in coal formation. Accurate data on coal formation can only be obtained by systematic biochemical studies. Analyses of the defined organic substances provide a better understanding of the conversion processes of the original substances. For example, the results of sterine analysis provide information on the mycoplancton, phytoplancton and zooplancton of the Upper Carboniferous. For some types of rock, the ratio of saponifiable to non-saponifiable constituents of the organic compounds yield information on stability under various geochemical conditions. The interactions between the various groups of microorganisms also play a major role in the solution of ecological problems.

  11. Histological preparation of developing vestibular otoconia for scanning electron microscopy

    Science.gov (United States)

    Huss, D.; Dickman, J. D.

    2003-01-01

    The unique nature of vestibular otoconia as calcium carbonate biominerals makes them particularly susceptible to chemical deformation during histological processing. We fixed and stored otoconia from all three otolith endorgans of embryonic, hatchling and adult Japanese quail in glutaraldehyde containing either phosphate or non-phosphate buffers for varying lengths of time and processed them for scanning electron microscopy. Otoconia from all age groups and otolith endorgans processed in 0.1 M phosphate buffer (pH 7.4) showed abnormal surface morphology when compared to acetone fixed controls. Otoconia processed in 0.1 M sodium cacodylate or HEPES buffered artificial endolymph (pH 7.4) showed normal morphology that was similar to controls. The degree of otoconial deformation was directly related to the time exposed to phosphate buffer. Short duration exposure produced particulate deformations while longer exposures resulted in fused otoconia that formed solid sheets. Otoconial surface deformation and fusing was independent of the glutaraldehyde component of the histological processing. These findings should help vestibular researchers to develop appropriate histological processing protocols in future studies of otoconia.

  12. Scanning electron microscopy and roughness study of dental composite degradation.

    Science.gov (United States)

    Soares, Luís Eduardo Silva; Cortez, Louise Ribeiro; Zarur, Raquel de Oliveira; Martin, Airton Abrahão

    2012-04-01

    Our aim was to test the hypothesis that the use of mouthwashes, consumption of soft drinks, as well as the type of light curing unit (LCU), would change the surface roughness (Ra) and morphology of a nanofilled composite resin (Z350® 3M ESPE). Samples (80) were divided into eight groups: Halogen LCU, group 1, saliva (control); group 2, Pepsi Twist®; group 3, Listerine®; group 4, Colgate Plax®; LED LCU, group 5, saliva; group 6, Pepsi Twist®; group 7, Listerine®; group 8, Colgate Plax®. Ra values were measured at baseline, and after 7 and 14 days. One specimen of each group was prepared for scanning electron microscopy analysis after 14 days. The data were subjected to multifactor analysis of variance at a 95% confidence followed by Tukey's honestly significant difference post-hoc test. All the treatments resulted in morphological changes in composite resin surface, and the most significant change was in Pepsi Twist® groups. The samples of G6 had the greatest increase in Ra. The immersion of nanofilled resin in mouthwashes with alcohol and soft drink increases the surface roughness. Polymerization by halogen LCU (reduced light intensity) associated with alcohol contained mouthwash resulted in significant roughness on the composite.

  13. Scanning electron microscopy of xiphinema, longidorus, and californidorus stylet morphology.

    Science.gov (United States)

    Cho, M R; Robbins, R T

    1990-04-01

    Stylet ultrastructure of five Xiphinema, four Longidorus, and three Californidorus species was compared by scanning electron microscopy. Morphological differences were seen in the odontophores and odontostyle bases between the genera and some of the species. All Xiphinema studied had well-developed odontophore flanges; the Longidorus species lacked flanges, except for weakly developed ones in L. diadecturus; and none of the Californidorus had flanges. Three sinuses were present in the odontophores of all species. The sinuses varied in length depending upon species. In Xiphinema and Californidorus the odontostyle bases had distinct overlapping collars, but in Longidorus the collars were absent except for L. diadecturus. The odontostyle-odontophore junction from a lateral view appeared as a slanted transverse line in all the species, but in a dorsal view of Xiphinema and Californidorus it was V-shaped. Dorsal longitudinal seams of the odontostyle and odontophore were observed in all the species. The dorsally located odontostyle aperture was ca. 1 mum from the anterior end in all species, except in one Longidorus sp. it was ca. 4 mum from the end.

  14. Electron microscopy, tissue culture,and immunology of ovarian carcinoma.

    Science.gov (United States)

    Ioachim, H L; Dorsett, B H; Sabbath, M; Barber, H R

    1975-10-01

    The ultrastructure of the major histologic types of ovarian carcinoma was investigated as part of a multilateral study of this tumor. The nuclear and nucleolar changes in size, shape, and structure correlated well with the degree of malignancy and tumor grading. Cytoplasmic organelles and intercellular junctions were abundant and fairly well differentiated even in ovarian carcinomas of higher grade and stage. Active processes of synthesis and secretion taking place in most of these tumors were suggested by the presence of a richly granulated endoplasmic reticulum, dilated cisternae, and numerous secretory granules. Seventy-eight different ovarian carcinomas of all histologic types were cultured in vitro for periods of up to 300 days, and their morphology in light and electron microscopy was compared to that of the original tumors. The cultures displayed a consistent pattern of growth which led to the conclusion that ovarian cancer cells in vitro preserve their salient features and are representative of the tumors of origin. Heterologous antisera raised with pooled extracts of various types of ovarian carcinomas reacted specifically in immunodiffusion and immunofluorescence tests only with ovarian carcinomas and not with normal ovaries, benigh ovarian tumors, and nonovarian malignant neoplasms, indicating the presence of a cross-reacting specific antigen for ovarian carcinomas. In other studies, autologous antibodies were isolated from antigen-antibody complexes recovered from peritoneal effusions of patients with ovarian carcinomas. These antibodies displayed a high degree of specificity against ovarian carcinoma cells when tested in immunofluorescence assays.

  15. Transmission electron microscopy (TEM) study of minerals in coal

    Energy Technology Data Exchange (ETDEWEB)

    Hsieh, Kuang-Chien

    1982-01-01

    Minerals in eight coals from different mines were characterized in the micron-size range by using analytical transmission electron microscopy. Specimens were thinned by ion-milling wafers cut from these coals; a cold stage cooled by liquid nitrogen was used to reduce thermal degradation of the minerals by the ion-beam. Different mineral compounds were observed in different coals. The major minerals are clays, sulfides, oxides, carbonates and some minor-element-bearing phosphates. Clays (kaolinite, illite and others) have been most commonly found as either flat sheets or round globules. Iron sulfide was mostly found in the No. 5 and No. 6 coals from Illinois, distributed as massive polycrystals, as clusters of single crystals (framboids) or as isolated single crystals with size range down to some 0.25 microns. Other sulfides and some oxides were found in other coals with particle size as small as some 200 angstroms. Quartz, titanium oxides and many other carbonates and phosphate compounds were also characterized. Brief TEM work in the organic mass of coal was also introduced to study the nature of the coal macerals.

  16. An overview on bioaerosols viewed by scanning electron microscopy

    Energy Technology Data Exchange (ETDEWEB)

    Wittmaack, K. [GSF-National Research Centre for Environment and Health, Institute of Radiation Protection, 85758 Neuherberg (Germany)]. E-mail: wittmaack@gsf.de; Wehnes, H. [GSF-National Research Centre for Environment and Health, Institute of Pathology, 85758 Neuherberg (Germany); Heinzmann, U. [GSF-National Research Centre for Environment and Health, Institute of Pathology, 85758 Neuherberg (Germany); Agerer, R. [Ludwig-Maximilians University Munich, Department Biology, Biodiversity Research: Mycology, Menzinger Stasse 67, 80638 Munich (Germany)

    2005-06-15

    Bioaerosols suspended in ambient air were collected with single-stage impactors at a semiurban site in southern Germany during late summer and early autumn. Sampling was mostly carried out at a nozzle velocity of 35 m/s, corresponding to a minimum aerodynamic diameter (cut-off diameter) of aerosol particles of 0.8 {mu}m. The collected particles, sampled for short periods ({approx}15 min) to avoid pile-up, were characterized by scanning electron microscopy (SEM). The observed bioaerosols include brochosomes, fungal spores, hyphae, insect scales, hairs of plants and, less commonly, bacteria and epicuticular wax. Brochosomes, which serve as a highly water repellent body coating of leafhoppers, are hollow spheroids with diameters around 400 nm, resembling C{sub 60} or footballs (soccer balls). They are usually airborne not as individuals but in the form of large clusters containing up to 10,000 individual species or even more. Various types of spores and scales were observed, but assignment turned out be difficult due to the large number of fungi and insects from which they may have originated. Pollens were observed only once. The absence these presumably elastic particles suggests that they are frequently lost, at the comparatively high velocities, due to bounce-off from the nonadhesive impaction surfaces.

  17. Histological preparation of developing vestibular otoconia for scanning electron microscopy

    Science.gov (United States)

    Huss, D.; Dickman, J. D.

    2003-01-01

    The unique nature of vestibular otoconia as calcium carbonate biominerals makes them particularly susceptible to chemical deformation during histological processing. We fixed and stored otoconia from all three otolith endorgans of embryonic, hatchling and adult Japanese quail in glutaraldehyde containing either phosphate or non-phosphate buffers for varying lengths of time and processed them for scanning electron microscopy. Otoconia from all age groups and otolith endorgans processed in 0.1 M phosphate buffer (pH 7.4) showed abnormal surface morphology when compared to acetone fixed controls. Otoconia processed in 0.1 M sodium cacodylate or HEPES buffered artificial endolymph (pH 7.4) showed normal morphology that was similar to controls. The degree of otoconial deformation was directly related to the time exposed to phosphate buffer. Short duration exposure produced particulate deformations while longer exposures resulted in fused otoconia that formed solid sheets. Otoconial surface deformation and fusing was independent of the glutaraldehyde component of the histological processing. These findings should help vestibular researchers to develop appropriate histological processing protocols in future studies of otoconia.

  18. Scanning electron microscopy applied to seed-borne fungi examination.

    Science.gov (United States)

    Alves, Marcelo de Carvalho; Pozza, Edson Ampélio

    2009-07-01

    The aim of this study was to test the standard scanning electron microscopy (SEM) as a potential alternative to study seed-borne fungi in seeds, by two different conditions of blotter test and water restriction treatment. In the blotter test, seeds were subjected to conditions that enabled pathogen growth and expression, whereas the water restriction method consisted in preventing seed germination during the incubation period, resulting in the artificial inoculation of fungi. In the first condition, seeds of common bean (Phaseolus vulgaris L.), maize (Zea mays L.), and cotton (Gossypium hirsutum L.) were submitted to the standard blotter test and then prepared and observed with SEM. In the second condition, seeds of cotton (G. hirsutum), soybean (Glycine max L.), and common bean (P. vulgaris L.) were, respectively, inoculated with Colletotrichum gossypii var. cephalosporioides, Colletotrichum truncatum, and Colletotrichum lindemuthianum by the water restriction technique, followed by preparation and observation with SEM. The standard SEM methodology was adopted to prepare the specimens. Considering the seeds submitted to the blotter test, it was possible to identify Fusarium sp. on maize, C. gossypii var. cephalosporioides, and Fusarium oxysporum on cotton, Aspergillus flavus, Penicillium sp., Rhizopus sp., and Mucor sp. on common bean. Structures of C. gossypii var. cephalosporioides, C. truncatum, and C. lindemuthianum were observed in the surface of inoculated seeds. (c) 2009 Wiley-Liss, Inc.

  19. TRANSMISSION ELECTRON MICROSCOPY STUDY OF HELIUM BEARING FUSION WELDS

    Energy Technology Data Exchange (ETDEWEB)

    Tosten, M; Michael Morgan, M

    2008-12-12

    A transmission electron microscopy (TEM) study was conducted to characterize the helium bubble distributions in tritium-charged-and-aged 304L and 21Cr-6Ni-9Mn stainless steel fusion welds containing approximately 150 appm helium-3. TEM foils were prepared from C-shaped fracture toughness test specimens containing {delta} ferrite levels ranging from 4 to 33 volume percent. The weld microstructures in the low ferrite welds consisted mostly of austenite and discontinuous, skeletal {delta} ferrite. In welds with higher levels of {delta} ferrite, the ferrite was more continuous and, in some areas of the 33 volume percent sample, was the matrix/majority phase. The helium bubble microstructures observed were similar in all samples. Bubbles were found in the austenite but not in the {delta} ferrite. In the austenite, bubbles had nucleated homogeneously in the grain interiors and heterogeneously on dislocations. Bubbles were not found on any austenite/austenite grain boundaries or at the austenite/{delta} ferrite interphase interfaces. Bubbles were not observed in the {delta} ferrite because of the combined effects of the low solubility and rapid diffusion of tritium through the {delta} ferrite which limited the amount of helium present to form visible bubbles.

  20. Electron beam heating effects during environmental scanning electron microscopy imaging of water condensation on superhydrophobic surfaces

    Science.gov (United States)

    Rykaczewski, K.; Scott, J. H. J.; Fedorov, A. G.

    2011-02-01

    Superhydrophobic surfaces (SHSs) show promise as promoters of dropwise condensation. Droplets with diameters below ˜10 μm account for the majority of the heat transferred during dropwise condensation but their growth dynamics on SHS have not been systematically studied. Due to the complex topography of the surface environmental scanning electron microscopy is the preferred method for observing the growth dynamics of droplets in this size regime. By studying electron beam heating effects on condensed water droplets we establish a magnification limit below which the heating effects are negligible and use this insight to study the mechanism of individual drop growth.

  1. High resolution measurements of the electron scattering for applications in electron microscopy and Monte-Carlo simulations of electron scattering

    CERN Document Server

    Berger, D

    2000-01-01

    scanning electron microscope is examined. By means of the scattering at mono-crystalline samples the influence of channeling (anomalous absorption and transmission) on backscattered electron spectra is shown. Captions are given in English language. This work presents high resolution measurements of the energy and complete angular distribution of the scattering of 20 keV electrons (energy resolution 0.55%). The examinations include take-off angles close to the target surface and non-perpendicular incidences of electrons partly for the first time. The results are of interest for the understanding of fundamental scattering processes, the interpretation of signals and new detector systems in electron microscopy and electron spectroscopy. Furthermore, they are used for the verification of electron scattering models and simulations. The applied compact electrostatic spectrometers with spherical and toroidal geometries are characterized and compared. High resolution spectra are obtained by deconvolution of the measu...

  2. Destructive effects induced by the electron beam in scanning electron microscopy

    Science.gov (United States)

    Popescu, M. C.; Bita, B. I.; Banu, M. A.; Tomescu, R. M.

    2016-12-01

    The Scanning Electron Microscopy has been validated by its impressive imaging and reliable measuring as an essential characterization tool for a variety of applications and research fields. This paper is a comprehensive study dedicated to the undesirable influence of the accelerated electron beam associated with the dielectric materials, sensitive structures or inappropriate sample manipulation. Depending on the scanning conditions, the electron beam may deteriorate the investigated sample due to the extended focusing or excessive high voltage and probe current applied on vulnerable configurations. Our aim is to elaborate an instructive material for improved SEM visualization capabilities by overcoming the specific limitations of the technique. Particular examination and measuring methods are depicted along with essential preparation and manipulation procedures in order to protect the integrity of the sample. Various examples are mentioned and practical solutions are described in respect to the general use of the electron microscope.

  3. EDITORIAL: Electron Microscopy and Analysis Group Conference 2011 (EMAG 2011)

    Science.gov (United States)

    Moebus, Guenter; Walther, Thomas; Brydson, Rik; Ozkaya, Dogan; MacLaren, Ian; Donnelly, Steve; Nellist, Pete; Li, Ziyou; Baker, Richard; Chiu, YuLung

    2012-07-01

    The biennial EMAG conference has established a strong reputation as a key event for the national and international electron microscopy community. In 2011 the meeting was held at The University of Birmingham, and I must first take this opportunity of thanking Birmingham for hosting the conference and for the excellent support we received from the local organisers. As a committee, we are delighted to see that enthusiasm for the EMAG conference series continues to be strong. We received more than 160 submitted abstracts, and 157 delegates attended the meeting. The scientific programme organiser, Ian MacLaren, put together an exciting programme. Plenary lectures were presented by Professor Knut Urban, Dr Frances Ross and Dr Richard Henderson. There were a further 10 invited speakers, from the UK, Continental Europe, Australia, the USA and Japan. The quality of the contributed oral and poster presentations was also very high. EMAG is keen to encourage student participation, and a winner and two runners-up were presented with prizes for the best oral and poster presentations from a student. I am always struck by the scientific quality of the oral and poster contributions and the vibrant discussions that occur both in the formal sessions and in the exhibition space at EMAG. I am convinced that a crucial part of maintaining that scientific quality is the opportunity that is offered of having a paper fully reviewed by two internationally selected referees and published in the Journal of Physics: Conference Series. For many students, this is the first fully reviewed paper they publish. I hope that you, like me, will be struck by the scientific quality of the 87 papers that follow, and that you will find them interesting and informative. Finally I must thank the platinum sponsors for their support of the meeting. These were Gatan, Zeiss, FEI, JEOL and Hitachi. I must also thank the European Microscopy Society for their generous sponsorship and support for the travel costs of

  4. Viral Infection at High Magnification: 3D Electron Microscopy Methods to Analyze the Architecture of Infected Cells

    Science.gov (United States)

    Romero-Brey, Inés; Bartenschlager, Ralf

    2015-01-01

    As obligate intracellular parasites, viruses need to hijack their cellular hosts and reprogram their machineries in order to replicate their genomes and produce new virions. For the direct visualization of the different steps of a viral life cycle (attachment, entry, replication, assembly and egress) electron microscopy (EM) methods are extremely helpful. While conventional EM has given important information about virus-host cell interactions, the development of three-dimensional EM (3D-EM) approaches provides unprecedented insights into how viruses remodel the intracellular architecture of the host cell. During the last years several 3D-EM methods have been developed. Here we will provide a description of the main approaches and examples of innovative applications. PMID:26633469

  5. Transmission Electron Microscopy of Iron Metal in Almahata Sitta Ureilite

    Science.gov (United States)

    Mikouchi, T.; Yubuta, K.; Sugiyama, K.; Aoyagi, Y.; Yasuhara, A.; Mihira, T.; Zolensky, M. E.; Goodrich, C. A.

    2013-01-01

    Almahata Sitta (AS) is a polymict breccia mainly composed of variable ureilite lithologies with small amounts of chondritic lithologies [1]. Fe metal is a common accessory phase in ureilites, but our earlier study on Fe metals in one of AS fragments (#44) revealed a unique mineralogy never seen in other ureilites [2,3]. In this abstract we report detailed transmission electron microscopy (TEM) on these metal grains to better understand the thermal history of ureilites. We prepared FIB sections of AS#44 by JEOL JIB-4000 from the PTS that was well characterized by SEM-EBSD in our earlier study [2]. The sections were then observed by STEM (JEOL JEM- 2100F). One of the FIB sections shows a submicron-sized symplectic intergrown texture composed of Fe metal (kamacite), Fe carbide (cohenite), Fe phosphide (schreibersite), and Fe sulfide (troilite). Each phase has an identical SAED pattern in spite of its complex texture, suggesting co-crystallization of all phases. This is probably caused by shock re-melting of pre-existing metal + graphite to form a eutectic-looking texture. The other FIB section is mostly composed of homogeneous Fe metal (93 wt% Fe, 5 wt% Ni, and 2 wt% Si), but BF-STEM images exhibited the presence of elongated lathy grains (approx. 2 microns long) embedded in the interstitial matrix. The SAED patterns from these lath grains could be indexed by alpha-Fe (bcc) while interstitial areas are gamma-Fe (fcc). The elongated alpha-Fe grains show tweed-like structures suggesting martensite transformation. Such a texture can be formed by rapid cooling from high temperature where gamma-Fe was stable. Subsequently alpha-Fe crystallized, but gamma-Fe remained in the interstitial matrix due to quenching from high temperature. This scenario is consistent with very rapid cooling history of ureilites suggested by silicate mineralogy.

  6. Transmission electron microscopy of subsolidus oxidation and weathering of olivine

    Science.gov (United States)

    Banfield, J.F.; Veblen, D.R.; Jones, B.F.

    1990-01-01

    Olivine crystals in basaltic andesites which crop out in the Abert Rim, south-central Oregon have been studied by high-resolution and analytical transmission electron microscopy. The observations reveal three distinct assemblages of alteration products that seem to correspond to three episodes of olivine oxidation. The olivine crystals contain rare, dense arrays of coherently intergrown Ti-free magnetite and inclusions of a phase inferred to be amorphous silica. We interpret this first assemblage to be the product of an early subsolidus oxidation event in the lava. The second olivine alteration assemblage contains complex ordered intergrowths on (001) of forsterite-rich olivine and laihunite (distorted olivine structure with Fe3+ charge balanced by vacancies). Based on experimental results for laihunite synthesis (Kondoh et al. 1985), these intergrowths probably formed by olivine oxidation between 400 and 800??C. The third episode of alteration involves the destruction of olivine by low-temperature hydrothermal alteration and weathering. Elongate etch-pits and channels in the margins of fresh olivine crystals contain semi-oriented bands of smectite. Olivine weathers to smectite and hematite, and subsequently to arrays of oriented hematite crystals. The textures resemble those reported by Eggleton (1984) and Smith et al. (1987). We find no evidence for a metastable phase intermediate between olivine and smectite ("M" - Eggleton 1984). The presence of laihunite exerts a strong control on the geometry of olivine weathering. Single laihunite layers and laihunite-forsteritic olivine intergrowths increase the resistance of crystals to weathering. Preferential development of channels between laihunite layers occurs where growth of laihunite produced compositional variations in olivine, rather than where coherency-strain is associated with laihunite-olivine interfaces. ?? 1990 Springer-Verlag.

  7. Molecular shape of Lumbricus terrestris erythrocruorin studied by electron microscopy and image analysis

    NARCIS (Netherlands)

    Boekema, Egbert J.; Heel, Marin van

    1989-01-01

    The molecular structure of erythrocruorin (hemoglobin) from Lumbricus terrestris has been studied by electron microscopy of negatively stained particles. Over 1000 molecular projections were selected from a number of electron micrographs and were then classified by multivariate statistical

  8. Visualization of Microbial Biomarkers by Scanning Electron Microscopy

    Science.gov (United States)

    Wainwright, Norman R.; Allen, Carlton C.; Child, Alice

    2001-01-01

    . Fortunately, many antimicrobial defense systems of higher organisms require sensitive detection to combat microbial pathogens. We employ here the primitive immune system of the evolutionarily ancient horseshoe crab, Limulus polyphemus. This species relies on multi-enzyme signal amplification detection of cell wall molecules and they can be applied to the development of useful detectors of life. An extension of this work includes the visualization of microbial signatures by labeling LAL components with chromogenic or electron dense markers. The protein Limulus Anti-LPS Factor (LALF) has an extremely high affinity for LPS. By coupling LALF binding with colloidal gold labels we demonstrate a correlation of the structures visible by electron microscopy with biochemical evidence of microbial cell wall materials. Pure silica particles were mixed with cultures of E. coli (10(exp 6) cfu/mL). Samples were washed sequentially with buffered saline, LALF, antibody to LALF and finally colloidal gold-labeled Protein A. Negative controls were not exposed to E. coli but received identical treatment otherwise. Samples were coated with carbon and imaged on a JEOL JSM-840 scanning electron microscope with LaB6 source in the back scatter mode with the JEOL annular back scatter detector. 20 nm-scale black spots in this contrast-reversed image originate from electrons back-scattered by gold atoms. Negative controls did not give any signal. Future work will expand application of this technique to soil simulants and mineralized rock samples.

  9. Visualization of Microbial Biomarkers by Scanning Electron Microscopy

    Science.gov (United States)

    Wainwright, Norman R.; Allen, Carlton C.; Child, Alice

    2001-01-01

    . Fortunately, many antimicrobial defense systems of higher organisms require sensitive detection to combat microbial pathogens. We employ here the primitive immune system of the evolutionarily ancient horseshoe crab, Limulus polyphemus. This species relies on multi-enzyme signal amplification detection of cell wall molecules and they can be applied to the development of useful detectors of life. An extension of this work includes the visualization of microbial signatures by labeling LAL components with chromogenic or electron dense markers. The protein Limulus Anti-LPS Factor (LALF) has an extremely high affinity for LPS. By coupling LALF binding with colloidal gold labels we demonstrate a correlation of the structures visible by electron microscopy with biochemical evidence of microbial cell wall materials. Pure silica particles were mixed with cultures of E. coli (10(exp 6) cfu/mL). Samples were washed sequentially with buffered saline, LALF, antibody to LALF and finally colloidal gold-labeled Protein A. Negative controls were not exposed to E. coli but received identical treatment otherwise. Samples were coated with carbon and imaged on a JEOL JSM-840 scanning electron microscope with LaB6 source in the back scatter mode with the JEOL annular back scatter detector. 20 nm-scale black spots in this contrast-reversed image originate from electrons back-scattered by gold atoms. Negative controls did not give any signal. Future work will expand application of this technique to soil simulants and mineralized rock samples.

  10. Simultaneous Correlative Light and Electron Microscopy of Samples in Liquid

    NARCIS (Netherlands)

    Liv, N.

    2014-01-01

    A combined use of fluorescence and light microscopy is a powerful approach to further increase our understanding in biological systems of structure-function relations at cellular and sub-cellular levels. The power of fluorescence microscopy (FM) is to spectrally resolve and visualize individual

  11. A charge coupled device camera with electron decelerator for intermediate voltage electron microscopy.

    Science.gov (United States)

    Downing, Kenneth H; Mooney, Paul E

    2008-04-01

    Electron microscopists are increasingly turning to intermediate voltage electron microscopes (IVEMs) operating at 300-400 kV for a wide range of studies. They are also increasingly taking advantage of slow-scan charge coupled device (CCD) cameras, which have become widely used on electron microscopes. Under some conditions, CCDs provide an improvement in data quality over photographic film, as well as the many advantages of direct digital readout. However, CCD performance is seriously degraded on IVEMs compared to the more conventional 100 kV microscopes. In order to increase the efficiency and quality of data recording on IVEMs, we have developed a CCD camera system in which the electrons are decelerated to below 100 kV before impacting the camera, resulting in greatly improved performance in both signal quality and resolution compared to other CCDs used in electron microscopy. These improvements will allow high-quality image and diffraction data to be collected directly with the CCD, enabling improvements in data collection for applications including high-resolution electron crystallography, single particle reconstruction of protein structures, tomographic studies of cell ultrastructure, and remote microscope operation. This approach will enable us to use even larger format CCD chips that are being developed with smaller pixels.

  12. Serial block face scanning electron microscopy--the future of cell ultrastructure imaging.

    Science.gov (United States)

    Hughes, Louise; Hawes, Chris; Monteith, Sandy; Vaughan, Sue

    2014-03-01

    One of the major drawbacks in transmission electron microscopy has been the production of three-dimensional views of cells and tissues. Currently, there is no one suitable 3D microscopy technique that answers all questions and serial block face scanning electron microscopy (SEM) fills the gap between 3D imaging using high-end fluorescence microscopy and the high resolution offered by electron tomography. In this review, we discuss the potential of the serial block face SEM technique for studying the three-dimensional organisation of animal, plant and microbial cells.

  13. Visualizing viral protein structures in cells using genetic probes for correlated light and electron microscopy.

    Science.gov (United States)

    Ou, Horng D; Deerinck, Thomas J; Bushong, Eric; Ellisman, Mark H; O'Shea, Clodagh C

    2015-11-15

    Structural studies of viral proteins most often use high-resolution techniques such as X-ray crystallography, nuclear magnetic resonance, single particle negative stain, or cryo-electron microscopy (EM) to reveal atomic interactions of soluble, homogeneous viral proteins or viral protein complexes. Once viral proteins or complexes are separated from their host's cellular environment, their natural in situ structure and details of how they interact with other cellular components may be lost. EM has been an invaluable tool in virology since its introduction in the late 1940's and subsequent application to cells in the 1950's. EM studies have expanded our knowledge of viral entry, viral replication, alteration of cellular components, and viral lysis. Most of these early studies were focused on conspicuous morphological cellular changes, because classic EM metal stains were designed to highlight classes of cellular structures rather than specific molecular structures. Much later, to identify viral proteins inducing specific structural configurations at the cellular level, immunostaining with a primary antibody followed by colloidal gold secondary antibody was employed to mark the location of specific viral proteins. This technique can suffer from artifacts in cellular ultrastructure due to compromises required to provide access to the immuno-reagents. Immunolocalization methods also require the generation of highly specific antibodies, which may not be available for every viral protein. Here we discuss new methods to visualize viral proteins and structures at high resolutions in situ using correlated light and electron microscopy (CLEM). We discuss the use of genetically encoded protein fusions that oxidize diaminobenzidine (DAB) into an osmiophilic polymer that can be visualized by EM. Detailed protocols for applying the genetically encoded photo-oxidizing protein MiniSOG to a viral protein, photo-oxidation of the fusion protein to yield DAB polymer staining, and

  14. 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.

  15. The New Electron Microscopy: Cells and Molecules in Three Dimensions | Poster

    Science.gov (United States)

    NCI recently announced the launch of the new National Cryo-Electron Microscopy Facility (NCEF) at the Frederick National Laboratory for Cancer Research (FNLCR). The launch comes while cryo-electron microscopy (cryo-EM) is enjoying the spotlight as a newly emerging, rapidly evolving technology with the potential to revolutionize the field of structural biology. Read more...

  16. Large-scale scanning transmission electron microscopy (nanotomy) of healthy and injured zebrafish brain

    NARCIS (Netherlands)

    J. Kuipers (Jeroen); R.D. Kalicharan (Ruby); Wolters, A.H.G. (Anouk H. G.); T.J. van Ham (Tjakko); B.N.G. Giepmans (Ben)

    2016-01-01

    textabstractLarge-scale 2D electron microscopy (EM), or nanotomy, is the tissue-wide application of nanoscale resolution electron microscopy. Others and we previously applied large scale EM to human skin pancreatic islets, tissue culture and whole zebrafish larvae1-7. Here we describe a universally

  17. Large-scale Scanning Transmission Electron Microscopy (Nanotomy) of Healthy and Injured Zebrafish Brain

    NARCIS (Netherlands)

    Kuipers, Jeroen; Kalicharan, Ruby D.; Wolters, Anouk H G; van Ham, Tjakko J; Giepmans, Ben N G

    2016-01-01

    Large-scale 2D electron microscopy (EM), or nanotomy, is the tissue-wide application of nanoscale resolution electron microscopy. Others and we previously applied large scale EM to human skin pancreatic islets, tissue culture and whole zebrafish larvae(1-7). Here we describe a universally applicable

  18. In-situ Liquid Electron Microscopy Setups for Investigation of Nanoscale Electrochemistry

    DEFF Research Database (Denmark)

    Jensen, Eric; Møller-Nilsen, Rolf Erling Robberstad; Canepa, Silvia

    2014-01-01

    of nanotoxicological studies, battery research and catalysis. Here we present two unique systems for in-situ electron microscopy. The first system is amonolithic chip for Transmission Electron Microscopy(2) (Figure 1a). The proof-of-conceptsystem verified the ability to separate the liquid from the vacuum while...

  19. The New Electron Microscopy: Cells and Molecules in Three Dimensions | Poster

    Science.gov (United States)

    NCI recently announced the launch of the new National Cryo-Electron Microscopy Facility (NCEF) at the Frederick National Laboratory for Cancer Research (FNLCR). The launch comes while cryo-electron microscopy (cryo-EM) is enjoying the spotlight as a newly emerging, rapidly evolving technology with the potential to revolutionize the field of structural biology. Read more...

  20. Seeing is believing : the impact of electron microscopy on autophagy research

    NARCIS (Netherlands)

    Eskelinen, Eeva-Liisa; Baba, Misuzu; Kovács, Attila L; Seglen, Per O; Reggiori, Fulvio

    2011-01-01

    Autophagy was first discovered by transmission electron microscopy more than 50 years ago. For decades, electron microscopy was the only way to reliably detect autophagic compartments in cells because no specific protein markers were known. In the 1970s, however, the introduction of biochemical meth

  1. Segmentation of scanning electron microscopy images from natural rubber samples with gold nanoparticles using starlet wavelets

    OpenAIRE

    de Siqueira, Alexandre Fioravante; Cabrera, Flavio Camargo [UNESP; Pagamisse, Aylton; Job,Aldo Eloizo

    2016-01-01

    Electronic microscopy has been used for morphology evaluation of different materials structures. However, microscopy results may be affected by several factors. Image processing methods can be used to correct and improve the quality of these results. In this article, we propose an algorithm based on starlets to perform the segmentation of scanning electron microscopy images. An application is presented in order to locate gold nanoparticles in natural rubber membranes. In this application, our...

  2. Correlative Analysis of Immunoreactivity in Confocal Laser-Scanning Microscopy and Scanning Electron Microscopy with Focused Ion Beam Milling

    Directory of Open Access Journals (Sweden)

    Takahiro eSonomura

    2013-02-01

    Full Text Available Three-dimensional reconstruction of ultrastructure of rat brain with minimal effort has recently been realized by scanning electron microscopy combined with focused ion beam milling (FIB-SEM. Because application of immunohistochemical staining to electron microscopy has a great advantage in that molecules of interest are specifically localized in ultrastructures, we here tried to apply immunocytochemistry to FIB-SEM and correlate immunoreactivity in confocal laser-scanning microcopy (CF-LSM with that in FIB-SEM. The dendrites of medium-sized spiny neurons in rat neostriatum were visualized with a recombinant viral vector, which labeled the infected neurons with membrane-targeted GFP in a Golgi stain-like fashion, and thalamostriatal afferent terminals were immunolabeled with Cy5 fluorescence for vesicular glutamate transporter 2 (VGluT2. After detecting the sites of terminals apposed to the dendrites in CF-LSM, GFP and VGluT2 immunoreactivities were further developed for electron microscopy by the immunogold/silver enhancement and immunoperoxidase/diaminobenzidine (DAB methods, respectively. In the contrast-inverted FIB-SEM images, silver precipitation and DAB deposits were observed as fine dark grains and diffuse dense profiles, respectively, indicating that these immunoreactivities were easily recognizable as in the images of transmission electron microscopy. In the sites of interest, some appositions were revealed to display synaptic specialization of asymmetric type. The present method is thus useful in the three-dimensional analysis of immunocytochemically differentiated synaptic connection in the central neural circuit.

  3. Backscattered Electron Microscopy as an Advanced Technique in Petrography.

    Science.gov (United States)

    Krinsley, David Henry; Manley, Curtis Robert

    1989-01-01

    Three uses of this method with sandstone, desert varnish, and granite weathering are described. Background information on this technique is provided. Advantages of this type of microscopy are stressed. (CW)

  4. Observation of the sweating in lipstick by scanning electron microscopy.

    Science.gov (United States)

    Seo, S Y; Lee, I S; Shin, H Y; Choi, K Y; Kang, S H; Ahn, H J

    1999-06-01

    The relationship between the wax matrix in lipstick and sweating has been investigated by observing the change of size and shape of the wax matrix due to sweating by Scanning Electron Microscopy (SEM). For observation by SEM, a lipstick sample was frozen in liquid nitrogen. The oil in the lipstick was then extracted in cold isopropanol (-70 degrees C) for 1-3 days. After the isopropanol was evaporated, the sample was sputtered with gold and examined by SEM. The change of wax matrix underneath the surface from fine, uniform structure to coarse, nonuniform structure resulted from the caking of surrounding wax matrix. The oil underneath the surface migrated to the surface of lipstick with sweating; consequently the wax matrix in that region was rearranged into the coarse matrix. In case of flamed lipstick, sweating was delayed and the wax matrix was much coarser than that of the unflamed one. The larger wax matrix at the surface region was good for including oil. The effect of molding temperature on sweating was also studied. As the molding temperature rose, sweating was greatly reduced and the size of the wax matrix increased. It was found that sweating was influenced by the compatibility of wax and oil. A formula consisting of wax and oil that have good compatibility has a tendency to reduce sweating and increase the size of the wax matrix. When pigments were added to wax and oil, the size of the wax matrix was changed, but in all cases sweating was increased due to the weakening of the binding force between wax and oil. On observing the thick membrane of wax at the surface of lipstick a month after molding it was also found that sweating was influenced by ageing. In conclusion, the structure of the wax matrix at the surface region of lipstick was changed with the process of flaming, molding temperature, compatibility of wax and oil, addition of pigment, and ageing. In most cases, as the size of the wax matrix was increased, sweating was reduced and delayed.

  5. Fast imaging with inelastically scattered electrons by off-axis chromatic confocal electron microscopy.

    Science.gov (United States)

    Zheng, Changlin; Zhu, Ye; Lazar, Sorin; Etheridge, Joanne

    2014-04-25

    We introduce off-axis chromatic scanning confocal electron microscopy, a technique for fast mapping of inelastically scattered electrons in a scanning transmission electron microscope without a spectrometer. The off-axis confocal mode enables the inelastically scattered electrons to be chromatically dispersed both parallel and perpendicular to the optic axis. This enables electrons with different energy losses to be separated and detected in the image plane, enabling efficient energy filtering in a confocal mode with an integrating detector. We describe the experimental configuration and demonstrate the method with nanoscale core-loss chemical mapping of silver (M4,5) in an aluminium-silver alloy and atomic scale imaging of the low intensity core-loss La (M4,5@840  eV) signal in LaB6. Scan rates up to 2 orders of magnitude faster than conventional methods were used, enabling a corresponding reduction in radiation dose and increase in the field of view. If coupled with the enhanced depth and lateral resolution of the incoherent confocal configuration, this offers an approach for nanoscale three-dimensional chemical mapping.

  6. Bright-field scanning confocal electron microscopy using a double aberration-corrected transmission electron microscope.

    Science.gov (United States)

    Wang, Peng; Behan, Gavin; Kirkland, Angus I; Nellist, Peter D; Cosgriff, Eireann C; D'Alfonso, Adrian J; Morgan, Andrew J; Allen, Leslie J; Hashimoto, Ayako; Takeguchi, Masaki; Mitsuishi, Kazutaka; Shimojo, Masayuki

    2011-06-01

    Scanning confocal electron microscopy (SCEM) offers a mechanism for three-dimensional imaging of materials, which makes use of the reduced depth of field in an aberration-corrected transmission electron microscope. The simplest configuration of SCEM is the bright-field mode. In this paper we present experimental data and simulations showing the form of bright-field SCEM images. We show that the depth dependence of the three-dimensional image can be explained in terms of two-dimensional images formed in the detector plane. For a crystalline sample, this so-called probe image is shown to be similar to a conventional diffraction pattern. Experimental results and simulations show how the diffracted probes in this image are elongated in thicker crystals and the use of this elongation to estimate sample thickness is explored.

  7. Characterization of gold nanoparticle films: Rutherford backscattering spectroscopy, scanning electron microscopy with image analysis, and atomic force microscopy

    Directory of Open Access Journals (Sweden)

    Pia C. Lansåker

    2014-10-01

    Full Text Available Gold nanoparticle films are of interest in several branches of science and technology, and accurate sample characterization is needed but technically demanding. We prepared such films by DC magnetron sputtering and recorded their mass thickness by Rutherford backscattering spectroscopy. The geometric thickness dg—from the substrate to the tops of the nanoparticles—was obtained by scanning electron microscopy (SEM combined with image analysis as well as by atomic force microscopy (AFM. The various techniques yielded an internally consistent characterization of the films. In particular, very similar results for dg were obtained by SEM with image analysis and by AFM.

  8. Spatiotemporal Observation of Electron-Impact Dynamics in Photovoltaic Materials Using 4D Electron Microscopy

    KAUST Repository

    Shaheen, Basamat

    2017-05-17

    Understanding light-triggered charge carrier dynamics near photovoltaic-material surfaces and at interfaces has been a key element and one of the major challenges for the development of real-world energy devices. Visualization of such dynamics information can be obtained using the one-of-a-kind methodology of scanning ultrafast electron microscopy (S-UEM). Here, we address the fundamental issue of how the thickness of the absorber layer may significantly affect the charge carrier dynamics on material surfaces. Time-resolved snapshots indicate that the dynamics of charge carriers generated by electron impact in the electron-photon dynamical probing regime is highly sensitive to the thickness of the absorber layer, as demonstrated using CdSe films of different thicknesses as a model system. This finding not only provides the foundation for potential applications of S-UEM to a wide range of devices in the fields of chemical and materials research, but also has impact on the use and interpretation of electron beam-induced current for optimization of photoactive materials in these devices.

  9. Simulation study of secondary electron images in scanning ion microscopy

    CERN Document Server

    Ohya, K

    2003-01-01

    The target atomic number, Z sub 2 , dependence of secondary electron yield is simulated by applying a Monte Carlo code for 17 species of metals bombarded by Ga ions and electrons in order to study the contrast difference between scanning ion microscopes (SIM) and scanning electron microscopes (SEM). In addition to the remarkable reversal of the Z sub 2 dependence between the Ga ion and electron bombardment, a fine structure, which is correlated to the density of the conduction band electrons in the metal, is calculated for both. The brightness changes of the secondary electron images in SIM and SEM are simulated using Au and Al surfaces adjacent to each other. The results indicate that the image contrast in SIM is much more sensitive to the material species and is clearer than that for SEM. The origin of the difference between SIM and SEM comes from the difference in the lateral distribution of secondary electrons excited within the escape depth.

  10. The Role of Electron Microscopy for the Diagnosis of Childhood Glomerular Diseases

    Directory of Open Access Journals (Sweden)

    Ahmad Ostadali Makhmalbaf

    2011-09-01

    Full Text Available Objective:Optimum diagnosis of glomerulopathies requires light microscopy, immunofluorescence and electron microcopy. In fact electron microscopy has a confirmatory role in glomerular diseases. It provides more information for patient management and can rule out other diseases. The goal of the present study is analysis the necessity of electron microscopy for the diagnosis of childhood glomerulopathies. Methods:134 cases of renal biopsy with some clinical data retrospectively were reviewed. The contribution of electron microscopy to the final diagnosis was graded as necessary - diagnosis could not be reached without it, supportive - it increased the level of confidence in the final diagnosis and noncontributory - the diagnosis dont need electron microscopy for confirmation. Findings:The contribution of electron microscopy to the final diagnosis was necessary in 51 cases (38%, supportive in 40 cases ( 30% and noncontributory in 43 cases (32%. Conclusion:In conclusion the results showed in about 68% of childhood glomerulopathies the ultrastructural study was necessary or supportive, so electron microscopy still remains an important tool in diagnosis of childhood glomerulopathies.

  11. Carbon contamination in scanning transmission electron microscopy and its impact on phase-plate applications.

    Science.gov (United States)

    Hettler, Simon; Dries, Manuel; Hermann, Peter; Obermair, Martin; Gerthsen, Dagmar; Malac, Marek

    2017-05-01

    We analyze electron-beam induced carbon contamination in a transmission electron microscope. The study is performed on thin films potentially suitable as phase plates for phase-contrast transmission electron microscopy. Electron energy-loss spectroscopy and phase-plate imaging is utilized to analyze the contamination. The deposited contamination layer is identified as a graphitic carbon layer which is not prone to electrostatic charging whereas a non-conductive underlying substrate charges. Several methods that inhibit contamination are evaluated and the impact of carbon contamination on phase-plate imaging is discussed. The findings are in general interesting for scanning transmission electron microscopy applications.

  12. Microscopy of electronic wave function; Microscopie de fonction d'onde electronique

    Energy Technology Data Exchange (ETDEWEB)

    Harb, M.

    2010-09-15

    This work of thesis aims to visualize, on a position sensitive detector, the spatial oscillations of slow electrons ({approx} meV) emitted by a threshold photoionization in the presence of an external electric field. The interference figure obtained represents the square magnitude of electronic wavefunction. This fundamental work allows us to have access to the electronic dynamics and thus to highlight several quantum mechanisms that occur at the atomic scale (field Coulomb, electron/electron interaction..). Despite the presence an electronic core in Li atom, we have succeeded, experimentally and for the first time, in visualizing the wave function associated with the quasi-discrete Stark states coupled to the ionization continuum. Besides, using simulations of wave packet propagation, based on the 'Split-operator' method, we have conducted a comprehensive study of the H, Li and Cs atoms while revealing the significant effects of the Stark resonances. A very good agreement, on and off resonances, was obtained between simulated and experimental results. In addition, we have developed a generalized analytical model to understand deeply the function of VMI (Velocity-Map Imaging) spectrometer. This model is based on the paraxial approximation; it is based on matrix optics calculation by making an analogy between the electronic trajectory and the light beam. An excellent agreement was obtained between the model predictions and the experimental results. (author)

  13. Resolution enhancement in transmission electron microscopy with 60-kV monochromated electron source

    Energy Technology Data Exchange (ETDEWEB)

    Morishita, Shigeyuki; Mukai, Masaki; Sawada, Hidetaka [JEOL Ltd., 3-1-2 Musashino, Akishima, Tokyo 196-8558 (Japan); Suenaga, Kazutomo [National Institute of Advanced Industrial Science and Technology (AIST), 1-1-1 Higashi, Tsukuba, Ibaraki 305-8565 (Japan)

    2016-01-04

    Transmission electron microscopy (TEM) at low accelerating voltages is useful to obtain images with low irradiation damage. For a low accelerating voltage, linear information transfer, which determines the resolution for observation of single-layered materials, is largely limited by defocus spread, which improves when a narrow energy spread is used in the electron source. In this study, we have evaluated the resolution of images obtained at 60 kV by TEM performed with a monochromated electron source. The defocus spread has been evaluated by comparing diffractogram tableaux from TEM images obtained under nonmonochromated and monochromated illumination. The information limits for different energy spreads were precisely measured by using diffractograms with a large beam tilt. The result shows that the information limit reaches 0.1 nm with an energy width of 0.10 eV. With this monochromated source and a higher-order aberration corrector, we have obtained images of single carbon atoms in a graphene sheet by TEM at 60 kV.

  14. Cryo scanning electron microscopy of Plasmodium falciparum-infected erythrocytes

    DEFF Research Database (Denmark)

    Hempel, Casper

    2017-01-01

    Plasmodium falciparum invades erythrocytes as an essential part of their life cycle. While living inside erythrocytes, the parasite remodels the cell's intracellular organization as well as its outer surface. Late trophozoite-stage parasites and schizonts introduce numerous small protrusions on t...... microscopy under cryogenic conditions allowing for high resolution and magnification of erythrocytes. This novel technique can be used for precise estimates of knob density and for studies on cytoadhesion....

  15. Cryo-electron Microscopy Structure of the Native Prototype Foamy Virus Glycoprotein and Virus Architecture.

    Directory of Open Access Journals (Sweden)

    Grégory Effantin

    2016-07-01

    Full Text Available Foamy viruses (FV belong to the genus Spumavirus, which forms a distinct lineage in the Retroviridae family. Although the infection in natural hosts and zoonotic transmission to humans is asymptomatic, FVs can replicate well in human cells making it an attractive gene therapy vector candidate. Here we present cryo-electron microscopy and (cryo-electron tomography ultrastructural data on purified prototype FV (PFV and PFV infected cells. Mature PFV particles have a distinct morphology with a capsid of constant dimension as well as a less ordered shell of density between the capsid and the membrane likely formed by the Gag N-terminal domain and the cytoplasmic part of the Env leader peptide gp18LP. The viral membrane contains trimeric Env glycoproteins partly arranged in interlocked hexagonal assemblies. In situ 3D reconstruction by subtomogram averaging of wild type Env and of a Env gp48TM- gp80SU cleavage site mutant showed a similar spike architecture as well as stabilization of the hexagonal lattice by clear connections between lower densities of neighboring trimers. Cryo-EM was employed to obtain a 9 Å resolution map of the glycoprotein in its pre-fusion state, which revealed extensive trimer interactions by the receptor binding subunit gp80SU at the top of the spike and three central helices derived from the fusion protein subunit gp48TM. The lower part of Env, presumably composed of interlaced parts of gp48TM, gp80SU and gp18LP anchors the spike at the membrane. We propose that the gp48TM density continues into three central transmembrane helices, which interact with three outer transmembrane helices derived from gp18LP. Our ultrastructural data and 9 Å resolution glycoprotein structure provide important new insights into the molecular architecture of PFV and its distinct evolutionary relationship with other members of the Retroviridae.

  16. Scanning Electron Microscopy with Samples in an Electric Field

    Science.gov (United States)

    Frank, Ludĕk; Hovorka, Miloš; Mikmeková, Šárka; Mikmeková, Eliška; Müllerová, Ilona; Pokorná, Zuzana

    2012-01-01

    The high negative bias of a sample in a scanning electron microscope constitutes the “cathode lens” with a strong electric field just above the sample surface. This mode offers a convenient tool for controlling the landing energy of electrons down to units or even fractions of electronvolts with only slight readjustments of the column. Moreover, the field accelerates and collimates the signal electrons to earthed detectors above and below the sample, thereby assuring high collection efficiency and high amplification of the image signal. One important feature is the ability to acquire the complete emission of the backscattered electrons, including those emitted at high angles with respect to the surface normal. The cathode lens aberrations are proportional to the landing energy of electrons so the spot size becomes nearly constant throughout the full energy scale. At low energies and with their complete angular distribution acquired, the backscattered electron images offer enhanced information about crystalline and electronic structures thanks to contrast mechanisms that are otherwise unavailable. Examples from various areas of materials science are presented.

  17. Center for Electron Microscopy, CEN-DTU; The building

    DEFF Research Database (Denmark)

    Horsewell, Andy

    Center for electron nanoscopy, CEN●DTU; The building Andy Horsewell Technical University of Denmark, DTU Materials Technology, Building 204, 2800 Lyngby ABSTRACT CEN●DTU, having been given[1] the opportunity to create a world-class facility with a unique suite of electron microscopes, is in full...... with an environmental cell, to provide in-situ observations of gas-solid interactions at high temperatures. 2 dual beam FIB-FEGSEMs, both with EDS and one with EBSD will allow 3D image, composition and crystallographic reconstruction at sub-nanometer resolution. Additional electron microscopes, making 7 in all...

  18. Theoretical study of ferroelectric nanoparticles using phase reconstructed electron microscopy

    DEFF Research Database (Denmark)

    Phatak, C.; Petford-Long, A. K.; Beleggia, Marco

    2014-01-01

    Ferroelectric nanostructures are important for a variety of applications in electronic and electro-optical devices, including nonvolatile memories and thin-film capacitors. These applications involve stability and switching of polarization using external stimuli, such as electric fields. We present...... a theoretical model describing how the shape of a nanoparticle affects its polarization in the absence of screening charges, and quantify the electron-optical phase shift for detecting ferroelectric signals with phase-sensitive techniques in a transmission electron microscope. We provide an example phase shift...

  19. Correlative analysis of immunoreactivity in confocal laser-scanning microscopy and scanning electron microscopy with focused ion beam milling.

    Science.gov (United States)

    Sonomura, Takahiro; Furuta, Takahiro; Nakatani, Ikuko; Yamamoto, Yo; Unzai, Tomo; Matsuda, Wakoto; Iwai, Haruki; Yamanaka, Atsushi; Uemura, Masanori; Kaneko, Takeshi

    2013-01-01

    Recently, three-dimensional reconstruction of ultrastructure of the brain has been realized with minimal effort by using scanning electron microscopy (SEM) combined with focused ion beam (FIB) milling (FIB-SEM). Application of immunohistochemical staining in electron microscopy (EM) provides a great advantage in that molecules of interest are specifically localized in ultrastructures. Thus, we applied immunocytochemistry for FIB-SEM and correlated this immunoreactivity with that in confocal laser-scanning microcopy (CF-LSM). Dendrites of medium-sized spiny neurons in the rat neostriatum were visualized using a recombinant viral vector, which labeled the infected neurons with membrane-targeted GFP in a Golgi stain-like fashion. Moreover, the thalamostriatal afferent terminals were immunolabeled with Cy5 fluorescence for vesicular glutamate transporter 2 (VGluT2). After detection of the sites of terminals apposed to the dendrites by using CF-LSM, GFP and VGluT2 immunoreactivities were further developed for EM by using immunogold/silver enhancement and immunoperoxidase/diaminobenzidine (DAB) methods, respectively. In contrast-inverted FIB-SEM images, silver precipitations and DAB deposits were observed as fine dark grains and diffuse dense profiles, respectively, indicating that these immunoreactivities were as easily recognizable as those in the transmission electron microscopy (TEM) images. Furthermore, in the sites of interest, some appositions displayed synaptic specializations of an asymmetric type. Thus, the present method was useful in the three-dimensional analysis of immunocytochemically differentiated synaptic connections in the central neural circuit.

  20. Microscopic techniques bridging between nanoscale and microscale with an atomically sharpened tip - field ion microscopy/scanning probe microscopy/ scanning electron microscopy.

    Science.gov (United States)

    Tomitori, Masahiko; Sasahara, Akira

    2014-11-01

    Over a hundred years an atomistic point of view has been indispensable to explore fascinating properties of various materials and to develop novel functional materials. High-resolution microscopies, rapidly developed during the period, have taken central roles in promoting materials science and related techniques to observe and analyze the materials. As microscopies with the capability of atom-imaging, field ion microscopy (FIM), scanning tunneling microscopy (STM), atomic force microscopy (AFM) and transmission electron microscopy (TEM) can be cited, which have been highly evaluated as methods to ultimately bring forward the viewpoint of reductionism in materials science. On one hand, there have been difficulties to derive useful and practical information on large (micro) scale unique properties of materials using these excellent microscopies and to directly advance the engineering for practical materials. To make bridges over the gap between an atomic scale and an industrial engineering scale, we have to develop emergence science step-by-step as a discipline having hierarchical structures for future prospects by combining nanoscale and microscale techniques; as promising ways, the combined microscopic instruments covering the scale gap and the extremely sophisticated methods for sample preparation seem to be required. In addition, it is noted that spectroscopic and theoretical methods should implement the emergence science.Fundamentally, the function of microscope is to determine the spatial positions of a finite piece of material, that is, ultimately individual atoms, at an extremely high resolution with a high stability. To define and control the atomic positions, the STM and AFM as scanning probe microscopy (SPM) have successfully demonstrated their power; the technological heart of SPM lies in an atomically sharpened tip, which can be observed by FIM and TEM. For emergence science we would like to set sail using the tip as a base. Meanwhile, it is significant

  1. Imaging of magnetic and electric fields by electron microscopy.

    Science.gov (United States)

    Zweck, Josef

    2016-10-12

    Nanostructured materials become more and more a part of our daily life, partly as self-assembled particles or artificially patterned. These nanostructures often possess intrinsic magnetic and/or electric fields which determine (at least partially) their physical properties. Therefore it is important to be able to measure these fields reliably on a nanometre scale. A rather common instrument for the investigation of these fields is the transmission electron microscope as it offers high spatial resolution. The use of an electron microscope to image electric and magnetic fields on a micron down to sub-nanometre scale is treated in detail for transmission electron microscopes (TEM) and scanning transmission electron microscopes (STEM). The formation of contrast is described for the most common imaging modes, the specific advantages and disadvantages of each technique are discussed and examples are given. In addition, the experimental requirements for the use of the techniques described are listed and explained.

  2. Imaging of magnetic and electric fields by electron microscopy

    Science.gov (United States)

    Zweck, Josef

    2016-10-01

    Nanostructured materials become more and more a part of our daily life, partly as self-assembled particles or artificially patterned. These nanostructures often possess intrinsic magnetic and/or electric fields which determine (at least partially) their physical properties. Therefore it is important to be able to measure these fields reliably on a nanometre scale. A rather common instrument for the investigation of these fields is the transmission electron microscope as it offers high spatial resolution. The use of an electron microscope to image electric and magnetic fields on a micron down to sub-nanometre scale is treated in detail for transmission electron microscopes (TEM) and scanning transmission electron microscopes (STEM). The formation of contrast is described for the most common imaging modes, the specific advantages and disadvantages of each technique are discussed and examples are given. In addition, the experimental requirements for the use of the techniques described are listed and explained.

  3. Nanomaterial datasets to advance tomography in scanning transmission electron microscopy

    OpenAIRE

    Levin, Barnaby D.A.; Padgett, Elliot; Chen, Chien-Chun; Scott, M C; Xu, Rui; Theis, Wolfgang; Jiang, Yi; Yang, Yongsoo; Ophus, Colin; Zhang, Haitao; Ha, Don-Hyung; Wang, Deli; Yu, Yingchao; Abruña, Hector D.; Robinson, Richard D.

    2016-01-01

    Electron tomography in materials science has flourished with the demand to characterize nanoscale materials in three dimensions (3D). Access to experimental data is vital for developing and validating reconstruction methods that improve resolution and reduce radiation dose requirements. This work presents five high-quality scanning transmission electron microscope (STEM) tomography datasets in order to address the critical need for open access data in this field. The datasets represent the cu...

  4. Lights Will Guide You : Sample Preparation and Applications for Integrated Laser and Electron Microscopy

    NARCIS (Netherlands)

    Karreman, M.A.

    2013-01-01

    Correlative microscopy is the combined use of two different forms of microscopy in the study of a specimen, allowing for the exploitation of the advantages of both imaging tools. The integrated Laser and Electron Microscope (iLEM), developed at Utrecht University, combines a fluorescence microscope

  5. CHROMATIN TEXTURE OF MELANOCYTIC NUCLEI - CORRELATION BETWEEN LIGHT AND ELECTRON-MICROSCOPY

    NARCIS (Netherlands)

    ABMAYR, W; STOLZ, W; KORHERR, S; WILD, W; SCHMOECKEL, C

    1987-01-01

    Cells of a benign pigmented mole and a malignant melanoma were used to compare electron microscopy (EM) and light microscopy (LM) with high-resolution TV-scanning and multivariate analysis methods. Special emphasis was placed on different kinds of chromatin texture features and their discriminating

  6. Transmission and scanning electron microscopy confirm that bone microstructure is similar in osteopenic and osteoporotic patients.

    Science.gov (United States)

    Gül, Orkun; Atik, O Sahap; Erdoğan, Deniz; Göktaş, Güleser; Elmas, Ciğdem

    2013-01-01

    The objective was to confirm the finding of "Bone microstructure is similar in osteopenic and osteoporotic patients with femoral neck fracture." obtained in previous "light microscopy study", which was new and important data. Fourteen patients (5 males, 9 females) who were admitted with proximal femoral fracture following low energy trauma (patients who participated in the light microscopy study) were included. The patients were divided into two groups based on the bone mineral density (BMD) measurement, including osteopenic group (n=7, mean age 69 years; range 63 to 74 years) and osteoporotic group (n=7, mean age 74.1 years; range 67 to 78 years). Cortical and trabecular bone samples were taken from the patients who underwent endoprosthesis during partial hip arthroplasty and these samples were analyzed using transmission electron microscopy and scanning electron microscopy evaluations which are more sophisticated higher resolution techniques. The mean cortical bone thickness was 3622.14 mm in osteopenic group and 2323.14 mm in osteoporotic group (pelectron microscopy and scanning electron microscopy evaluations revealed similar findings for both groups. Although a significant difference in cortical thickness was found between the groups, transmission and scanning electron microscopy confirmed that bone microstructure shared similar characteristics in osteopenic and osteoporotic patients with low-energy femoral neck fracture, as it was in previous light microscopy study.

  7. Investigation of the ultrastructure of yeast using cryo-immuno-electron microscopy

    NARCIS (Netherlands)

    Mari, Muriel; Griffith, Janice; Reggiori, Fulvio

    2010-01-01

    Yeast has been a crucial model system for the investigation of a multitude of cellular processes and many approaches have been used to study these, including genetics, biochemistry and fluorescence microscopy approaches. The morphological analyses of these organisms by electron microscopy (EM),

  8. In situ transmission electron microscopy of light-induced photocatalytic reactions

    DEFF Research Database (Denmark)

    Cavalca, Filippo; Laursen, Anders Bo; Kardynal, Beata

    2012-01-01

    Transmission electron microscopy (TEM) makes it possible to obtain insight into the structure, composition and reactivity of photocatalysts, which are of fundamental interest for sustainable energy research. Such insight can be used for further material optimization. Here, we combine conventional...

  9. Proton induced X-ray emission and electron microscopy analysis of induced mutants of sorghum

    CSIR Research Space (South Africa)

    Mbambo, Z

    2014-01-01

    Full Text Available of elements in preferential accumulation tissues and entire changes in cellular localization. Transmission and scanning electron microscopy of the mutants resolved changes in size, shape, ultra-structure and packed cell volumes of protein- and starch bodies...

  10. Detection of silver nanoparticles inside marine diatom Thalassiosira pseudonana by electron microscopy and focused ion beam.

    Directory of Open Access Journals (Sweden)

    César Pascual García

    Full Text Available In the following article an electron/ion microscopy study will be presented which investigates the uptake of silver nanoparticles (AgNPs by the marine diatom Thalassiosira pseudonana, a primary producer aquatic species. This organism has a characteristic silica exoskeleton that may represent a barrier for the uptake of some chemical pollutants, including nanoparticles (NPs, but that presents a technical challenge when attempting to use electron-microscopy (EM methods to study NP uptake. Here we present a convenient method to detect the NPs interacting with the diatom cell. It is based on a fixation procedure involving critical point drying which, without prior slicing of the cell, allows its inspection using transmission electron microscopy. Employing a combination of electron and ion microscopy techniques to selectively cut the cell where the NPs were detected, we are able to demonstrate and visualize for the first time the presence of AgNPs inside the cell membrane.

  11. Transmission electron microscopy of unstained hybrid Au nanoparticles capped with PPAA (plasma-poly-allylamine)

    DEFF Research Database (Denmark)

    Gontard, Lionel C.; Fernández, Asunción; Dunin-Borkowski, Rafal E.;

    2014-01-01

    Hybrid (organic shell-inorganic core) nanoparticles have important applications in nanomedicine. Although the inorganic components of hybrid nanoparticles can be characterized readily using conventional transmission electron microscopy (TEM) techniques, the structural and chemical arrangement of ...

  12. Scanning electron microscopy of acrothoracican cypris larvae (Crustacea, Thecostraca, Cirripedia, Acrothoracica, Lithoglyptidae)

    NARCIS (Netherlands)

    Kolbasov, Gregory A.; Høeg, Jens T.; Elfimov, Alexei S.

    1999-01-01

    Scanning electron microscopy was used to provide a full morphological description of cypris morphology in the acrothoracican species Lithoglyptes milis and L. habei (Lithoglyptidae). Special attention was given to lattice organs, antennules, thorax, thoracopods, abdomen, and furcal rami. Cypris larv

  13. Automated magnification calibration in transmission electron microscopy using Fourier analysis of replica images.

    NARCIS (Netherlands)

    Laak, J.A.W.M. van der; Dijkman, H.B.P.M.; Pahlplatz, M.M.M.

    2006-01-01

    The magnification factor in transmission electron microscopy is not very precise, hampering for instance quantitative analysis of specimens. Calibration of the magnification is usually performed interactively using replica specimens, containing line or grating patterns with known spacing. In the pre

  14. Quantitative detection of gold nanoparticles on individual, unstained cancer cells by scanning electron microscopy

    NARCIS (Netherlands)

    Hartsuiker, L.; Es, van P.; Petersen, W.; Leeuwen, van 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

  15. 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

  16. Electron microscopy observations of surface morphologies and particle arrangement behaviors of magnetic fluids

    Institute of Scientific and Technical Information of China (English)

    沈辉; 徐雪青; 王伟

    2003-01-01

    The surface morphology of quasi-periodic stripe-shaped patterns of magnetite fluids was observed in applied perpendicular magnetic fields by means of scanning electron microscopy. The nanoparticles of the magnetite fluids are arranged in oriental quasilinear chains in applied perpendicular magnetic fields as observed using transmission electron microscopy. This arrangement results from particle-particle interactions and particle-carrier liquids interactions, which are eventually controlled by the magnetic fields distribution.

  17. Low voltage scanning electron microscopy of interplanetary dust particles

    Science.gov (United States)

    Blake, D. F.; Bunch, T. E.; Reilly, T. W.; Brownlee, D. E.

    1987-01-01

    The resolution of available low-voltage SEM (LVSEM) models used in the characterization of interplanetary dust particles (IDPs) is limited by a number of factors including energy spread in the electron source, beam brightness, scanning electron detector geometry, and various lens aberrations. This paper describes an improved model of LVSEM which offers an increased resolution at low voltage. The improvements include a cold cathode FE source which has an extremely low inherent energy spread and high brightness, a second condenser lens to converge the beam and maintain an optimum aperture half-angle, and a detector optimized for low-voltage scanning-electron collection. To reduce lens aberrations, the specimen is immersed in the objective lens field. The features of several IDP samples observed using the images obtained with this LVSEM model are described.

  18. Low voltage scanning electron microscopy of interplanetary dust particles

    Science.gov (United States)

    Blake, D. F.; Bunch, T. E.; Reilly, T. W.; Brownlee, D. E.

    1987-01-01

    The resolution of available low-voltage SEM (LVSEM) models used in the characterization of interplanetary dust particles (IDPs) is limited by a number of factors including energy spread in the electron source, beam brightness, scanning electron detector geometry, and various lens aberrations. This paper describes an improved model of LVSEM which offers an increased resolution at low voltage. The improvements include a cold cathode FE source which has an extremely low inherent energy spread and high brightness, a second condenser lens to converge the beam and maintain an optimum aperture half-angle, and a detector optimized for low-voltage scanning-electron collection. To reduce lens aberrations, the specimen is immersed in the objective lens field. The features of several IDP samples observed using the images obtained with this LVSEM model are described.

  19. Structural studies of glasses by transmission electron microscopy and electron diffraction

    Energy Technology Data Exchange (ETDEWEB)

    Kashchieva, E.P. [University of Chemical Technology and Metallurgy, Sofia (Bulgaria)

    1997-07-01

    The purpose of this work is to present information about the applications of transmission electron microscopy (TEM) and electron diffraction (ED) for structural investigations of glasses. TEM investigations have been carried out on some binary and on a large number of ternary borate-telluride systems where glass-forming oxides, oxides of transitional elements and modified oxides of elements from I, II and III groups in the periodic table, are used as third component. The large experimental data given by TEM method allows the fine classification of the micro-heterogeneities. A special case of micro-heterogeneous structure with technological origin occurs near the boundary between the 2 immiscible liquids obtained at macro-phase separation. TEM was also used for the direct observation of the glass structure and we have studied the nano-scale structure of borate glasses obtained at slow and fast cooling of the melts. The ED possesses advantages for analysis of amorphous thin films or micro-pastilles and it is a very useful technique for study in materials containing simultaneously light and heavy elements. A comparison between the possibilities of the 3 diffraction techniques (X-ray diffraction, neutron diffraction and ED) is presented.

  20. 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.

  1. 40 CFR Appendix A to Subpart E of... - Interim Transmission Electron Microscopy Analytical Methods-Mandatory and Nonmandatory-and...

    Science.gov (United States)

    2010-07-01

    ..., Subpt. E, App. A Appendix A to Subpart E of Part 763—Interim Transmission Electron Microscopy Analytical... completion of response actions. This unit is mandatory. II. Mandatory Transmission Electron Microscopy Method... sample. 17. PCM—Phase contrast microscopy. 18. SAED—Selected area electron diffraction. 19....

  2. Scanning electron microscopy of erythropoietin-stimulated bone marrow

    Energy Technology Data Exchange (ETDEWEB)

    Leblond, P.F. (Hospital of St. Sacrement, Quebec); Chamberlain, J.K.; Weed, R.I.

    1975-01-01

    This work describes and illustrates the scanning electron microscopic modifications observed in the femoral bone marrow of normal mice 72 hours after a single injection of partly purified sheep erythropoietin and of mice afflicted with a chronic congenital hemolytic anemia analogous to the disease Hereditary Spherocytosis in man. In acordance with previous transmission electron microscopic studies, the observations are consistent with an effect of erythropoietin both on the frequency of cell migration across the normally intact marrow sinus endothelium and on the morphology of sinus adventitial cells. It is suggested that these ultrastructural modifications may be responsible for the greater patency of the marrow-blood barrier under erythropoietin stimulation.

  3. Image simulations of kinked vortices for transmission electron microscopy

    DEFF Research Database (Denmark)

    Beleggia, Marco; Pozzi, G.; Tonomura, A.

    2010-01-01

    We present an improved model of kinked vortices in high-Tc superconductors suitable for the interpretation of Fresnel or holographic observations carried out with a transmission electron microscope. A kinked vortex is composed of two displaced half-vortices, perpendicular to the film plane...... observations of high-Tc superconducting films, where the Fresnel contrast associated with some vortices showed a dumbbell like appearance. Here, we show that under suitable conditions the JV segment may reveal itself in Fresnel imaging or holographic phase mapping in a transmission electron microscope....

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

    Science.gov (United States)

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

    2012-12-01

    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 0.5Å 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.

  5. A quantum mechanical scheme to reduce radiation damage in electron microscopy

    CERN Document Server

    Okamoto, Hiroshi; Fink, Hans-Werner

    2015-01-01

    We show that radiation damage to unstained biological specimens is not an intractable problem in electron microscopy. When a structural hypothesis of a specimen is available, quantum mechanical principles allow us to verify the hypothesis with a very low electron dose. Realization of such a concept requires precise control of the electron wave front. Based on a diffractive electron optical implementation, we demonstrate the feasibility of this new method by both experimental and numerical investigations.

  6. Transmission electron microscopy characterization of photocatalysts for water splitting

    DEFF Research Database (Denmark)

    Cavalca, Filippo; Laursen, Anders Bo; Dahl, Søren

    , it is necessary to understand the fundamentals of their reaction mechanisms, chemical behavior, structure and morphology before, during and after reaction using in situ investigations. Here, we focus on the in situ characterization of photocatalysts [1] in an environmental transmission electron microscope (ETEM...

  7. Stratification of centrifuged amoeba nuclei investigated by electron microscopy

    Science.gov (United States)

    Breyer, E. P.; Daniels, E. W.

    1968-01-01

    Study establishes a relationship between radioresistance and the nucleolar stratification characteristics of various amoeba species. Two species of fresh water amoeba are studied with the electron microscope. The report discusses the nature of nucleolar layers and their possible relationship to the differences in radiosensitivity of the two amoeba species.

  8. Early studies of placental ultrastructure by electron microscopy

    DEFF Research Database (Denmark)

    Carter, A M; Enders, A C

    2016-01-01

    many other scientists to Washington University in St. Louis. Work on human placental ultrastructure was initiated at Cambridge and Kyoto whilst domestic animals were initially studied by Björkman in Stockholm and electron micrographs of bat placenta were published by Wimsatt of Cornell University...

  9. Bulk sensitive hard x-ray photoemission electron microscopy

    Energy Technology Data Exchange (ETDEWEB)

    Patt, M., E-mail: m.patt@fz-juelich.de; Wiemann, C. [Peter Grünberg Institute (PGI-6) and JARA-FIT, Research Center Jülich, D-52425 Jülich (Germany); Weber, N.; Escher, M.; Merkel, M. [Focus GmbH, Neukirchner Str. 2, D-65510 Hünstetten (Germany); Gloskovskii, A.; Drube, W. [DESY Photon Science, Deutsches Elektronen-Synchrotron, D-22603 Hamburg (Germany); Schneider, C. M. [Peter Grünberg Institute (PGI-6) and JARA-FIT, Research Center Jülich, D-52425 Jülich (Germany); Fakultät f. Physik and Center for Nanointegration Duisburg-Essen (CeNIDE), Universität Duisburg-Essen, D-47048 Duisburg (Germany)

    2014-11-15

    Hard x-ray photoelectron spectroscopy (HAXPES) has now matured into a well-established technique as a bulk sensitive probe of the electronic structure due to the larger escape depth of the highly energetic electrons. In order to enable HAXPES studies with high lateral resolution, we have set up a dedicated energy-filtered hard x-ray photoemission electron microscope (HAXPEEM) working with electron kinetic energies up to 10 keV. It is based on the NanoESCA design and also preserves the performance of the instrument in the low and medium energy range. In this way, spectromicroscopy can be performed from threshold to hard x-ray photoemission. The high potential of the HAXPEEM approach for the investigation of buried layers and structures has been shown already on a layered and structured SrTiO{sub 3} sample. Here, we present results of experiments with test structures to elaborate the imaging and spectroscopic performance of the instrument and show the capabilities of the method to image bulk properties. Additionally, we introduce a method to determine the effective attenuation length of photoelectrons in a direct photoemission experiment.

  10. Invited Talk: Electron Microscopy of Quantum Dots for Display Applications

    OpenAIRE

    Fern, G; Silver, J.; Ireland, T; Howkins, A; Hobson, PH; Coe-Sullivan, S

    2015-01-01

    CdSe/ZnCdS core/shell Quantum dots with high quantum yield (~84%) were used in this experiment. For the first time the red filtered cathodoluminescence images are shown along with their corresponding electron energy loss spectrum map, and high angle annular dark field image of the corresponding particles is shown.

  11. Low impact to fixed cell processing aiming transmission electron microscopy

    Science.gov (United States)

    Barth, Ortrud Monika; da Silva, Marcos Alexandre Nunes; Barreto-Vieira, Debora Ferreira

    2016-01-01

    In cell culture, cell structures suffer strong impact due to centrifugation during processing for electron microscope observation. In order to minimise this effect, a new protocol was successfully developed. Using conventional reagents and equipments, it took over one week, but cell compression was reduced to none or the lowest deformation possible. PMID:27276186

  12. Scanning electron microscopy of the male genitalia of Sarcophagidae (Diptera

    Directory of Open Access Journals (Sweden)

    Hugo de Souza Lopes

    1990-03-01

    Full Text Available The male genitalia of nine species of Sarcophagidae (Diptera - Goniophyto honsuensis Rohdendorf, 1962, Tricharaea brevicornis (Wiedemann, 1830, Chaetoravinia derelicta (Walker, 1852, Austrohartigia spinigena (Rondani, 1864, Chrysagria duodecimpunctata Townsend, 1935, Boettcheria bisetosa Parker, 1914, Lipoptilocnema lanei Townsend, 1934, L. crispina (Lopes, 1938 and Euboettcheria alvarengai Lopes & Tibana, 1982 - were examined by scanning electron microscope (SEM and the main morphological features are descirbed.

  13. Scanning electron microscopy analysis of experimental bone hacking trauma.

    Science.gov (United States)

    Alunni-Perret, Veronique; Muller-Bolla, Michèle; Laugier, Jean-Pierre; Lupi-Pégurier, Laurence; Bertrand, Marie-France; Staccini, Pascal; Bolla, Marc; Quatrehomme, Gérald

    2005-07-01

    The authors report on their macro- and microscopy study of bone lesions made by a sharp force instrument (a single blade knife), and a sharp-blunt instrument classified as a chopping weapon (a hatchet). The aim of this work was to attempt to identify the instrument by analyzing the general class characteristics of the cuts. Each weapon was used on human bones. The results indicate that macroscopic analysis is more problematic. The microscopic analysis assessed that characteristics examined were effective in distinguishing sharp from sharp-blunt injury to the bone. The microscope facilitates analysis unachievable with macroscopic methods, some three-dimensional characteristics not visible to the naked eye being clearly defined with its use. Emphasis has been placed on the value of SEM as an anthropologist's tool in bone lesion injuries.

  14. Integrating electron microscopy into nanoscience and materials engineering programs

    Science.gov (United States)

    Cormia, Robert D.; Oye, Michael M.; Nguyen, Anh; Skiver, David; Shi, Meng; Torres, Yessica

    2014-10-01

    Preparing an effective workforce in high technology is the goal of both academic and industry training, and has been the engine that drives innovation and product development in the United States for over a century. During the last 50 years, technician training has comprised a combination of two-year academic programs, internships and apprentice training, and extensive On-the-Job Training (OJT). Recently, and especially in Silicon Valley, technicians have four-year college degrees, as well as relevant hands-on training. Characterization in general, and microscopy in particular, is an essential tool in process development, manufacturing and QA/QC, and failure analysis. Training for a broad range of skills and practice is challenging, especially for community colleges. Workforce studies (SRI/Boeing) suggest that even four year colleges often do not provide the relevant training and experience in laboratory skills, especially design of experiments and analysis of data. Companies in high-tech further report difficulty in finding skilled labor, especially with industry specific experience. Foothill College, in partnership with UCSC, SJSU, and NASA-Ames, has developed a microscopy training program embedded in a research laboratory, itself a partnership between university and government, providing hands-on experience in advanced instrumentation, experimental design and problem solving, with real-world context from small business innovators, in an environment called `the collaboratory'. The program builds on AFM-SEM training at Foothill, and provides affordable training in FE-SEM and TEM through a cost recovery model. In addition to instrument and engineering training, the collaboratory also supports academic and personal growth through a multiplayer social network of students, faculty, researchers, and innovators.

  15. Quantitating morphological changes in biological samples during scanning electron microscopy sample preparation with correlative super-resolution microscopy.

    Science.gov (United States)

    Zhang, Ying; Huang, Tao; Jorgens, Danielle M; Nickerson, Andrew; Lin, Li-Jung; Pelz, Joshua; Gray, Joe W; López, Claudia S; Nan, Xiaolin

    2017-01-01

    Sample preparation is critical to biological electron microscopy (EM), and there have been continuous efforts on optimizing the procedures to best preserve structures of interest in the sample. However, a quantitative characterization of the morphological changes associated with each step in EM sample preparation is currently lacking. Using correlative EM and superresolution microscopy (SRM), we have examined the effects of different drying methods as well as osmium tetroxide (OsO4) post-fixation on cell morphology during scanning electron microscopy (SEM) sample preparation. Here, SRM images of the sample acquired under hydrated conditions were used as a baseline for evaluating morphological changes as the sample went through SEM sample processing. We found that both chemical drying and critical point drying lead to a mild cellular boundary retraction of ~60 nm. Post-fixation by OsO4 causes at least 40 nm additional boundary retraction. We also found that coating coverslips with adhesion molecules such as fibronectin prior to cell plating helps reduce cell distortion from OsO4 post-fixation. These quantitative measurements offer useful information for identifying causes of cell distortions in SEM sample preparation and improving current procedures.

  16. Electron microscopy localization and characterization of functionalized composite organic-inorganic SERS nanoparticles on leukemia cells

    Energy Technology Data Exchange (ETDEWEB)

    Koh, Ai Leen [Department of Materials Science and Engineering, Stanford University, Durand Building Room 139, 496 Lomita Mall, Stanford, CA 94305 (United States); Stanford Nanocharacterization Laboratory, Stanford University, Stanford, CA 94305 (United States); Department of Mechanical Engineering, Stanford University, Stanford, CA 94305 (United States)], E-mail: alkoh@stanford.edu; Shachaf, Catherine M.; Elchuri, Sailaja; Nolan, Garry P. [Baxter Laboratory in Genetic Pharmacology, Department of Microbiology and Immunology, Stanford University, Stanford, CA 94305 (United States); Sinclair, Robert [Department of Materials Science and Engineering, Stanford University, Durand Building Room 139, 496 Lomita Mall, Stanford, CA 94305 (United States); Stanford Nanocharacterization Laboratory, Stanford University, Stanford, CA 94305 (United States)

    2008-12-15

    We demonstrate the use of electron microscopy as a powerful characterization tool to identify and locate antibody-conjugated composite organic-inorganic nanoparticle (COINs) surface enhanced Raman scattering (SERS) nanoparticles on cells. U937 leukemia cells labeled with antibody CD54-conjugated COINs were characterized in their native, hydrated state using wet scanning electron microscopy (SEM) and in their dehydrated state using high-resolution SEM. In both cases, the backscattered electron (BSE) detector was used to detect and identify the silver constituents in COINs due to its high sensitivity to atomic number variations within a specimen. The imaging and analytical capabilities in the SEM were further complemented by higher resolution transmission electron microscopy (TEM) images and scanning Auger electron spectroscopy (AES) data to give reliable and high-resolution information about nanoparticles and their binding to cell surface antigens.

  17. Nanomaterial datasets to advance tomography in scanning transmission electron microscopy

    CERN Document Server

    Levin, Barnaby D A; Chen, Chien-Chun; Scott, M C; Xu, Rui; Theis, Wolfgang; Jiang, Yi; Yang, Yongsoo; Ophus, Colin; Zhang, Haitao; Ha, Don-Hyung; Wang, Deli; Yu, Yingchao; Abruna, Hector D; Robinson, Richard D; Ercius, Peter; Kourkoutis, Lena F; Miao, Jianwei; Muller, David A; Hovden, Robert

    2016-01-01

    Electron tomography in materials science has flourished with the demand to characterize nanoscale materials in three dimensions (3D). Access to experimental data is vital for developing and validating reconstruction methods that improve resolution and reduce radiation dose requirements. This work presents five high-quality scanning transmission electron microscope (STEM) tomography datasets in order to address the critical need for open access data in this field. The datasets represent the current limits of experimental technique, are of high quality, and contain materials with structural complexity. Included are tomographic series of a hyperbranched Co2P nanocrystal, platinum nanoparticles on a carbon nanofibre imaged over the complete 180{\\deg} tilt range, a platinum nanoparticle and a tungsten needle both imaged at atomic resolution by equal slope tomography, and a through-focal tilt series of PtCu nanoparticles. A volumetric reconstruction from every dataset is provided for comparison and development of p...

  18. Quantification of interfacial segregation by analytical electron microscopy

    CERN Document Server

    Muellejans, H

    2003-01-01

    The quantification of interfacial segregation by spatial difference and one-dimensional profiling is presented in general where special attention is given to the random and systematic uncertainties. The method is demonstrated for an example of Al-Al sub 2 O sub 3 interfaces in a metal-ceramic composite material investigated by energy-dispersive X-ray spectroscopy and electron energy loss spectroscopy in a dedicated scanning transmission electron microscope. The variation of segregation measured at different interfaces by both methods is within the uncertainties, indicating a constant segregation level and interfacial phase formation. The most important random uncertainty is the counting statistics of the impurity signal whereas the specimen thickness introduces systematic uncertainties (via k factor and effective scan width). The latter could be significantly reduced when the specimen thickness is determined explicitly. (orig.)

  19. A Nanoaquarium for in situ Electron Microscopy in Liquid Media

    CERN Document Server

    Grogan, Joseph M

    2010-01-01

    The understanding of many nanoscale processes occurring in liquids such as colloidal crystal formation, aggregation, nanowire growth, electrochemical deposition, and biological interactions would benefit greatly from real-time, in situ imaging with the nanoscale resolution of transmission electron microscopes (TEMs) and scanning transmission electron microscopes (STEMs). However, these imaging tools cannot readily be used to observe processes occurring in liquid media without addressing two experimental hurdles: sample thickness and sample evaporation in the high vacuum microscope chamber. To address these challenges, we have developed a nano-Hele-Shaw cell, dubbed the nanoaquarium. The device consists of a hermetically-sealed, 100 nm tall, liquid-filled chamber sandwiched between two freestanding, 50 nm thick, silicon nitride membranes. Embedded electrodes are integrated into the device. This fluid dynamics video features particle motion and aggregation during in situ STEM of nanoparticles suspended in liqui...

  20. Molybdenum work function determined by electron emission microscopy.

    Science.gov (United States)

    Jacobson, D. L.; Campbell, A. E.

    1971-01-01

    A polycrystalline molybdenum sample was recrystallized and thermally stabilized. Quantitative measurements of the emission from each individual grain were obtained with an electron emission microscope. The effective work function for each grain was then calculated. The crystallographic orientation of each grain was determined by Laue back-reflection techniques. A polar plot of effective work function vs crystallographic orientation for the sample was constructed to provide a correlation between effective work function and crystallographic orientation.

  1. Electron microscopy study of the iron meteorite Santa Catharina

    Science.gov (United States)

    Zhang, J.; Williams, D. B.; Goldstein, J. I.; Clarke, R. S., Jr.

    1990-01-01

    A characterization of the microstructural features of Santa Catharina (SC) from the millimeter to submicron scale is presented. The same specimen was examined using an optical microscope, a scanning electron microscope, an electron probe microanalyzer, and an analytical electron microscope. Findings include the fact that SC metal nodules may have different bulk Ni values, leading to different microstructures upon cooling; that SC USNM 6293 is the less corroded sample, as tetrataenite exists as less than 10 nm ordered domains throughout the entire fcc matrix (it is noted that this structure is the same as that of the Twin City meteorite and identical to clear taenite II in the retained taenite regions of the octahedrites); that SC USNM 3043 has a more complicated microstructure due to corrosion; and that the low Ni phase of the cloudy zone was selectively corroded in some areas and formed the dark regions, indicating that the SC meteorite corrosion process was electrochemical in nature and may involve Cl-containing akaganeite.

  2. Environmental Transmission Electron Microscopy of catalysts for the methanol synthesis

    DEFF Research Database (Denmark)

    Duchstein, Linus Daniel Leonhard

    Everywhere around the world, natural resources like crude oil are becoming less and harder to extract. It is therefore necessary to find alternatives to secure our future transportation in a sustainable way. This can be done e.g. through chemical conversion of lignocelluloses into bio-alcohol thr......Everywhere around the world, natural resources like crude oil are becoming less and harder to extract. It is therefore necessary to find alternatives to secure our future transportation in a sustainable way. This can be done e.g. through chemical conversion of lignocelluloses into bio...... atmosphere and temperature was investigated. CuNi was exposed to the electron beam for 3 different intensities and 3 different temperatures while the oxidation state of the Cu2+ was measured by energy electron loss spectroscopy. It turns out that the electron beam does have an influence but it does not seem...... to scale with the beam current density but foremost with the exposure time. The ETEM shows phase and chemical changes during the reaction....

  3. Electron tomography of HEK293T cells using scanning electron microscope-based scanning transmission electron microscopy.

    Science.gov (United States)

    You, Yun-Wen; Chang, Hsun-Yun; Liao, Hua-Yang; Kao, Wei-Lun; Yen, Guo-Ji; Chang, Chi-Jen; Tsai, Meng-Hung; Shyue, Jing-Jong

    2012-10-01

    Based on a scanning electron microscope operated at 30 kV with a homemade specimen holder and a multiangle solid-state detector behind the sample, low-kV scanning transmission electron microscopy (STEM) is presented with subsequent electron tomography for three-dimensional (3D) volume structure. Because of the low acceleration voltage, the stronger electron-atom scattering leads to a stronger contrast in the resulting image than standard TEM, especially for light elements. Furthermore, the low-kV STEM yields less radiation damage to the specimen, hence the structure can be preserved. In this work, two-dimensional STEM images of a 1-μm-thick cell section with projection angles between ±50° were collected, and the 3D volume structure was reconstructed using the simultaneous iterative reconstructive technique algorithm with the TomoJ plugin for ImageJ, which are both public domain software. Furthermore, the cross-sectional structure was obtained with the Volume Viewer plugin in ImageJ. Although the tilting angle is constrained and limits the resulting structural resolution, slicing the reconstructed volume generated the depth profile of the thick specimen with sufficient resolution to examine cellular uptake of Au nanoparticles, and the final position of these nanoparticles inside the cell was imaged.

  4. 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.

  5. The importance of transmission electron microscopy analysis of spermatozoa: Diagnostic applications and basic research.

    Science.gov (United States)

    Moretti, Elena; Sutera, Gaetano; Collodel, Giulia

    2016-06-01

    This review is aimed at discussing the role of ultrastructural studies on human spermatozoa and evaluating transmission electron microscopy as a diagnostic tool that can complete andrology protocols. It is clear that morphological sperm defects may explain decreased fertilizing potential and acquire particular value in the field of male infertility. Electron microscopy is the best method to identify systematic or monomorphic and non-systematic or polymorphic sperm defects. The systematic defects are characterized by a particular anomaly that affects the vast majority of spermatozoa in a semen sample, whereas a heterogeneous combination of head and tail defects found in variable percentages are typically non-systematic or polymorphic sperm defects. A correct diagnosis of these specific sperm alterations is important for choosing the male infertility's therapy and for deciding to turn to assisted reproduction techniques. Transmission electron microscopy (TEM) also represents a valuable method to explore the in vitro effects of different compounds (for example drugs with potential spermicidal activity) on the morphology of human spermatozoa. Finally, TEM used in combination with immunohistochemical techniques, integrates structural and functional aspects that provide a wide horizon in the understanding of sperm physiology and pathology. transmission electron microscopy: TEM; World Health Organization: WHO; light microscopy: LM; motile sperm organelle morphology examination: MSOME; intracytoplasmic morphologically selected sperm injection: IMSI; intracytoplasmic sperm injection: ICSI; dysplasia of fibrous sheath: DFS; primary ciliary dyskinesia: PCD; outer dense fibers: ODF; assisted reproduction technologies: ART; scanning electron microscopy: SEM; polyvinylpirrolidone: PVP; tert-butylhydroperoxide: TBHP.

  6. Channelling and related effects in electron microscopy: The current status

    Energy Technology Data Exchange (ETDEWEB)

    Krishnan, K.M.

    1989-05-01

    Channelling or Borrmann effect in electron diffraction has been developed into a versatile, high spatial resolution, crystallographic technique with demonstrated applicability in solving a variety of materials problems. In general, either the characteristic x-ray emissions or the electron energy-loss intensities are monitored as a function of the orientation of the incident beam. The technique, as formulated in the planar geometry has found wide applications in specific site occupancy and valence measurements, determination of small atomic displacements and crystal polarity studies. For site occupancy studies, the appropriate orientations in most cases can be determined by inspection and the analysis carried out according to a simple classification of the crystal structure discussed in this paper. Concentration levels as low as 0.1 wt% can be easily detected. The reciprocity principle may be used to advantage in all these studies, if electron energy-loss spectra are monitored, as both the channelling of the incoming beam and the blocking of the outgoing beam are included in the formulation and analysis. The formulation in the axial geometry is an useful alternative, particularly for monatomic crystals. Localization effects are important if, either the experiment is performed in the axial geometry or if low atomic number elements (z < 11) are detected. In general, the sensitivity to L-shells is lower compared to K-shell excitations. Other experimental parameters to be considered include temperature of the sample, the acceleration voltage and parallelism of the incident beam. Any detrimental effects of channelling on conventional microanalysis can be minimized either by tilting the crystal to an orientation where no lower order diffraction vectors are excited or by using a convergent probe such that a large range of incident beam orientations are averaged in the analysis. 49 refs., 9 figs.

  7. Transmission Electron Microscopy of Magnetite Plaquettes in Orgueil

    Science.gov (United States)

    Chan, Q. H. S.; Han, J.; Zolensky, M.

    2016-01-01

    Magnetite sometimes takes the form of a plaquette - barrel-shaped stack of magnetite disks - in carbonaceous chondrites (CC) that show evidence of aqueous alteration. The asymmetric nature of the plaquettes caused Pizzarello and Groy to propose magnetite plaquettes as a naturally asymmetric mineral that can indroduce symmetry-breaking in organic molecules. Our previous synchrotron X-ray computed microtomography (SXRCT) and electron backscatter diffraction (EBSD) analyses of the magnetite plaquettes in fifteen CCs indicate that magnetite plaquettes are composed of nearly parallel discs, and the crystallographic orientations of the discs change around a rotational axis normal to the discs surfaces. In order to further investigate the nanostructures of magnetite plaquettes, we made two focused ion beam (FIB) sections of nine magnetite plaquettes from a thin section of CI Orgueil for transmission electron microscope (TEM) analysis. The X-ray spectrum imaging shows that the magnetite discs are purely iron oxide Fe3O4 (42.9 at% Fe and 57.1 at% O), which suggest that the plaquettes are of aqueous origin as it is difficult to form pure magnetite as a nebular condensate. The selected area electron diffraction (SAED) patterns acquired across the plaquettes show that the magnetite discs are single crystals. SEM and EBSD analyses suggest that the planar surfaces of the magnetite discs belong to the {100} planes of the cubic inverse spinel structure, which are supported by our TEM observations. Kerridge et al. suggested that the epitaxial relationship between magnetite plaquette and carbonate determines the magnetite face. However, according to our TEM observation, the association of magnetite with porous networks of phyllosilicate indicates that the epitaxial relationship with carbonate is not essential to the formation of magnetite plaquettes. It was difficult to determine the preferred rotational orientation of the plaquettes due to the symmetry of the cubic structure

  8. Scanning Electron Microscopy of Lagochilascaris minor Leiper, 1909 (Nematoda: Ascarididae

    Directory of Open Access Journals (Sweden)

    Lanfredi Reinalda Marisa

    1998-01-01

    Full Text Available Lagochilascaris minor Leiper, 1909 is a parasitic nematode with its biological cycle still unknown, even though it was found in humans, domestic and silvatic animals. Adult worms, collected by surgical drainage from a human patient from the State of Pará, Brazil, were micrographed using a scanning electron microscope. Morphological aspects of males and females such as cephalic structures, caudal papillae and cuticular patterns were analyzed and compared with the previous descriptions adding new data for the identification of this species.

  9. [Scanning electron microscopy of heat-damaged bone tissue].

    Science.gov (United States)

    Harsanyl, L

    1977-02-01

    Parts of diaphyses of bones were exposed to high temperature of 200-1300 degrees C. Damage to the bone tissue caused by the heat was investigated. The scanning electron microscopic picture seems to be characteristic of the temperature applied. When the bones heated to the high temperature of 700 degrees C characteristic changes appear on the periostal surface, higher temperatura on the other hand causes damage to the compact bone tissue and can be observed on the fracture-surface. Author stresses the importance of this technique in the legal medicine and anthropology.

  10. Electron microscopy analysis of structural changes within white etching areas

    DEFF Research Database (Denmark)

    Diederichs, Annika Martina; Schwedt, A.; Mayer, J.

    2016-01-01

    In the present work, crack networks with white etching areas (WEAs) in cross-sections of bearings were investigated by a complementary use of SEM and TEM with the focus on the use of orientation contrast imaging and electron backscatter diffraction (EBSD). Orientation contrast imaging was used...... observed within WEAs. Using EBSD analysis, evidence was obtained that WEA formation and accompanying crack growth are without relation microstructural features. In addition, an inhomogeneous chemical structure of WEA as a result of carbide dissolution is revealed by analytical investigations....

  11. 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

    2016-10-21

    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 AlxGa1-xAs, 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.

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

    NARCIS (Netherlands)

    Rührig, M.; Porthun, S.; Lodder, J.C.; McVitie, 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

  13. U-10Mo Sample Preparation and Examination using Optical and Scanning Electron Microscopy

    Energy Technology Data Exchange (ETDEWEB)

    Prabhakaran, Ramprashad [Pacific Northwest National Lab. (PNNL), Richland, WA (United States); Joshi, Vineet V. [Pacific Northwest National Lab. (PNNL), Richland, WA (United States); Rhodes, Mark A. [Pacific Northwest National Lab. (PNNL), Richland, WA (United States); Schemer-Kohrn, Alan L. [Pacific Northwest National Lab. (PNNL), Richland, WA (United States); Guzman, Anthony D. [Pacific Northwest National Lab. (PNNL), Richland, WA (United States); Lavender, Curt A. [Pacific Northwest National Lab. (PNNL), Richland, WA (United States)

    2016-10-01

    The purpose of this document is to provide guidelines to prepare specimens of uranium alloyed with 10 weight percent molybdenum (U-10Mo) for optical metallography and scanning electron microscopy. This document also provides instructions to set up an optical microscope and a scanning electron microscope to analyze U-10Mo specimens and to obtain the required information.

  14. Anisotropic Shape Changes of Silica Nanoparticles Induced in Liquid with Scanning Transmission Electron Microscopy

    NARCIS (Netherlands)

    Zecevic, J.; Hermannsdorfer, Justus; Schuh, Tobias; de Jong, Krijn P.; de Jonge, Niels

    2017-01-01

    Liquid-phase transmission electron microscopy (TEM) is used for in-situ imaging of nanoscale processes taking place in liquid, such as the evolution of nanoparticles during synthesis or structural changes of nanomaterials in liquid environment. Here, it is shown that the focused electron beam of

  15. Scanning Electron Microscopy (SEM) Procedure for HE Powders on a LEO 438VP System

    Energy Technology Data Exchange (ETDEWEB)

    Zaka, Fowzia [Lawrence Livermore National Lab. (LLNL), Livermore, CA (United States). Energetic Materials Center

    2016-03-21

    This method describes the characterization of HE powders by Scanning Electron Microscopy (SEM). HE particles are dispersed onto an aluminum standard SEM specimen mount. Electron micrographs are collected at various magnifications (150 to 10,000 X) depending on HE particle size.

  16. Scanning Electron Microscopy (SEM) Procedure for HE Powders on a LEO 438VP System

    Energy Technology Data Exchange (ETDEWEB)

    Zaka, Fowzia [Lawrence Livermore National Lab. (LLNL), Livermore, CA (United States). Energetic Materials Center

    2016-03-08

    This method describes the characterization of HE powders by Scanning Electron Microscopy (SEM). HE particles are dispersed onto an aluminum standard SEM specimen mount. Electron micrographs are collected at various magnifications (150 to 10,000 X) depending on HE particle size.

  17. Direct observations of the MOF (UiO-66) structure by transmission electron microscopy

    KAUST Repository

    Zhu, Liangkui

    2013-01-01

    As a demonstration of ab initio structure characterizations of nano metal organic framework (MOF) crystals by high resolution transmission electron microscopy (HRTEM) and electron diffraction tomography methods, a Zr-MOF (UiO-66) structure was determined and further confirmed by Rietveld refinements of powder X-ray diffraction. HRTEM gave direct imaging of the channels. © 2013 The Royal Society of Chemistry.

  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. U-10Mo Sample Preparation and Examination using Optical and Scanning Electron Microscopy

    Energy Technology Data Exchange (ETDEWEB)

    Prabhakaran, Ramprashad [Pacific Northwest National Lab. (PNNL), Richland, WA (United States); Joshi, Vineet V. [Pacific Northwest National Lab. (PNNL), Richland, WA (United States); Rhodes, Mark A. [Pacific Northwest National Lab. (PNNL), Richland, WA (United States); Schemer-Kohrn, Alan L. [Pacific Northwest National Lab. (PNNL), Richland, WA (United States); Guzman, Anthony D. [Pacific Northwest National Lab. (PNNL), Richland, WA (United States); Lavender, Curt A. [Pacific Northwest National Lab. (PNNL), Richland, WA (United States)

    2016-03-30

    The purpose of this document is to provide guidelines to prepare specimens of uranium alloyed with 10 weight percent molybdenum (U-10Mo) for optical metallography and scanning electron microscopy. This document also provides instructions to set up an optical microscope and a scanning electron microscope to analyze U-10Mo specimens and to obtain the required information.

  20. Integrated Raman and electron microscopy : correlative chemical specificity and nanoscale resolution

    NARCIS (Netherlands)

    Timmermans, Frank Jan

    2017-01-01

    This thesis describes the integration of a Raman microscope in a focused ion beam - scanning electron microscope (FIB-SEM). Raman micro-spectroscopy enables chemical specific characterization, while electron microscopy enables high resolution imaging. The Raman - SEM combination thus enables the

  1. Nanomaterial datasets to advance tomography in scanning transmission electron microscopy.

    Science.gov (United States)

    Levin, Barnaby D A; Padgett, Elliot; Chen, Chien-Chun; Scott, M C; Xu, Rui; Theis, Wolfgang; Jiang, Yi; Yang, Yongsoo; Ophus, Colin; Zhang, Haitao; Ha, Don-Hyung; Wang, Deli; Yu, Yingchao; Abruña, Hector D; Robinson, Richard D; Ercius, Peter; Kourkoutis, Lena F; Miao, Jianwei; Muller, David A; Hovden, Robert

    2016-06-07

    Electron tomography in materials science has flourished with the demand to characterize nanoscale materials in three dimensions (3D). Access to experimental data is vital for developing and validating reconstruction methods that improve resolution and reduce radiation dose requirements. This work presents five high-quality scanning transmission electron microscope (STEM) tomography datasets in order to address the critical need for open access data in this field. The datasets represent the current limits of experimental technique, are of high quality, and contain materials with structural complexity. Included are tomographic series of a hyperbranched Co2P nanocrystal, platinum nanoparticles on a carbon nanofibre imaged over the complete 180° tilt range, a platinum nanoparticle and a tungsten needle both imaged at atomic resolution by equal slope tomography, and a through-focal tilt series of PtCu nanoparticles. A volumetric reconstruction from every dataset is provided for comparison and development of post-processing and visualization techniques. Researchers interested in creating novel data processing and reconstruction algorithms will now have access to state of the art experimental test data.

  2. Progress and new advances in simulating electron microscopy datasets using MULTEM.

    Science.gov (United States)

    Lobato, I; Van Aert, S; Verbeeck, J

    2016-09-01

    A new version of the open source program MULTEM is presented here. It includes a graphical user interface, tapering truncation of the atomic potential, CPU multithreading functionality, single/double precision calculations, scanning transmission electron microscopy (STEM) simulations using experimental detector sensitivities, imaging STEM (ISTEM) simulations, energy filtered transmission electron microscopy (EFTEM) simulations, STEM electron energy loss spectroscopy (EELS) simulations along with other improvements in the algorithms. We also present a mixed channeling approach for the calculation of inelastic excitations, which allows one to considerably speed up time consuming EFTEM/STEM-EELS calculations.

  3. In-situ reduction of promoted cobalt oxide supported on alumina by environmental transmission electron microscopy

    DEFF Research Database (Denmark)

    Dehghan, Roya; Hansen, Thomas Willum; Wagner, Jakob Birkedal

    2011-01-01

    Reduction of 12wt.%Co/0.5wt.%Re/α-Al2O3 Fischer–Tropsch catalyst has been studied in-situ in an environmental transmission electron microscope. Reduction of Co3O4 to metallic cobalt was observed dynamically at 360 °C under 3.4 mbar H2. Structural and morphological changes were observed by high...... resolution transmission electron microscopy and scanning transmission electron microscopy imaging. The cobalt particles were mainly face centred cubic while some hexagonal close packed particles were also found. Reoxidation of the sample upon cooling to room temperature, still under flowing H2, underlines...

  4. Sub-micron imaging of buried integrated circuit structures using scanning confocal electron microscopy.

    Energy Technology Data Exchange (ETDEWEB)

    Frigo, S. P.; Levine, Z.; Zaluzec, N. J.; Materials Science Division; Northern Arizona Univ.; NIST

    2002-09-09

    Two-dimensional images of model integrated circuit components were collected using the technique of scanning confocal electron microscopy. For structures embedded about 5 {mu}m below the surface of a silicon oxide dielectric, a lateral resolution of 76{+-}9 nm was measured. Elemental mapping via x-ray emission spectrometry is demonstrated. A parallax analysis of images taken for various tilt angles to the electron beam allowed determination of the spacing between two wiring planes. The results show that scanning confocal electron microscopy is capable of probing buried structures at resolutions that will be necessary for the inspection of next-generation integrated circuit technology.

  5. Ballistic-Electron-Emission Microscopy Techniques for Nanometer-scale Characterization of Interfaces

    Science.gov (United States)

    Bell, L. D.; Grunthaner, F. J.; Hecht, M. H.; Manion, S. J.; Milliken, A. M.; Kaiser, W. J.

    1993-01-01

    Semiconductor interface properties are among the most important phenomena in materials science and technology. The study of metal/semiconductor Schottky barrier interfaces has been the primary focus of a large research and development community for decades. Throughout the long history of interface investigation, the study of interface defect electronic properties have been seriously hindered by the fundamental experimental difficulty of probing subsurface structures. A new method, Ballistic-Electron-Emission Microscopy (BEEM), has been developed which not only enables spectroscopic probing of subsurface interface properties, but also, provides nanometer-resolution imaging capabilities. BEEM employs Scanning Tunneling Microscopy (STM) and a unique spatially localized ballistic electron spectroscopy method...

  6. Electron microscopy approach for the visualization of the epithelial and endothelial glycocalyx.

    Science.gov (United States)

    Chevalier, L; Selim, J; Genty, D; Baste, J M; Piton, N; Boukhalfa, I; Hamzaoui, M; Pareige, P; Richard, V

    2017-06-01

    This study presents a methodological approach for the visualization of the glycocalyx by electron microscopy. The glycocalyx is a three dimensional network mainly composed of glycolipids, glycoproteins and proteoglycans associated with the plasma membrane. Since less than a decade, the epithelial and endothelial glycocalyx proved to play an important role in physiology and pathology, increasing its research interest especially in vascular functions. Therefore, visualization of the glycocalyx requires reliable techniques and its preservation remains challenging due to its fragile and dynamic organization, which is highly sensitive to the different process steps for electron microscopy sampling. In this study, chemical fixation was performed by perfusion as a good alternative to conventional fixation. Additional lanthanum nitrate in the fixative enhances staining of the glycocalyx in transmission electron microscopy bright field and improves its visualization by detecting the elastic scattered electrons, thus providing a chemical contrast. Copyright © 2017 Elsevier Masson SAS. All rights reserved.

  7. Thermal Properties of Materials Characterized by Scanning Electron-Acoustic Microscopy

    Institute of Scientific and Technical Information of China (English)

    GAO Chun-Ming; ZHANG Shu-Yi; ZHANG Zhong-Ning; SHUI Xiu-Ji; JIANG Tao

    2005-01-01

    @@ A modified technique of scanning electron-acoustic microscopy is employed to determine thermal diffusivity of materials. Using the dependence of the electron-acoustic signal on modulation frequency of the electron beam,the thermal diffusivity of materials is characterized based on a simplified thermoelastic theory. The thermal diffusivities of several metals characterized by the modified scanning electron-acoustic microscopy are in good agreement with the referential values of the corresponding materials, which proves that the scanning electronacoustic microscopy can be used to characterize the thermal diffusivity of materials effectively. In addition, for micro-inhomogeneous materials, such as biological tissues, the macro-effective (average) thermal diffusivities are characterized by the technique.

  8. The application of Graphene as a sample support in Transmission Electron Microscopy

    CERN Document Server

    Pantelic, R S; Kaiser, U; Stahlberg, H

    2012-01-01

    Transmission electron microscopy has witnessed rampant development and surging point resolution over the past few years. The improved imaging performance of modern electron microscopes shifts the bottleneck for image contrast and resolution to sample preparation. Hence, it is increasingly being realized that the full potential of electron microscopy will only be realized with the optimization of current sample preparation techniques. Perhaps the most recognized issues are background signal and noise contributed by sample supports, sample charging and instability. Graphene provides supports of single atom thickness, extreme physical stability, periodic structure, and ballistic electrical conductivity. As an increasing number of applications adapting graphene to their benefit emerge, we discuss the unique capabilities afforded by the use of graphene as a sample support for electron microscopy.

  9. Scanning electron microscopy of cells and tissues under fully hydrated conditions.

    Science.gov (United States)

    Thiberge, Stephan; Nechushtan, Amotz; Sprinzak, David; Gileadi, Opher; Behar, Vered; Zik, Ory; Chowers, Yehuda; Michaeli, Shulamit; Schlessinger, Joseph; Moses, Elisha

    2004-03-09

    A capability for scanning electron microscopy of wet biological specimens is presented. A membrane that is transparent to electrons protects the fully hydrated sample from the vacuum. The result is a hybrid technique combining the ease of use and ability to see into cells of optical microscopy with the higher resolution of electron microscopy. The resolution of low-contrast materials is approximately 100 nm, whereas in high-contrast materials the resolution can reach 10 nm. Standard immunogold techniques and heavy-metal stains can be applied and viewed in the fluid to improve the contrast. Images present a striking combination of whole-cell morphology with a wealth of internal details. A possibility for direct inspection of tissue slices transpires, imaging only the external layer of cells. Simultaneous imaging with photons excited by the electrons incorporates data on material distribution, indicating a potential for multilabeling and specific scintillating markers.

  10. Kinematics of gold nanoparticles manipulation in situ transmission electron microscopy

    Energy Technology Data Exchange (ETDEWEB)

    Alducin, Diego; Casillas, Gilberto; Mendoza-Santoyo, Fernando; Ponce, Arturo; José-Yacamán, Miguel, E-mail: miguel.yacaman@utsa.edu [University of Texas at San Antonio, Department of Physics and Astronomy (United States)

    2015-05-15

    Nanostructured materials such as nanoparticles, nanotubes, and nanowires are subject to different forces regimes compared with their macroscopic counterparts. In this work, we report the experimental manipulation of an individual gold nanoparticle (96 nm) capped with PVP considering forces surrounding the nanoparticle such as adhesion, friction, and the external load in real time, and how the differences between these forces produce distinct motions. Combining a scanning probe tool within a transmission electron microscope, we manipulated a gold nanoparticle and recorded the sliding and rolling kinematic motions. Our observations show quantitatively the adhesion force, maximum rolling resistance, and friction coefficients of the probe and the surface of the capped particle as well as particle and substrate surface.

  11. Ultrastructure of emulsions - a comparative electron microscopy study

    DEFF Research Database (Denmark)

    Jensen, Louise Helene Søgaard

    without off flavors. But the highly unsaturated nature of the fatty acids renders them especially subjective to oxidation; a process which creates undesired off flavors and deteriorates the nutritional value. It is therefore of interest to protect the fish oil against oxidation. One method suggested to do...... is added. The Nanomega project, which is a cooperation between the National Food Institute, the Center for Electron Nanoscopy and the Department of Mechanical Engineering, all at the Technical University of Denmark, has dealt mainly with pure oil in water emulsions to describe the oxidation without...... as a consequence of whether the emulsion has been processed at high pressure or not. The results suggest a structural similarity with the native casein micelle and a similar response to high pressure conditions, but diverging slightly in the resulting structure. This deviation can probably be ascribed to missing...

  12. Some strategies for quantitative scanning Auger electron microscopy

    Science.gov (United States)

    Browning, R.; Peacock, D. C.; Prutton, M.

    1985-01-01

    The general applicability of power law forms of the background in electron spectra is pointed out and exploited for background removal from under Auger peaks. This form of B(E) is found to be extremely sensitive to instrumental alignment and to fault-free construction - an observation which can be used to set up analyser configurations in an accurate way. Also, differences between N(E) and B(E) can be used to derive a spectrometer transmission function T(E). The questions of information density in an energy-analysing spatially-resolving instrument are addressed after reliable instrumental characterization has been established. Strategies involving ratio histograms, showing the population distribution of the ratio of a pair of Auger peak heights, composition scatter diagrams and windowed imaging are discussed and illustrated.

  13. Some strategies for quantitative scanning Auger electron microscopy

    Science.gov (United States)

    Browning, R.; Peacock, D. C.; Prutton, M.

    1985-01-01

    The general applicability of power law forms of the background in electron spectra is pointed out and exploited for background removal from under Auger peaks. This form of B(E) is found to be extremely sensitive to instrumental alignment and to fault-free construction - an observation which can be used to set up analyser configurations in an accurate way. Also, differences between N(E) and B(E) can be used to derive a spectrometer transmission function T(E). The questions of information density in an energy-analysing spatially-resolving instrument are addressed after reliable instrumental characterization has been established. Strategies involving ratio histograms, showing the population distribution of the ratio of a pair of Auger peak heights, composition scatter diagrams and windowed imaging are discussed and illustrated.

  14. Moessbauer spectroscopy and scanning electron microscopy of the Murchison meteorite

    Science.gov (United States)

    Brown, Christopher L.; Oliver, Frederick W.; Hammond, Ernest C., Jr.

    1989-01-01

    Meteorites provide a wealth of information about the solar system's formation, since they have similar building blocks as the Earth's crust but have been virtually unaltered since their formation. Some stony meteorites contain minerals and silicate inclusions, called chondrules, in the matrix. Utilizing Moessbauer spectroscopy, we identified minerals in the Murchison meteorite, a carbonaceous chondritic meteorite, by the gamma ray resonance lines observed. Absorption patterns of the spectra were found due to the minerals olivine and phyllosilicate. We used a scanning electron microscope to describe the structure of the chondrules in the Murchison meteorite. The chondrules were found to be deformed due to weathering of the meteorite. Diameters varied in size from 0.2 to 0.5 mm. Further enhancement of the microscopic imagery using a digital image processor was used to describe the physical characteristics of the inclusions.

  15. Scanning electron microscopy of human cortical bone failure surfaces.

    Science.gov (United States)

    Braidotti, P; Branca, F P; Stagni, L

    1997-02-01

    Undecalcified samples extracted from human femoral shafts are fractured by bending and the fracture surfaces are examined with a scanning electron microscope (SEM). The investigation is performed on both dry and wet (hydrated with a saline solution) specimens. SEM micrographs show patterns in many respects similar to those observed in fractography studies of laminated fiber-reinforced synthetic composites. In particular, dry and wet samples behave like brittle and ductile matrix laminates, respectively. An analysis carried out on the basis of the mechanisms that dominate the fracture process of laminates shows that a reasonable cortical bone model is that of a laminated composite material whose matrix is composed of extracellular noncollagenous calcified proteins, and the reinforcement is constituted by the calcified collagen fiber system.

  16. 48-Channel electron detector for photoemission spectroscopy and microscopy

    Science.gov (United States)

    Gregoratti, L.; Barinov, A.; Benfatto, E.; Cautero, G.; Fava, C.; Lacovig, P.; Lonza, D.; Kiskinova, M.; Tommasini, R.; Mähl, S.; Heichler, W.

    2004-01-01

    We show that it is possible to use a multichannel electron detector in a zone plate based photoemission spectromicroscopy in a snap shot mode to reduce the total acquisition time for a given counting time by 50% relative to the standard scanning mode while preserving the feature of the spectra. We describe the result of tests performed at Elettra using its microbeam (150 nm) together with a 48-channel detector designed for the PHOIBOS 100 analyzer optimized for extremely small x-ray sources. We also give a short summary of the technical features of the detector and describe one possible calibration procedure for its use in the snap shot mode. We show initial results from using this device to perform chemical maps of surfaces at a resolution of 150 nm.

  17. Survey of high voltage electron microscopy worldwide in 1998.

    Energy Technology Data Exchange (ETDEWEB)

    Allen, C. W.

    1998-03-05

    High voltage TEMs were introduced commercially thirty years ago, with the installations of 500 kV Hitachi instruments at the Universities of Nagoya and Tokyo. Since that time 53 commercial instruments, having maximum accelerating potentials of 0.5-3.5 MV, will have been delivered by the end of 1998. Table 1 summarizes the sites and some information regarding those HVEMS which are available in 1998. This corrects, updates and expands an earlier report of this sort [2]. There have been three commercial HVEM manufacturers: AEI (UK), Hitachi and JEOL (Japan). The proportion of the total number of HVEMS produced by each manufacturer is similar to that reflected in Table 1: AEI and Kratos/AEI (12), Hitachi (20) and JEOL (21). The term Kratos/AEI refers to instruments delivered after the takeover of AEI by Grates in the late 1970's. In Table 1 only maximum accelerating potentials are listed, which is generally also the design value for which the resolution for imaging was optimized. It is important to realize that in many applications, especially those studying irradiation effects, much lower voltages may be employed somewhat routinely to minimize atom displacements by the incident electron beam during analysis. These minimum values range from 100 kV for the AEI and Kratos/AEI instruments to typically 400 kV for the current generation of atomic resolution instruments, the latter being well above the thresholds for displacement in light elements such as Al and Si and for displacement of anions in many ceramic materials such as the high Tc superconductors, for example. An additional potential problem is electron-induced sputtering and differential sputtering (unequal sputtering rates in multicomponent materials), especially when accurate elemental microanalysis is being attempted. These same issues may arise for intermediate voltage TEMs as well, of course.

  18. Improved Hilbert phase contrast for transmission electron microscopy

    Energy Technology Data Exchange (ETDEWEB)

    Koeck, Philip J.B.

    2015-07-15

    Hilbert phase contrast has been recognized as a means of recording high resolution images with high contrast using a transmission electron microscope. This imaging mode could be used to image typical phase objects such as unstained biological molecules or cryo sections of biological tissue. According to the original proposal by (Danev et al., 2002) the Hilbert phase plate applies a phase shift of π to approximately half the focal plane (for example the right half excluding the central beam) and an image is recorded at Gaussian focus. After correction for the inbuilt asymmetry of differential phase contrast this image will have an almost perfect contrast transfer function (close to 1) from the lowest spatial frequency up to a maximum resolution determined by the wave length and spherical aberration of the microscope. In this paper I present theory and simulations showing that this maximum spatial frequency can be increased considerably almost without loss of contrast by using a Hilbert phase plate of half the thickness, leading to a phase shift of π/2, and recording images at Scherzer defocus. The maximum resolution can be improved even more by imaging at extended Scherzer defocus, though at the cost of contrast loss at lower spatial frequencies. - Highlights: • In this paper I present theory and simulations for a Hilbert phase plate that phase shifts the electron wave by π/2 instead of π while images are recorded close to Scherzer defocus instead of Gaussian focus. • I show that the point resolution for this new imaging mode is considerably higher without loss of contrast. • An additional advantage lies in the reduced thickness of the phase plate which leads to reduced inelastic scattering in the phase plate and less noise.

  19. A rad-hard CMOS active pixel sensor for electron microscopy

    Energy Technology Data Exchange (ETDEWEB)

    Battaglia, Marco [Department of Physics, University of California at Berkeley, CA 94720 (United States); Lawrence Berkeley National Laboratory, Berkeley, CA 94720 (United States)], E-mail: MBattaglia@lbl.gov; Contarato, Devis; Denes, Peter; Doering, Dionisio [Lawrence Berkeley National Laboratory, Berkeley, CA 94720 (United States); Giubilato, Piero [Lawrence Berkeley National Laboratory, Berkeley, CA 94720 (United States); INFN, Sezione di Padova, I-35131 (Italy); Dipartimento di Fisica, Universita degli Studi, Padova I-35131 (Italy); Kim, Tae Sung [Lawrence Berkeley National Laboratory, Berkeley, CA 94720 (United States); Mattiazzo, Serena [INFN, Sezione di Padova, I-35131 (Italy); Dipartimento di Fisica, Universita degli Studi, Padova I-35131 (Italy); Radmilovic, Velimir [Lawrence Berkeley National Laboratory, Berkeley, CA 94720 (United States); Zalusky, Sarah [Department of Physics, University of California at Berkeley, CA 94720 (United States); Lawrence Berkeley National Laboratory, Berkeley, CA 94720 (United States)

    2009-01-11

    Monolithic CMOS pixel sensors offer unprecedented opportunities for fast nano-imaging through direct electron detection in transmission electron microscopy. We present the design and a full characterisation of a CMOS pixel test structure able to withstand doses in excess of 1 Mrad. Data collected with electron beams at various energies of interest in electron microscopy are compared to predictions of simulation and to 1.5 GeV electron data to disentagle the effect of multiple scattering. The point spread function measured with 300 keV electrons is (8.1{+-}1.6){mu}m for 10{mu}m pixel and (10.9{+-}2.3){mu}m for 20{mu}m pixels, respectively, which agrees well with the values of 8.4 and 10.5{mu}m predicted by our simulation.

  20. PREFACE: Electron Microscopy and Analysis Group Conference (EMAG2015)

    Science.gov (United States)

    MacLaren, Ian

    2015-10-01

    2015 marked a new venture for the EMAG group of the Institute of Physics in that the conference was held in conjunction with the MMC2015 conference at the wonderful Manchester Central conference centre. As anyone who was there would be able to confirm, this went exceptionally well and was a really vibrant and top quality conference. The oral sessions were filled with good talks, the poster sessions were very lively, and there was a good balance between oral sessions with a specifically "EMAG" identity, and the integration into a larger conference with the ability to switch between up to six parallel sessions covering physical sciences, techniques, and life sciences. The large conference also attracted a wide range of exhibitors, and this is essential for the ongoing success of all of our work, in a field that is very dependent on continued technical innovation and on collaborations between academic researchers and commercial developers of microscopes, holders, detectors, spectrometers, sample preparation equipment, and software, among other things. As has long been the case at EMAG, all oral and poster presenters were invited to submit papers for consideration for the proceedings. As ever, these papers were independently reviewed by other conference attendees, with the aim of continuing the long tradition of the EMAG proceedings being a top quality, peer-reviewed publication, worthy of reference in future years. Whilst I recognise that not all presenters were able to submit papers to the proceedings (for instance due to the need not to prejudice publication in some other journals, or due to avoiding duplicate publication of data), we are gratified that our presenters submitted as many papers as they did. The 41 papers included provide an interesting snapshot of many of the areas covered in the conference presentations, including functional materials, coatings, 3D microscopy, FIB and SEM, nanomaterials, magnetic and structural materials, advances in EM techniques

  1. Magnetic dynamics studied by high-resolution electron spectroscopy and time-resolved electron microscopy

    Science.gov (United States)

    Jayaraman, Rajeswari

    Future information technology requires an increased magnetically encoded data density and novel electromagnetic modes of data transfer. While to date magnetic properties are observed and characterized mostly statically, the need emerges to monitor and capture their fast dynamics. In this talk, I will focus on the spin dynamics i.e. spin wave excitations and the dynamics of a new topological distribution of spins termed ``skyrmions''. Wave packets of spin waves offer the unique capability to transport a quantum bit, the spin, without the transport of charge or mass. Here, large wave-vector spin waves are of particular interest as they admit spin localization within a few nanometers. By using our recently developed electron energy loss spectrometer, we could study such spin waves in ultrathin films with an unprecedented energy resolution of 4 meV. By virtue of the finite penetration depth of low energy electrons, spin waves localized at interfaces between a substrate and a thin capping layer can be been studied yielding information about the exchange coupling between atoms at the interface. The quantization of spin waves with wave vectors perpendicular to the film gives rise to standing modes to which EELS has likewise access. Such studies when carried out as function of the film thickness again yield information on the layer dependence of the exchange coupling. Magnetic skyrmions are promising candidates as information carriers in logic or storage devices. Currently, little is known about the influence of disorder, defects, or external stimuli on the spatial distribution and temporal evolution of the skyrmion lattice. In this talk, I will describe the dynamical role of disorder in a large and flat thin film of Cu2OSeO3, exhibiting a skyrmion phase in an insulating material. We image up to 70,000 skyrmions by means of cryo-Lorentz Transmission Electron Microscopy as a function of the applied magnetic field. In the skyrmion phase, dislocations are shown to cause the

  2. EM∩IM: software for relating ion mobility mass spectrometry and electron microscopy data.

    Science.gov (United States)

    Degiacomi, Matteo T; Benesch, Justin L P

    2016-01-07

    We present EM∩IM, software that allows the calculation of collision cross-sections from electron density maps obtained for example by means of transmission electron microscopy. This allows the assessment of structures other than those described by atomic coordinates with ion mobility mass spectrometry data, and provides a new means for contouring and validating electron density maps. EM∩IM thereby facilitates the use of data obtained in the gas phase within structural biology studies employing diverse experimental methodologies.

  3. Visualization of newt aragonitic otoconial matrices using transmission electron microscopy

    Science.gov (United States)

    Steyger, P. S.; Wiederhold, M. L.

    1995-01-01

    Otoconia are calcified protein matrices within the gravity-sensing organs of the vertebrate vestibular system. These protein matrices are thought to originate from the supporting or hair cells in the macula during development. Previous studies of mammalian calcitic, barrel-shaped otoconia revealed an organized protein matrix consisting of a thin peripheral layer, a well-defined organic core and a flocculent matrix inbetween. No studies have reported the microscopic organization of the aragonitic otoconial matrix, despite its protein characterization. Pote et al. (1993b) used densitometric methods and inferred that prismatic (aragonitic) otoconia have a peripheral protein distribution, compared to that described for the barrel-shaped, calcitic otoconia of birds, mammals, and the amphibian utricle. By using tannic acid as a negative stain, we observed three kinds of organic matrices in preparations of fixed, decalcified saccular otoconia from the adult newt: (1) fusiform shapes with a homogenous electron-dense matrix; (2) singular and multiple strands of matrix; and (3) more significantly, prismatic shapes outlined by a peripheral organic matrix. These prismatic shapes remain following removal of the gelatinous matrix, revealing an internal array of organic matter. We conclude that prismatic otoconia have a largely peripheral otoconial matrix, as inferred by densitometry.

  4. Scanning electron microscopy investigations regarding Adonis vernalis L. flower morphology

    Directory of Open Access Journals (Sweden)

    Irina Neta GOSTIN

    2009-11-01

    Full Text Available The floral morphology of Adonis vernalis L. was observed with a scanning electron microscope (SEM. The investigations are important to clarify some taxonomical problems and also could provide useful diagnostic elements for the identification of this medicinal plant in powdered materials. All floral organs are initiated spirally and centripetally and develop centripetally. The petals (8-12 are shorter than the sepals (5-6 in early developmental stages. The petals are disposed on spiral (with 3-4 whorls. The stamens (numerous are unbranched and reach maturity centripetally; they are free of the perianth. The anther walls consisting of a single layer epidermis in the anther wall surrounding the sporagenous tissue, one row of endothecium, two to four rows of middle layer and one row of tapetum layer. In the anther walls, the tapetal cells, by glandular type, persist later in ontogenesis. Pollen grains are tricolpate with echinate surface. The gynoecium is multiple, apocarpous with distinct carpels. The carpels are ascidiate from the beginning. At the base of each carpel, numerousness short, unicellular, trichomes are present. The stigma differentiates as two crests along the ventral slit of the ovary. Each carpel contains a single ovule inside the ovary cavity. The mature ovule is anatropous, with two integuments. It is almost parallel to the funicle.

  5. Electron Microscopy and Image Analysis for Selected Materials

    Science.gov (United States)

    Williams, George

    1999-01-01

    This particular project was completed in collaboration with the metallurgical diagnostics facility. The objective of this research had four major components. First, we required training in the operation of the environmental scanning electron microscope (ESEM) for imaging of selected materials including biological specimens. The types of materials range from cyanobacteria and diatoms to cloth, metals, sand, composites and other materials. Second, to obtain training in surface elemental analysis technology using energy dispersive x-ray (EDX) analysis, and in the preparation of x-ray maps of these same materials. Third, to provide training for the staff of the metallurgical diagnostics and failure analysis team in the area of image processing and image analysis technology using NIH Image software. Finally, we were to assist in the sample preparation, observing, imaging, and elemental analysis for Mr. Richard Hoover, one of NASA MSFC's solar physicists and Marshall's principal scientist for the agency-wide virtual Astrobiology Institute. These materials have been collected from various places around the world including the Fox Tunnel in Alaska, Siberia, Antarctica, ice core samples from near Lake Vostoc, thermal vents in the ocean floor, hot springs and many others. We were successful in our efforts to obtain high quality, high resolution images of various materials including selected biological ones. Surface analyses (EDX) and x-ray maps were easily prepared with this technology. We also discovered and used some applications for NIH Image software in the metallurgical diagnostics facility.

  6. Scanning and Transmission Electron Microscopy of High Temperature Materials

    Science.gov (United States)

    1994-01-01

    Software and hardware updates to further extend the capability of the electron microscope were carried out. A range of materials such as intermetallics, metal-matrix composites, ceramic-matrix composites, ceramics and intermetallic compounds, based on refractory elements were examined under this research. Crystal structure, size, shape and volume fraction distribution of various phases which constitute the microstructures were examined. Deformed materials were studied to understand the effect of interfacial microstructure on the deformation and fracture behavior of these materials. Specimens tested for a range of mechanical property requirements, such as stress rupture, creep, low cycle fatigue, high cycle fatigue, thermomechanical fatigue, etc. were examined. Microstructural and microchemical stability of these materials exposed to simulated operating environments were investigated. The EOIM Shuttle post-flight samples were also examined to understand the influence of low gravity processing on microstructure. In addition, fractographic analyses of Nb-Zr-W, titanium aluminide, molybdenum silicide and silicon carbide samples were carried out. Extensive characterization of sapphire fibers in the fiber-reinforced composites made by powder cloth processing was made. Finally, pressure infiltration casting of metal-matrix composites was carried out.

  7. Electron microscopy of human fascia lata: focus on telocytes.

    Science.gov (United States)

    Dawidowicz, Joanna; Szotek, Sylwia; Matysiak, Natalia; Mielańczyk, Łukasz; Maksymowicz, Krzysztof

    2015-10-01

    From the histological point of view, fascia lata is a dense connective tissue. Although extracellular matrix is certainly the most predominant fascia's feature, there are also several cell populations encountered within this structure. The aim of this study was to describe the existence and characteristics of fascia lata cell populations viewed through a transmission electron microscope. Special emphasis was placed on telocytes as a particular interstitial cell type, recently discovered in a wide variety of tissues and organs such as the heart, skeletal muscles, skin, gastrointestinal tract, uterus and urinary system. The conducted study confirmed the existence of a telocyte population in fascia lata samples. Those cells fulfil main morphological criteria of telocytes, namely, the presence of very long, thin cell processes (telopodes) extending from a relatively small cell body. Aside from telocytes, we have found fibroblasts, mast cells and cells with features of myofibroblastic differentiation. This is the first time it has been shown that telocytes exist in human fascia. Currently, the exact role of those cells within the fascia is unknown and definitely deserves further attention. One can speculate that fascia lata telocytes likewise telocytes in other organs may be involved in regeneration, homeostasis and intracellular signalling.

  8. Electron microscopy of human fascia lata: focus on telocytes

    Science.gov (United States)

    Dawidowicz, Joanna; Szotek, Sylwia; Matysiak, Natalia; Mielańczyk, Łukasz; Maksymowicz, Krzysztof

    2015-01-01

    From the histological point of view, fascia lata is a dense connective tissue. Although extracellular matrix is certainly the most predominant fascia’s feature, there are also several cell populations encountered within this structure. The aim of this study was to describe the existence and characteristics of fascia lata cell populations viewed through a transmission electron microscope. Special emphasis was placed on telocytes as a particular interstitial cell type, recently discovered in a wide variety of tissues and organs such as the heart, skeletal muscles, skin, gastrointestinal tract, uterus and urinary system. The conducted study confirmed the existence of a telocyte population in fascia lata samples. Those cells fulfil main morphological criteria of telocytes, namely, the presence of very long, thin cell processes (telopodes) extending from a relatively small cell body. Aside from telocytes, we have found fibroblasts, mast cells and cells with features of myofibroblastic differentiation. This is the first time it has been shown that telocytes exist in human fascia. Currently, the exact role of those cells within the fascia is unknown and definitely deserves further attention. One can speculate that fascia lata telocytes likewise telocytes in other organs may be involved in regeneration, homeostasis and intracellular signalling. PMID:26311620

  9. Nuclear uptake of ultrasmall gold-doxorubicin conjugates imaged by fluorescence lifetime imaging microscopy (FLIM) and electron microscopy

    Science.gov (United States)

    Zhang, Xuan; Shastry, Sathvik; Bradforth, Stephen E.; Nadeau, Jay L.

    2014-11-01

    Fluorescence lifetime imaging microscopy (FLIM) has been used to image free and encapsulated doxorubicin (Dox) uptake into cells, since interaction of Dox with DNA leads to a characteristic lifetime change. However, none of the reported Dox conjugates were able to enter cell nuclei. In this work, we use FLIM to show nuclear uptake of 2.7 nm mean diameter Au nanoparticles conjugated to Dox. The pattern of labelling differed substantially from what was seen with free Dox, with slower nuclear entry and stronger cytoplasmic labelling at all time points. As the cells died, the pattern of labelling changed further as intracellular structures disintegrated, consistent with association of Au-Dox to membranes. The patterns of Au distribution and intracellular structure changes were confirmed using electron microscopy, and indicate different mechanisms of cytotoxicity with stable Au-Dox conjugates compared to Dox alone. Such conjugates are promising tools for overcoming resistance in Dox-resistant cancers.Fluorescence lifetime imaging microscopy (FLIM) has been used to image free and encapsulated doxorubicin (Dox) uptake into cells, since interaction of Dox with DNA leads to a characteristic lifetime change. However, none of the reported Dox conjugates were able to enter cell nuclei. In this work, we use FLIM to show nuclear uptake of 2.7 nm mean diameter Au nanoparticles conjugated to Dox. The pattern of labelling differed substantially from what was seen with free Dox, with slower nuclear entry and stronger cytoplasmic labelling at all time points. As the cells died, the pattern of labelling changed further as intracellular structures disintegrated, consistent with association of Au-Dox to membranes. The patterns of Au distribution and intracellular structure changes were confirmed using electron microscopy, and indicate different mechanisms of cytotoxicity with stable Au-Dox conjugates compared to Dox alone. Such conjugates are promising tools for overcoming resistance in

  10. Environmental scanning electron microscopy of hydrated conditioned/etched dentine.

    Science.gov (United States)

    de Wet, F A; van der Vyver, P J; Eick, J D; Dusevich, V M

    2000-11-01

    Various etchants/conditioners are used during dental treatment to affect or remove the smear layer. The purpose of this study was to evaluate the effect of different treatments on moist dentine, using a field emission environmental scanning electron microscope (FE-ESEM). Twenty freshly extracted, human molar teeth were utilised. The roots and pulps were removed, and the crowns horizontally sectioned with a low speed diamond saw (Isomet) (with cooling in a saline solution) in order to expose superficial dentine. A smear layer was created on these surfaces by using 600 grit silicone carbide paper. Test surfaces were then treated in one of the following ways: 1. 37% phosphoric acid liquid 2. 37% phosphoric acid gel 3. NRC (non-rinse conditioner) without rinsing 4. NRC with rinsing. Shallow grooves were cut on the untreated sides, using a thin diamond bur. This enabled the samples to be split in half when pressure was applied in the grooves. Samples were maintained moist throughout specimen preparation. Samples were examined in the FE-ESEM (Philips XL 30) in such a way that the effect of the treatment could be viewed occlusally, as well as perpendicular to the treated interface. Phosphoric acid liquid and gel removed the smear layer, and demineralised the dentine for approximately 5-10 micrometers. NRC penetrated the smear layer and modified it to a lesser degree. However, washing of the NRC treated surface removed part of the smear layer, and opened up some dentinal tubules. Excellent resolution was possible with the FE-ESEM in both the wet and dry modes.

  11. Scanning electron microscopy of hair treated in hard water.

    Science.gov (United States)

    Srinivasan, Gautham; Chakravarthy Rangachari, Srinivas

    2016-06-01

    Hardness of water is determined by the amount of calcium carbonate (CaCO3 ) and magnesium sulfate (MgSO4 ) dissolved in it. Hardness of water used for washing hair may damage the hair. The objective of this study is to observe the surface changes due to hard water usage and compare the thickness of hair between hard and soft water treated samples. Ten to 15 hair strands of length 15-20 cm, which were lost during combing, were obtained from 15 healthy female volunteers. Each hair sample was cut into two equal halves to obtain two sets per volunteer. Each hair sample was wrapped around a glass rod. One set of 15 samples was washed with hard water, and the other set was washed with distilled water for 10 minutes on alternate days and air-dried. This procedure was carried out for 30 days. The surface of hair treated in hard and soft water was examined under a scanning electron microscope. The CaCO3 and MgSO4 content of hard and distilled water samples were determined as 212.5 ppm of CaCO3 and 10 ppm of CaCO3 respectively. The mean calcium deposition in hard and distilled water treated hair was determined as 0.804% and 0.26%, respectively. The mean magnesium deposition in hard and distilled water treated hair was determined as 0.34% and 0.078%, respectively. The mean thickness of hair treated in hard water and distilled water were 72.78 and 78.14 μm, respectively. The surface of hard water treated hair has a ruffled appearance with higher mineral deposition and decreased thickness when compared with the surface of distilled water treated hair. © 2015 The International Society of Dermatology.

  12. Probing Electronic States of Magnetic Semiconductors Using Atomic Scale Microscopy & Spectroscopy

    Science.gov (United States)

    2013-12-01

    N000140710348 Final Report as of December 2013 Objective: This project was focused on the application of the scanning tunneling microscopy (STM...magnetic atoms on the surface of a superconductor can be used as a versatile platform for creating a topological superconductor . These initial...the application of the scanning tunneling microscopy (STM) to understand the electronic structure of magnetically doped semiconductors and to develop

  13. Interactive stereo electron microscopy enhanced with virtual reality

    Energy Technology Data Exchange (ETDEWEB)

    Bethel, E.Wes; Bastacky, S.Jacob; Schwartz, Kenneth S.

    2001-12-17

    An analytical system is presented that is used to take measurements of objects perceived in stereo image pairs obtained from a scanning electron microscope (SEM). Our system operates by presenting a single stereo view that contains stereo image data obtained from the SEM, along with geometric representations of two types of virtual measurement instruments, a ''protractor'' and a ''caliper''. The measurements obtained from this system are an integral part of a medical study evaluating surfactant, a liquid coating the inner surface of the lung which makes possible the process of breathing. Measurements of the curvature and contact angle of submicron diameter droplets of a fluorocarbon deposited on the surface of airways are performed in order to determine surface tension of the air/liquid interface. This approach has been extended to a microscopic level from the techniques of traditional surface science by measuring submicrometer rather than millimeter diameter droplets, as well as the lengths and curvature of cilia responsible for movement of the surfactant, the airway's protective liquid blanket. An earlier implementation of this approach for taking angle measurements from objects perceived in stereo image pairs using a virtual protractor is extended in this paper to include distance measurements and to use a unified view model. The system is built around a unified view model that is derived from microscope-specific parameters, such as focal length, visible area and magnification. The unified view model ensures that the underlying view models and resultant binocular parallax cues are consistent between synthetic and acquired imagery. When the view models are consistent, it is possible to take measurements of features that are not constrained to lie within the projection plane. The system is first calibrated using non-clinical data of known size and resolution. Using the SEM, stereo image pairs of grids and spheres of

  14. Morphological aspects of Angiostrongylus costaricensis by light and scanning electron microscopy.

    Science.gov (United States)

    Rebello, Karina M; Menna-Barreto, Rubem F S; Chagas-Moutinho, Vanessa A; Mota, Ester M; Perales, Jonas; Neves-Ferreira, Ana Gisele C; Oliveira-Menezes, Aleksandra; Lenzi, Henrique

    2013-09-01

    Angiostrongylus costaricensis is a parasitic nematode that can cause severe gastrointestinal disease, known as abdominal angiostrongiliasis, in humans. This paper presents the characterization of first- and third-stage larvae and male and female adult worms of A. costaricensis by scanning electron and light microscopy. Several novel anatomical structures were identified by scanning electron microscopy, including details of the cuticular striations of the spicules in male worms and a protective flap of the cuticle covering the vulvar aperture in female worms. Other taxonomic features revealed by light microscopy include the gubernaculum and the esophageal-intestinal valve. The use of two microscopy techniques allowed a detailed characterization of the morphology of this nematode. A number of previously identified taxonomic features, such as the striated nature of the spicules and the lateral alae were confirmed; however, the use of scanning electron microscopy resulted in a reassessment of the correct number of papillae distributed around the oral opening and behind the cloacal opening. These observations, in combination with light microscopy-based characterization of the gubernaculum and esophageal valves, have allowed a more detailed description of this nematode taxonomy.

  15. Advanced transmission electron microscopy studies in low-energy ion implanted Si Semiconductors; Junctions; Silicon

    CERN Document Server

    Wang, T S

    2002-01-01

    As the dimensions of semiconductor devices shrink down to 0.1 mu m and beyond, low energy ion implantation is required to introduce shallower junctions to match such small devices. In this work, transmission electron microscopy (TEM) is employed to analyse low energy implanted junctions with both structural and chemical analyses. High resolution transmission electron microscopy (HRTEM) has been employed to observe Si crystal damage and amorphization due to low energy B sup + /As sup + ion implantations, and also, defect formation/annihilation during rapid thermal annealing (RTA). The damage effects due to different implant temperatures between 300 deg C and -150 deg C are also discussed. Since knowledge of the distribution of low energy ion implanted dopants in Si is extremely important for semiconductor device processing, energy filtered transmission electron microscopy (EFTEM) has been employed to determine implanted B distributions in Si while Z-contrast imaging and X-ray analytical mapping techniques are ...

  16. Nanoscale electron tomography and atomic scale high-resolution electron microscopy of nanoparticles and nanoclusters:A short survey

    Institute of Scientific and Technical Information of China (English)

    John Meurig Thomasn; Paul A. Midgley; Caterina Ducati; Rowan K. Leary

    2013-01-01

    The outstanding merits of scanning transmission electron tomography as a technique for the investigation of the internal structure and morphology of nanoparticle and nanocluster materials are summarized with the aid of numerous typical illustrations. Reference is made also to the significant advances that have arisen in probing ultrastructural characteristics of nanoscale solids using aberration-corrected (AC) electron microscopy (EM). Information of a unique kind may be retrieved by combining the imaging and analytical power of ACEM.

  17. Cryogenic-temperature electron microscopy direct imaging of carbon nanotubes and graphene solutions in superacids.

    Science.gov (United States)

    Kleinerman, O; Parra-Vasquez, A Nicholas G; Green, M J; Behabtu, N; Schmidt, J; Kesselman, E; Young, C C; Cohen, Y; Pasquali, M; Talmon, Y

    2015-07-01

    Cryogenic electron microscopy (cryo-EM) is a powerful tool for imaging liquid and semiliquid systems. While cryogenic transmission electron microscopy (cryo-TEM) is a standard technique in many fields, cryogenic scanning electron microscopy (cryo-SEM) is still not that widely used and is far less developed. The vast majority of systems under investigation by cryo-EM involve either water or organic components. In this paper, we introduce the use of novel cryo-TEM and cryo-SEM specimen preparation and imaging methodologies, suitable for highly acidic and very reactive systems. Both preserve the native nanostructure in the system, while not harming the expensive equipment or the user. We present examples of direct imaging of single-walled, multiwalled carbon nanotubes and graphene, dissolved in chlorosulfonic acid and oleum. Moreover, we demonstrate the ability of these new cryo-TEM and cryo-SEM methodologies to follow phase transitions in carbon nanotube (CNT)/superacid systems, starting from dilute solutions up to the concentrated nematic liquid-crystalline CNT phases, used as the 'dope' for all-carbon-fibre spinning. Originally developed for direct imaging of CNTs and graphene dissolution and self-assembly in superacids, these methodologies can be implemented for a variety of highly acidic systems, paving a way for a new field of nonaqueous cryogenic electron microscopy. © 2015 The Authors Journal of Microscopy © 2015 Royal Microscopical Society.

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

    Science.gov (United States)

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

    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. © 2013 Published by Elsevier B.V.

  19. Correlation of live-cell imaging with volume scanning electron microscopy.

    Science.gov (United States)

    Lucas, Miriam S; Günthert, Maja; Bittermann, Anne Greet; de Marco, Alex; Wepf, Roger

    2017-01-01

    Live-cell imaging is one of the most widely applied methods in live science. Here we describe two setups for live-cell imaging, which can easily be combined with volume SEM for correlative studies. The first procedure applies cell culture dishes with a gridded glass support, which can be used for any light microscopy modality. The second approach is a flow-chamber setup based on Ibidi μ-slides. Both live-cell imaging strategies can be followed up with serial blockface- or focused ion beam-scanning electron microscopy. Two types of resin embedding after heavy metal staining and dehydration are presented making best use of the particular advantages of each imaging modality: classical en-bloc embedding and thin-layer plastification. The latter can be used only for focused ion beam-scanning electron microscopy, but is advantageous for studying cell-interactions with specific substrates, or when the substrate cannot be removed. En-bloc embedding has diverse applications and can be applied for both described volume scanning electron microscopy techniques. Finally, strategies for relocating the cell of interest are discussed for both embedding approaches and in respect to the applied light and scanning electron microscopy methods. Copyright © 2017 Elsevier Inc. All rights reserved.

  20. Magnetic-field-modulated written bits in TbFeCo thin films: Transmission electron microscopy Lorentz and scanning electron microscopy with polarization analysis studies

    Science.gov (United States)

    Aeschlimann, M.; Scheinfein, M.; Unguris, J.; Greidanus, F. J. A. M.; Klahn, S.

    1990-11-01

    Domains written thermomagnetically in TbFeCo thin films are studied with Lorentz transmission electron microscopy (TEM) and scanning electron microscopy with polarization analysis (SEMPA). Four different rare-earth/transition-metal compositions TbxFeyCo1-x-y are examined. The domain structures observed with both techniques are similar even though TEM Lorentz only detects the transverse component of the net magnetic field along the electron's trajectory through the sample, while SEMPA detects the surface electron-spin polarization (magnetization). We find that the magnetic contrast in the SEMPA measurements is proportional to the magnetization of the transition-metal (TM) subnetwork which is antiferromagnetically coupled to the rare-earth (RE) subnetwork. This allows high-contrast SEMPA images to be acquired even when the system is magnetically compensated (Ms=‖MRE-MTM‖=0). The surface magnetization can be explained by assuming that the surface of the TbFeCo alloy consists of an outermost thin oxide layer followed by an Fe-rich subsurface layer. The importance of the demagnetizing field on the switching and domain nucleation process for thermomagnetically written bits is examined.

  1. Magnetic-field-modulated written bits in TbFeCo thin films: Transmission electron microscopy Lorentz and scanning electron microscopy with polarization analysis studies

    Energy Technology Data Exchange (ETDEWEB)

    Aeschlimann, M.; Scheinfein, M.; Unguris, J. (National Institute of Standards and Technology, Gaithersburg, Maryland 20899 (USA) Greidanus, F.J.A.M. (Philips Research Laboratories, P.O. Box 80.000, 5600 JA, Eindhoven (The Netherlands)))

    1990-11-01

    Domains written thermomagnetically in TbFeCo thin films are studied with Lorentz transmission electron microscopy (TEM) and scanning electron microscopy with polarization analysis (SEMPA). Four different rare-earth/transition-metal compositions Tb{sub {ital x}}Fe{sub {ital y}}Co{sub 1{minus}{ital x}{minus}{ital y}} are examined. The domain structures observed with both techniques are similar even though TEM Lorentz only detects the transverse component of the net magnetic field along the electron's trajectory through the sample, while SEMPA detects the surface electron-spin polarization (magnetization). We find that the magnetic contrast in the SEMPA measurements is proportional to the magnetization of the transition-metal (TM) subnetwork which is antiferromagnetically coupled to the rare-earth (RE) subnetwork. This allows high-contrast SEMPA images to be acquired even when the system is magnetically compensated ({ital M}{sub {ital s}}={vert bar}{ital M}{sub RE}{minus}{ital M}{sub TM}{vert bar}=0). The surface magnetization can be explained by assuming that the surface of the TbFeCo alloy consists of an outermost thin oxide layer followed by an Fe-rich subsurface layer. The importance of the demagnetizing field on the switching and domain nucleation process for thermomagnetically written bits is examined.

  2. Transmission Electron Microscopy of a CMSX-4 Ni-Base Superalloy Produced by Selective Electron Beam Melting

    Directory of Open Access Journals (Sweden)

    Alireza B. Parsa

    2016-10-01

    Full Text Available In this work, the microstructures of superalloy specimens produced using selective electron beam melting additive manufacturing were characterized. The materials were produced using a CMSX-4 powder. Two selective electron beam melting processing strategies, which result in higher and lower effective cooling rates, are described. Orientation imaging microscopy, scanning transmission electron microscopy and conventional high resolution transmission electron microscopy are used to investigate the microstructures. Our results suggest that selective electron beam melting processing results in near equilibrium microstructures, as far as γ′ volume fractions, the formation of small amounts of TCP phases and the partitioning behavior of the alloy elements are concerned. As expected, higher cooling rates result in smaller dendrite spacings, which are two orders of magnitude smaller than observed during conventional single crystal casting. During processing, columnar grains grow in <100> directions, which are rotated with respect to each other. There are coarse γ/γ′ microstructures in high angle boundary regions. Dislocation networks form low angle boundaries. A striking feature of the as processed selective electron beam melting specimens is their high dislocation density. From a fundamental point of view, this opens new possibilities for the investigation of elementary dislocation processes which accompany solidification.

  3. 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.

  4. Structure and Stability of Pt-Y Alloy Particles for Oxygen Reduction Studied by Electron Microscopy

    DEFF Research Database (Denmark)

    Deiana, Davide; Wagner, Jakob Birkedal; Hansen, Thomas Willum

    2015-01-01

    Platinum-yttrium alloy nanoparticles show both a high activity and stability for the oxygen reduction reaction. The catalysts were prepared by magnetron sputter aggregation and mass filtration providing a model catalyst system with a narrow size distribution. The structure and stability...... of nanostructured Pt-Y alloy catalysts were studied using transmission electron microscopy techniques. Using elemental X-ray mapping and high-resolution electron microscopy, the specific compositional structure and distribution of the individual nanoparticles was unraveled and the stability assessed. Studying...... the catalyst after reaction and after aging tests shows the development of a core-shell type structure after being exposed to reaction conditions....

  5. A diagnostic mystery solved by electron microscopy: a case of an "atypical" lymphoproliferative disorder.

    Science.gov (United States)

    Lee, Sophie; Graham, Linda M; Chan, George; Ruskova, Anna

    2012-10-01

    An elderly woman with a previous diagnosis of atypical chronic lymphocytic leukemia (CLL) was noted to have a strikingly abnormal blood film, with the lymphocytes displaying numerous large cytoplasmic granules. This appearance had not been described before in the literature to the best of the authors' knowledge. After a series of investigations, electron microscopy was eventually performed, which demonstrated that the abnormal granules were composed of immunoglobulin crystals. The immunofixation study confirmed that they were monoclonal IgM paraprotein. These results led to a change of diagnosis to lymphoplasmacytic lymphoma. This report illustrates how electron microscopy can be used as a valuable additional diagnostic tool in difficult cases.

  6. Scanning electron microscopy of the human endolymphatic sac: a preliminary report.

    Science.gov (United States)

    Galey, F R; House, W F

    1980-04-01

    Scanning electron microscopy has been used to examine and compare one normal endolymphatic sac with one endolymphatic sac from a patient with Meniere's disease. The surgical procedure for obtaining these specimens and their preparation for scanning electron microscopy are described. The luminal surface of the rugose portion of both specimens was lined with two populations of epithelial cells: one with a dome-shaped apical surface, the other with a flattened polygonal surface. The surface of dome-shaped cells in both specimens was covered with microvilli. Neither specimen had observable loss of epithelial integrity or fibrosis.

  7. Transmission Electron Microscopy Analysis of Skin Lesions from Sporotrichosis Epidemic in Rio de Janeiro, Brazil

    Science.gov (United States)

    Porto Ferreira, Cassio; Oliveira de Almeida, Ana Cristina; Corte-Real, Suzana

    2015-01-01

    Transmission electron microscopy can yield useful information in a range of scientific fields; it is capable of imaging at a significantly higher resolution than light microscopes and has been a very useful tool in the identification of morphological changes of the dermis as well as assessment of changes in the extracellular matrix. Our aim is to characterize by electron microscopy the cellular profile of lesions caused by Sporothrix schenckii from the sporotrichosis epidemic in its zoonotic form that occurs in Rio de Janeiro, Brazil. PMID:25653392

  8. Electron microscopy of Mg/TiO{sub 2} photocatalyst morphology for deep desulfurization of diesel

    Energy Technology Data Exchange (ETDEWEB)

    Yin, Yee Cia, E-mail: gabrielle.ciayin@gmail.com [Department of Chemical Engineering, Universiti Teknologi PETRONAS, 31750 Tronoh, Perak (Malaysia); Kait, Chong Fai, E-mail: chongfaikait@petronas.com.my; Fatimah, Hayyiratul, E-mail: hayyiratulfatimah@yahoo.com; Wilfred, Cecilia, E-mail: cecili@petronas.com.my [Department of Fundamental and Applied Sciences, Universiti Teknologi PETRONAS, 31750 Tronoh, Perak (Malaysia)

    2015-07-22

    A series of Mg/TiO{sub 2} photocatalysts were prepared and characterized using Field Emission Scanning Electron Microscopy (FESEM) and High-Resolution Transmission Electron Microscopy (HRTEM). The average particle sizes of the photocatalysts were ranging from 25.7 to 35.8 nm. Incorporation of Mg on TiO{sub 2} did not lead to any surface lattice distortion to TiO{sub 2}. HRTEM data indicated the presence of MgO and Mg(OH){sub 2} mixture at low Mg loading while at higher Mg loading, the presence of lamellar Mg-oxyhydroxide intermediates and Mg(OH){sub 2}.

  9. Structure and Stability of Pt-Y Alloy Particles for Oxygen Reduction Studied by Electron Microscopy

    DEFF Research Database (Denmark)

    Deiana, Davide; Wagner, Jakob Birkedal; Hansen, Thomas Willum

    2015-01-01

    Platinum-yttrium alloy nanoparticles show both a high activity and stability for the oxygen reduction reaction. The catalysts were prepared by magnetron sputter aggregation and mass filtration providing a model catalyst system with a narrow size distribution. The structure and stability...... of nanostructured Pt-Y alloy catalysts were studied using transmission electron microscopy techniques. Using elemental X-ray mapping and high-resolution electron microscopy, the specific compositional structure and distribution of the individual nanoparticles was unraveled and the stability assessed. Studying...... the catalyst after reaction and after aging tests shows the development of a core-shell type structure after being exposed to reaction conditions....

  10. The detection and influence of food soils on microorganisms on stainless steel using scanning electron microscopy and epifluorescence microscopy.

    Science.gov (United States)

    Whitehead, Kathryn A; Smith, Lindsay A; Verran, Joanna

    2010-07-31

    A range of food soils and components (complex [meat extract, fish extract, and cottage cheese extract]; oils [cholesterol, fish oil, and mixed fatty acids]; proteins [bovine serum albumin (BSA), fish peptones, and casein]; and carbohydrates [glycogen, starch, and lactose]) were deposited onto 304 2B finish stainless steel surfaces at different concentrations (10-0.001%). Scanning electron microscopy (SEM) and epifluorescence microscopy were used to visualise the cell and food soil distribution across the surface. Epifluorescence microscopy was also used to quantify the percentage of a field covered by cells or soil. At 10% concentration, most soils, with the exception of BSA and fish peptone were easily visualised using SEM, presenting differences in gross soil morphology and distribution. When soil was stained with acridine orange and visualised by epifluorescence microscopy, the limit of detection of the method varied between soils, but some (meat, cottage cheese and glycogen) were detected at the lowest concentrations used (0.001%). The decrease in soil concentration did not always relate to the surface coverage measurement. When 10% food soil was applied to a surface with Escherichia coli and compared, cell attachment differed depending on the nature of the soil. The highest percentage coverage of cells was observed on surfaces with fish extract and related products (fish peptone and fish oil), followed by carbohydrates, meat extract/meat protein, cottage cheese/casein and the least to the oils (cholesterol and mixed fatty acids). Cells could not be clearly observed in the presence of some food soils using SEM. Findings demonstrate that food soils heterogeneously covered stainless steel surfaces in differing patterns. The pattern and amount of cell attachment was related to food soil type rather than to the amount of food soil detected. This work demonstrates that in the study of conditioning film and cell retention on the hygienic properties of surfaces, SEM

  11. Effects of Topography in Nano-Structured Thin Films : A Lorentz Transmission Electron Microscopy and Electron Holography Study

    NARCIS (Netherlands)

    Hosson, Jeff Th.M. De; Raedt, Hans A. De

    2003-01-01

    This paper aims at applying advanced transmission electron microscopy (TEM) to functional materials, such as ultra-soft magnetic films for high-frequency inductors, to reveal the structure-property relationship. The ultimate goal is to delineate a more quantitative way to obtain information of the

  12. Introduction to 3D reconstruction of macromolecules using single particle electron microscopy

    Institute of Scientific and Technical Information of China (English)

    Oscar LLORCA

    2005-01-01

    Single-particle electron microscopy has now reached maturity, becoming a commonly used method in the examination ofmacromolecular structure. Using a small amount of purified protein, isolated molecules are observed under the electron microscope and the data collected can be averaged into a 3D reconstruction.Single-particle electron microscopy is an appropriate tool for the analysis of proteins that can only be obtained in modest quantities, like many of the large complexes currently of interest in biomedicine. Whilst the use of electron microscopy expands, new methods are being developed and improved to deal with further challenges, such as reaching higher resolutions and the combination of information at different levels of structural detail. More importantly, present methodology is still not robust enough when studying certain "tricky" proteins like those displaying extensive conformational flexibility and a great deal of user expertise is required, posing a threat to the consistency of the final structure. This mini review describes a brief outline of the methods currently used in the 3D analysis of macromolecules using single-particle electron microscopy, intended for those first approaching this field. A summary of methods, techniques, software, and some recent work is presented. The spectacular improvements to the technique in recent years, its advantages and limitations compared to other structural methods,and its future developments are discussed.

  13. SEM, TEM and SLEEM (scanning low energy electron microscopy) of CB2 steel after creep testing

    Science.gov (United States)

    Kasl, J.; Mikmeková, Š.; Jandová, D.

    2014-03-01

    The demand to produce electrical power with higher efficiency and with lower environmental pollution is leading to the use of new advanced materials in the production of power plant equipment. To understand the processes taking place in parts produced from these materials during their operation under severe conditions (such as high temperature, high stress, and environmental corrosion) requires detailed evaluation of their substructure. It is usually necessary to use transmission electron microscopy (TEM). However, this method is very exacting and time-consuming. So there is an effort to use new scanning electron microscopy techniques instead of TEM. One of them is scanning low energy electron microscopy (SLEEM). This paper deals with an assessment of the possibility to use SLEEM for describing the substructure of creep resistant steel CB2 after long-term creep testing. In the SLEEM images more information is contained about the microstructure of the material in comparison with standard scanning electron microscopy. Study of materials using slow and very slow electrons opens the way to better understanding their microstructures.

  14. A novel approach to scanning electron microscopy at ambient atmospheric pressure.

    Science.gov (United States)

    Ominami, Yusuke; Kawanishi, Shinsuke; Ushiki, Tatsuo; Ito, Sukehiro

    2015-04-01

    Scanning electron microscopy (SEM) for observing samples at ambient atmospheric pressure is introduced in this study. An additional specimen chamber with a small window is inserted in the main specimen chamber, and the window is separated with a thin membrane or diaphragm allowing electron beam propagation. Close proximity of the sample to the membrane enables the detection of back-scattered electrons sufficient for imaging. In addition to the empirical imaging data, a probability analysis of the un-scattered fraction of the incident electron beam further supports the feasibility of atmospheric SEM imaging over a controlled membrane-sample distance.

  15. Fluorescent Nanodiamond-Gold Hybrid Particles for Multimodal Optical and Electron Microscopy Cellular Imaging.

    Science.gov (United States)

    Liu, Weina; Naydenov, Boris; Chakrabortty, Sabyasachi; Wuensch, Bettina; Hübner, Kristina; Ritz, Sandra; Cölfen, Helmut; Barth, Holger; Koynov, Kaloian; Qi, Haoyuan; Leiter, Robert; Reuter, Rolf; Wrachtrup, Jörg; Boldt, Felix; Scheuer, Jonas; Kaiser, Ute; Sison, Miguel; Lasser, Theo; Tinnefeld, Philip; Jelezko, Fedor; Walther, Paul; Wu, Yuzhou; Weil, Tanja

    2016-10-12

    There is a continuous demand for imaging probes offering excellent performance in various microscopy techniques for comprehensive investigations of cellular processes by more than one technique. Fluorescent nanodiamond-gold nanoparticles (FND-Au) constitute a new class of "all-in-one" hybrid particles providing unique features for multimodal cellular imaging including optical imaging, electron microscopy, and, and potentially even quantum sensing. Confocal and optical coherence microscopy of the FND-Au allow fast investigations inside living cells via emission, scattering, and photothermal imaging techniques because the FND emission is not quenched by AuNPs. In electron microscopy, transmission electron microscopy (TEM) and scanning transmission electron microscopy (STEM) analysis of FND-Au reveals greatly enhanced contrast due to the gold particles as well as an extraordinary flickering behavior in three-dimensional cellular environments originating from the nanodiamonds. The unique multimodal imaging characteristics of FND-Au enable detailed studies inside cells ranging from statistical distributions at the entire cellular level (micrometers) down to the tracking of individual particles in subcellular organelles (nanometers). Herein, the processes of endosomal membrane uptake and release of FNDs were elucidated for the first time by the imaging of individual FND-Au hybrid nanoparticles with single-particle resolution. Their convenient preparation, the availability of various surface groups, their flexible detection modalities, and their single-particle contrast in combination with the capability for endosomal penetration and low cytotoxicity make FND-Au unique candidates for multimodal optical-electronic imaging applications with great potential for emerging techniques, such as quantum sensing inside living cells.

  16. Possibilities and limitations of advanced transmission electron microscopy for carbon-based nanomaterials

    Directory of Open Access Journals (Sweden)

    Xiaoxing Ke

    2015-07-01

    Full Text Available A major revolution for electron microscopy in the past decade is the introduction of aberration correction, which enables one to increase both the spatial resolution and the energy resolution to the optical limit. Aberration correction has contributed significantly to the imaging at low operating voltages. This is crucial for carbon-based nanomaterials which are sensitive to electron irradiation. The research of carbon nanomaterials and nanohybrids, in particular the fundamental understanding of defects and interfaces, can now be carried out in unprecedented detail by aberration-corrected transmission electron microscopy (AC-TEM. This review discusses new possibilities and limits of AC-TEM at low voltage, including the structural imaging at atomic resolution, in three dimensions and spectroscopic investigation of chemistry and bonding. In situ TEM of carbon-based nanomaterials is discussed and illustrated through recent reports with particular emphasis on the underlying physics of interactions between electrons and carbon atoms.

  17. Possibilities and limitations of advanced transmission electron microscopy for carbon-based nanomaterials.

    Science.gov (United States)

    Ke, Xiaoxing; Bittencourt, Carla; Van Tendeloo, Gustaaf

    2015-01-01

    A major revolution for electron microscopy in the past decade is the introduction of aberration correction, which enables one to increase both the spatial resolution and the energy resolution to the optical limit. Aberration correction has contributed significantly to the imaging at low operating voltages. This is crucial for carbon-based nanomaterials which are sensitive to electron irradiation. The research of carbon nanomaterials and nanohybrids, in particular the fundamental understanding of defects and interfaces, can now be carried out in unprecedented detail by aberration-corrected transmission electron microscopy (AC-TEM). This review discusses new possibilities and limits of AC-TEM at low voltage, including the structural imaging at atomic resolution, in three dimensions and spectroscopic investigation of chemistry and bonding. In situ TEM of carbon-based nanomaterials is discussed and illustrated through recent reports with particular emphasis on the underlying physics of interactions between electrons and carbon atoms.

  18. Correlative light/electron microscopy for the investigation of microbial mats from Black Sea Cold Seeps.

    Science.gov (United States)

    Wrede, Christoph; Heller, Christina; Reitner, Joachim; Hoppert, Michael

    2008-05-01

    In several fields of cell biology, correlative microscopy is applied to compare the structure of objects at high resolution under the electron microscope with low resolution light microscopy images of the same sample. It is, however, difficult to prepare samples and marker systems that are applicable for both microscopic techniques for the same specimen at the same time. In our studies, we used microbial mats from Cold Seep communities for a simple and rapid correlative microscopy method. The mats consist of bacterial and archaeal microorganisms, coupling reverse methanogenesis to the reduction of sulfate. The reverse methanogenic pathway also generates carbonates that precipitate inside the mat and may be the main reason for the formation of a microbial reef. The mat shows highly differentiated aggregates of various organisms, tightly interconnected by extracellular polysaccharides. In order to investigate the role of EPS as adhesive mucilage for the biofilm and as a precipitation matrix for carbonate minerals, samples were embedded in a hydrophilic resin (Lowicryl K4 M). Sections were suitable for light as well as electron microscopy in combination with lectins, either labeled with a fluorescent marker or with colloidal gold. This allows lectin mapping at low resolution for light microscopy in direct comparison with a highly resolved electron microscopic image.

  19. Topographic and electronic contrast of the graphene moir´e on Ir(111) probed by scanning tunneling microscopy and noncontact atomic force microscopy

    NARCIS (Netherlands)

    Sun, Z.; Hämäläinen, K.; Sainio, K.; Lahtinen, J.; Vanmaekelbergh, D.A.M.; Liljeroth, P.

    2011-01-01

    Epitaxial graphene grown on transition-metal surfaces typically exhibits a moir´e pattern due to the lattice mismatch between graphene and the underlying metal surface. We use both scanning tunneling microscopy (STM) and atomic force microscopy (AFM) to probe the electronic and topographic contrast

  20. A Simple Transmission Electron Microscopy Method for Fast Thickness Characterization of Suspended Graphene and Graphite Flakes.

    Science.gov (United States)

    Rubino, Stefano; Akhtar, Sultan; Leifer, Klaus

    2016-02-01

    We present a simple, fast method for thickness characterization of suspended graphene/graphite flakes that is based on transmission electron microscopy (TEM). We derive an analytical expression for the intensity of the transmitted electron beam I 0(t), as a function of the specimen thickness t (tgraphene/graphite, the method we propose has the advantage of being simple and fast, requiring only the acquisition of bright-field images.

  1. Joint denoising and distortion correction of atomic scale scanning transmission electron microscopy images

    OpenAIRE

    Berkels, Benjamin; Wirth, Benedikt

    2016-01-01

    Nowadays, modern electron microscopes deliver images at atomic scale. The precise atomic structure encodes information about material properties. Thus, an important ingredient in the image analysis is to locate the centers of the atoms shown in micrographs as precisely as possible. Here, we consider scanning transmission electron microscopy (STEM), which acquires data in a rastering pattern, pixel by pixel. Due to this rastering combined with the magnification to atomic scale, movements of th...

  2. Gold nanoparticle-protein arrays improve resolution for cryo-electron microscopy

    OpenAIRE

    Hu, Minghui; Qian, Luping; Briñas, Raymond P.; Lymar, Elena S.; Kuznetsova, Larisa; Hainfeld, James F.

    2007-01-01

    Cryo-electron microscopy single particle analysis shows limited resolution due to poor alignment precision of noisy images taken under low electron exposure. Certain advantages can be obtained by assembling proteins into two-dimensional (2D) arrays since protein particles are locked into repetitive orientations, thus improving alignment precision. We present a labeling method to prepare protein 2D arrays using gold nanoparticles (NPs) interconnecting genetic tag sites on proteins. As an examp...

  3. Possibilities and limitations of advanced transmission electron microscopy for carbon-based nanomaterials

    OpenAIRE

    Xiaoxing Ke; Carla Bittencourt; Gustaaf Van Tendeloo

    2015-01-01

    A major revolution for electron microscopy in the past decade is the introduction of aberration correction, which enables one to increase both the spatial resolution and the energy resolution to the optical limit. Aberration correction has contributed significantly to the imaging at low operating voltages. This is crucial for carbon-based nanomaterials which are sensitive to electron irradiation. The research of carbon nanomaterials and nanohybrids, in particular the fundamental understanding...

  4. Analysis of acute brain slices by electron microscopy: a correlative light-electron microscopy workflow based on Tokuyasu cryo-sectioning.

    Science.gov (United States)

    Loussert Fonta, Celine; Leis, Andrew; Mathisen, Cliff; Bouvier, David S; Blanchard, Willy; Volterra, Andrea; Lich, Ben; Humbel, Bruno M

    2015-01-01

    Acute brain slices are slices of brain tissue that are kept vital in vitro for further recordings and analyses. This tool is of major importance in neurobiology and allows the study of brain cells such as microglia, astrocytes, neurons and their inter/intracellular communications via ion channels or transporters. In combination with light/fluorescence microscopies, acute brain slices enable the ex vivo analysis of specific cells or groups of cells inside the slice, e.g. astrocytes. To bridge ex vivo knowledge of a cell with its ultrastructure, we developed a correlative microscopy approach for acute brain slices. The workflow begins with sampling of the tissue and precise trimming of a region of interest, which contains GFP-tagged astrocytes that can be visualised by fluorescence microscopy of ultrathin sections. The astrocytes and their surroundings are then analysed by high resolution scanning transmission electron microscopy (STEM). An important aspect of this workflow is the modification of a commercial cryo-ultramicrotome to observe the fluorescent GFP signal during the trimming process. It ensured that sections contained at least one GFP astrocyte. After cryo-sectioning, a map of the GFP-expressing astrocytes is established and transferred to correlation software installed on a focused ion beam scanning electron microscope equipped with a STEM detector. Next, the areas displaying fluorescence are selected for high resolution STEM imaging. An overview area (e.g. a whole mesh of the grid) is imaged with an automated tiling and stitching process. In the final stitched image, the local organisation of the brain tissue can be surveyed or areas of interest can be magnified to observe fine details, e.g. vesicles or gold labels on specific proteins. The robustness of this workflow is contingent on the quality of sample preparation, based on Tokuyasu's protocol. This method results in a reasonable compromise between preservation of morphology and maintenance of

  5. Correlative super-resolution fluorescence and electron microscopy of the nuclear pore complex with molecular resolution.

    Science.gov (United States)

    Löschberger, Anna; Franke, Christian; Krohne, Georg; van de Linde, Sebastian; Sauer, Markus

    2014-10-15

    Here, we combine super-resolution fluorescence localization microscopy with scanning electron microscopy to map the position of proteins of nuclear pore complexes in isolated Xenopus laevis oocyte nuclear envelopes with molecular resolution in both imaging modes. We use the periodic molecular structure of the nuclear pore complex to superimpose direct stochastic optical reconstruction microscopy images with a precision of <20 nm on electron micrographs. The correlative images demonstrate quantitative molecular labeling and localization of nuclear pore complex proteins by standard immunocytochemistry with primary and secondary antibodies and reveal that the nuclear pore complex is composed of eight gp210 (also known as NUP210) protein homodimers. In addition, we find subpopulations of nuclear pore complexes with ninefold symmetry, which are found occasionally among the more typical eightfold symmetrical structures.

  6. Segmentation of scanning electron microscopy images from natural rubber samples with gold nanoparticles using starlet wavelets.

    Science.gov (United States)

    de Siqueira, Alexandre Fioravante; Cabrera, Flávio Camargo; Pagamisse, Aylton; Job, Aldo Eloizo

    2014-01-01

    Electronic microscopy has been used for morphology evaluation of different materials structures. However, microscopy results may be affected by several factors. Image processing methods can be used to correct and improve the quality of these results. In this article, we propose an algorithm based on starlets to perform the segmentation of scanning electron microscopy images. An application is presented in order to locate gold nanoparticles in natural rubber membranes. In this application, our method showed accuracy greater than 85% for all test images. Results given by this method will be used in future studies, to computationally estimate the density distribution of gold nanoparticles in natural rubber samples and to predict reduction kinetics of gold nanoparticles at different time periods.

  7. Microtubule organization within mitotic spindles revealed by serial block face scanning electron microscopy and image analysis.

    Science.gov (United States)

    Nixon, Faye M; Honnor, Thomas R; Clarke, Nicholas I; Starling, Georgina P; Beckett, Alison J; Johansen, Adam M; Brettschneider, Julia A; Prior, Ian A; Royle, Stephen J

    2017-05-15

    Serial block face scanning electron microscopy (SBF-SEM) is a powerful method to analyze cells in 3D. Here, working at the resolution limit of the method, we describe a correlative light-SBF-SEM workflow to resolve microtubules of the mitotic spindle in human cells. We present four examples of uses for this workflow that are not practical by light microscopy and/or transmission electron microscopy. First, distinguishing closely associated microtubules within K-fibers; second, resolving bridging fibers in the mitotic spindle; third, visualizing membranes in mitotic cells, relative to the spindle apparatus; and fourth, volumetric analysis of kinetochores. Our workflow also includes new computational tools for exploring the spatial arrangement of microtubules within the mitotic spindle. We use these tools to show that microtubule order in mitotic spindles is sensitive to the level of TACC3 on the spindle. © 2017. Published by The Company of Biologists Ltd.

  8. Direct observation of defect structure in protein crystals by atomic force and transmission electron microscopy

    Energy Technology Data Exchange (ETDEWEB)

    Devaud, G. (Department of Physics, University of Colorado, Boulder, Colorado 80309 (United States)); Furcinitti, P.S. (Department of Molecular, Cellular and Developmental Biology, University of Colorado, Boulder, Colorado 80309 (United States)); Fleming, J.C.; Lyon, M.K.; Douglas, K. (Department of Physics, University of Colorado, Boulder, Colorado 80309 (United States))

    1992-09-01

    We have examined the structure of S-layers isolated from {ital Sulfolobus} {ital acidocaldarius} using atomic force microscopy (AFM) and transmission electron microscopy (TEM). From the AFM images, we were able to directly observe individual dimers of the crystal, defects in the crystal structure, and twin boundaries. We have identified two types of boundaries, one defined by a mirror plane and the other by a glide plane. This work shows that twin boundaries are highly structured regions that are directly related to the organization of units within each crystal domain. Projection maps from TEM images have shown that there are significant differences in the final average maps, depending on which side of the sample is adsorbed to the carbon support film. Comparison of AFM images to TEM projection maps has allowed us to relate high magnification views obtained by AFM to the relatively high resolution information obtained by electron microscopy and image processing.

  9. Morphologic differences observed by scanning electron microscopy according to the reason for pseudophakic IOL explantation

    DEFF Research Database (Denmark)

    Fernandez-Buenaga, Roberto; Alio, Jorge L.; Ramirez, Jose M.

    2015-01-01

    Purpose To compare variations in surface morphology, as studied by scanning electron microscopy (SEM), of explanted intraocular lenses (IOLs) concerning the cause leading to the explantation surgery. Methods In this prospective multicenter study, explanted IOLs were analyzed by SEM and energy-dis...

  10. Scanning electron microscopy of the oral apparatus and buccopharyngeal cavity of Atelognathus salai larvae (Anura, Neobatrachia

    Directory of Open Access Journals (Sweden)

    Dinorah D. Echeverría

    2006-09-01

    Full Text Available The aim of this study is to describe the horny structures of the buccal apparatus and buccopharyngeal cavity of A. salai by means ofscanning electron microscopy (SEM, and to compare them to those of the other known species of Atelognathus and related genera.

  11. Electron Microscopy Characterization of Aerosols Collected at Mauna Loa Observatory During Asian Dust Storm Event

    Science.gov (United States)

    Atmospheric aerosol particles have a significant influence on global climate due to their ability to absorb and scatter incoming solar radiation. Size, composition, and morphology affect a particle’s radiative properties and these can be characterized by electron microscopy. Lo...

  12. Calibration-free quantitative surface topography reconstruction in scanning electron microscopy

    NARCIS (Netherlands)

    Faber, E.T.; Martinez-Martinez, D.; Mansilla, C.; Ocelik, V.; De Hosson, J. Th. M.

    2015-01-01

    This work presents a new approach to obtain reliable surface topography reconstructions from 2D Scanning Electron Microscopy (SEM) images. In this method a set of images taken at different tilt angles are compared by means of digital image correlation (DlC). It is argued that the strength of the met

  13. Dental wax impressions of plant tissues for viewing with scanning electron microscopy (SEM).

    Science.gov (United States)

    Beermann, Anke; Hülskamp, Martin

    2010-09-01

    Scanning electron microscopy (SEM) is a valuable method for examining surface structures. Taking wax impressions of plant structures, such as leaves, is a nondestructive procedure that makes it possible to view changes in surface structures over time, such as during development. This protocol describes a method for making dental wax impressions of plant tissues.

  14. Surface sensitivity effects with local probe scanning Auger-scanning electron microscopy

    NARCIS (Netherlands)

    Van Agterveld, DTL; Palasantzas, G; De Hosson, JTM; Bentley, J; Allen, C; Dahmen, U; Petrov,

    2001-01-01

    Ultra-high-vacuum segregation studies on in-situ fractured Cu-Sb alloys were performed in terms of nanometer scale scanning Auger/Electron microscopy. S contamination leads to the formation Of Cu2S precipitates which, upon removal due to fracture, expose pits with morphology that depends on the prec

  15. Cryogenic transmission electron microscopy (cryo-TEM) for studying the morphology of colloidal drug delivery systems

    DEFF Research Database (Denmark)

    Kuntsche, Judith; Horst, Jennifer C; Bunjes, Heike

    2011-01-01

    Cryogenic transmission electron microscopy (cryo-TEM) has evolved into an indispensable tool for the characterization of colloidal drug delivery systems. It can be applied to study the size, shape and internal structure of nanoparticulate carrier systems as well as the overall colloidal composition...

  16. Assignment of two ultrastructures formed by a mixture of hexonamides using autoradiography and electron microscopy

    NARCIS (Netherlands)

    Boettcher, Christoph; Boekema, Egbert J.; Fuhrhop, Juergen-H.

    1990-01-01

    The combined application of autoradiography and electron microscopy allowed the assignment of molecular components to individual micellar fibres in a mixed gel. Resolution was of the order of 0·1 µm. As a result, it was shown that bimolecular sheets of N-dodecyl-L-mannonamide (= L-Man-12) completely

  17. Developments in application of light and scanning electron microscopy techniques for cell wall degradation studies.

    NARCIS (Netherlands)

    Engels, F.M.

    1996-01-01

    The results of recent technological developments in light and scanning electron microscopy closely used for research on forage cell wall degradation in ruminants, are reviewed. The indigestibility of forages by rumen microorganisms used to be ascribed mainly to an overall presence of lignin in the p

  18. Advances in Transmission Electron Microscopy : In Situ Straining and In Situ Compression Experiments on Metallic Glasses

    NARCIS (Netherlands)

    De Hosson, Jeff Th. M.

    In the field of transmission electron microscopy (TEM), fundamental and practical reasons still remain that hamper a straightforward correlation between microscopic structural information and deformation mechanisms in materials. In this article, it is argued that one should focus in particular on in

  19. High-Resolution Transmission Electron Microscopy Observation of Colloidal Nanocrystal Growth Mechanisms using Graphene Liquid Cells

    Energy Technology Data Exchange (ETDEWEB)

    Yuk, Jong Min; Park, Jungwon; Ercius, Peter; Kim, Kwanpyo; Hellebusch, Danny J.; Crommie, Michael F.; Lee, Jeong Yong; Zettl, A.; Alivisatos, A. Paul

    2011-12-12

    We introduce a new type of liquid cell for in-situ electron microscopy based upon entrapment of a liquid film between layers of graphene. We employ this cell to achieve high-resolution imaging of colloidal platinum nanocrystal growth. The ability to directly image and resolve critical steps at atomic resolution provides new insights into nanocrystal coalescence and reshaping during growth.

  20. Transmission Electron Microscopy Specimen Preparation Method for Multiphase Porous Functional Ceramics

    DEFF Research Database (Denmark)

    Zhang, Wei; Kuhn, Luise Theil; Jørgensen, Peter Stanley

    2013-01-01

    An optimum method is proposed to prepare thin foil transmission electron microscopy (TEM) lamellae of multiphase porous functional ceramics: prefilling the pore space of these materials with an epoxy resin prior to focused ion beam milling. Several advantages of epoxy impregnation are demonstrate...