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Sample records for electron microscopy apfim

  1. Electron Microscopy.

    ERIC Educational Resources Information Center

    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. Scanning ultrafast electron microscopy.

    PubMed

    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.

  3. Scanning ultrafast electron microscopy

    PubMed Central

    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

  4. Conventional transmission electron microscopy

    PubMed Central

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

    2014-01-01

    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. PMID:24482357

  5. Electronic detectors for electron microscopy.

    PubMed

    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.

  6. Bridging fluorescence microscopy and electron microscopy

    PubMed Central

    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 breakthrough in fluorescence microscopy in biology is the ability to follow specific targets on or in living cells, revealing dynamic localization and/or function of target molecules. One of the inherent limitations of fluorescence microscopy is the resolution. Several efforts are undertaken to overcome this limit. The traditional and most well-known way to achieve higher resolution imaging is by electron microscopy. Moreover, electron microscopy reveals organelles, membranes, macromolecules, and thus aids in the understanding of cellular complexity and localization of molecules of interest in relation to other structures. With the new probe development, a solid bridge between fluorescence microscopy and electron microscopy is being built, even leading to correlative imaging. This connection provides several benefits, both scientifically as well as practically. Here, I summarize recent developments in bridging microscopy. PMID:18575880

  7. Combined atom-probe and electron microscopy characterization of fine scale structures in aged primary coolant pipe stainless steel

    SciTech Connect

    Bentley, J.; Miller, M.K.

    1986-01-01

    The capabilities and complementary nature of atom probe field-ion microscopy (APFIM) and analytical electron microscopy (AEM) for the characterization of fine-scale microstructures are illustrated by examination of the changes that occur after long term thermal aging of cast CF 8 and CF 8M duplex stainless steels. In material aged at 300 or 400/sup 0/C for up to 70,000 h, the ferrite had spinodally decomposed into a modulated fine-scaled interconnected network consisting of an iron-rich ..cap alpha.. phase and a chromium-enriched ..cap alpha..' phase with periodicities of between 2 and 9 nm. G-phase precipitates 2 to 10 nm in diameter were also observed in the ferrite at concentrations of more than 10/sup 21/ m/sup -3/. The reported degradation in mechanical properties is most likely a consequence of the spinodal decomposition in the ferrite.

  8. Silver stain for electron microscopy

    NASA Technical Reports Server (NTRS)

    Corbett, R. L.

    1972-01-01

    Ammoniacal silver stain used for light microscopy was adapted advantageously for use with very thin biological sections required for electron microscopy. Silver stain can be performed in short time, has more contrast, and is especially useful for low power electron microscopy.

  9. Soil microstructure and electron microscopy

    NASA Technical Reports Server (NTRS)

    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.

  10. Electronic Blending in Virtual Microscopy

    ERIC Educational Resources Information Center

    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…

  11. Materials science through electron microscopy

    NASA Astrophysics Data System (ADS)

    Fujita, Hiroshi

    1992-03-01

    Electron microscopy has greatly contributed as a powerful tool in both the characterization and identification of materials in the atomic scale. In these contributions, the most important advantage is it's ability for dynamic study of phenomena, i.e., in situ experiments. This research has been carried out using high voltage electron microscopes, but some results have been obtained with high resolution electron microscopes under critical conditions. Electron microscopy has been improved further to become an indispensable ?Micro-Laboratory? in which formation of various advance materials can also be carried out precisely in the atomic scale. Electron beam science and engineering is a typical example in this research field, and detailed processes of crystalline-amorphous transition and electron irradiation induced foreign atom implantation have been clarified by this method. Recently, new applications to the research fields of non-linear material behavior, such as the behavior of atom clusters and the role of electric dipoles on diffusion, have been carried out.

  12. Four-dimensional electron microscopy.

    PubMed

    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.

  13. Electron microscopy of electromagnetic waveforms

    NASA Astrophysics Data System (ADS)

    Ryabov, A.; Baum, P.

    2016-07-01

    Rapidly changing electromagnetic fields are the basis of almost any photonic or electronic device operation. We report how electron microscopy can measure collective carrier motion and fields with subcycle and subwavelength resolution. A collimated beam of femtosecond electron pulses passes through a metamaterial resonator that is previously excited with a single-cycle electromagnetic pulse. If the probing electrons are shorter in duration than half a field cycle, then time-frozen Lorentz forces distort the images quasi-classically and with subcycle time resolution. A pump-probe sequence reveals in a movie the sample’s oscillating electromagnetic field vectors with time, phase, amplitude, and polarization information. This waveform electron microscopy can be used to visualize electrodynamic phenomena in devices as small and fast as available.

  14. APFIM characterization of 15Kh2MFA Cr-Mo-V and 15Kh2NMFA Ni-Cr-Mo-V type steels

    NASA Astrophysics Data System (ADS)

    Miller, M. K.; Jayaram, R.; Othen, P. J.; Brauer, G.

    1994-03-01

    A microstructural characterization of 15Kh2MFA Cr-Mo-V and 15Kh2NMFA Ni-Cr-Mo-V type steels that are used in the pressure vessels of Russian VVER 440 and VVER 1000 nuclear reactors, respectively, has been performed with the use of the techniques of atom-probe field-ion microscopy (APFIM) and transmission electron microscopy. The microstructure of these materials was found to be tempered martensite and bainite. A high number density of coarse (≈ 50 to ≈ 500 nm) blocky M 7C 3 carbides and some inclusions were observed. In addition to these coarse carbides, some finer (≈ 10 nm diameter) approximately spherical MC carbides were also observed in the VVER 440 steel. Field-ion microscopy has revealed that the lath boundaries in both unirradiated VVER 440 and VVER 1000 reactor steels are decorated with an ultrathin semicontinuous film of molybdenum-carbonitride precipitates. Atom-probe analysis has revealed a high enrichment of phosphorus at the lath boundaries.

  15. Dynamic imaging with electron microscopy

    ScienceCinema

    Campbell, Geoffrey; McKeown, Joe; Santala, Melissa

    2016-07-12

    Livermore researchers have perfected an electron microscope to study fast-evolving material processes and chemical reactions. By applying engineering, microscopy, and laser expertise to the decades-old technology of electron microscopy, the dynamic transmission electron microscope (DTEM) team has developed a technique that can capture images of phenomena that are both very small and very fast. DTEM uses a precisely timed laser pulse to achieve a short but intense electron beam for imaging. When synchronized with a dynamic event in the microscope's field of view, DTEM allows scientists to record and measure material changes in action. A new movie-mode capability, which earned a 2013 R&D 100 Award from R&D Magazine, uses up to nine laser pulses to sequentially capture fast, irreversible, even one-of-a-kind material changes at the nanometer scale. DTEM projects are advancing basic and applied materials research, including such areas as nanostructure growth, phase transformations, and chemical reactions.

  16. Dynamic imaging with electron microscopy

    SciTech Connect

    Campbell, Geoffrey; McKeown, Joe; Santala, Melissa

    2014-02-20

    Livermore researchers have perfected an electron microscope to study fast-evolving material processes and chemical reactions. By applying engineering, microscopy, and laser expertise to the decades-old technology of electron microscopy, the dynamic transmission electron microscope (DTEM) team has developed a technique that can capture images of phenomena that are both very small and very fast. DTEM uses a precisely timed laser pulse to achieve a short but intense electron beam for imaging. When synchronized with a dynamic event in the microscope's field of view, DTEM allows scientists to record and measure material changes in action. A new movie-mode capability, which earned a 2013 R&D 100 Award from R&D Magazine, uses up to nine laser pulses to sequentially capture fast, irreversible, even one-of-a-kind material changes at the nanometer scale. DTEM projects are advancing basic and applied materials research, including such areas as nanostructure growth, phase transformations, and chemical reactions.

  17. Electron Microscopy of Intracellular Protozoa.

    DTIC Science & Technology

    1982-08-01

    the erythrocytes infected with P. falciparum. Scannning electron microscopy demonstrated numerous cone-shaped knobs evenly distributed over the entire...Mystromys albicaudatus and are being used as an excellent model of American cutaneous leishmaniasis In anti-leishmanial drug screen tests at WRAIR1 s...available liquid media for rapid cultivation. J Parasitol 1978, 76:309- .316. - 16 - I- ~. . . ... .. . . " " :" "". . REFERENCES (cont’d) 17. Cohn ZA

  18. Spectroscopic imaging in electron microscopy

    SciTech Connect

    Pennycook, Stephen J; Colliex, C.

    2012-01-01

    In the scanning transmission electron microscope, multiple signals can be simultaneously collected, including the transmitted and scattered electron signals (bright field and annular dark field or Z-contrast images), along with spectroscopic signals such as inelastically scattered electrons and emitted photons. In the last few years, the successful development of aberration correctors for the electron microscope has transformed the field of electron microscopy, opening up new possibilities for correlating structure to functionality. Aberration correction not only allows for enhanced structural resolution with incident probes into the sub-angstrom range, but can also provide greater probe currents to facilitate mapping of intrinsically weak spectroscopic signals at the nanoscale or even the atomic level. In this issue of MRS Bulletin, we illustrate the power of the new generation of electron microscopes with a combination of imaging and spectroscopy. We show the mapping of elemental distributions at atomic resolution and also the mapping of electronic and optical properties at unprecedented spatial resolution, with applications ranging from graphene to plasmonic nanostructures, and oxide interfaces to biology.

  19. Atomic level microstructural characterization by APFIM

    SciTech Connect

    Miller, M.K.

    1996-10-01

    Atom probe field ion microscopy has been used to characterize Ni aluminides in addition to changes in microstructure of pressure vessel steels as a result of exposure to neutron irradiation. Ultrafine intragranular Cu precipitates and P segregation to grain and lath boundaries have been quantified in the pressure vessel steels. In boron-doped Ni{sub 3}Al, the B additions were found to segregate to dislocations, low angle boundaries, antiphase boundaries, stacking faults, and grain boundaries. In boron-doped NiAl, B segregation to grain boundaries and ultrafine MB{sub 2} precipitates were observed. In Mo-doped NiAl, enrichments of Mo, C, N/Si, B, and Fe were observed at the grain boundaries together with Mo precipitates and low Mo matrix solubility.

  20. Synthetic incoherence for electron microscopy.

    PubMed

    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.

  1. Liquid Cell Transmission Electron Microscopy.

    PubMed

    Liao, Hong-Gang; Zheng, Haimei

    2016-05-27

    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.

  2. Liquid Cell Transmission Electron Microscopy

    NASA Astrophysics Data System (ADS)

    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.

  3. Epoxy Resins in Electron Microscopy

    PubMed Central

    Finck, Henry

    1960-01-01

    A method of embedding biological specimens in araldite 502 (Ciba) has been developed for materials available in the United States. Araldite-embedded tissues are suitable for electron microscopy, but the cutting qualities of the resin necessitates more than routine attention during microtomy. The rather high viscosity of araldite 502 also seems to be an unnecessary handicap. The less viscous epoxy epon 812 (Shell) produces specimens with improved cutting qualities, and has several features—low shrinkage and absence of specimen damage during cure, minimal compression of sections, relative absence of electron beam-induced section damage, etc.—which recommends it as a routine embedding material. The hardness of the cured resin can be easily adjusted by several methods to suit the materials embedded in it. Several problems and advantages of working with sections of epoxy resins are also discussed. PMID:13822825

  4. Electron microscopy and forensic practice

    NASA Astrophysics Data System (ADS)

    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).

  5. Picosecond Fresnel transmission electron microscopy

    NASA Astrophysics Data System (ADS)

    Schliep, Karl B.; Quarterman, P.; Wang, Jian-Ping; Flannigan, David J.

    2017-05-01

    We report the demonstration of picosecond Fresnel imaging with an ultrafast transmission electron microscope (UEM). By operating with a low instrument repetition rate (5 kHz) and without objective-lens excitation, the picosecond demagnetization of an FePt film, via in situ, femtosecond laser excitation, is directly imaged. The dynamics are quantified and monitored as a time-dependent change in the degree of electron coherence within the magnetic domain walls. The relative coherence of conventional (thermionic) Fresnel transmission electron microscopy is also directly compared to that of Fresnel UEM through the domain-wall size. Further, the robustness and reversibility of the domain-wall dynamics are illustrated by repeating the picosecond image scans at defocus values having the same magnitude but different signs (e.g., +25 mm vs. -25 mm). Control experiments and approaches to identifying and isolating systematic errors and sources of artifacts are also described. This work, and continued future developments also described here, opens the way to direct correlation of transient structure, morphology, and magnetic dynamics in magnetic thin films and spintronic devices.

  6. Analytical transmission electron microscopy in materials science

    SciTech Connect

    Fraser, H.L.

    1980-01-01

    Microcharacterization of materials on a scale of less than 10 nm has been afforded by recent advances in analytical transmission electron microscopy. The factors limiting accurate analysis at the limit of spatial resolution for the case of a combination of scanning transmission electron microscopy and energy dispersive x-ray spectroscopy are examined in this paper.

  7. Fast electron microscopy via compressive sensing

    SciTech Connect

    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.

  8. Atomic resolution 3D electron diffraction microscopy

    SciTech Connect

    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.

  9. Ultrafast Science Opportunities with Electron Microscopy

    SciTech Connect

    Durr, Hermann

    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.

  10. The future of electron microscopy

    SciTech Connect

    Zhu, Yimei; Durr, Hermann

    2015-04-01

    Seeing is believing. So goes the old adage and seen evidence is undoubtedly satisfying because it can be interpreted easily, though not always correctly. For centuries, humans have developed such instruments as telescopes that observe the heavens and microscopes that reveal bacteria and viruses. The 2014 Nobel Prize in Chemistry was awarded to Eric Betzig, Stefan Hell, and William Moerner for their foundational work on superresolution fluorescence microscopy in which they overcame the Abbe diffraction limit for the resolving power of conventional light microscopes. (See Physics Today, December 2014, page 18.) That breakthrough enabled discoveries in biological research and testifies to the importance of modern microscopy.

  11. The future of electron microscopy

    DOE PAGES

    Zhu, Yimei; Durr, Hermann

    2015-04-01

    Seeing is believing. So goes the old adage and seen evidence is undoubtedly satisfying because it can be interpreted easily, though not always correctly. For centuries, humans have developed such instruments as telescopes that observe the heavens and microscopes that reveal bacteria and viruses. The 2014 Nobel Prize in Chemistry was awarded to Eric Betzig, Stefan Hell, and William Moerner for their foundational work on superresolution fluorescence microscopy in which they overcame the Abbe diffraction limit for the resolving power of conventional light microscopes. (See Physics Today, December 2014, page 18.) That breakthrough enabled discoveries in biological research and testifiesmore » to the importance of modern microscopy.« less

  12. Techniques in electron microscopy of animal tissue.

    PubMed

    Cheville, N F; Stasko, J

    2014-01-01

    Technical improvements in electron microscopy, both instrumental and preparative, permit increasingly accurate analyses. Digital images for transmission electron microscopy (TEM) can be processed by software programs that automate tasks and create custom tools that allow for image enhancement for brightness, contrast and coloration; for creation of rectangular, ellipsoidal or irregular area selections; and for measurement of mean area and standard deviation. Sample preparation remains a source of error since organelles and spatial arrangements of macromolecules rapidly change after anoxia. Guidelines for maintaining consistency in preparation, examination and interpretation are presented for different electron microscopy (EM) modalities.

  13. Probing the Proton with Electron Microscopy

    NASA Astrophysics Data System (ADS)

    Friedman, Jerome I.

    2014-01-01

    This article is written as a tribute and memorial to Dr. Akira Tonomura who was an outstanding experimental physicist and a friend. Early in his career, he opened a new era in electron microscopy by demonstrating in 1968 that electron holography, proposed by Gabor in 1949, was possible; and later he developed Lorentz "phase" microscopy, which allows one to generate real-space, real-time images. All through his career, he perfected these designs into superb instruments that he employed to investigate fundamental questions in physics. Dr. Tonomura set world standards for electron microscopy.

  14. Fluorescence-integrated transmission electron microscopy images: integrating fluorescence microscopy with transmission electron microscopy.

    PubMed

    Sims, Paul A; Hardin, Jeff D

    2007-01-01

    This chapter describes high-pressure freezing (HPF) techniques for correlative light and electron microscopy on the same sample. Laser scanning confocal microscopy (LSCM) is exploited for its ability to collect fluorescent, as well as transmitted and back scattered light (BSL) images at the same time. Fluorescent information from a whole mount (preembedding) or from thin sections (post-embedding) can be displayed as a color overlay on transmission electron microscopy (TEM) images. Fluorescence-integrated TEM (F-TEM) images provide a fluorescent perspective to TEM images. The pre-embedding method uses a thin two-part agarose pad to immobilize live Caenorhabditis elegans embryos for LSCM, HPF, and TEM. Pre-embedding F-TEM images display fluorescent information collected from a whole mount of live embryos onto all thin sections collected from that sample. In contrast, the postembedding method uses HPF and freeze substitution with 1% paraformaldehyde in 95% ethanol followed by low-temperature embedding in methacrylate resin. This procedure preserves the structure and function of green fluorescent protein (GFP) as determined by immunogold labeling of GFP, when compared with GFP expression, both demonstrated in the same thin section.

  15. Transmission electron microscopy: Imaging of materials

    SciTech Connect

    Thomas, G.

    1988-10-01

    This report was an invited paper for a symposium and only covers general aspects of transmission electron microscopy. A history, and examples of work done on ceramics and alloys are covered. 6 refs., 44 figs. (JL)

  16. Low voltage transmission electron microscopy of graphene.

    PubMed

    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.

  17. Transmission electron microscopy of composites

    NASA Technical Reports Server (NTRS)

    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.

  18. Electron Microscopy of Natural and Epitaxial Diamond

    NASA Technical Reports Server (NTRS)

    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.

  19. Electron Microscopy of Natural and Epitaxial Diamond

    NASA Technical Reports Server (NTRS)

    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.

  20. Analysis of cytokinesis by electron microscopy.

    PubMed

    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.

  1. Environmental scanning electron microscopy in cell biology.

    PubMed

    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.

  2. Perspectives on in situ electron microscopy

    DOE PAGES

    Zheng, Haimei; Zhu, Yimei

    2017-03-29

    In situ transmission electron microscopy (TEM) with the ability to reveal materials dynamic processes with high spatial and temporal resolution has attracted significant interest. The recent advances in in situ methods, including liquid and gas sample environment, pump-probe ultrafast microscopy, nanomechanics and ferroelectric domain switching the aberration corrected electron optics as well as fast electron detector has opened new opportunities to extend the impact of in situ TEM in broad areas of research ranging from materials science to chemistry, physics and biology. Here in this paper, we highlight the development of liquid environment electron microscopy and its applications in themore » study of colloidal nanoparticle growth, electrochemical processes and others; in situ study of topological vortices in ferroelectric and ferromagnetic materials. At the end, perspectives of future in situ TEM are provided.« less

  3. Scanning Electron Microscopy Sample Preparation and Imaging.

    PubMed

    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.

  4. TEM (transmission electron microscopy), APFIM (atom-probe field ion microscopy), and SANS (small-angle neutron scattering) examination of aged duplex stainless steel components from some decommissioned reactors

    SciTech Connect

    Chung, H.M.; Chopra, O.K.

    1987-12-01

    The present investigation indicates that the primary embrittlement processes of the CF-8 grade cast stainless steel components during extended reactor service are spinodal decomposition of the ferrite phase and M/sub 23/C/sub 6/ carbide precipitation on the austenite-ferrite boundaries. The ferrite hardness measured for the Shippingport reactor valves appears to reflect the different extent of spinodal decomposition among the different valves which contain slightly different Cr contents. G-phase precipitation was minimal compared to that during accelerated aging of CF-8 steel in the laboratory (i.e., near 400/degree/C). This indicates that the activation energy may be strongly influenced by the synergism among the G-phase precipitation, carbide formation, and spinodal decomposition. 13 refs., 2 figs.

  5. Electron Microscopy of the Cell

    PubMed Central

    Leeson, T. S.

    1965-01-01

    The use of the electron microscope has added much to our knowledge of the cell. The fine structure of the component parts of the nucleus and the cytoplasm is described, and their functions are indicated. The nature and structural modifications of the plasma membrane are illustrated with particular reference to function. To illustrate the interrelationships of the nucleus and cytoplasm, the theory of protein secretion is discussed, the secretion of a particular protein or polypeptide being determined by a particular nucleotide sequence in the desoxyribonucleic acid of a chromosome, that is, by a gene. This information is transferred from nucleus to cytoplasm. It is in the cytoplasm that the majority of the work is performed while the nucleus directs the work of the cell. ImagesFig. 2Fig. 3Fig. 4Fig. 5Fig. 6Fig. 7Fig. 8Fig. 9Fig. 10Fig. 11Fig. 12Fig. 13Fig. 14Fig. 15Fig. 16Fig. 17Fig. 18Fig. 19Fig. 20Fig. 21Fig. 22Fig. 23Fig. 24Fig. 25Fig. 26 PMID:5829410

  6. Advanced Electron Microscopy in Materials Physics

    SciTech Connect

    Zhu, Y.; Jarausch, K.

    2009-06-01

    Aberration correction has opened a new frontier in electron microscopy by overcoming the limitations of conventional round lenses, providing sub-angstrom-sized probes and extending information limits. The imaging and analytical performance of these corrector-equipped microscopes affords an unprecedented opportunity to study structure-property relationships of matter at the atomic scale. This new generation of microscopes is able to retrieve high-quality structural information comparable to neutron and synchrotron x-ray experiments, but with local atomic resolution. These advances in instrumentation are accelerating the research and development of various functional materials ranging from those for energy generation, conversion, transportation and storage to those for catalysis and nano-device applications. The dramatic improvements in electron-beam illumination and detection also present a host of new challenges for the interpretation and optimization of experiments. During 7-9 November 2007, a workshop, entitled 'Aberration Corrected Electron Microscopy in Material Physics', was convened at the Center for Functional Nanomaterials, Brookhaven National Laboratories (BNL) to address these opportunities and challenges. The workshop was co-sponsored by Hitachi High Technologies, a leader in electron microscopy instrumentation, and BNL's Institute of Advanced Electron Microscopy, a leader in materials physics research using electron microscopy. The workshop featured presentations by internationally prominent scientists working at the frontiers of electron microscopy, both on developing instrumentation and applying it in materials physics. The meeting, structured to stimulate scientific exchanges and explore new capabilities, brought together {approx}100 people from over 10 countries. This special issue complies many of the advances in instrument performance and materials physics reported by the invited speakers and attendees at the workshop.

  7. Active Pixel Sensors for electron microscopy

    NASA Astrophysics Data System (ADS)

    Denes, P.; Bussat, J.-M.; Lee, Z.; Radmillovic, V.

    2007-09-01

    The technology used for monolithic CMOS imagers, popular for cell phone cameras and other photographic applications, has been explored for charged particle tracking by the high-energy physics community for several years. This technology also lends itself to certain imaging detector applications in electron microscopy. We have been developing such detectors for several years at Lawrence Berkeley National Laboratory, and we and others have shown that this technology can offer excellent point-spread function, direct detection and high readout speed. In this paper, we describe some of the design constraints peculiar to electron microscopy and summarize where such detectors could play a useful role.

  8. Optical microscopy versus scanning electron microscopy in urolithiasis.

    PubMed

    Marickar, Y M Fazil; Lekshmi, P R; Varma, Luxmi; Koshy, Peter

    2009-10-01

    Stone analysis is incompletely done in many clinical centers. Identification of the stone component is essential for deciding future prophylaxis. X-ray diffraction, Fourier transform infrared spectroscopy, and scanning electron microscopy (SEM) still remains a distant dream for routine hospital work. It is in this context that optical microscopy is suggested as an alternate procedure. The objective of this article was to assess the utility of an optical microscope which gives magnification of up to 40x and gives clear picture of the surface of the stones. In order to authenticate the morphological analysis of urinary stones, SEM and elemental distribution analysis were performed. A total of 250 urinary stones of different compositions were collected from stone clinic, photographed, observed under an optical microscope, and optical photographs were taken at different angles. Twenty-five representative samples among these were gold sputtered to make them conductive and were fed into the SEM machine. Photographs of the samples were taken at different angles at magnifications up to 4,000. Elemental distribution analysis (EDAX) was done to confirm the composition. The observations of the two studies were compared. The different appearances of the stones under optical illuminated microscopy were mostly standardized appearances, namely bosselations of pure whewellite, spiculations of weddellite, bright yellow colored appearance of uric acid, and dirty white amorphous appearance of phosphates. SEM and EDAX gave clearer pictures and gave added confirmation of the stone composition. From the references thus obtained, it was possible to confirm the composition by studying the optical microscopic pictures. Higher magnification capacity of the SEM and the EDAX patterns are useful to give reference support for performing optical microscopy work. After standardization, routine analysis can be performed with optical microscopy. The advantage of the optical microscope is that, it

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

    PubMed Central

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

    2016-01-01

    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. PMID:26580231

  10. Scanning electron microscopy study of Trichomonas gallinae.

    PubMed

    Tasca, Tiana; De Carli, Geraldo A

    2003-12-01

    A scanning electron microscopy (SEM) study of Trichomonas gallinae (Rivolta, 1878), provided more information about the morphology of this flagellated protozoan. SEM showed the morphological features of the trophozoites; the emergence of the anterior flagella, the structure of the undulating membrane, the position and shape of the pelta, axostyle and posterior flagellum. Of special interest were the pseudocyst forms.

  11. Photon-induced near field electron microscopy

    NASA Astrophysics Data System (ADS)

    Park, Sang Tae; Zewail, Ahmed H.

    2013-09-01

    Ultrafast electron microscopy in the space and time domains utilizes a pulsed electron probe to directly map structural dynamics of nanomaterials initiated by an optical pump pulse, in imaging, di raction, spectroscopy, and their combinations. It has demonstrated its capability in the studies of phase transitions, mechanical vibrations, and chemical reactions. Moreover, electrons can directly interact with photons via the near eld component of light scattering by nanostructures, and either gain or lose light quanta discretely in energy. By energetically selecting those electrons that exchanged photon energies, we can map this photon-electron interaction, and the technique is termed photon-induced near eld electron microscopy (PINEM). Here, we give an account of the theoretical understanding of PINEM. Experimentally, nanostructures such as a sphere, cylinder, strip, and triangle have been investigated. Theoretically, time-dependent Schrodinger and Dirac equations for an electron under light are directly solved to obtain analytical solutions. The interaction probability is expressed by the mechanical work done by an optical wave on a traveling electron, which can be evaluated analytically by the near eld components of the Rayleigh scattering for small spheres and thin cylinders, and numerically by the discrete dipole approximation for other geometries. Application in visualization of plasmon elds is discussed.

  12. Application of Electron Diffraction to Biological Electron Microscopy

    PubMed Central

    Glaeser, Robert M.; Thomas, Gareth

    1969-01-01

    Three methods by which electron diffraction may be applied to problems in electron microscopy are discussed from a fundamental point of view, and experimental applications with biological specimens are demonstrated for each case. It is shown that wide-angle electron diffraction provides valuable information for evaluating specimen damage that can occur either during specimen preparation or while in the electron beam. Dark-field electron microscopy can be used both to enhance the image contrast and to provide highly restricted and therefore highly specific information about the object. Low-angle electron diffraction provides quantitative information about the object structure in the range from 20 A to ∼ 1000 A. Lowangle electron diffraction also demonstrates the important role of Fourier contrast with biological specimens, which are usually characterized by structural features with dimensions of 20 A or larger. ImagesFigure 1Figure 2Figure 5Figure 6Figure 7Figure 8Figure 9Figure 10Figure 11Figure 13 PMID:4896898

  13. Mudrocks examined by backscattered electron microscopy

    NASA Technical Reports Server (NTRS)

    Pye, K.; Krinsley, D.

    1983-01-01

    A method of studying mudrocks is developed using backscattered electrons (BSE) in scanning electron microscopy. Commercially available detectors are utilized to mix the BSE and secondary electron signals in order to obtain the optimum image for a particular material. Thin sections or polished rock chip surfaces are examined with BSE which provides both the atomic number contrast and topographic contrast. This technique provides very detailed information about the form and composition of individual grains in the mudrock thin sections and can be used in studies of the source, mode of deposition, diagenesis, and tectonic deformational history of mudrocks.

  14. Mudrocks examined by backscattered electron microscopy

    NASA Technical Reports Server (NTRS)

    Pye, K.; Krinsley, D.

    1983-01-01

    A method of studying mudrocks is developed using backscattered electrons (BSE) in scanning electron microscopy. Commercially available detectors are utilized to mix the BSE and secondary electron signals in order to obtain the optimum image for a particular material. Thin sections or polished rock chip surfaces are examined with BSE which provides both the atomic number contrast and topographic contrast. This technique provides very detailed information about the form and composition of individual grains in the mudrock thin sections and can be used in studies of the source, mode of deposition, diagenesis, and tectonic deformational history of mudrocks.

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

    PubMed

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

    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.

  16. DNA base identification by electron microscopy.

    PubMed

    Bell, David C; Thomas, W Kelley; Murtagh, Katelyn M; Dionne, Cheryl A; Graham, Adam C; Anderson, Jobriah E; Glover, William R

    2012-10-01

    Advances in DNA sequencing, based on fluorescent microscopy, have transformed many areas of biological research. However, only relatively short molecules can be sequenced by these technologies. Dramatic improvements in genomic research will require accurate sequencing of long (>10,000 base-pairs), intact DNA molecules. Our approach directly visualizes the sequence of DNA molecules using electron microscopy. This report represents the first identification of DNA base pairs within intact DNA molecules by electron microscopy. By enzymatically incorporating modified bases, which contain atoms of increased atomic number, direct visualization and identification of individually labeled bases within a synthetic 3,272 base-pair DNA molecule and a 7,249 base-pair viral genome have been accomplished. This proof of principle is made possible by the use of a dUTP nucleotide, substituted with a single mercury atom attached to the nitrogenous base. One of these contrast-enhanced, heavy-atom-labeled bases is paired with each adenosine base in the template molecule and then built into a double-stranded DNA molecule by a template-directed DNA polymerase enzyme. This modification is small enough to allow very long molecules with labels at each A-U position. Image contrast is further enhanced by using annular dark-field scanning transmission electron microscopy (ADF-STEM). Further refinements to identify additional base types and more precisely determine the location of identified bases would allow full sequencing of long, intact DNA molecules, significantly improving the pace of complex genomic discoveries.

  17. Aberration corrected Lorentz scanning transmission electron microscopy.

    PubMed

    McVitie, S; McGrouther, D; McFadzean, S; MacLaren, D A; O'Shea, K J; Benitez, M J

    2015-05-01

    We present results from an aberration corrected scanning transmission electron microscope which has been customised for high resolution quantitative Lorentz microscopy with the sample located in a magnetic field free or low field environment. We discuss the innovations in microscope instrumentation and additional hardware that underpin the imaging improvements in resolution and detection with a focus on developments in differential phase contrast microscopy. Examples from materials possessing nanometre scale variations in magnetisation illustrate the potential for aberration corrected Lorentz imaging as a tool to further our understanding of magnetism on this lengthscale.

  18. Multi-pass transmission electron microscopy

    DOE PAGES

    Juffmann, Thomas; Koppell, Stewart A.; Klopfer, Brannon B.; ...

    2017-05-10

    Feynman once asked physicists to build better electron microscopes to be able to watch biology at work. While electron microscopes can now provide atomic resolution, electron beam induced specimen damage precludes high resolution imaging of sensitive materials, such as single proteins or polymers. Here, we use simulations to show that an electron microscope based on a multi-pass measurement protocol enables imaging of single proteins, without averaging structures over multiple images. While we demonstrate the method for particular imaging targets, the approach is broadly applicable and is expected to improve resolution and sensitivity for a range of electron microscopy imaging modalities,more » including, for example, scanning and spectroscopic techniques. The approach implements a quantum mechanically optimal strategy which under idealized conditions can be considered interaction-free.« less

  19. Image Restoration in Cryo-electron Microscopy

    PubMed Central

    Penczek, Pawel A.

    2011-01-01

    Image restoration techniques are used to obtain, given experimental measurements, the best possible approximation of the original object within the limits imposed by instrumental conditions and noise level in the data. In molecular electron microscopy, we are mainly interested in linear methods that preserve the respective relationships between mass densities within the restored map. Here, we describe the methodology of image restoration in structural electron microscopy, and more specifically, we will focus on the problem of the optimum recovery of Fourier amplitudes given electron microscope data collected under various defocus settings. We discuss in detail two classes of commonly used linear methods, the first of which consists of methods based on pseudoinverse restoration, and which is further subdivided into mean-square error, chi-square error, and constrained based restorations, where the methods in the latter two subclasses explicitly incorporates non-white distribution of noise in the data. The second class of methods is based on the Wiener filtration approach. We show that the Wiener filter-based methodology can be used to obtain a solution to the problem of amplitude correction (or “sharpening”) of the electron microscopy map that makes it visually comparable to maps determined by X-ray crystallography, and thus amenable to comparable interpretation. Finally, we present a semi-heuristic Wiener filter-based solution to the problem of image restoration given sets of heterogeneous solutions. We conclude the chapter with a discussion of image restoration protocols implemented in commonly used single particle software packages. PMID:20888957

  20. Frontiers of in situ electron microscopy

    DOE PAGES

    Zheng, Haimei; Zhu, Yimei; Meng, Shirley Ying

    2015-01-01

    In situ transmission electron microscopy (TEM) has become an increasingly important tool for materials characterization. It provides key information on the structural dynamics of a material during transformations and the correlation between structure and properties of materials. With the recent advances in instrumentation, including aberration corrected optics, sample environment control, the sample stage, and fast and sensitive data acquisition, in situ TEM characterization has become more and more powerful. In this article, a brief review of the current status and future opportunities of in situ TEM is included. It also provides an introduction to the six articles covered by inmore » this issue of MRS Bulletin explore the frontiers of in situ electron microscopy, including liquid and gas environmental TEM, dynamic four-dimensional TEM, nanomechanics, ferroelectric domain switching studied by in situ TEM, and state-of-the-art atomic imaging of light elements (i.e., carbon atoms) and individual defects.« less

  1. 4D electron microscopy: principles and applications.

    PubMed

    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

  2. Biological cryo‐electron microscopy in China

    PubMed Central

    2016-01-01

    Abstract Cryo‐electron microscopy (cryo‐EM) plays an increasingly more important role in structural biology. With the construction of an arm of the Chinese National Protein Science Facility at Tsinghua University, biological cryo‐EM has entered a phase of rapid development in China. This article briefly reviews the history of biological cryo‐EM in China, describes its current status, comments on its impact on the various biological research fields, and presents future outlook. PMID:27534377

  3. Biological cryo-electron microscopy in China.

    PubMed

    Wang, Hong-Wei; Lei, Jianlin; Shi, Yigong

    2017-01-01

    Cryo-electron microscopy (cryo-EM) plays an increasingly more important role in structural biology. With the construction of an arm of the Chinese National Protein Science Facility at Tsinghua University, biological cryo-EM has entered a phase of rapid development in China. This article briefly reviews the history of biological cryo-EM in China, describes its current status, comments on its impact on the various biological research fields, and presents future outlook.

  4. Correlative photoactivated localization and scanning electron microscopy.

    PubMed

    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.

  5. Scanning electron microscopy of superficial white onychomycosis*

    PubMed Central

    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

  6. SESAM: exploring the frontiers of electron microscopy.

    PubMed

    Koch, Christoph T; Sigle, Wilfried; Höschen, Rainer; Rühle, Manfred; Essers, Erik; Benner, Gerd; Matijevic, Marko

    2006-12-01

    We report on the sub-electron-volt-sub-angstrom microscope (SESAM), a high-resolution 200-kV FEG-TEM equipped with a monochromator and an in-column MANDOLINE filter. We report on recent results obtained with this instrument, demonstrating its performance (e.g., 87-meV energy resolution at 10-s exposure time, or a transmissivity of the energy filter of T1 ev = 11,000 nm2). New opportunities to do unique experiments that may advance the frontiers of microscopy in areas such as energy-filtered TEM, spectroscopy, energy-filtered electron diffraction and spectroscopic profiling are also discussed.

  7. Phase-contrast scanning transmission electron microscopy.

    PubMed

    Minoda, Hiroki; Tamai, Takayuki; Iijima, Hirofumi; Hosokawa, Fumio; Kondo, Yukihito

    2015-06-01

    This report introduces the first results obtained using phase-contrast scanning transmission electron microscopy (P-STEM). A carbon-film phase plate (PP) with a small center hole is placed in the condenser aperture plane so that a phase shift is introduced in the incident electron waves except those passing through the center hole. A cosine-type phase-contrast transfer function emerges when the phase-shifted scattered waves interfere with the non-phase-shifted unscattered waves, which passed through the center hole before incidence onto the specimen. The phase contrast resulting in P-STEM is optically identical to that in phase-contrast transmission electron microscopy that is used to provide high contrast for weak phase objects. Therefore, the use of PPs can enhance the phase contrast of the STEM images of specimens in principle. The phase shift resulting from the PP, whose thickness corresponds to a phase shift of π, has been confirmed using interference fringes displayed in the Ronchigram of a silicon single crystal specimen. The interference fringes were found to abruptly shift at the edge of the PP hole by π. © The Author 2015. Published by Oxford University Press on behalf of The Japanese Society of Microscopy. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

  8. Electron microscopy of Crotalaria pulmonary hypertension

    PubMed Central

    Kay, J. M.; Smith, Paul; Heath, Donald

    1969-01-01

    The lungs of 11 rats fed on Crotalaria spectabilis seeds for periods ranging from 12 to 61 days were examined by both light and electron microscopy. The findings were compared with those obtained from nine control rats given a normal diet. Eight of the 11 test rats showed morphological evidence of pulmonary arterial hypertension in the form of right ventricular hypertrophy; the exceptions were rats killed after receiving the Crotalaria diet for 12, 22, and 29 days respectively. On light microscopy, all the test rats showed exudative lesions in the lungs consisting of eosinophilic alveolar coagulum, intra-alveolar haemorrhage, interstitial fibrosis, and a proliferation of mast cells. Enlarged and proliferated cells were seen to line the alveolar walls or lie free within the alveolar spaces. Electron microscopy showed these cells to be enlarged granular pneumocytes containing enlarged, electron-dense, lamellar secretory inclusions. Scanty macrophages were also seen in the alveolar spaces, in which excessive numbers of myelin figures and lattices were seen: these structures resembled phospholipid membranes and were probably related to pulmonary surfactant. We think that proliferation of granular pneumocytes is a non-specific reaction of the alveolar walls to injury. The alveolar-capillary wall showed interstitial oedema with the formation of intraluminal endothelial vesicles, probably representing the early ultrastructural phase of pulmonary oedema, and more likely to be an effect of the pulmonary hypertension than its cause. Images PMID:5348317

  9. Embedment-free section electron microscopy.

    PubMed

    Kondo, Hisatake

    2006-08-01

    Because of potential hindrance of clear viewing in epoxy sections of biological entities having an electron density similar to and lower than that of epoxy resin, the author has stressed that the embedment-free section electron microscopy is necessary for re-examination and/or clarification of biological specimen structures, and that the embedment-free electron microscopy is reliably done by using water-soluble polyethylene glycol (PEG) as a transient embedding media and by critical point-drying of embedment-free sections after de-embedment of PEG by immersion of semithin sections into water. With the embedment-free electron microscopy, the author has presented five major findings: the appearance of microtrabecular lattices with different compactnesses in various cells and in intracellular domains of a given cell, the faithful reproduction of microtrabecula-like strand lattices in vitro with increasing compactnesses from artificial protein solutions at correspondingly increasing concentrations, the appearance of more compact lattices from gelated gelatin than from solated gelatin at a given concentration in vitro, the changeability in compactnesses of the microtrabecular lattices by hyper- or hypotonic shock treatments of cells, and the confined appearance of an intracellular protein in the centripetal demilune of centrifuged ganglion cells which is occupied with the microtrabecular lattices of a substantial compactness. From these findings, several conclusions are drawn: individual strands themselves of the microtrabeculae are meaningless, the appearance of microtrabeculae represents the presence of proteins at a certain concentration, and it is therefore likely that the aqueous cytoplasm is equivalent to the aqueous solution. In addition, it is possible that the appearance of two contiguous lattice domains exhibiting different compactnesses in a given cell may represent the occurrence of a contiguity of sol to gel states of cytoplasmic domains. It is thus

  10. Scanning electron microscopy of lichen sclerosus*

    PubMed Central

    de Almeida, Hiram Larangeira; Bicca, Eduardo de Barros Coelho; Breunig, Juliano de Avelar; Rocha, Nara Moreira; Silva, Ricardo Marques e

    2013-01-01

    Lichen sclerosus is an acquired inflammatory condition characterized by whitish fibrotic plaques, with a predilection for the genital skin. We performed scanning electron microscopy of the dermis from a lesion of lichen sclerosus. Normal collagen fibers could be easily found in deeper layers of the specimen, as well as the transition to pathologic area, which seems homogenized. With higher magnifications in this transitional area collagen fibers are adherent to each other, and with very high magnifications a pearl chain aspect became evident along the collagen fibers. In the superficial dermis this homogenization is even more evident, collagen fibers are packed together and round structures are also observed. Rupture of collagen fibers and inflammatory cells were not found. These autoimmune changes of the extracellular matrix lead to the aggregation of immune complexes and/or changed matrix proteins along the collagen fibers, the reason why they seem hyalinized when examined by light microscopy. PMID:23739707

  11. Mechanisms of decoherence in electron microscopy.

    PubMed

    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.

  12. Immunogold Labeling for Scanning Electron Microscopy.

    PubMed

    Goldberg, Martin W; Fišerová, Jindřiška

    2016-01-01

    Scanning electron microscopes are useful biological tools that can be used to image the surface of whole organisms, tissues, cells, cellular components, and macromolecules. Processes and structures that exist at surfaces can be imaged in pseudo, or real 3D at magnifications ranging from about 10× to 1,000,000×. Therefore a whole multicellular organism, such as a fly, or a single protein embedded in one of its cell membranes can be visualized. In order to identify that protein at high resolution, or to see and quantify its distribution at lower magnifications, samples can be labeled with antibodies. Any surface that can be exposed can potentially be studied in this way. Presented here is a generic method for immunogold labeling for scanning electron microscopy, using two examples of specimens: isolated nuclear envelopes and the cytoskeleton of mammalian culture cells. Various parameters for sample preparation, fixation, immunogold labeling, drying, metal coating, and imaging are discussed so that the best immunogold scanning electron microscopy results can be obtained from different types of specimens.

  13. Immunogold labelling for scanning electron microscopy.

    PubMed

    Goldberg, Martin W; Fiserova, Jindriska

    2010-01-01

    Scanning electron microscopes are useful biological tools that can be used to image the surface of whole organisms, tissues, cells, cellular components and macromolecules. Processes and structures that exist at surfaces can be imaged in pseudo or real 3D at magnifications of anything from about x10 to x1,000,000. Therefore a whole multicellular organism, such as a fly, or a single protein embedded in one of its cell membranes can be visualised. In order to identify that protein at high resolution, or to see and quantify its distribution at lower magnifications, samples can be labelled with antibodies. Any surface that can be exposed can potentially be studied in this way. Presented here is a generic method for immunogold labelling for scanning electron microscopy, using two examples of specimens: isolated nuclear envelopes and the cytoskeleton of mammalian culture cells. Various parameters for sample preparation, fixation, immunogold labelling, drying, metal coating and imaging are discussed so that the best immunogold scanning electron microscopy results can be obtained from different types of specimens.

  14. Sewage coliphages studied by electron microscopy.

    PubMed Central

    Ackermann, H W; Nguyen, T M

    1983-01-01

    Sewage was enriched with 35 Escherichia coli strains, and sediments of enrichment cultures were studied in the electron microscope. They contained up to 10 varieties of morphologically different particles. T-even-type phages predominated in 14 samples. Thirteen phages were enriched, representing the families Myoviridae (seven), Styloviridae (two), Podoviridae (three), and Microviridae (one). Twelve of these corresponded to known enterobacterial phage species, namely, 121, K19, FC3-9, O1, 9266, T2, 16-19, kappa, beta 4, N4, T7, and phi X174. Cubic RNA phages and filamentous phages were not detected. Types 121 and 9266 have previously been observed only in Romania and South Africa. Identification by morphology is usually simple. Our investigative technique is qualitative and will not detect all phages present. Most enrichment strains are polyvalent, and electron microscopy is always required for phage identification. In a general way, electron microscopy seems to be the method of choice for investigation of phage geography and ecology. Images PMID:6847179

  15. Scanning Electron Microscopy of the Presbylarynx.

    PubMed

    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.

  16. Scanning electron microscopy study of Tritrichomonas augusta.

    PubMed

    Borges, Fernanda P; Wiltuschnig, Renata C M; Tasca, Tiana; De Carli, Geraldo A

    2004-09-01

    Tritrichomonas augusta is a flagellated protozoan that parasitizes amphibians and reptiles. According to scanning electron microscopy (SEM), the cell shape of T. augusta varies from slender pyriform to ovoidal. Our data show the morphological features of the trophozoites: the emergence of the anterior flagella, the structure of the undulating membrane and the position and shape of the pelta, axostyle and posterior flagellum. In addition, herein we describe spherical forms which are probably pseudocysts. The description of the external structure of T. augusta, as demonstrated by SEM, contributes to the understanding of the biology of this parasite.

  17. Electron Microscopy of Young Candida albicans Chlamydospores

    PubMed Central

    Miller, Sara E.; Spurlock, Ben O.; Michaels, G. E.

    1974-01-01

    One- to three-day-old cultures of Candida albicans bearing chlamydospores were grown and harvested by a special technique, free of agar, and prepared for ultramicrotomy and electron microscopy. These young chlamydospores exhibited a subcellular structure similar to that of the yeast phase, e.g., cytoplasmic membrane, ribosomes, and mitochondria. Other structural characteristics unique to chlamydospores were a very thick, layered cell wall, the outer layer of which was continuous with the outer layer of the suspensor cell wall and was covered by hair-like projections; membrane bound organelles; and large lipoid inclusions. Only young chlamydospores less than 3 to 4 days old exhibited these ultrastructural characteristics. Images PMID:4368664

  18. Feature Adaptive Sampling for Scanning Electron Microscopy

    PubMed Central

    Dahmen, Tim; Engstler, Michael; Pauly, Christoph; Trampert, Patrick; de Jonge, Niels; Mücklich, Frank; Slusallek, Philipp

    2016-01-01

    A new method for the image acquisition in scanning electron microscopy (SEM) was introduced. The method used adaptively increased pixel-dwell times to improve the signal-to-noise ratio (SNR) in areas of high detail. In areas of low detail, the electron dose was reduced on a per pixel basis, and a-posteriori image processing techniques were applied to remove the resulting noise. The technique was realized by scanning the sample twice. The first, quick scan used small pixel-dwell times to generate a first, noisy image using a low electron dose. This image was analyzed automatically, and a software algorithm generated a sparse pattern of regions of the image that require additional sampling. A second scan generated a sparse image of only these regions, but using a highly increased electron dose. By applying a selective low-pass filter and combining both datasets, a single image was generated. The resulting image exhibited a factor of ≈3 better SNR than an image acquired with uniform sampling on a Cartesian grid and the same total acquisition time. This result implies that the required electron dose (or acquisition time) for the adaptive scanning method is a factor of ten lower than for uniform scanning. PMID:27150131

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

    SciTech Connect

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

    2016-07-25

    Abstract

    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 400μm, 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 andin situchemical and electrochemical processes.

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

    PubMed

    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.

  1. Scanning electron microscopy studies of bacterial cultures

    NASA Astrophysics Data System (ADS)

    Swinger, Tracy; Blust, Brittni; Calabrese, Joseph; Tzolov, Marian

    2012-02-01

    Scanning electron microscopy is a powerful tool to study the morphology of bacteria. We have used conventional scanning electron microscope to follow the modification of the bacterial morphology over the course of the bacterial growth cycle. The bacteria were fixed in vapors of Glutaraldehyde and ruthenium oxide applied in sequence. A gold film of about 5 nm was deposited on top of the samples to avoid charging and to enhance the contrast. We have selected two types of bacteria Alcaligenes faecalis and Kocuria rhizophila. Their development was carefully monitored and samples were taken for imaging in equal time intervals during their cultivation. These studies are supporting our efforts to develop an optical method for identification of the Gram-type of bacterial cultures.

  2. Quantitative characterization of electron detectors for transmission electron microscopy

    PubMed Central

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

    2013-01-01

    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 F416 scintillator-based cameras. We compare the results from our new method with published curves. PMID:24189638

  3. Characterization of nanomaterials with transmission electron microscopy

    NASA Astrophysics Data System (ADS)

    Anjum, D. 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. High voltage electron microscopy of lunar samples

    NASA Technical Reports Server (NTRS)

    Fernandez-Moran, H.

    1973-01-01

    Lunar pyroxenes from Apollo 11, 12, 14, and 15 were investigated. The iron-rich and magnesium-rich pyroxene specimens were crushed to a grain size of ca. 50 microns and studied by a combination of X-ray and electron diffraction, electron microscopy, 57 Fe Mossbauer spectroscopy and X-ray crystallography techniques. Highly ordered, uniform electron-dense bands, corresponding to exsolution lamellae, with average widths of ca. 230A to 1000A dependent on the source specimen were observed. These were?qr separated by wider, less-dense interband spacings with average widths of ca. 330A to 3100A. In heating experiments, splitting of the dense bands into finer structures, leading finally to obliteration of the exsolution lamellae was recorded. The extensive exsolution is evidence for significantly slower cooling rates, or possibly annealing, at temperatures in the subsolidus range, adding evidence that annealing of rock from the surface of the moon took place at ca. 600 C. Correlation of the band structure with magnetic ordering at low temperatures and iron clustering within the bands was studied.

  5. Improved methods for high resolution electron microscopy

    SciTech Connect

    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.

  6. Improved methods for high resolution electron microscopy

    NASA Astrophysics Data System (ADS)

    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 C44H90 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.

  7. Electric fields in Scanning Electron Microscopy simulations

    NASA Astrophysics Data System (ADS)

    Arat, K. T.; Bolten, J.; Klimpel, T.; Unal, N.

    2016-03-01

    The electric field distribution and charging effects in Scanning Electron Microscopy (SEM) were studied by extending a Monte-Carlo based SEM simulator by a fast and accurate multigrid (MG) based 3D electric field solver. The main focus is on enabling short simulation times with maintaining sufficient accuracy, so that SEM simulation can be used in practical applications. The implementation demonstrates a gain in computation speed, when compared to a Gauss-Seidel based reference solver is roughly factor of 40, with negligible differences in the result (~10-6 𝑉). In addition, the simulations were compared with experimental SEM measurements using also complex 3D sample, showing that i) the modelling of e-fields improves the simulation accuracy, and ii) multigrid method provide a significant benefit in terms of simulation time.

  8. Scanning electron microscopy of tinea nigra.

    PubMed

    Guarenti, Isabelle Maffei; Almeida, Hiram Larangeira de; Leitão, Aline Hatzenberger; Rocha, Nara Moreira; Silva, Ricardo Marques E

    2014-01-01

    Tinea nigra is a rare superficial mycosis caused by Hortaea werneckii. This infection presents as asymptomatic brown to black maculae mostly in palmo-plantar regions. We performed scanning electron microscopy of a superficial shaving of a tinea nigra lesion. The examination of the outer surface of the sample showed the epidermis with corneocytes and hyphae and elimination of fungal filaments. The inner surface of the sample showed important aggregation of hyphae among keratinocytes, which formed small fungal colonies. The ultrastructural findings correlated with those of dermoscopic examination - the small fungal aggregations may be the dark spicules seen on dermoscopy - and also allowed to document the mode of dissemination of tinea nigra, showing how hyphae are eliminated on the surface of the lesion.

  9. ELECTRON MICROSCOPY OF PLASMOLYSIS IN ESCHERICHIA COLI.

    PubMed

    COTA-ROBLES, E H

    1963-03-01

    Cota-Robles, Eugene H. (University of California, Riverside). Electron microscopy of plasmolysis in Escherichia coli. J. Bacteriol. 85:499-503. 1963.-Escherichia coli cells plasmolyzed in 0.35 m sucrose reveal plasmolysis at one tip of a cell or in the center of dividing cells in which protoplast partition has been complete. Central plasmolysis reveals that protoplast separation can be completed before the invagination of the cell wall is complete. These studies support the concept that these cells divide by constriction. The strength of the union between cell wall and cytoplasm is not uniform around the entire cell. It is strongest along the sides of these rod-shaped cells and weakest at one tip of the single cell. Thus, a single cell generally forms one cup-shaped vacuole in which the cytoplasm has collapsed away from one tip of the cell.

  10. ELECTRON MICROSCOPY OF PLASMOLYSIS IN ESCHERICHIA COLI

    PubMed Central

    Cota-Robles, Eugene H.

    1963-01-01

    Cota-Robles, Eugene H. (University of California, Riverside). Electron microscopy of plasmolysis in Escherichia coli. J. Bacteriol. 85:499–503. 1963.—Escherichia coli cells plasmolyzed in 0.35 m sucrose reveal plasmolysis at one tip of a cell or in the center of dividing cells in which protoplast partition has been complete. Central plasmolysis reveals that protoplast separation can be completed before the invagination of the cell wall is complete. These studies support the concept that these cells divide by constriction. The strength of the union between cell wall and cytoplasm is not uniform around the entire cell. It is strongest along the sides of these rod-shaped cells and weakest at one tip of the single cell. Thus, a single cell generally forms one cup-shaped vacuole in which the cytoplasm has collapsed away from one tip of the cell. Images PMID:14042923

  11. Scanning electron microscopy of molluscum contagiosum*

    PubMed Central

    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

  12. Hexamethyldisilazane for scanning electron microscopy of Gastrotricha.

    PubMed

    Hochberg, R; Litvaitis, M K

    2000-01-01

    We evaluated treatment with hexamethyldisilazane (HMDS) as an alternative to critical-point drying (CPD) for preparing microscopic Gastrotricha for scanning electron microscopy (SEM). We prepared large marine (2 mm) and small freshwater (100 microm) gastrotrichs using HMDS as the primary dehydration solvent and compared the results to earlier investigations using CPD. The results of HMDS dehydration are similar to or better than CPD for resolution of two important taxonomic features: cuticular ornamentation and patterns of ciliation. The body wall of both sculpted (Lepidodermella) and smooth (Dolichodasys) gastrotrichs retained excellent morphology as did the delicate sensory and locomotory cilia. The only unfavorable result of HMDS dehydration was an occasional coagulation of gold residue when the solvent had not fully evaporated before sputter-coating. We consider HMDS an effective alternative for preparing of gastrotrichs for SEM because it saves time and expense compared to CPD.

  13. Digital position determination system for electron microscopy.

    PubMed

    Hohmann-Marriott, Martin F; Sharp, William P; Roberson, Robert W; Blankenship, Robert E

    2005-06-01

    The precise determination of object positions within a specimen grid is important for many applications in electron microscopy. For example, real-time position determination is necessary for current statistical approaches and the efficient mapping and relocation of objects. Unfortunately, precise real-time position determination is not available on many older electron microscopes with manual stage controls. This report demonstrates the cost-effective and flexible implementation of a digital position determination system that can be adapted to many hand-operated electron microscopes. A customized solution that includes the hardware and software to accomplish position determination is presented. Lists of required parts, instructions for building the hardware, and descriptions of the developed programs are included. Two LED-photodiode assemblies detect x and y movements via an optical wheel that is in physical contact with the mechanical x and y stage control elements. These detector assemblies are interfaced with an integrated circuit that converts movement information into serial port-compatible signals, which are interpreted by a computer with specialized software. Two electron microscopes, a Philips CM12 (S)TEM and a Philips 201 TEM, were equipped with the described digital position determination system. The position fidelity and position fidelity after reloading of grids were determined for both microscopes. The determined position deviation was 1.06 microm in the x axis and 0.565 microm in the y axis for the Philips CM12 (S)TEM, and 0.303 microm in the x axis and 0.545 microm in the y axis for the Philips 201 TEM. After reloading and computational realigning, the determined average position variation was 2.66 microm in the x axis and 2.61 microm in the y axis for the Philips CM12 (S)TEM, and 1.13 microm in the x axis and 1.27 microm in the y axis for the Philips 201 TEM.

  14. Ultrafast electron microscopy integrated with a direct electron detection camera

    PubMed Central

    Lee, Young Min; Kim, Young Jae; Kim, Ye-Jin; Kwon, Oh-Hoon

    2017-01-01

    In the past decade, we have witnessed the rapid growth of the field of ultrafast electron microscopy (UEM), which provides intuitive means to watch atomic and molecular motions of matter. Yet, because of the limited current of the pulsed electron beam resulting from space-charge effects, observations have been mainly made to periodic motions of the crystalline structure of hundreds of nanometers or higher by stroboscopic imaging at high repetition rates. Here, we develop an advanced UEM with robust capabilities for circumventing the present limitations by integrating a direct electron detection camera for the first time which allows for imaging at low repetition rates. This approach is expected to promote UEM to a more powerful platform to visualize molecular and collective motions and dissect fundamental physical, chemical, and materials phenomena in space and time. PMID:28529964

  15. Electronic cameras for low-light microscopy.

    PubMed

    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.

  16. Electron microscopy: Phase transition singled out

    SciTech Connect

    Browning, Nigel D.

    2013-04-23

    One of the fundamental challenges within nanotechnology is to understand and control how nanoscale properties are initiated, evolve, and eventually terminate as a system moves from an individual nanostructure towards the meso- and macro-scale ensembles that are used in most applications. The ability to directly observe individual nanostructures and characterize their structure and composition has long been within the purview of transmission electron microscopy (TEM). The almost ubiquitous application of spherical aberration correction in TEM and in scanning-TEM (STEM) that has occurred over the last 10 years, now means it is possible to routinely characterize such nanostructures with both atomic resolution and sensitivity [1,2]. The development of temporal resolution in the TEM and the ability to study fast dynamics, on the other hand, has only recently come to the forefront of instrumentation development and is currently defined by two different approaches in the use of photoemission sources: the single shot s-ns dynamic TEM (DTEM) [3] and the stroboscopic ps-fs 4-D EM [4]. In the case of the DTEM, the goal is to observe the longer timescale irreversible structural changes that occur during nucleation and growth phenomena (here the single shot approach means there are enough electrons in a single pulsed beam to form a complete image). The 4-D EM focuses on a stroboscopic approach with the goal of studying very rapid reversible effects that occur during phase transitions (here an image is composed of thousands of pump-probe events each occurring with exactly the same time signature, with an individual pulse containing only a few electrons).

  17. The role of electron microscopy for the diagnosis of glomerulopathies.

    PubMed

    Sementilli, Angelo; Moura, Luiz Antonio; Franco, Marcello Fabiano

    2004-05-06

    Electron microscopy has been used for the morphological diagnosis of glomerular diseases for more than three decades and its value has been widely emphasized. However, recent reports have analyzed the routine use of electron microscopy critically. Its use in other areas of diagnosis such as tumor diseases has declined considerably; in addition, in view of the unavoidable financial pressure for the reduction of costs due to investigations and diagnostic routines, the selection of cases for electron microscopy has been quite rigorous. To identify the glomerular diseases that depend on electron microscopy for a final diagnosis, by means of reviewing renal biopsies performed over a 12-year period. Prospective Hospital Ana Costa, Hospital Guilherme Alvaro and Serviço de Anatomia Patológica de Santos, Santos, São Paulo, Brazil. 200 consecutive renal biopsies obtained from private hospitals and the teaching hospital from 1979 to 1991 were studied. All cases were analyzed via light microscopy, immunofluorescence and electron microscopy. The diagnosis was first made via light microscopy plus immunofluorescence and then via electron microscopy. Electron microscopy was diagnostic or essential for diagnosis in 10.0% of the cases, corresponding to 3.4% of primary glomerulopathies and 100% of hereditary glomerulopathies. Electron microscopy was contributory (useful) to the diagnosis in 5.5% of the cases, confirming the preliminary diagnosis formulated on the basis of clinical and laboratory data and light microscopy plus immunofluorescence findings. We obtained a 7.5% rate of discordant immunofluorescence, which was considered as such when negative immunofluorescence findings were not confirmed by electron microscopy. The final diagnosis with the use of light microscopy plus immunofluorescence alone was 77.0%. It was possible to diagnose with certainty a great percentage of glomerulopathies (82.5-90% of the cases) based on the light microscopy and immunofluorescence findings

  18. Advanced electron microscopy characterization of multimetallic nanoparticles

    NASA Astrophysics Data System (ADS)

    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

  19. Metal particles in a ceramic matrix--scanning electron microscopy and transmission electron microscopy characterization.

    PubMed

    Konopka, K

    2006-09-01

    This paper is concerned with ceramic matrix (Al(2)O(3)) composites with introduced metal particles (Ni, Fe). The composites were obtained via sintering of powders under very high pressure (2.5 GPa). Scanning electron microscopy and transmission electron microscopy were chosen as the tools for the identification and description of the shape, size and distribution of the metal particles. The Al(2)O(3)-Ni composite contained agglomerates of the Ni particles surrounded by ceramic grains and nanometre-size Ni particles located inside the ceramic grains and at the ceramic grain boundaries. In the Al(2)O(3)-Fe composite, the Fe particles were mostly surrounded by ceramic grains. Moreover, holes left by the Fe particles were found. The high pressure used in the fabrication of the composites changed the shape of the metal and ceramic powder grains via plastic deformation.

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

    PubMed

    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.

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

    PubMed

    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.

  2. Simplifying Electron Beam Channeling in Scanning Transmission Electron Microscopy (STEM).

    PubMed

    Wu, Ryan J; Mittal, Anudha; Odlyzko, Michael L; Mkhoyan, K Andre

    2017-08-01

    Sub-angstrom scanning transmission electron microscopy (STEM) allows quantitative column-by-column analysis of crystalline specimens via annular dark-field images. The intensity of electrons scattered from a particular location in an atomic column depends on the intensity of the electron probe at that location. Electron beam channeling causes oscillations in the STEM probe intensity during specimen propagation, which leads to differences in the beam intensity incident at different depths. Understanding the parameters that control this complex behavior is critical for interpreting experimental STEM results. In this work, theoretical analysis of the STEM probe intensity reveals that intensity oscillations during specimen propagation are regulated by changes in the beam's angular distribution. Three distinct regimes of channeling behavior are observed: the high-atomic-number (Z) regime, in which atomic scattering leads to significant angular redistribution of the beam; the low-Z regime, in which the probe's initial angular distribution controls intensity oscillations; and the intermediate-Z regime, in which the behavior is mixed. These contrasting regimes are shown to exist for a wide range of probe parameters. These results provide a new understanding of the occurrence and consequences of channeling phenomena and conditions under which their influence is strengthened or weakened by characteristics of the electron probe and sample.

  3. Analytical electron microscopy of thin films

    NASA Astrophysics Data System (ADS)

    Malac, Marek

    An analytical transmission electron microscope (ATEM) yields an impressive amount of information from a single instrument. The chemical composition of small areas of a sample is often obtained by energy dispersive x-ray microanalysis (EDX). EDX is routinely used both in research and industry to obtain fractions of heavier elements (Z > 11). To allow quantitative EDX analysis of samples containing light elements (B, C, N, O, F, Mg and Si) we developed, fabricated and characterized a set of three calibration samples. These calibration specimens allow users to obtain experimental Cliff-Lorimer factors with 10% to 15% accuracy and are sufficiently stable during storage, as well as under electron beam irradiation. Quantitative electron energy-loss spectroscopy (EELS) was employed to characterize these samples. The good light-element sensitivity of EELS makes it a suitable method for chemical analysis of biological samples in ATEM. It is desirable to probe the detection limits of EELS and energy filtering transmission electron microscopy (EFTEM) as well as determine what physical processes underlying these limits. We find that a TEM/EELS system is capable quantifying of 2000 ppm of boron with about 10% accuracy and 1 mum resolution. EFTEM mapping using Gatan Image filter is capable of mapping 5000 ppm of boron with 66 nm pixel size. The minimum detectable fraction (MDF) was limited by detector gain-variations and beam-shot noise. Spatial (EFTEM or TEM/EELS) mapping of low boron concentrations is important for boron-neutron capture therapy (BNCT), a method of cancer treatment. The high spatial resolution of TEM imaging and chemical analysis was applied to study microscopic mechanism of growth of thin films deposited onto oblique (rotating) substrate. The structure of these films can vary between arrays of columns (stationary substrate), helices (slowly-rotated substrate) or pillars (fast-rotated substrate). These structures (columns, pillars, helices) are composed of many

  4. Electron microscopy: Ultrastable gold substrates for electron cryomicroscopy.

    PubMed

    Russo, Christopher J; Passmore, Lori A

    2014-12-12

    Despite recent advances, the structures of many proteins cannot be determined by electron cryomicroscopy because the individual proteins move during irradiation. This blurs the images so that they cannot be aligned with each other to calculate a three-dimensional density. Much of this movement stems from instabilities in the carbon substrates used to support frozen samples in the microscope. Here we demonstrate a gold specimen support that nearly eliminates substrate motion during irradiation. This increases the subnanometer image contrast such that α helices of individual proteins are resolved. With this improvement, we determine the structure of apoferritin, a smooth octahedral shell of α-helical subunits that is particularly difficult to solve by electron microscopy. This advance in substrate design will enable the solution of currently intractable protein structures.

  5. [Scanning electron microscopy of Paragonimus proliferus].

    PubMed

    Zhou, Ben-jiang

    2005-10-30

    To identify the species of Paragonimus proliferus with scanning electron microscopy (SEM) based on the surface structure of excysted metacercariae, adult worms and eggs. Crabs were collected from the endemic area of P. proliferus and excysted metacercariae were separated. Adult worms at different ages and eggs were obtained from the experimentally infected rats. After being fixed by 2.5% glutardialdehyde and 1% osmic acid, alcohol dehydration, gilded by ion spatter, the specimens were observed under SEM by STEREOSCAN-100. The cuticular spines of excysted metacercariae distributed in single pattern, bayonet-shaped or scale-shaped. There were 6 dome-shape papillae around the rim of the ventral sucker symmetrically arranged. The cuticular spines of different age adult worms distributed in group pattern, relatively denser and more regularly arranged in the anterior part than the posterior part of the worm body. The shape and arrangement of the cuticular spines on adult worms at different ages were basically uniform. The surface of eggshell including the operculum was generally smooth. The shell rim joining the operculum was thick and prominent. A knot-like prominence was observed at the aboperculum end. The cuticular spines of both excysted metacercariae and adult worms of P. proliferus show its own characteristics, but the size and shape of the cuticular spines among individuals or different parts of the same specimen show certain differences.

  6. Imaging Cytoskeleton Components by Electron Microscopy

    PubMed Central

    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

  7. Electron microscopy in the investigation of asthenozoospermia.

    PubMed

    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.

  8. Microwave energy fixation for electron microscopy.

    PubMed Central

    Login, G. R.; Dvorak, A. M.

    1985-01-01

    We have demonstrated that microwave energy (MW) can be used in conjunction with chemical cross-linking agents in order to rapidly fix cell suspensions and tissue blocks for electron microscopy in 7-9 seconds. The optimal MW fixation method involved immersing tissues up to 1 cu cm in dilute aldehyde fixation and immediately irradiating the specimens in a conventional microwave oven for 9 seconds to 50 C. Ultrastructural preservation of samples irradiated by MW energy was comparable to that of the control samples immersed in aldehyde fixative for 2 hours at 25 C. Stereologic analysis showed that tissue blocks fixed by the MW fixation method did not cause organelles such as liver mitochondria and salivary gland granules to shrink or to swell. Potential applications for this new fixation technology include the investigation of rapid intracellular processes (eg, vesicular transport) and preservation of proteins that are difficult to demonstrate with routine fixation methods (eg, antigens and enzymes). Images Figure 4 Figure 5 Figure 2 Figure 3 Figure 6 Figure 7 Figure 8 Figure 9 Figure 10 Figure 11 PMID:3927740

  9. Imaging Cytoskeleton Components by Electron Microscopy.

    PubMed

    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.

  10. Electron microscopy and theoretical modeling of cochleates.

    PubMed

    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.

  11. Scanning electron microscopy of softened enamel.

    PubMed

    Eisenburger, M; Shellis, R P; Addy, M

    2004-01-01

    After exposing enamel specimens to 0.3% citric acid at pH 3.2 for various times, the acid was titrated to pH 7 before rinsing the specimens in water. After freeze-drying the specimens were examined by scanning electron microscopy. This procedure eliminates artefacts due to drying and mineral precipitation. The results showed that the outer region of softened enamel is much more delicate than previously thought, even after short (5- to 20-min) etching times. Mineral was lost from both prism boundaries and the prism bodies, resulting in a surface presenting thin, separate crystal bundles. In further studies, replicas of subsurface pores, created by resin impregnation, showed the softening depth to be several times greater than is suggested by techniques based on removing the softened enamel by physical forces. The results point to a need for improved methods of measuring softening depth. More importantly, it appears that the outer region of the softened layer remaining after an erosive challenge might be too fragile to resist frictional forces in vivo. Copyright 2004 S. Karger AG, Basel

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

    PubMed

    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.

  13. Probing Structural and Electronic Dynamics with Ultrafast Electron Microscopy

    SciTech Connect

    Plemmons, DA; Suri, PK; Flannigan, DJ

    2015-05-12

    In this Perspective, we provide an overview,of the field of ultrafast electron microscopy (UEM). We begin by briefly discussing the emergence of methods for probing ultrafast structural dynamics and the information that can be obtained. Distinctions are drawn between the two main types a probes for femtosecond (fs) dynamics fast electrons and X-ray photons and emphasis is placed on hour the nature of charged particles is exploited in ultrafast electron-based' experiments:. Following this, we describe the versatility enabled by the ease with which electron trajectories and velocities can be manipulated with transmission electron microscopy (TEM): hardware configurations, and we emphasize how this is translated to the ability to measure scattering intensities in real, reciprocal, and energy space from presurveyed and selected rianoscale volumes. Owing to decades of ongoing research and development into TEM instrumentation combined with advances in specimen holder technology, comprehensive experiments can be conducted on a wide range of materials in various phases via in situ methods. Next, we describe the basic operating concepts, of UEM, and we emphasize that its development has led to extension of several of the formidable capabilities of TEM into the fs domain, dins increasing the accessible temporal parameter spade by several orders of magnitude. We then divide UEM studies into those conducted in real (imaging), reciprocal (diffraction), and energy (spectroscopy) spate. We begin each of these sections by providing a brief description of the basic operating principles and the types of information that can be gathered followed by descriptions of how these approaches are applied in UM, the type of specimen parameter space that can be probed, and an example of the types of dynamics that can be resolved. We conclude with an Outlook section, wherein we share our perspective on some future directions of the field pertaining to continued instrument development and

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

    PubMed Central

    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

  15. 3D correlative light and electron microscopy of cultured cells using serial blockface scanning electron microscopy.

    PubMed

    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.

  16. Preparing Fission Yeast for Electron Microscopy.

    PubMed

    Giddings, Thomas H; Morphew, Mary K; McIntosh, J Richard

    2017-01-03

    Freezing samples while simultaneously subjecting them to a rapid increase in pressure, which inhibits ice crystal formation, is a reliable method for cryofixing fission yeast. The procedure consists simply of harvesting cells and loading them into a high-pressure freezer (HPF), and then operating the device. If equipment for high-pressure freezing is not available, fission yeast can be frozen by plunging a monolayer of cells into a liquid cryogen, usually ethane or propane. Unlike the HPF, where relatively large volumes of cells can be frozen in a single run, plunge freezing requires cells to be dispersed in a layer <20 µm thick. Unless frozen cells are to be imaged in the vitreous state, they must be fixed, dehydrated, and embedded for subsequent study by transmission electron microscopy; warming frozen cells without fixation badly damages cell structure. Fixation is best accomplished by freeze-substitution, a process in which frozen water is removed from samples by a water-miscible solvent that is liquid at a temperature low enough to prevent the cellular water from recrystallizing. Low concentrations of chemical fixatives and stains are generally added to this solvent such that they permeate the cells as the water is replaced. The activity of these additives is quite limited at the low temperatures required for minimizing ice crystal formation, but they are in the right place to react effectively as the cells warm up. Step-by-step protocols for HPF, plunge freezing, and freeze-substitution are provided here. © 2017 Cold Spring Harbor Laboratory Press.

  17. THE ELECTRON MICROSCOPY OF THE CHOROID PLEXUS

    PubMed Central

    Maxwell, David S.; Pease, Daniel C.

    1956-01-01

    1. The choroid plexus of the rat has been studied in detail by electron microscopy. Samples from the frog, rabbit, and cat have also been examined without noting significant differences. 2. The surface of the ependymal epithelium is covered by pedicels of variable size. There is reason for thinking of these structures as labile. They may actually pinch off and contribute to the secretory product. In any case, the surface area is vastly increased by their presence. Polypoid border seems an apt term to apply to this type of surface. 3. There is also a great expansion of the basal surface of ependymal cells. In the vicinity of cell junctions this surface is deeply infolded, and continuous with elaborate interdigitations of the lateral intercellular surfaces. Analogous infolding of the basal cell surface is known to exist in other epithelia also noted for their water transport (kidney tubules, salivary gland, and ciliary body). 4. Pretreatment of rats with diamox, an agent known to block cerebro-spinal fluid production, did not produce an important morphological change in the features of the ependyma, or any other part of the choroid plexus. 5. Capillaries of the choroid plexus have a very attenuated endothelium. This is seen to be fenestrated. It is thought this probably represents the condition in life, and is not simply a fixation artefact. 6. Pial cells tend to interpose sheets of cytoplasm between the capillaries and ependyma. The sheets are not continuous, however, and so would not constitute a serious diffusion barrier. These cells belong to the reticuloendothelial system, and undergo shape changes, and probably increase in number, when the system is stimulated by the repeated injection of trypan blue. PMID:13357511

  18. Ballistic-electron-emission Microscopy of Semiconductor Heterostructures

    NASA Technical Reports Server (NTRS)

    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.

  19. Ballistic-electron-emission Microscopy of Semiconductor Heterostructures

    NASA Technical Reports Server (NTRS)

    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.

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

    PubMed

    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.

  1. [Environmental scanning electron microscopy for biofilm detection in tonsils].

    PubMed

    Ramírez-Camacho, Rafael; González-Tallón, Ana Isabel; Gómez, David; Trinidad, Almudena; Ibáñez, Andrés; García-Berrocal, José Ramón; Verdaguer, José María; González-García, José Angel; San Román, Julio

    2008-01-01

    To describe an environmental scanning electron microscopic method for the study of biofilms in clinical samples. A comparison with standard scanning electron microscopy is performed. Nine patients with a past history of recurrent tonsillitis underwent tonsillectomy. Samples from each patient were obtained for both conventional and environmental scanning electron microscopy. The tonsils removed from 2 patients with sleep apnoea syndrome were used as controls. Eight of nine tonsils had biofilms on their surface. Scanning electron microscopy showed accumulations of bacteria covered by fibrillar structures resulting from the sample dehydration process. Environmental scanning electron microscopy provided a view of bacteria embedded in a homogeneous, amorphous substance that was preserved during the examination. Environmental scanning electron microscopy permits the imaging of wet systems at different degrees of dehydration. It therefore allows researchers to observe biofilms in their natural hydrated state.

  2. Fully Hydrated Yeast Cells Imaged with Electron Microscopy

    PubMed Central

    Peckys, Diana B.; Mazur, Peter; Gould, Kathleen L.; de Jonge, Niels

    2011-01-01

    We demonstrate electron microscopy of fully hydrated eukaryotic cells with nanometer resolution. Living Schizosaccaromyces pombe cells were loaded in a microfluidic chamber and imaged in liquid with scanning transmission electron microscopy (STEM). The native intracellular (ultra)structures of wild-type cells and three different mutants were studied without prior labeling, fixation, or staining. The STEM images revealed various intracellular components that were identified on the basis of their shape, size, location, and mass density. The maximal achieved spatial resolution in this initial study was 32 ± 8 nm, an order of magnitude better than achievable with light microscopy on pristine cells. Light-microscopy images of the same samples were correlated with the corresponding electron-microscopy images. Achieving synergy between the capabilities of light and electron microscopy, we anticipate that liquid STEM will be broadly applied to explore the ultrastructure of live cells. PMID:21575587

  3. Fully hydrated yeast cells imaged with electron microscopy.

    PubMed

    Peckys, Diana B; Mazur, Peter; Gould, Kathleen L; de Jonge, Niels

    2011-05-18

    We demonstrate electron microscopy of fully hydrated eukaryotic cells with nanometer resolution. Living Schizosaccharomyces pombe cells were loaded in a microfluidic chamber and imaged in liquid with scanning transmission electron microscopy (STEM). The native intracellular (ultra)structures of wild-type cells and three different mutants were studied without prior labeling, fixation, or staining. The STEM images revealed various intracellular components that were identified on the basis of their shape, size, location, and mass density. The maximal achieved spatial resolution in this initial study was 32 ± 8 nm, an order of magnitude better than achievable with light microscopy on pristine cells. Light-microscopy images of the same samples were correlated with the corresponding electron-microscopy images. Achieving synergy between the capabilities of light and electron microscopy, we anticipate that liquid STEM will be broadly applied to explore the ultrastructure of live cells. Copyright © 2011 Biophysical Society. Published by Elsevier Inc. All rights reserved.

  4. Electron Microscopy for Rapid Diagnosis of Emerging Infectious Agents1

    PubMed Central

    Gelderblom, Hans R.

    2003-01-01

    Diagnostic electron microscopy has two advantages over enzyme-linked immunosorbent assay and nucleic acid amplification tests. After a simple and fast negative stain preparation, the undirected, “open view” of electron microscopy allows rapid morphologic identification and differential diagnosis of different agents contained in the specimen. Details for efficient sample collection, preparation, and particle enrichment are given. Applications of diagnostic electron microscopy in clinically or epidemiologically critical situations as well as in bioterrorist events are discussed. Electron microscopy can be applied to many body samples and can also hasten routine cell culture diagnosis. To exploit the potential of diagnostic electron microscopy fully, it should be quality controlled, applied as a frontline method, and be coordinated and run in parallel with other diagnostic techniques. PMID:12643823

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

    NASA Astrophysics Data System (ADS)

    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

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

    PubMed

    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.

  7. Recent developments and applications of electron microscopy to heterogeneous catalysis.

    PubMed

    Yang, Judith C; Small, Matthew W; Grieshaber, Ross V; Nuzzo, Ralph G

    2012-12-21

    Transmission electron microscopy (TEM) and scanning transmission electron microscopy (STEM) are popular and powerful techniques used to characterize heterogeneous catalysts. Rapid developments in electron microscopy--especially aberration correctors and in situ methods--permit remarkable capabilities for visualizing both morphologies and atomic and electronic structures. The purpose of this review is to summarize the significant developments and achievements in this field with particular emphasis on the characterization of catalysts. We also highlight the potential and limitations of the various methods, describe the need for synergistic and complementary tools when characterizing heterogeneous catalysts, and conclude with an outlook that also envisions future needs in the field.

  8. Correlative light and volume electron microscopy: using focused ion beam scanning electron microscopy to image transient events in model organisms.

    PubMed

    Bushby, Andrew J; Mariggi, Giovanni; Armer, Hannah E J; Collinson, Lucy M

    2012-01-01

    The study of a biological event within a live model organism has become routine through the use of fluorescent labeling of specific proteins in conjunction with laser confocal imaging. These methods allow 3D visualization of temporal events that can elucidate biological function but cannot resolve the tissue organization, extracellular and subcellular details of the tissues. Here, we present a method for correlating electron microscopy image data with the light microscopy data from the same sample volume to reveal the 3D structural information: "correlative light and volume electron microscopy." The methods for live video confocal microscopy, fixation and embedding of the tissue for electron microscopy, the focused ion beam scanning electron microscopy method for sequentially slicing and imaging the volume of interest, and the treatment of the resulting 3D dataset are presented. The method is illustrated with data collected during the angiogenesis of blood vessels in a transgenic zebrafish embryo. Copyright © 2012 Elsevier Inc. All rights reserved.

  9. Graphene-enabled electron microscopy and correlated super-resolution microscopy of wet cells.

    PubMed

    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.

  10. Graphene-enabled electron microscopy and correlated super-resolution microscopy of wet cells

    PubMed Central

    Wojcik, Michal; Hauser, Margaret; Li, Wan; Moon, Seonah; Xu, Ke

    2015-01-01

    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. PMID:26066680

  11. Silicon Nitride Windows for Electron Microscopy of Whole Cells

    PubMed Central

    Ring, E. A.; Peckys, D. B.; Dukes, M. J.; Baudoin, J. P.; de Jonge, N.

    2012-01-01

    Summary Silicon microchips with thin electron transparent silicon nitride windows provide a sample support that accommodates both light-, and electron microscopy of whole eukaryotic cells in vacuum or liquid, with minimum sample preparation steps. The windows are robust enough that cellular samples can be cultured directly onto them, with no addition of a supporting film, and no need to embed or section the sample, as is typically required in electron microscopy. By combining two microchips, a microfluidic chamber can be constructed for the imaging of samples in liquid in the electron microscope. We provide microchip design specifications, a fabrication outline, instructions on how to prepare them for biological samples, and examples of images obtained using different light-, and electron microscopy modalities. The use of these microchips is particularly advantageous for correlative light-, and electron microscopy. PMID:21770941

  12. Correlative video-light-electron microscopy: development, impact and perspectives.

    PubMed

    Rizzo, Riccardo; Parashuraman, Seetharaman; Luini, Alberto

    2014-08-01

    Green fluorescent protein (GFP)-based video microscopy can provide profound insight into biological processes by generating information on the 'history,' or dynamics, of the cellular structures involved in such processes in live cells. A crucial limitation of this approach, however, is that many such structures may not be resolved by light microscopy. Like more recent super-resolution techniques, correlative video-light-electron microscopy (CLEM) was developed to overcome this limitation. CLEM integrates GFP-based video microscopy and electron microscopy through a series of ancillary techniques, such as proper fixation, hybrid labeling and retracing, and so provides sufficient resolution as well as, crucially, cellular 'context' to the fluorescent dynamic structures of interest. CLEM 'multiplies' the power of video microscopy and is having an important impact in several areas cell and developmental biology. Here, we discuss potential, limitations and perspectives of correlative approaches aimed at integrating the unique insight generated by video microscopy with information from other forms of imaging.

  13. Grain size quantification by optical microscopy, electron backscatter diffraction, and magnetic force microscopy.

    PubMed

    Chen, Hansheng; Yao, Yin; Warner, Jacob A; Qu, Jiangtao; Yun, Fan; Ye, Zhixiao; Ringer, Simon P; Zheng, Rongkun

    2017-06-13

    Quantification of microstructure, especially grain size, in polycrystalline materials is a vital aspect to understand the structure-property relationships in these materials. In this paper, representative characterization techniques for determining the grain size, including optical microscopy (OM), electron backscatter diffraction (EBSD) in the scanning electron microscopy (SEM), and atomic force microscopy/magnetic force microscopy (AFM/MFM), are thoroughly evaluated in comparison, illustrated by rare-earth sintered Nd-Fe-B permanent magnets. Potential applications and additional information achieved by using aforementioned characterization techniques have been discussed and summarized. Copyright © 2017 Elsevier Ltd. All rights reserved.

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

    PubMed

    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. Correlative Light Electron Microscopy: Connecting Synaptic Structure and Function

    PubMed Central

    Begemann, Isabell; Galic, Milos

    2016-01-01

    Many core paradigms of contemporary neuroscience are based on information obtained by electron or light microscopy. Intriguingly, these two imaging techniques are often viewed as complementary, yet separate entities. Recent technological advancements in microscopy techniques, labeling tools, and fixation or preparation procedures have fueled the development of a series of hybrid approaches that allow correlating functional fluorescence microscopy data and ultrastructural information from electron micrographs from a singular biological event. As correlative light electron microscopy (CLEM) approaches become increasingly accessible, long-standing neurobiological questions regarding structure-function relation are being revisited. In this review, we will survey what developments in electron and light microscopy have spurred the advent of correlative approaches, highlight the most relevant CLEM techniques that are currently available, and discuss its potential and limitations with respect to neuronal and synapse-specific applications. PMID:27601992

  16. Ion-induced electron emission microscopy

    DOEpatents

    Doyle, Barney L.; Vizkelethy, Gyorgy; Weller, Robert A.

    2001-01-01

    An ion beam analysis system that creates multidimensional maps of the effects of high energy ions from an unfocussed source upon a sample by correlating the exact entry point of an ion into a sample by projection imaging of the secondary electrons emitted at that point with a signal from a detector that measures the interaction of that ion within the sample. The emitted secondary electrons are collected in a strong electric field perpendicular to the sample surface and (optionally) projected and refocused by the electron lenses found in a photon emission electron microscope, amplified by microchannel plates and then their exact position is sensed by a very sensitive X Y position detector. Position signals from this secondary electron detector are then correlated in time with nuclear, atomic or electrical effects, including the malfunction of digital circuits, detected within the sample that were caused by the individual ion that created these secondary electrons in the fit place.

  17. Image Resolution in Scanning Transmission Electron Microscopy

    SciTech Connect

    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.

  18. Electron Microscopy of Ebola Virus-Infected Cells.

    PubMed

    Noda, Takeshi

    2017-01-01

    Ebola virus (EBOV) replicates in host cells, where both viral and cellular components show morphological changes during the process of viral replication from entry to budding. These steps in the replication cycle can be studied using electron microscopy (EM), including transmission electron microscopy (TEM) and scanning electron microscopy (SEM), which is one of the most useful methods for visualizing EBOV particles and EBOV-infected cells at the ultrastructural level. This chapter describes conventional methods for EM sample preparation of cultured cells infected with EBOV.

  19. Cryo-scanning electron microscopy and light microscopy for the study of fungi interactions.

    PubMed

    Sempere, F; Santamarina, M P

    2011-03-01

    The application of the cryo-scanning electron microscopy and light microscopy for the study of the interactions at different environmental conditions between Penicillium oxalicum and Fusarium verticillioides is described. A dual microculture was developed for the light microscopy analysis of the interaction. The microscope and macroscopic examinations were compared. Analysis of Petri plates revealed that F. verticillioides was a competitor for space and nutrients while P. oxalicum was a mycoparasite under the microscopic observations.

  20. Phase contrast in high resolution electron microscopy

    DOEpatents

    Rose, H.H.

    1975-09-23

    This patent relates to a device for developing a phase contrast signal for a scanning transmission electron microscope. The lens system of the microscope is operated in a condition of defocus so that predictable alternate concentric regions of high and low electron density exist in the cone of illumination. Two phase detectors are placed beneath the object inside the cone of illumination, with the first detector having the form of a zone plate, each of its rings covering alternate regions of either higher or lower electron density. The second detector is so configured that it covers the regions of electron density not covered by the first detector. Each detector measures the number of electrons incident thereon and the signal developed by the first detector is subtracted from the signal developed by the record detector to provide a phase contrast signal. (auth)

  1. Soft X-ray contact microscopy and transmission electron microscopy: Comparative study of biological samples

    NASA Astrophysics Data System (ADS)

    Limongi, T.; Palladino, L.; Bernieri, E.; Tomassetti, G.; Reale, L.; Flora, F.; Cesare, P.; Ercole, C.; Aimola, P.; Ragnelli, A. M.

    2003-03-01

    Isolated cellular organelles (mitochondria, chloroplasts) and cultured bacteria were analysed both by soft X-ray contact microscopy (SXCM), and by transmission electron microscopy (TEM) after negative staining. For each sample, a comparison was performed between images obtained with either technique, with the aim of facilitating the interpretation of SXCM images. The validity and the limits of this comparative approach are discussed.

  2. Structure of Wet Specimens in Electron Microscopy

    ERIC Educational Resources Information Center

    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)

  3. Structure of Wet Specimens in Electron Microscopy

    ERIC Educational Resources Information Center

    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)

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

    PubMed

    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. © 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  5. Nanowire growth kinetics in aberration corrected environmental transmission electron microscopy

    SciTech Connect

    Chou, Yi -Chia; Panciera, Federico; Reuter, Mark C.; Stach, Eric A.; Ross, Frances M.

    2016-03-15

    Here, we visualize atomic level dynamics during Si nanowire growth using aberration corrected environmental transmission electron microscopy, and compare with lower pressure results from ultra-high vacuum microscopy. We discuss the importance of higher pressure observations for understanding growth mechanisms and describe protocols to minimize effects of the higher pressure background gas.

  6. Nanowire growth kinetics in aberration corrected environmental transmission electron microscopy

    DOE PAGES

    Chou, Yi -Chia; Panciera, Federico; Reuter, Mark C.; ...

    2016-03-15

    Here, we visualize atomic level dynamics during Si nanowire growth using aberration corrected environmental transmission electron microscopy, and compare with lower pressure results from ultra-high vacuum microscopy. We discuss the importance of higher pressure observations for understanding growth mechanisms and describe protocols to minimize effects of the higher pressure background gas.

  7. Diagnostic applications of scanning electron microscopy and microanalysis in pathology.

    PubMed

    Abraham, J L

    1979-08-01

    Microanalytical technology developed within the last decade provides important information in diagnostic pathology. Scanning electron microscopy, including backscattered electron imaging and energy dispersive X-ray analysis should become at least as valuable as polarized light microscopy, histochemistry and conventional transmission electron microscopy. Other as yet less available techniques such as the ion microprobe and laser Raman microprobe are also valuable. The pathologist should consider the use of microanalytic techniques in any disease process in which endogenous or exogenous materials may be present in the tissues, in the same manner in which one would perform stains for microorganisms. Cases are presented illustrating the tissue preparation and results of scanning electron microscopy and energy dispersive X-ray analysis in diagnosis.

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

    SciTech Connect

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

    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.

  9. Entanglement-assisted electron microscopy based on a flux qubit

    SciTech Connect

    Okamoto, Hiroshi; Nagatani, Yukinori

    2014-02-10

    A notorious problem in high-resolution biological electron microscopy is radiation damage caused by probe electrons. Hence, acquisition of data with minimal number of electrons is of critical importance. Quantum approaches may represent the only way to improve the resolution in this context, but all proposed schemes to date demand delicate control of the electron beam in highly unconventional electron optics. Here we propose a scheme that involves a flux qubit based on a radio-frequency superconducting quantum interference device, inserted in a transmission electron microscope. The scheme significantly improves the prospect of realizing a quantum-enhanced electron microscope for radiation-sensitive specimens.

  10. Scanning Transmission Electron Microscopy at High Resolution

    PubMed Central

    Wall, J.; Langmore, J.; Isaacson, M.; Crewe, A. V.

    1974-01-01

    We have shown that a scanning transmission electron microscope with a high brightness field emission source is capable of obtaining better than 3 Å resolution using 30 to 40 keV electrons. Elastic dark field images of single atoms of uranium and mercury are shown which demonstrate this fact as determined by a modified Rayleigh criterion. Point-to-point micrograph resolution between 2.5 and 3.0 Å is found in dark field images of micro-crystallites of uranium and thorium compounds. Furthermore, adequate contrast is available to observe single atoms as light as silver. Images PMID:4521050

  11. Atmospheric pressure scanning transmission electron microscopy.

    PubMed

    de Jonge, Niels; Bigelow, Wilbur C; Veith, Gabriel M

    2010-03-10

    Scanning transmission electron microscope (STEM) images of gold nanoparticles at atmospheric pressure have been recorded through a 0.36 mm thick mixture of CO, O2, and He. This was accomplished using a reaction cell consisting of two electron-transparent silicon nitride membranes. Gold nanoparticles of a full width at half-maximum diameter of 1.0 nm were visible above the background noise, and the achieved edge resolution was 0.4 nm in accordance with calculations of the beam broadening.

  12. Data Management For Quantitative Electron Microscopy

    NASA Astrophysics Data System (ADS)

    Cavallari, Vittorio

    1986-05-01

    A set of computer-assisted procedures was developed for sampling and processing quantitative information from electron images. Data are manually collected using a digitizing tablet, and subsequently elaborated with a I B M- compatible personal computer. Programs presented here deals with stereological calculations, numerical taxonomy and basic statistics applied to tumor cell samples.

  13. Scanning Electron and Phase-Contrast Microscopy of Bacterial Spores

    PubMed Central

    Bulla, L. A.; Julian, G. St.; Rhodes, R. A.; Hesseltine, C. W.

    1969-01-01

    The three-dimensional immages of free and intrasporangial spores produced by scanning electron microscopy show surface structures not visible by phase-contrast microscopy. Although fine surface detail is not elucidated by scanning electron microscopy, this technique does afford a definitive picture of the general shape of spores. Spores of Bacillus popilliae, B. lentimorbus, B. thuringiensis, B. alvei, B. cereus, and Sarcina ureae have varying patterns of surface ridge formation, whereas spores of B. larvae, B. subtilis, and B. licheniformis have relatively smooth surfaces. Images PMID:4907010

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

    PubMed

    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.

  15. Electron Microscopy of Biological Materials at the Nanometer Scale

    NASA Astrophysics Data System (ADS)

    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.

  16. Preparation of Xenopus laevis retinal cryosections for electron microscopy.

    PubMed

    Tam, Beatrice M; Yang, Lee Ling; Bogėa, Tami H; Ross, Bradford; Martens, Garnet; Moritz, Orson L

    2015-07-01

    Transmission electron microscopy is the gold standard for examination of photoreceptor outer segment morphology and photoreceptor outer segment abnormalities in transgenic animal models of retinal disease. Small vertebrates such as zebrafish and Xenopus laevis tadpoles have been used to generate retinal disease models and to study outer segment processes such as protein trafficking, and their breeding capabilities facilitate experiments involving large numbers of animals and conditions. However, electron microscopy processing and analysis of these very small eyes can be challenging. Here we present a methodology that facilitates processing of X. laevis tadpole eyes for electron microscopy by introducing an intermediate cryosectioning step. This method reproducibly provides a well-oriented tissue block that can be sectioned with minimal effort by a non-expert, and also allows retroactive analysis of samples collected on slides for light microscopy.

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

    PubMed

    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.

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

    PubMed

    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.

  19. Quantitative analytical electron microscopy of multiphase alloys.

    PubMed

    Prybylowski, J; Ballinger, R; Elliott, C

    1989-02-01

    In this paper, we present a technique for analysis of composition gradients, using an analytical electron microscope, within the primary phase of a two-phase alloy for the case where the second-phase particle size is similar to the size of the irradiated volume. If the composition difference between the two phases is large, the detected compositional fluctuations associated with varying phase fractions may mask any underlying composition gradient of the primary phase. The analysis technique was used to determine grain boundary chromium concentration gradients in a nickel-base superalloy, alloy X-750. The technique may also be of use in other alloy systems.

  20. Preparation of nematodes for scanning electron microscopy.

    PubMed

    Green, C D; Stone, A R; Turner, R H; Clark, S A

    1975-01-01

    Nematodes from the orders Tlyenchida and Rhabditida were fixed and processed in several different ways for examination with the scanning electron microscope (SEM). Four processes produced good preparations of fixed nematodes. Drying from acetone was the simplest of these techniques and most useful for regions of the tylenchid nematodes supported by skeletal tissue. Critical point drying, a more complicated procedure, gave good preparations, but they required special care in processing. Nematodes infiltrated with glycerol and a conducting agent were the most life-like but were difficult to examine. Specimens infiltrated with an epoxy resin looked natural and this was the most promising process tried.

  1. Noninvasive electron microscopy with interaction-free quantum measurements

    SciTech Connect

    Putnam, William P.; Yanik, Mehmet Fatih

    2009-10-15

    We propose the use of interaction-free quantum measurements with electrons to eliminate sample damage in electron microscopy. This might allow noninvasive molecular-resolution imaging. We show the possibility of such measurements in the presence of experimentally measured quantum decoherence rates and using a scheme based on existing charged particle trapping techniques.

  2. Photon-induced near-field electron microscopy.

    PubMed

    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.

  3. Ultra-low voltage scanning electron microscopy

    SciTech Connect

    Joy, D.C.; Joy, C.S.

    1996-12-31

    An interesting new opportunity is to perform imaging in the ultra-low energy region between 1eV and 500eV. Over this energy range significant changes in the details of electron-solid interactions take place offering the chance of novel contrast modes, and the rapid fall in the electron beam range leads to the condition where the penetration of the incident beam into the sample is effectively limited to 1 or 2 nanometers. The practical problem is that of achieving useful levels of resolution and acceptable signal to noise ratios in the image. At energies below 1keV chromatic aberration dominates the probe formation in conventional instruments even when using an FEG source. However, the use of optimized retarding field optics essentially maintains chromatic aberration independent of landing energy down to very low values. Figure (1) shows an example of the performance that can be achieved on a commercial instrument - an Hitachi S-4500 - modified to operate in this mode, in this case at 50eV landing energy. The resolution of the image is judged from edge sharpness and detail to be significantly better than 0.1{mu}m and, from experimental observation, this performance is apparently limited by residual astigmatism caused by uncorrected sample charging rather than by fundamental aberrations in the probe forming optics. Comparable, if somewhat lower resolution, ages have been achieved on this, and other FEG SEMs, at energies as low as 1eV.

  4. Electron microscopy methods in studies of cultural heritage sites

    NASA Astrophysics Data System (ADS)

    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.

  5. A Correlative Optical Microscopy and Scanning Electron Microscopy Approach to Locating Nanoparticles in Brain Tumors

    PubMed Central

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

    2014-01-01

    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. PMID:25464144

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

    PubMed

    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.

  7. Scanning electron microscopy of bacteria Tetrasphaera duodecadis.

    PubMed

    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.

  8. Applications of cryogenics in electron microscopy

    NASA Technical Reports Server (NTRS)

    Fernandez-Moran, H.

    1973-01-01

    Description of research and development efforts which resulted in a high-voltage cryoelectron microscope system capable of consistent operation at 1.8 to 4.2 K. Attention is given to the design and operation of superconducting objective lenses providing enhanced resolution during longer exposure times at lower beam intensities (thus reducing radiation damage of specimens). A specific system described combines a closed-cycle superfluid helium refrigerator integrated with a modified 200 kV electron microscope. Consistent resolutions of 8 to 16 A are attained with significantly reduced radiation damage, contamination, and thermal noise in prolonged vibration-free examination of specimens at temperatures from 1.8 to 4.2 K. Applications in specific disciplines are discussed, including membrane ultrastructure, cryobiology, microelectronics, and general superconductivity research.

  9. A new epoxy embedment for electron microscopy.

    PubMed

    FREEMAN, J A; SPURLOCK, B O

    1962-06-01

    A new epoxy embedding mixture has been developed utilizing Maraglas 655 and Cardolite NC-513 with benzyldimethylamine (BDMA) as a curing agent. This epoxy mixture permits cellular preservation comparable to that obtained with Epon 812, ease of preparation of tissues, a wide range of miscibility, low viscosity, and, most important, ease of sectioning on a Porter-Blum microtome. In contrast to Epon-812-embedded tissues, Maraglas-Cardolite-embedded tissues can be sectioned in large dimensions with ease and consistent results without "chatter." No background granularity is detectable with high magnification study of Maraglas-Cardolite-embedded tissues. This epoxy is readily stained with lead hydroxide and is relatively stable in the electron beam.

  10. Concepts, facts and artifacts in electron microscopy.

    PubMed

    Sjöstrand, F S

    2005-12-16

    This communication illustrates how the electron microscope has contributed to biochemistry by revealing how multienzyme systems in mitochondria are structurally organized to secure high speed ATP synthesis and has extended physiology to the molecular level. Ribonucleoprotein complexes form a gel in the cytoplasm determining the conditions for translation... Photoreceptor stimulation involves two phases, trapping of light by a light reflecting cylinder formed by the outer segment disks and energy transduction by bleaching of photopigment molecules changing the charge of the outer segment disks driving the photoreceptor toward hyperpolarization. Revealing the synaptic connections between retinal neurons extends neurophysiology to the level of information processing by neural circuits, which are designed for high speed processing. Spatial brightness contrast enhancement is eliminated in connection with macular degeneration, which leads to partial blindness, revealing the importance of contrast enhancement for vision.

  11. A NEW EPOXY EMBEDMENT FOR ELECTRON MICROSCOPY

    PubMed Central

    Freeman, James A.; Spurlock, Ben O.

    1962-01-01

    A new epoxy embedding mixture has been developed utilizing Maraglas 655 and Cardolite NC-513 with benzyldimethylamine (BDMA) as a curing agent. This epoxy mixture permits cellular preservation comparable to that obtained with Epon 812, ease of preparation of tissues, a wide range of miscibility, low viscosity, and, most important, ease of sectioning on a Porter-Blum microtome. In contrast to Epon-812-embedded tissues, Maraglas-Cardolite-embedded tissues can be sectioned in large dimensions with ease and consistent results without "chatter." No background granularity is detectable with high magnification study of Maraglas-Cardolite-embedded tissues. This epoxy is readily stained with lead hydroxide and is relatively stable in the electron beam. PMID:13894888

  12. Electron microscopy of biomaterials based on hydroxyapatite

    SciTech Connect

    Suvorova, E. I. Klechkovskaya, V. V.; Komarov, V. F.; Severin, A. V.; Melikhov, I. V.; Buffat, P. A.

    2006-10-15

    Three types of biomaterials based on hydroxyapatite are synthesized and investigated. Hydroxyapatite nanocrystals or microcrystals precipitated from low-temperature aqueous solutions serve as the initial material used for preparing spherical porous granules approximately 300-500 {mu}m in diameter. Sintering of hydroxyapatite crystals at a temperature of 870 deg. C for 2 h or at 1000 deg. C (for 3 h) + 1200 deg. C (for 2 h) brings about the formation of solid ceramics with different internal structures. According to the electron microscopic data, the ceramic material prepared at 870 deg. C is formed by agglomerated hydroxyapatite nanocrystals, whereas the ceramics sintered at 1200 deg. C (with a bending strength of the order of 100 MPa) are composed of crystal blocks as large as 2 {mu}m. It is established that all the biomaterials have a single-phase composition and consist of the hydroxyapatite with a structure retained up to a temperature of 1200 deg. C.

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

    PubMed

    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.

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

    SciTech Connect

    Timmermans, F. J.; Otto, C.

    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.

  15. Ultra-high resolution electron microscopy.

    PubMed

    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.

  16. Ultra-high resolution electron microscopy

    NASA Astrophysics Data System (ADS)

    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.

  17. Ultra-high resolution electron microscopy

    DOE PAGES

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

    2016-12-23

    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. Here 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 tomore » 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.« less

  18. Ultra-high resolution electron microscopy

    SciTech Connect

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

    2016-12-23

    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. Here 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.

  19. Sad State of Phage Electron Microscopy. Please Shoot the Messenger

    PubMed Central

    Ackermann, Hans-W.

    2013-01-01

    Two hundred and sixty publications from 2007 to 2012 were classified according to the quality of electron micrographs; namely as good (71); mediocre (21); or poor (168). Publications were from 37 countries; appeared in 77 journals; and included micrographs produced with about 60 models of electron microscopes. The quality of the micrographs was not linked to any country; journal; or electron microscope. Main problems were poor contrast; positive staining; low magnification; and small image size. Unsharp images were frequent. Many phage descriptions were silent on virus purification; magnification control; even the type of electron microscope and stain used. The deterioration in phage electron microscopy can be attributed to the absence of working instructions and electron microscopy courses; incompetent authors and reviewers; and lenient journals. All these factors are able to cause a gradual lowering of standards. PMID:27694773

  20. Persistent misconceptions about incoherence in electron microscopy.

    PubMed

    Van Dyck, D

    2011-06-01

    Incoherence in electron microscopic imaging occurs when during the observation the microscope and the object are subject to fluctuations. In order to speed up the computer simulation of the images, approximations are used that are considered as valid. In this paper we will question the validity of these approximations and show that in specific cases they can lead to erroneous results. It is shown in particular in the case of one single vibrating atom that the thermal diffuse scattering that causes the signal in HAADF STEM is not only dependent on Z but also on the mean square displacement of the atom so that it can even be large for light atoms in soft matter, provided the right HAADF aperture is used. In HREM imaging the diffuse scattering leaks out of the coherent (elastic) wave and is redistributed in the background. This might explain the mismatch in elastic contrast (Stobbs factor) especially for crystals with a thickness beyond the extinction distance, where also the HAADF signal saturates and the elastic (coherent) component vanishes. Copyright © 2011 Elsevier B.V. All rights reserved.

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

    PubMed

    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.

  2. Photon gating in four-dimensional ultrafast electron microscopy

    PubMed Central

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

    2015-01-01

    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. PMID:26438835

  3. Transmission Electron Microscopy of Itokawa Regolith Grains

    NASA Technical Reports Server (NTRS)

    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

  4. Chromosome observation by scanning electron microscopy using ionic liquid.

    PubMed

    Dwiranti, Astari; Lin, Linyen; Mochizuki, Eiko; Kuwabata, Susumu; Takaoka, Akio; Uchiyama, Susumu; Fukui, Kiichi

    2012-08-01

    Electron microscopy has been used to visualize chromosome since it has high resolution and magnification. However, biological samples need to be dehydrated and coated with metal or carbon before observation. Ionic liquid is a class of ionic solvent that possesses advantageous properties of current interest in a variety of interdisciplinary areas of science. By using ionic liquid, biological samples need not be dehydrated or metal-coated, because ionic liquid behaves as the electronically conducting material for electron microscopy. The authors have investigated chromosome using ionic liquid in conjunction with electron microscopy and evaluated the factors that affect chromosome visualization. Experimental conditions used in the previous studies were further optimized. As a result, prewarmed, well-mixed, and low concentration (0.5∼1.0%) ionic liquid provides well-contrasted images, especially when the more hydrophilic and the higher purity ionic liquid is used. Image contrast and resolution are enhanced by the combination of ionic liquid and platinum blue staining, the use of an indium tin oxide membrane, osmium tetroxide-coated coverslip, or aluminum foil as substrate, and the adjustment of electron acceleration voltage. The authors conclude that the ionic-liquid method is useful for the visualization of chromosome by scanning electron microscopy without dehydration or metal coating.

  5. Focused ion beam scanning electron microscopy in biology.

    PubMed

    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.

  6. Evaluations of carbon nanotube field emitters for electron microscopy

    NASA Astrophysics Data System (ADS)

    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.

  7. Laboratory design for high-performance electron microscopy

    SciTech Connect

    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.

  8. Microscopy with slow electrons: from LEEM to XPEEM

    ScienceCinema

    Bauer, Ernst [Arizona State University, Phoenix, Arizona, United States

    2016-07-12

    The short penetration and escape depth of electrons with energies below 1 keV make them ideally suited for the study of surfaces and ultrathin films. The combination of the low energy electrons and the high lateral resolution of a microscope produces a powerful method for the characterization of nanostructures on bulk samples, in particular if the microscope is equipped with an imaging energy filter and connected to a synchrotron radiation source. Comprehensive characterization by imaging, diffraction, and spectroscope of the structural, chemical, and magnetic properties is then possible. The Talk will describe the various imaging techniques in using reflected and emitted electrons in low-energy electron microscopy (LEEM) and x-ray photoemission electron microscopy (XPEEM), with an emphasis on magnetic materials with spin-polarized LEEM and x-ray magnetic circular dichroism PEEM. The talk with end with an outlook on future possibilities.

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

    SciTech Connect

    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.

  10. Imaging Hydrated Microbial Extracellular Polymers: Comparative Analysis by Electron Microscopy

    SciTech Connect

    Dohnalkova, Alice; Marshall, Matthew J.; Arey, Bruce W.; Williams, Kenneth H.; Buck, Edgar C.; Fredrickson, Jim K.

    2011-02-01

    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 cryo-electron microscopy methods providing imaging in the close-to-natural hydrated state. In situ, we observed an irreversible transformation of bacterial extracellular polymers during the traditional dehydration-based sample preparation that resulted in the collapse of hydrated gel-like EPS 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.

  11. Atomic resolution imaging of graphene by transmission electron microscopy

    NASA Astrophysics Data System (ADS)

    Robertson, Alex W.; Warner, Jamie H.

    2013-05-01

    The atomic structure of a material influences its electronic, chemical, magnetic and mechanical properties. Characterising carbon nanomaterials, such as fullerenes, nanotubes and graphene, at the atomic level is challenging due to their chemical reactivity and low atomic mass. Transmission electron microscopy and scanning probe microscopy are two of the leading methods for imaging graphene at the atomic level. Here, we report on recent advances in atomic resolution imaging of graphene using aberration-corrected high resolution transmission electron microscopy and how it has revealed many of the structural deviations from the pristine monolayer form. Structures in graphene such as vacancy defects, edges, grain boundaries, linear chains, impurity dopants, layer number, layer stacking and bond rotations are explored.

  12. The CryoCapsule: Simplifying correlative light to electron microscopy

    PubMed Central

    Heiligenstein, Xavier; Heiligenstein, Jérôme; Delevoye, Cédric; Hurbain, Ilse; Bardin, Sabine; Paul-Gilloteaux, Perrine; Sengmanivong, Lucie; Régnier, Gilles; Salamero, Jean; Antony, Claude; Raposo, Graca

    2014-01-01

    Correlating complementary multiple scale images of the same object is a straightforward means to decipher biological processes. Light and electron microscopy are the most commonly used imaging techniques, yet despite their complementarity, the experimental procedures available to correlate them are technically complex. We designed and manufactured a new device adapted to many biological specimens, the CryoCapsule, that simplifies the multiple sample preparation steps, which at present separate live cell fluorescence imaging from contextual high-resolution electron microscopy, thus opening new strategies for full correlative light to electron microscopy. We tested the biological application of this highly optimized tool on three different specimens: the in-vitro Xenopus laevis mitotic spindle, melanoma cells over-expressing YFP-langerin sequestered in organized membranous subcellular organelles and a pigmented melanocytic cell in which the endosomal system was labeled with internalized fluorescent transferrin. PMID:24533564

  13. Electron microscopy studies of potassium sodium niobate ceramics.

    PubMed

    Jenko, Darja; Bencan, Andreja; Malic, Barbara; Holc, Janez; Kosec, Marija

    2005-12-01

    Using electron microscopy, K0.5Na0.5NbO3 (KNN) ceramics sintered at 1030 degrees C for 8 h and 1100 degrees C for 2 and 24 h was studied. The scanning electron microscopy and X-ray spectrometry revealed that the materials consisted of a matrix phase in which the (Na+K)/Nb ratio corresponded closely to the nominal composition and a small amount of Nb-rich secondary phase. A bimodal microstructure of cube-shaped grains was revealed in the fracture and thermally-etched surfaces of the KNN. In the ceramics sintered at 1100 degrees C, the larger grains (up to 30 mum across), contained angular trapped pores. The transmission electron microscopy analysis revealed that the crystal planes of the grains bordering the intragranular pore faces were of the {100} family with respect to the simple perovskite cell. Ferroelectric domains were observed in the grains of this material.

  14. Simultaneous Correlative Scanning Electron and High-NA Fluorescence Microscopy

    PubMed Central

    Liv, Nalan; Zonnevylle, A. Christiaan; Narvaez, Angela C.; Effting, Andries P. J.; Voorneveld, Philip W.; Lucas, Miriam S.; Hardwick, James C.; Wepf, Roger A.; Kruit, Pieter; Hoogenboom, Jacob P.

    2013-01-01

    Correlative light and electron microscopy (CLEM) is a unique method for investigating biological structure-function relations. With CLEM protein distributions visualized in fluorescence can be mapped onto the cellular ultrastructure measured with electron microscopy. Widespread application of correlative microscopy is hampered by elaborate experimental procedures related foremost to retrieving regions of interest in both modalities and/or compromises in integrated approaches. We present a novel approach to correlative microscopy, in which a high numerical aperture epi-fluorescence microscope and a scanning electron microscope illuminate the same area of a sample at the same time. This removes the need for retrieval of regions of interest leading to a drastic reduction of inspection times and the possibility for quantitative investigations of large areas and datasets with correlative microscopy. We demonstrate Simultaneous CLEM (SCLEM) analyzing cell-cell connections and membrane protrusions in whole uncoated colon adenocarcinoma cell line cells stained for actin and cortactin with AlexaFluor488. SCLEM imaging of coverglass-mounted tissue sections with both electron-dense and fluorescence staining is also shown. PMID:23409024

  15. Studying Arabidopsis chloroplast structural organisation using transmission electron microscopy.

    PubMed

    Hyman, Stefan; Jarvis, R Paul

    2011-01-01

    Chloroplasts, as well as other, non-photosynthetic types of plastid, are characteristic structures within plant cells. They are relatively large organelles (typically 1-5 μm in diameter), and so can readily be analysed by electron microscopy. Chloroplast structure is remarkably complex, comprising at least six distinct sub-organellar compartments, and is sensitive to developmental changes, environmental effects, and genetic lesions. Transmission electron microscopy (TEM), therefore, represents a powerful technique for monitoring the effects of various changing parameters or treatments on the development and differentiation of these important organelles. We describe a method for the analysis of Arabidopsis plant material by TEM, primarily for the assessment of plastid ultrastructure.

  16. Electron microscopy of myocardial tissue. A nine year review

    PubMed Central

    Mudhar, H; Wagner, B; Suvarna, S

    2001-01-01

    Aim—To review and reassess the role of this department's experience with routine electron microscopy of myocardial tissues. Methods—A nine year series of myocardial samples that underwent electron microscopy analysis were audited. Fifty nine samples were derived from 46 male and 13 female subjects with an age range of 15–90 years (mean, 50.6). Forty two samples were endomyocardial specimens, with 13 being derived from explanted hearts, and four from necropsies. Two cases were from transplanted hearts. These were all reviewed in a blinded fashion, by all three authors separately, in terms of the myocardium at the ultrastructural level. Subsequently, the interpretations/diagnoses were cross compared with the light microscopy and clinical data results. Results—Four cases of amyloid were identified; in addition, one case of granulomatous inflammation and one case of basophilic degeneration were seen, although all these had been evident on light microscopy. One case of possible mitochondrial myopathy was found. A total of 18 cases revealed changes of a presumed non-specific type including glycogen, lipid, and mitochondrial accumulations. Varying types of degeneration involving myofibres were seen together with variations in interstitial fibrosis and occasional cytoplasmic inclusions. Conclusion—Overall, although interesting, the electron microscopy of myocardial tissue added little to the understanding of the patient's disease, with only one case showing changes not found at light microscopy or with other investigations. Further study might shed light on the "non-specific" ultrastructural findings encountered. Key Words: electron microscopy • myocardial tissue • mitochondrial myopathy PMID:11304852

  17. Direct investigation of subsurface interface electronic structure by ballistic-electron-emission microscopy

    NASA Technical Reports Server (NTRS)

    Kaiser, W. J.; Bell, L. D.

    1988-01-01

    A new technique for spectroscopic investigation of subsurface interface electronic structure has been developed. The method, ballistic-electron-emission microscopy (BEEM), is based on scanning tunneling microscopy. BEEM makes possible, for the first time, direct imaging of subsurface interface properties with nanometer spatial resolution. The first application of BEEM to subsurface Schottky-barrier interfaces is reported.

  18. Direct investigation of subsurface interface electronic structure by ballistic-electron-emission microscopy

    NASA Technical Reports Server (NTRS)

    Kaiser, W. J.; Bell, L. D.

    1988-01-01

    A new technique for spectroscopic investigation of subsurface interface electronic structure has been developed. The method, ballistic-electron-emission microscopy (BEEM), is based on scanning tunneling microscopy. BEEM makes possible, for the first time, direct imaging of subsurface interface properties with nanometer spatial resolution. The first application of BEEM to subsurface Schottky-barrier interfaces is reported.

  19. Modulated structures in calcian dolomite: A study by electron microscopy

    NASA Astrophysics Data System (ADS)

    van Tendeloo, G.; Wenk, H. R.; Gronsky, R.

    1985-11-01

    Calcian dolomite from the Devonian Lost Burro formation has been investigated with electron microscopy techniques. Electron diffraction shows evidence for “c” and “d” type reflections which may occur independently and are indicative of ordered superstructures. High resolution electron microscopy combined with selected area optical diffraction is the basis for models to explain the superstructures in calcian dolomite. It is proposed that “c” reflections are due to ordered substitution of Mg by Ca in basal cation layers. “d” reflections result when the rhombohedral stacking of basal layers is interrupted by intercalation of additional Ca layers. During electron irradiation at 1 MeV the Mg-Ca distribution becomes disordered and the crystal structure attains calcite symmetry. The arrangement of CO3 groups remains ordered.

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

    DOE PAGES

    Vecchione, T.; Denes, P.; Jobe, R. K.; ...

    2017-03-15

    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 in this paper, 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-μm spatial resolution and less than 20 analogue-to-digital converter count RMS pixelmore » 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.« less

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

    NASA Astrophysics Data System (ADS)

    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-μ m 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. Standardless atom counting in scanning transmission electron microscopy.

    PubMed

    LeBeau, James M; Findlay, Scott D; Allen, Leslie J; Stemmer, Susanne

    2010-11-10

    We demonstrate that high-angle annular dark-field imaging in scanning transmission electron microscopy allows for quantification of the number and location of all atoms in a three-dimensional, crystalline, arbitrarily shaped specimen without the need for a calibration standard. We show that the method also provides for an approach to directly measure the finite effective source size of a scanning transmission electron microscope.

  3. Staining and embedding the whole mouse brain for electron microscopy.

    PubMed

    Mikula, Shawn; Binding, Jonas; Denk, Winfried

    2012-12-01

    The development of methods for imaging large contiguous volumes with the electron microscope could allow the complete mapping of a whole mouse brain at the single-axon level. We developed a method based on prolonged immersion that enables staining and embedding of the entire mouse brain with uniform myelin staining and a moderate preservation of the tissue's ultrastructure. We tested the ability to follow myelinated axons using serial block-face electron microscopy.

  4. Enhanced imaging in low dose electron microscopy using electron counting

    PubMed Central

    McMullan, G.; Clark, A.T.; Turchetta, R.; Faruqi, A.R.

    2009-01-01

    We compare the direct electron imaging performance at 120 keV of a monolithic active pixel sensor (MAPS) operated in a conventional integrating mode with the performance obtained when operated in a single event counting mode. For the combination of sensor and incident electron energy used here, we propose a heuristic approach with which to process the single event images in which each event is renormalised to have an integrated weight of unity. Using this approach we find enhancements in the Nyquist frequency modulation transfer function (MTF) and detective quantum efficiency (DQE) over the corresponding integrating mode values by factors of 8 and 3, respectively. PMID:19647366

  5. Optimising electron microscopy experiment through electron optics simulation.

    PubMed

    Kubo, Y; Gatel, C; Snoeck, E; Houdellier, F

    2017-04-01

    We developed a new type of electron trajectories simulation inside a complete model of a modern transmission electron microscope (TEM). Our model incorporates the precise and real design of each element constituting a TEM, i.e. the field emission (FE) cathode, the extraction optic and acceleration stages of a 300kV cold field emission gun, the illumination lenses, the objective lens, the intermediate and projection lenses. Full trajectories can be computed using magnetically saturated or non-saturated round lenses, magnetic deflectors and even non-cylindrical symmetry elements like electrostatic biprism. This multi-scale model gathers nanometer size components (FE tip) with parts of meter length (illumination and projection systems). We demonstrate that non-trivial TEM experiments requiring specific and complex optical configurations can be simulated and optimized prior to any experiment using such model. We show that all the currents set in all optical elements of the simulated column can be implemented in the real column (I2TEM in CEMES) and used as starting alignment for the requested experiment. We argue that the combination of such complete electron trajectory simulations in the whole TEM column with automatic optimization of the microscope parameters for optimal experimental data (images, diffraction, spectra) allows drastically simplifying the implementation of complex experiments in TEM and will facilitate the development of advanced use of the electron microscope in the near future. Copyright © 2017 Elsevier B.V. All rights reserved.

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

    PubMed

    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.

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

    PubMed Central

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

    2009-01-01

    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. PMID:19497671

  8. Studying localized corrosion using liquid cell transmission electron microscopy.

    PubMed

    Chee, See Wee; Pratt, Sarah H; Hattar, Khalid; Duquette, David; Ross, Frances M; Hull, Robert

    2015-01-04

    Localized corrosion of Cu and Al thin films exposed to aqueous NaCl solutions was studied using liquid cell transmission electron microscopy (LCTEM). We demonstrate that potentiostatic control can be used to initiate pitting and that local compositional changes, due to focused ion beam implantation of Au(+) ions, can modify the corrosion susceptibility of Al films.

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

    NASA Astrophysics Data System (ADS)

    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.

  10. A national facility for biological cryo-electron microscopy

    SciTech Connect

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

    2015-01-01

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

  11. Detection of parvoviruses in wolf feces by electron microscopy

    USGS Publications Warehouse

    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.

  12. Scanning electron microscopy analysis of corrosion degradation on tinplate substrates.

    PubMed

    Zumelzu, E; Cabezas, C; Vera, A

    2003-01-01

    The degradation of electrolytic tinplate used in food containers was analysed and evaluated, using scanning electron microscopy and electrochemical measurements of microcorrosion and ion dissolution by atomic absorption to prevent food contamination caused by metal traces and to increase the durability of such tinplates.

  13. Automated data collection in single particle electron microscopy

    PubMed Central

    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

  14. Microstress contrast in scanning electron acoustic microscopy of ceramics

    NASA Technical Reports Server (NTRS)

    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.

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

    PubMed

    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.

  16. Scanning electron microscopy image representativeness: morphological data on nanoparticles.

    PubMed

    Odziomek, Katarzyna; Ushizima, Daniela; Oberbek, Przemyslaw; Kurzydłowski, Krzysztof Jan; Puzyn, Tomasz; Haranczyk, Maciej

    2017-01-01

    A sample of a nanomaterial contains a distribution of nanoparticles of various shapes and/or sizes. A scanning electron microscopy image of such a sample often captures only a fragment of the morphological variety present in the sample. In order to quantitatively analyse the sample using scanning electron microscope digital images, and, in particular, to derive numerical representations of the sample morphology, image content has to be assessed. In this work, we present a framework for extracting morphological information contained in scanning electron microscopy images using computer vision algorithms, and for converting them into numerical particle descriptors. We explore the concept of image representativeness and provide a set of protocols for selecting optimal scanning electron microscopy images as well as determining the smallest representative image set for each of the morphological features. We demonstrate the practical aspects of our methodology by investigating tricalcium phosphate, Ca3 (PO4 )2 , and calcium hydroxyphosphate, Ca5 (PO4 )3 (OH), both naturally occurring minerals with a wide range of biomedical applications. © 2016 The Authors Journal of Microscopy © 2016 Royal Microscopical Society.

  17. Automated data collection in single particle electron microscopy.

    PubMed

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

    2016-02-01

    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.

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

    ERIC Educational Resources Information Center

    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…

  19. Microstress contrast in scanning electron acoustic microscopy of ceramics

    NASA Technical Reports Server (NTRS)

    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.

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

    ERIC Educational Resources Information Center

    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…

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

    ERIC Educational Resources Information Center

    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…

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

    ERIC Educational Resources Information Center

    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…

  3. Nano-fEM: protein localization using photo-activated localization microscopy and electron microscopy.

    PubMed

    Watanabe, Shigeki; Richards, Jackson; Hollopeter, Gunther; Hobson, Robert J; Davis, Wayne M; Jorgensen, Erik M

    2012-12-03

    Mapping the distribution of proteins is essential for understanding the function of proteins in a cell. Fluorescence microscopy is extensively used for protein localization, but subcellular context is often absent in fluorescence images. Immuno-electron microscopy, on the other hand, can localize proteins, but the technique is limited by a lack of compatible antibodies, poor preservation of morphology and because most antigens are not exposed to the specimen surface. Correlative approaches can acquire the fluorescence image from a whole cell first, either from immuno-fluorescence or genetically tagged proteins. The sample is then fixed and embedded for electron microscopy, and the images are correlated (1-3). However, the low-resolution fluorescence image and the lack of fiducial markers preclude the precise localization of proteins. Alternatively, fluorescence imaging can be done after preserving the specimen in plastic. In this approach, the block is sectioned, and fluorescence images and electron micrographs of the same section are correlated (4-7). However, the diffraction limit of light in the correlated image obscures the locations of individual molecules, and the fluorescence often extends beyond the boundary of the cell. Nano-resolution fluorescence electron microscopy (nano-fEM) is designed to localize proteins at nano-scale by imaging the same sections using photo-activated localization microscopy (PALM) and electron microscopy. PALM overcomes the diffraction limit by imaging individual fluorescent proteins and subsequently mapping the centroid of each fluorescent spot (8-10). We outline the nano-fEM technique in five steps. First, the sample is fixed and embedded using conditions that preserve the fluorescence of tagged proteins. Second, the resin blocks are sectioned into ultrathin segments (70-80 nm) that are mounted on a cover glass. Third, fluorescence is imaged in these sections using the Zeiss PALM microscope. Fourth, electron dense structures are

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

    PubMed

    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.

  5. Seeing in 4D with electrons: Development of ultrafast electron microscopy at Caltech

    NASA Astrophysics Data System (ADS)

    Spencer Baskin, J.; Zewail, Ahmed H.

    2014-02-01

    The vision to develop 4D electron microscopy, a union of the capabilities of electron microscopy with ultrafast techniques to capture clearly defined images of the nanoscale structure of a material at each step in the course of its chemical or physical transformations, has been pursued at Caltech for the last decade. In this contribution, we will give a brief overview of the capabilities of three currently active Caltech 4D microscopy laboratories. Ongoing work is illustrated by a description of the most recent application of photon-induced near-field electron microscopy (PINEM), a field made possible only by the development of the 4D ultrafast electron microscopy (UEM). An appendix gives the various applications made so far and the historic roots of the development at Caltech. xml:lang="fr"

  6. Environmental scanning electron microscopy gold immunolabeling in cell biology.

    PubMed

    Rosso, Francesco; Papale, Ferdinando; Barbarisi, Alfonso

    2013-01-01

    Immunogold labeling (IGL) technique has been utilized by many authors in combination with scanning electron microscopy (SEM) and transmission electron microscopy (TEM) to obtain the identification/localization of receptors and antigens, both in cells and tissues. Environmental scanning electron microscopy (ESEM) represents an important tool in biomedical research, since it does not require any severe processing of the sample, lowering the risk of generating artifacts and interfere with the IGL procedure. The absence of metal coating could yield further advantages for our purpose as the labeling detection is based on the atomic number difference between nanogold spheres and the biological material. Using the gaseous secondary electron detector, compositional contrast is easily revealed by the backscattered electron component of the signal. In spite of this fact, only few published papers present a combination of ESEM and IGL. Hereby we present our method, optimized to improve the intensity and the specificity of the labeling signal, in order to obtain a semiquantitative evaluation of the labeling signal.In particular, we used a combination of IGL and ESEM to detect the presence of a protein on the cell surface. To achieve this purpose, we chose as an experimental system 3T3 Swiss albino mouse fibroblasts and galectin-3.

  7. The genetic encoded toolbox for electron microscopy and connectomics.

    PubMed

    Shigemoto, Ryuichi; Joesch, Maximilian

    2017-08-11

    Developments in bioengineering and molecular biology have introduced a palette of genetically encoded probes for identification of specific cell populations in electron microscopy. These probes can be targeted to distinct cellular compartments, rendering them electron dense through a subsequent chemical reaction. These electron densities strongly increase the local contrast in samples prepared for electron microscopy, allowing three major advances in ultrastructural mapping of circuits: genetic identification of circuit components, targeted imaging of regions of interest and automated analysis of the tagged circuits. Together, the gains from these advances can decrease the time required for the analysis of targeted circuit motifs by over two orders of magnitude. These genetic encoded tags for electron microscopy promise to simplify the analysis of circuit motifs and become a central tool for structure-function studies of synaptic connections in the brain. We review the current state-of-the-art with an emphasis on connectomics, the quantitative analysis of neuronal structures and motifs. For further resources related to this article, please visit the WIREs website. © 2017 Wiley Periodicals, Inc.

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

    PubMed

    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. Aberration-corrected Electron Microscopy Imaging for Nanoelectronics Applications

    NASA Astrophysics Data System (ADS)

    Kisielowski, C.; Specht, P.; Alloyeau, D.; Erni, R.; Ramasse, Q.

    2009-09-01

    This paper addresses advances in electron microscopy that were accomplished over the past years with the incorporation of new electron optical components such as aberration correctors, monochromators or high brightness guns. Many of these developments are currently pursued within the DoE's TEAM project. As a result electron microscopy has reached 50 pm resolution. In this paper it is shown how the resolution improvement has helped to boost signal to noise ratios enabling a detection of single atoms across the Periodic Table of Elements. The described achievements allow for investigations of single point defects in nanoelectronic devices even if printed on single sheets of carbon atoms (graphene). Further it is now possible to access depth information from single projections with a precision that has reached interatomic distances.

  10. Imaging plasmodesmata with high-resolution scanning electron microscopy.

    PubMed

    Barton, Deborah A; Overall, Robyn L

    2015-01-01

    High-resolution scanning electron microscopy (HRSEM) is an effective tool to investigate the distribution of plasmodesmata within plant cell walls as well as to probe their complex, three-dimensional architecture. It is a useful alternative to traditional transmission electron microscopy (TEM) in which plasmodesmata are sectioned to reveal their internal substructures. Benefits of adopting an HRSEM approach to studies of plasmodesmata are that the specimen preparation methods are less complex and time consuming than for TEM, many plasmodesmata within a large region of tissue can be imaged in a single session, and three-dimensional information is readily available without the need for reconstructing TEM serial sections or employing transmission electron tomography, both of which are lengthy processes. Here we describe methods to prepare plant samples for HRSEM using pre- or postfixation extraction of cellular material in order to visualize plasmodesmata embedded within plant cell walls.

  11. Quantitative high-resolution transmission electron microscopy of single atoms.

    PubMed

    Gamm, Björn; Blank, Holger; Popescu, Radian; Schneider, Reinhard; Beyer, André; Gölzhäuser, Armin; Gerthsen, Dagmar

    2012-02-01

    Single atoms can be considered as the most basic objects for electron microscopy to test the microscope performance and basic concepts for modeling image contrast. In this work high-resolution transmission electron microscopy was applied to image single platinum, molybdenum, and titanium atoms in an aberration-corrected transmission electron microscope. The atoms are deposited on a self-assembled monolayer substrate that induces only negligible contrast. Single-atom contrast simulations were performed on the basis of Weickenmeier-Kohl and Doyle-Turner form factors. Experimental and simulated image intensities are in quantitative agreement on an absolute intensity scale, which is provided by the vacuum image intensity. This demonstrates that direct testing of basic properties such as form factors becomes feasible.

  12. Biological imaging with 4D ultrafast electron microscopy.

    PubMed

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

    2010-06-01

    Advances in the imaging of biological structures with transmission electron microscopy continue to reveal information at the nanometer length scale and below. The images obtained are static, i.e., time-averaged over seconds, and the weak contrast is usually enhanced through sophisticated specimen preparation techniques and/or improvements in electron optics and methodologies. Here we report the application of the technique of photon-induced near-field electron microscopy (PINEM) to imaging of biological specimens with femtosecond (fs) temporal resolution. In PINEM, the biological structure is exposed to single-electron packets and simultaneously irradiated with fs laser pulses that are coincident with the electron pulses in space and time. By electron energy-filtering those electrons that gained photon energies, the contrast is enhanced only at the surface of the structures involved. This method is demonstrated here in imaging of protein vesicles and whole cells of Escherichia coli, both are not absorbing the photon energy, and both are of low-Z contrast. It is also shown that the spatial location of contrast enhancement can be controlled via laser polarization, time resolution, and tomographic tilting. The high-magnification PINEM imaging provides the nanometer scale and the fs temporal resolution. The potential of applications is discussed and includes the study of antibodies and immunolabeling within the cell.

  13. Biological imaging with 4D ultrafast electron microscopy

    PubMed Central

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

    2010-01-01

    Advances in the imaging of biological structures with transmission electron microscopy continue to reveal information at the nanometer length scale and below. The images obtained are static, i.e., time-averaged over seconds, and the weak contrast is usually enhanced through sophisticated specimen preparation techniques and/or improvements in electron optics and methodologies. Here we report the application of the technique of photon-induced near-field electron microscopy (PINEM) to imaging of biological specimens with femtosecond (fs) temporal resolution. In PINEM, the biological structure is exposed to single-electron packets and simultaneously irradiated with fs laser pulses that are coincident with the electron pulses in space and time. By electron energy-filtering those electrons that gained photon energies, the contrast is enhanced only at the surface of the structures involved. This method is demonstrated here in imaging of protein vesicles and whole cells of Escherichia coli, both are not absorbing the photon energy, and both are of low-Z contrast. It is also shown that the spatial location of contrast enhancement can be controlled via laser polarization, time resolution, and tomographic tilting. The high-magnification PINEM imaging provides the nanometer scale and the fs temporal resolution. The potential of applications is discussed and includes the study of antibodies and immunolabeling within the cell. PMID:20479261

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

    PubMed

    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.

  15. Human enamel structure studied by high resolution electron microscopy

    SciTech Connect

    Wen, S.L. )

    1989-01-01

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

  16. Immuno-electron microscopy of primary cell cultures from genetically modified animals in liquid by atmospheric scanning electron microscopy.

    PubMed

    Kinoshita, Takaaki; Mori, Yosio; Hirano, Kazumi; Sugimoto, Shinya; Okuda, Ken-ichi; Matsumoto, Shunsuke; Namiki, Takeshi; Ebihara, Tatsuhiko; Kawata, Masaaki; Nishiyama, Hidetoshi; Sato, Mari; Suga, Mitsuo; Higashiyama, Kenichi; Sonomoto, Kenji; Mizunoe, Yoshimitsu; Nishihara, Shoko; Sato, Chikara

    2014-04-01

    High-throughput immuno-electron microscopy is required to capture the protein-protein interactions realizing physiological functions. Atmospheric scanning electron microscopy (ASEM) allows in situ correlative light and electron microscopy of samples in liquid in an open atmospheric environment. Cells are cultured in a few milliliters of medium directly in the ASEM dish, which can be coated and transferred to an incubator as required. Here, cells were imaged by optical or fluorescence microscopy, and at high resolution by gold-labeled immuno-ASEM, sometimes with additional metal staining. Axonal partitioning of neurons was correlated with specific cytoskeletal structures, including microtubules, using primary-culture neurons from wild type Drosophila, and the involvement of ankyrin in the formation of the intra-axonal segmentation boundary was studied using neurons from an ankyrin-deficient mutant. Rubella virus replication producing anti-double-stranded RNA was captured at the host cell's plasma membrane. Fas receptosome formation was associated with clathrin internalization near the surface of primitive endoderm cells. Positively charged Nanogold clearly revealed the cell outlines of primitive endoderm cells, and the cell division of lactic acid bacteria. Based on these experiments, ASEM promises to allow the study of protein interactions in various complexes in a natural environment of aqueous liquid in the near future.

  17. Ultrafast electron microscopy in materials science, biology, and chemistry

    SciTech Connect

    King, Wayne E.; Campbell, Geoffrey H.; Frank, Alan; Reed, Bryan; Schmerge, John F.; Siwick, Bradley J.; Stuart, Brent C.; Weber, Peter M.

    2005-06-01

    The use of pump-probe experiments to study complex transient events has been an area of significant interest in materials science, biology, and chemistry. While the emphasis has been on laser pump with laser probe and laser pump with x-ray probe experiments, there is a significant and growing interest in using electrons as probes. Early experiments used electrons for gas-phase diffraction of photostimulated chemical reactions. More recently, scientists are beginning to explore phenomena in the solid state such as phase transformations, twinning, solid-state chemical reactions, radiation damage, and shock propagation. This review focuses on the emerging area of ultrafast electron microscopy (UEM), which comprises ultrafast electron diffraction (UED) and dynamic transmission electron microscopy (DTEM). The topics that are treated include the following: (1) The physics of electrons as an ultrafast probe. This encompasses the propagation dynamics of the electrons (space-charge effect, Child's law, Boersch effect) and extends to relativistic effects. (2) The anatomy of UED and DTEM instruments. This includes discussions of the photoactivated electron gun (also known as photogun or photoelectron gun) at conventional energies (60-200 keV) and extends to MeV beams generated by rf guns. Another critical aspect of the systems is the electron detector. Charge-coupled device cameras and microchannel-plate-based cameras are compared and contrasted. The effect of various physical phenomena on detective quantum efficiency is discussed. (3) Practical aspects of operation. This includes determination of time zero, measurement of pulse-length, and strategies for pulse compression. (4) Current and potential applications in materials science, biology, and chemistry. UEM has the potential to make a significant impact in future science and technology. Understanding of reaction pathways of complex transient phenomena in materials science, biology, and chemistry will provide fundamental

  18. Microfabricated high-bandpass foucault aperture for electron microscopy

    DOEpatents

    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.

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

    PubMed

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

    PubMed

    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.

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

    PubMed

    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.

    PubMed

    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. A fast image simulation algorithm for scanning transmission electron microscopy.

    PubMed

    Ophus, Colin

    2017-01-01

    Image simulation for scanning transmission electron microscopy at atomic resolution for samples with realistic dimensions can require very large computation times using existing simulation algorithms. We present a new algorithm named PRISM that combines features of the two most commonly used algorithms, namely the Bloch wave and multislice methods. PRISM uses a Fourier interpolation factor f that has typical values of 4-20 for atomic resolution simulations. We show that in many cases PRISM can provide a speedup that scales with f(4) compared to multislice simulations, with a negligible loss of accuracy. We demonstrate the usefulness of this method with large-scale scanning transmission electron microscopy image simulations of a crystalline nanoparticle on an amorphous carbon substrate.

  4. In situ transmission electron microscopy for magnetic nanostructures

    NASA Astrophysics Data System (ADS)

    Ngo, Duc-The; Theil Kuhn, Luise

    2016-12-01

    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 that allows understanding of both physical structure and micromagnetic structure of the thin samples at nanoscale. Among TEM techniques, in situ TEM is the state-of-the-art approach for imaging such structures in dynamic experiments, reconstructing a real-time nanoscale picture of the properties-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.

  5. Atom probe field-ion microscopy and related topics: A bibliography, 1988

    SciTech Connect

    Miller, M.K.; Hawkins, A.R.

    1989-10-01

    This bibliography includes references related to the following topics: field-ion microscopy (FIM), field emission microscopy (FEM), atom probe field-ion microscopy (APFIM), and liquid metal ion sources (LMIS). Technique-orientated studies and applications are included. The references contained in this document were compiled from a variety of sources including computer searches and personal lists of publications. To reduce the length of this document, the references have been reduced to the minimum necessary to locate the articles.

  6. Atom probe field ion microscopy and related topics: A bibliography 1989

    SciTech Connect

    Miller, M.K.; Hawkins, A.R.; Russell, K.F.

    1990-12-01

    This bibliography includes references related to the following topics: atom probe field ion microscopy (APFIM), field ion spectroscopy (FIM), field emission microscopy (FEM), liquid metal ion sources (LMIS), scanning tunneling microscopy (STM), and theory. Technique-orientated studies and applications are included. This bibliography covers the period 1989. The references contained in this document were compiled from a variety of sources including computer searches and personal lists of publications.

  7. Practical aspects of monochromators developed for transmission electron microscopy

    PubMed Central

    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

  8. Studying localized corrosion using liquid cell transmission electron microscopy

    DOE PAGES

    Chee, See Wee; Pratt, Sarah H.; Hattar, Khalid; ...

    2014-11-07

    Using liquid cell transmission electron microscopy (LCTEM), localized corrosion of Cu and Al thin films immersed in aqueous NaCl solutions was studied. We demonstrate that potentiostatic control can be used to initiate pitting and that local compositional changes, due to focused ion beam implantation of Au+ ions, can modify the corrosion susceptibility of Al films. Likewise, a discussion on strategies to control the onset of pitting is also presented.

  9. Studying localized corrosion using liquid cell transmission electron microscopy

    SciTech Connect

    Chee, See Wee; Pratt, Sarah H.; Hattar, Khalid; Duquette, David; Ross, Frances M.; Hull, Robert

    2014-11-07

    Using liquid cell transmission electron microscopy (LCTEM), localized corrosion of Cu and Al thin films immersed in aqueous NaCl solutions was studied. We demonstrate that potentiostatic control can be used to initiate pitting and that local compositional changes, due to focused ion beam implantation of Au+ ions, can modify the corrosion susceptibility of Al films. Likewise, a discussion on strategies to control the onset of pitting is also presented.

  10. Three-dimensional cryo-electron microscopy on intermediate filaments.

    PubMed

    Kirmse, Robert; Bouchet-Marquis, Cédric; Page, Cynthia; Hoenger, Andreas

    2010-01-01

    Together with microtubules and actin filaments (F-actin), intermediate filaments (IFs) form the cytoskeleton of metazoan cells. However, unlike the other two entities that are extremely conserved, IFs are much more diverse and are grouped into five different families. In contrast to microtubules and F-actin, IFs do not exhibit a polarity, which may be the reason that no molecular motors travel along them. The molecular structure of IFs is less well resolved than that of the other cytoskeletal systems. This is partially due to their functional variability, tissue-specific expression, and their intrinsic structural properties. IFs are composed mostly of relatively smooth protofibrils formed by antiparallel arranged α-helical coiled-coil bundles flanked by small globular domains at either end. These features make them difficult to study by various electron microscopy methods or atomic force microscopy (AFM). Furthermore, the elongated shape of monomeric or dimeric IF units interferes with the formation of highly ordered three-dimensional (3-D) crystals suitable for atomic resolution crystallography. So far, most of the data we currently have on IF macromolecular structures come from electron microscopy of negatively stained samples, and fragmented α-helical coiled-coil units solved by X-ray diffraction. In addition, AFM allows the observation of the dynamic states of IFs in solution and delivers a new view into the assembly properties of IFs. Here, we discuss the applicability of cryo-electron microscopy (cryo-EM) and cryo-electron tomography (cryo-ET) for the field. Both methods are strongly related and have only recently been applied to IFs. However, cryo-EM revealed distinct new features within IFs that have not been seen before, and cryo-ET adds a 3-D view of IFs revealing the path and number of protofilaments within the various IF assemblies.

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

    PubMed Central

    Nagy, Gabor; Pinczes, Gyula; Pinter, Gabor; Pocsi, Istvan; Prokisch, Jozsef; Banfalvi, Gaspar

    2016-01-01

    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. PMID:27376279

  12. ENHANCEMENT OF CELL BOUNDARIES IN TRANSMISSION ELECTRON MICROSCOPY IMAGES

    PubMed Central

    Tasdizen, Tolga; Whitaker, Ross; Marc, Robert; Jones, Bryan

    2009-01-01

    Transmission electron microscopy (TEM) is an important modality for the analysis of cellular structures in neurobiology. The computational analysis of neurons entail their segmentation and reconstruction from TEM images. This problem is complicated by the heavily textured nature of cellular TEM images and typically low signal-to-noise ratios. In this paper, we propose a new partial differential equation for enhancing the contrast and continuity of cell membranes in TEM images. PMID:19169423

  13. Staining of Tissue Sections for Electron Microscopy with Heavy Metals

    PubMed Central

    Watson, Michael L.

    1958-01-01

    Heavy metals may be incorporated from solution into tissue sections for electron microscopy. The resulting increase in density of the tissue provides greatly enhanced contrast with minimal distortion. Relative densities of various structures are found to depend on the heavy metal ions present and on the conditions of staining. Certain hitherto unobserved details are revealed and some sort of specificity exists, although the factors involved are not yet understood. PMID:13563554

  14. Reciprocity relations in transmission electron microscopy: A rigorous derivation.

    PubMed

    Krause, Florian F; Rosenauer, Andreas

    2017-01-01

    A concise derivation of the principle of reciprocity applied to realistic transmission electron microscopy setups is presented making use of the multislice formalism. The equivalence of images acquired in conventional and scanning mode is thereby rigorously shown. The conditions for the applicability of the found reciprocity relations is discussed. Furthermore the positions of apertures in relation to the corresponding lenses are considered, a subject which scarcely has been addressed in previous publications.

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

    PubMed Central

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

    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 preserving long-standing expertise and to encourage new investigators and clinicians to include EM studies of liver cells and tissue in their projects. PMID:20556830

  16. Free-standing graphene by scanning transmission electron microscopy.

    PubMed

    Song, F Q; Li, Z Y; Wang, Z W; He, L; Han, M; Wang, G H

    2010-11-01

    Free-standing graphene sheets have been imaged by scanning transmission electron microscopy (STEM). We show that the discrete numbers of graphene layers enable an accurate calibration of STEM intensity to be performed over an extended thickness and with single atomic layer sensitivity. We have applied this calibration to carbon nanoparticles with complex structures. This leads to the direct and accurate measurement of the electron mean free path. Here, we demonstrate potentials using graphene sheets as a novel mass standard in STEM-based mass spectrometry.

  17. Transmission Electron Microscopy Study of InN Nanorods

    SciTech Connect

    Liliental-Weber, Z.; Li, X.; Kryliouk, Olga; Park, H.J.; Mangum,J.; Anderson, T.

    2006-07-13

    InN nanorods were grown on a, c-, and r-plane of sapphire and also on Si (111) and GaN (0001) by non-catalytic, template-free hydride metal-organic vapor phase epitaxy and studied by transmission electron microscopy, electron energy loss (EELS) and photoluminescence (PL) at room temperature. These nanocrystals have different shapes and different faceting depending on the substrate used and their crystallographic orientation. EELS measurements have confirmed the high purity of these crystals. The observed PL peak was in the range of 0.9-0.95 eV. The strongest PL intensity was observed for the nanocrystals with the larger diameters.

  18. Transmission electron microscopy of a model crystalline organic, theophylline

    NASA Astrophysics Data System (ADS)

    Cattle, J.; S'ari, M.; Hondow, N.; Abellán, P.; Brown, A. P.; Brydson, R. M. D.

    2015-10-01

    We report on the use of transmission electron microscopy (TEM) to analyse the diffraction patterns of the model crystalline organic theophylline to investigate beam damage in relation to changing accelerating voltage, sample temperature and TEM grid support films. We find that samples deposited on graphene film grids have the longest lifetimes when also held at -190 °C and imaged at 200 kV accelerating voltage. Finally, atomic lattice images are obtained in bright field STEM by working close to the estimated critical electron dose for theophylline.

  19. System and method for compressive scanning electron microscopy

    DOEpatents

    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.

  20. Experiments in electron microscopy: from metals to nerves

    NASA Astrophysics Data System (ADS)

    Unwin, Nigel

    2015-04-01

    Electron microscopy has advanced remarkably as a tool for biological structure research since the development of methods to examine radiation-sensitive unstained specimens and the introduction of cryo-techniques. Structures of biological molecules at near-atomic resolution can now be obtained from images of single particles as well as crystalline arrays. It has also become possible to analyze structures of molecules in their functional context, i.e. in their natural membrane or cellular setting, and in an ionic environment like that in living tissue. Electron microscopy is thus opening ways to answer definitively questions about physiological mechanisms. Here I recall a number of experiments contributing to, and benefiting from the technical advances that have taken place. I begin—in the spirit of this crystallography series—with some biographical background, and then sketch the path to an analysis by time-resolved microscopy of the opening mechanism of an ion channel (nicotinic acetylcholine receptor). This analysis illustrates how electron imaging can be combined with freeze-trapping to illuminate a transient biological event: in our case, chemical-to-electrical transduction at the nerve-muscle synapse.

  1. Navigating 3D electron microscopy maps with EM-SURFER.

    PubMed

    Esquivel-Rodríguez, Juan; Xiong, Yi; Han, Xusi; Guang, Shuomeng; Christoffer, Charles; Kihara, Daisuke

    2015-05-30

    The Electron Microscopy DataBank (EMDB) is growing rapidly, accumulating biological structural data obtained mainly by electron microscopy and tomography, which are emerging techniques for determining large biomolecular complex and subcellular structures. Together with the Protein Data Bank (PDB), EMDB is becoming a fundamental resource of the tertiary structures of biological macromolecules. To take full advantage of this indispensable resource, the ability to search the database by structural similarity is essential. However, unlike high-resolution structures stored in PDB, methods for comparing low-resolution electron microscopy (EM) density maps in EMDB are not well established. We developed a computational method for efficiently searching low-resolution EM maps. The method uses a compact fingerprint representation of EM maps based on the 3D Zernike descriptor, which is derived from a mathematical series expansion for EM maps that are considered as 3D functions. The method is implemented in a web server named EM-SURFER, which allows users to search against the entire EMDB in real-time. EM-SURFER compares the global shapes of EM maps. Examples of search results from different types of query structures are discussed. We developed EM-SURFER, which retrieves structurally relevant matches for query EM maps from EMDB within seconds. The unique capability of EM-SURFER to detect 3D shape similarity of low-resolution EM maps should prove invaluable in structural biology.

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

    PubMed

    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.

  3. An APFIM/AEM Study of Phase Decompositions in Fe-Ni Alloys at Low Temperatures

    NASA Technical Reports Server (NTRS)

    Zhang, J.; Miller, M. K.; Williams, D. B.; Goldstein, J. I.

    1992-01-01

    A combined atom probe field ion microscopy and analytical electron microscopy characterization has been performed on laboratory aged martensitic and austenitic specimens of FeNi and FeNiP alloys. These techniques revealed that the martensitic 24.1 and 28.6 at.% Ni alloys decomposed during aging for 1 year at 300 C to form face centered cubic precipitates of approx. 56 at.% Ni in a body centered cubic matrix containing approx. 20 at.% Ni. Some thin platelets were observed in the field ion micrographs of the austenitic Fe-42.9 at. % Ni alloy and the Fe-43.2 at.% Ni-0.44 at.% P alloy after aging at 400 and 350 C. Atom probe analysis revealed phosphorus clustering in the ternary alloy aged at 300 C.

  4. High-resolution transmission electron microscopy: the ultimate nanoanalytical technique.

    PubMed

    Thomas, John Meurig; Midgley, Paul A

    2004-06-07

    To be able to determine the elemental composition and morphology of individual nanoparticles consisting of no more than a dozen or so atoms that weigh a few zeptograms (10(-21) g) is but one of the attainments of modern electron microscopy. With slightly larger specimens (embracing a few unit cells of the structure) their symmetry, crystallographic phase, unit-cell dimension, chemical composition and often the valence state (from parallel electron spectroscopic measurements) of the constituent atoms may also be determined using a scanning beam of electrons of ca. 0.5 nm diameter. Nowadays electron crystallography, which treats the digital data of electron diffraction (ED) and high-resolution transmission electron microscope (HRTEM) images of minute (ca. 10(-18)g) specimens in a quantitatively rigorous manner, solves hitherto unknown structures just as X-ray diffraction does with bulk single crystals. In addition, electron tomography (see cover photograph and its animation) enables a three-dimensional picture of the internal structure of minute objects, such as nanocatalysts in a single pore, as well as structural faults such as micro-fissures, to be constructed with a resolution of 1 nm from an angular series of two-dimensional (projected) images. Very recently (since this article was first written) a new meaning has been given to electron crystallography as a result of the spatio-temporal resolution of surface phenomena achieved on a femtosecond timescale.

  5. Axial ion-electron emission microscopy of IC radiation hardness

    NASA Astrophysics Data System (ADS)

    Doyle, B. L.; Vizkelethy, G.; Walsh, D. S.; Swenson, D.

    2002-05-01

    A new system for performing radiation effects microscopy (REM) has been developed at Sandia National Laboratory in Albuquerque. This system combines two entirely new concepts in accelerator physics and nuclear microscopy. A radio frequency quadrupole (RFQ) linac is used to boost the energy of ions accelerated by a conventional Tandem Van de Graaff-Pelletron to velocities of 1.9 MeV/amu. The electronic stopping power for heavy ions is near a maximum at this velocity, and their range is ˜20 μm in Si. These ions therefore represent the most ionizing form of radiation in nature, and are nearly ideal for performing single event effects testing of integrated circuits. Unfortunately, the energy definition of the RFQ-boosted ions is rather poor (˜ a few %), which makes problematic the focussing of such ions to the submicron spots required for REM. To circumvent this problem, we have invented ion electron emission microscopy (IEEM). One can perform REM with the IEEM system without focussing or scanning the ion beam. This is because the position on the sample where each ion strikes is determined by projecting ion-induced secondary electrons at high magnification onto a single electron position sensitive detector. This position signal is then correlated with each REM event. The IEEM system is now mounted along the beam line in an axial geometry so that the ions pass right through the electron detector (which is annular), and all of the electrostatic lenses used for projection. The beam then strikes the sample at normal incidence which results in maximum ion penetration and removes a parallax problem experienced in an earlier system. Details of both the RFQ-booster and the new axial IEEM system are given together with some of the initial results of performing REM on Sandia-manufactured radiation hardened integrated circuits.

  6. Electron microscopy study of antioxidant interaction with bacterial cells

    NASA Astrophysics Data System (ADS)

    Plotnikov, Oleg P.; Novikova, Olga V.; Konnov, Nikolai P.; Korsukov, Vladimir N.; Gunkin, Ivan F.; Volkov, Uryi P.

    2000-10-01

    To maintain native microorganisms genotype and phenotype features a lyophylization technique is widely used. However in this case cells are affected by influences of vacuum and low temperature that cause a part of the cells population to be destruction. Another factor reduced microorganisms vitality is formation of reactive oxygen forms that damage certain biological targets (such as DNA, membranes etc.) Recently to raise microorganism's resistance against adverse condition natural and synthetic antioxidants are used. Antioxidant- are antagonists of free radicals. Introduction of antioxidants in protective medium for lyophylization increase bacteria storage life about 2,0-4,8 fold in comparison with reference samples. In the article the main results of our investigation of antioxidants interaction with microorganism cells is described. As bacteria cells we use vaccine strain yersinia pestis EV, that were grown for 48 h at 28 degree(s)C on the Hottinger agar (pH 7,2). Antioxidants are inserted on the agar surface in specimen under test. To investigate a localization of antioxidants for electron microscopy investigation, thallium organic antioxidants were used. The thallium organic compounds have an antioxidant features if thallium is in low concentration (about 1(mu) g/ml). The localization of the thallium organic antioxidants on bacteria Y. pestis EV is visible in electron microscopy images, thallium being heavy metal with high electron density. The negatively stained bacteria and bacteria thin sections with thallium organic compounds were investigated by means of transmission electron microscopy. The localization of the thallium organic compounds is clearly visible in electron micrographs as small dark spots with size about 10-80nm. Probably mechanisms of interaction of antioxidants with bacteria cells are discussed.

  7. A simple energy filter for low energy electron microscopy/photoelectron emission microscopy instruments.

    PubMed

    Tromp, R M; Fujikawa, Y; Hannon, J B; Ellis, A W; Berghaus, A; Schaff, O

    2009-08-05

    Addition of an electron energy filter to low energy electron microscopy (LEEM) and photoelectron emission microscopy (PEEM) instruments greatly improves their analytical capabilities. However, such filters tend to be quite complex, both electron optically and mechanically. Here we describe a simple energy filter for the existing IBM LEEM/PEEM instrument, which is realized by adding a single scanning aperture slit to the objective transfer optics, without any further modifications to the microscope. This energy filter displays a very high energy resolution ΔE/E = 2 × 10(-5), and a non-isochromaticity of ∼0.5 eV/10 µm. The setup is capable of recording selected area electron energy spectra and angular distributions at 0.15 eV energy resolution, as well as energy filtered images with a 1.5 eV energy pass band at an estimated spatial resolution of ∼10 nm. We demonstrate the use of this energy filter in imaging and spectroscopy of surfaces using a laboratory-based He I (21.2 eV) light source, as well as imaging of Ag nanowires on Si(001) using the 4 eV energy loss Ag plasmon.

  8. [Electron microscopy of the surfaces of bacillary spores].

    PubMed

    Smirnova, T A; Zubasheva, M V; Shevliagina, N V; Nikolaenko, M A; Azizbekian, R R

    2013-01-01

    The surface structures of the spores of Bacillus. cereus, B. thuringiensis, and Brevibacillus laterosporus were studied by transmission and scanning electron microscopy. Platinum deposition and negative staining with uranyl acetate revealed appendages and exosporium in B. thuringiensis and B. cereus. The exosporium structure was visualized by negative staining and ultrathin sectioning. For staining the exosporium polysaccharide, Alcian blue was used during fixation. The results obtained show the differences in structural organization of appendages and exosporium in different strains. Canoe-shaped inclusions were revealed in all Br. laterosporus strains, while strain IGM16-92 had a fibrillar capsule as well. Electron microscopy using a dual beam scanning electron microscope Quanta 200 3D provided the information of the spore surface relief without sample treatment (fixation and dehydration). The spores of Br. laterosporus strains had folded surface, unlike the smooth surface of B. cereus and B. thuringiensis spores. Diversity of external spore structures was shown within a species, which may be used for detection of bacteria at the strain level. Optimized procedures for visualization of spore surface by different electron microscopic techniques were discussed.

  9. 4D multiple-cathode ultrafast electron microscopy

    PubMed Central

    Baskin, John Spencer; Liu, Haihua; Zewail, Ahmed H.

    2014-01-01

    Four-dimensional multiple-cathode ultrafast electron microscopy is developed to enable the capture of multiple images at ultrashort time intervals for a single microscopic dynamic process. The dynamic process is initiated in the specimen by one femtosecond light pulse and probed by multiple packets of electrons generated by one UV laser pulse impinging on multiple, spatially distinct, cathode surfaces. Each packet is distinctly recorded, with timing and detector location controlled by the cathode configuration. In the first demonstration, two packets of electrons on each image frame (of the CCD) probe different times, separated by 19 picoseconds, in the evolution of the diffraction of a gold film following femtosecond heating. Future elaborations of this concept to extend its capabilities and expand the range of applications of 4D ultrafast electron microscopy are discussed. The proof-of-principle demonstration reported here provides a path toward the imaging of irreversible ultrafast phenomena of materials, and opens the door to studies involving the single-frame capture of ultrafast dynamics using single-pump/multiple-probe, embedded stroboscopic imaging. PMID:25006261

  10. High-temporal-resolution electron microscopy for imaging ultrafast electron dynamics

    NASA Astrophysics Data System (ADS)

    Hassan, M. Th.; Baskin, J. S.; Liao, B.; Zewail, A. H.

    2017-07-01

    Ultrafast electron microscopy (UEM) has been demonstrated as an effective table-top technique for imaging the temporally evolving dynamics of matter with a subparticle spatial resolution on the timescale of atomic motion. However, imaging the faster motion of electron dynamics in real time has remained beyond reach. Here we demonstrate more than an order of magnitude (16 times) enhancement in the typical temporal resolution of UEM by generating isolated ∼30 fs electron pulses, accelerated at 200 keV, via the optical-gating approach, with sufficient intensity to probe efficiently the electronic dynamics of matter. Moreover, we investigate the feasibility of attosecond optical gating to generate isolated subfemtosecond electron pulses and attain the desired temporal resolution in electron microscopy to establish 'attomicroscopy' to allow the imaging of electron motion in the act.

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

    NASA Astrophysics Data System (ADS)

    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

  12. Atomic force microscopy and scanning electron microscopy study of MgO(110) surface faceting

    NASA Astrophysics Data System (ADS)

    Giese, D. R.; Lamelas, F. J.; Owen, H. A.; Plass, R.; Gajdardziska-Josifovska, M.

    2000-06-01

    Phosphoric- and nitric-acid etching of the MgO(110) surface generates vicinal faceting in both the <001> and <110> directions. Vacuum annealing (to 1000°C) does not introduce thermal faceting, and does not alter the chemical-etch morphology. Three types of acid-induced faceting (early-stage pits, later-stage grooves, and inverted trapezoidal pyramids) are seen as a function of etching time. Facet-angle analysis by atomic force microscopy (AFM) and scanning electron microscopy (SEM) shows the etch morphology to be vicinal, with angles in the range of 9° to 23°, not the low-energy {100} planes expected from minimization of surface energy.

  13. A study of hydrogenated carbon fibers by scanning electron microscopy and confocal laser scanning microscopy.

    PubMed

    de la Cal, Antonio Madroñero; Aguado-Serrano, Juan; Rojas-Cervantes, Maria Luisa; Adame, Elena V Rosa; Marron, Belen Sarmiento; Rosende, Africa Castro; Nevshupa, Roman

    2009-06-01

    The hydrogen absorption process is studied in carbonaceous fibers produced from a mixture of methane and hydrogen. The absorption of the hydrogen was examined in two types of fibers, in "as-grown" state and after a process of desorption during an annealing to 1.473 K under vacuum. Later to its production process, the fibers withstand an oxidation in air to 973 K. The fibers were examined by means of scanning electron microscopy (SEM) and confocal microscopy by reflection. Differences in the behavior during the oxidation were observed between the fibers in as-grown state and those subjected to a further annealing. It could be verified that the fibers were really constituted by two different phases. In one of the phases, the storage of the hydrogen absorbed took place, whereas in the other phase there was no alteration. The process of annealing prior to the absorption of the hydrogen has an appreciable effect on the desorption rate of the hydrogen.

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

    NASA Astrophysics Data System (ADS)

    Tong, Yongpeng; Li, Changming; Liang, Feng; Chen, Jianmin; Zhang, Hong; Liu, Guoqing; Sun, Huibin; Luong, John H. T.

    2008-12-01

    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 2O 3 and TiO 2) on a T lymphoblastic leukemia Jurkat cell line. The increased calcium ion (from CaCl 2) in the culture medium stimulated the accumulation of BaP and EB inside the cell, leading to cell death. ZnO, Al 2O 3 and TiO 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 2O 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.

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

    PubMed

    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.

  16. Big Data Analytics for Scanning Transmission Electron Microscopy Ptychography

    SciTech Connect

    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. In this paper, 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. Finally, 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.

  17. Big Data Analytics for Scanning Transmission Electron Microscopy Ptychography

    DOE PAGES

    Jesse, S.; Chi, M.; Belianinov, A.; ...

    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. In this paper, 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 andmore » 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. Finally, 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.« less

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

    PubMed

    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.

  19. Big Data Analytics for Scanning Transmission Electron Microscopy Ptychography

    NASA Astrophysics Data System (ADS)

    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.

  20. Big Data Analytics for Scanning Transmission Electron Microscopy Ptychography

    PubMed Central

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

    2016-01-01

    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. PMID:27211523

  1. Time resolved electron microscopy for in situ experiments

    SciTech Connect

    Campbell, Geoffrey H. McKeown, Joseph T.; Santala, Melissa K.

    2014-12-15

    Transmission electron microscopy has functioned for decades as a platform for in situ observation of materials and processes with high spatial resolution. Yet, the dynamics often remain elusive, as they unfold too fast to discern at these small spatial scales under traditional imaging conditions. Simply shortening the exposure time in hopes of capturing the action has limitations, as the number of electrons will eventually be reduced to the point where noise overtakes the signal in the image. Pulsed electron sources with high instantaneous current have successfully shortened exposure times (thus increasing the temporal resolution) by about six orders of magnitude over conventional sources while providing the necessary signal-to-noise ratio for dynamic imaging. We describe here the development of this new class of microscope and the principles of its operation, with examples of its application to problems in materials science.

  2. Electron Microscopy of Chloramphenicol-treated Escherichia coli

    PubMed Central

    Morgan, Councilman; Rosenkranz, Herbert S.; Carr, Howard S.; Rose, Harry M.

    1967-01-01

    Thin sections of Escherichia coli were examined by electron microscopy at sequential intervals after addition and then removal of chloramphenicol. The first changes, occurring at 1 hr after exposure to the drug, were disappearance of the ribosomes and aggregation of the nuclear material toward the center of the bacteria. At 2 hr, aggregates of abnormal cytoplasmic granules first appeared and subsequently increased in size. By 23 hr, amorphous, electron-dense material had accumulated within, and at the periphery of, the nuclear matrix. With the removal of chloramphenicol, the bacteria became normal in appearance, passing through a series of stages that were sequential but not synchronous. At 145 min after removal of chloramphenicol, bacteria were encountered in the process of abnormal division. The influence of deoxyribonucleic acid and ribonucleic acid synthesis, and of energy metabolism, upon the changes seen electron microscopically in chloramphenicol-treated cells, was investigated by selectively inhibiting these functions with hydroxyurea, azauracil, and sodium azide, respectively. Images PMID:5337775

  3. Introduction to high-resolution cryo-electron microscopy.

    PubMed

    Czarnocki-Cieciura, Mariusz; Nowotny, Marcin

    2016-01-01

    For many years two techniques have dominated structural biology - X-ray crystallography and NMR spectroscopy. Traditional cryo-electron microscopy of biological macromolecules produced macromolecular reconstructions at resolution limited to 6-10 Å. Recent development of transmission electron microscopes, in particular the development of direct electron detectors, and continuous improvements in the available software, have led to the "resolution revolution" in cryo-EM. It is now possible to routinely obtain near-atomic-resolution 3D maps of intact biological macromolecules as small as ~100 kDa. Thus, cryo-EM is now becoming the method of choice for structural analysis of many complex assemblies that are unsuitable for structure determination by other methods.

  4. Microbial Nanowire Electronic Structure Probed by Scanning Tunneling Microscopy

    NASA Astrophysics Data System (ADS)

    Veazey, Joshua P.; Lampa-Pastirk, Sanela; Reguera, Gemma; Tessmer, Stuart H.

    2010-03-01

    Complex molecules produced by living organisms provide laboratories for interesting physical properties. The study of such interesting physics, likewise, gives new insight into intriguing biological processes. We have studied the pilus nanowires expressed by the bacterium, Geobacter sulfurreducens, using high resolution scanning tunneling microscopy (STM). G. sulfurreducens is a metal reducing bacterium that has evolved electrically conductive pili to efficiently transfer electrons across large distances.footnotetextG. Reguera, K.D. McCarthy, T. Mehta, J.S. Nicoll, M.T. Tuominen, and D.R. Lovley, Nature 435, 1098 (2005) Here we employ the electronic sensitivity of STM to resolve the molecular substructure and the local electronic density of states (LDOS) along the nanowire, in an effort to elucidate the mechanism of conduction. We observe LDOS dependent upon the location of the tip above the nanowire.

  5. Rapid processing of biopsy specimens for examination by electron microscopy.

    PubMed

    Todd, W J; Burgdorfer, W

    1982-01-01

    A rapid method for embedding biopsies and other small specimens for examination by electron microscopy is described. This method is based on an embedding medium consisting of vinyl cyclohexane dioxide and cyanoacrylate which infiltrates into small specimens and polymerizes to form a cured specimen block within 15 minutes. Fixation, dehydration, infiltration and curing of specimens are all accomplished with 1 hour. The embedded specimens are relatively easy to section, stain by commonly employed electron-dense stains and are very stable under the electron beam. All specimens studied including blood, bacteria, and biopsy samples retained the salient ultrastructural features commonly preserved during lengthy methods of embedding. This method is designed for diagnostic laboratories that require rapid results.

  6. Effects of instrument imperfections on quantitative scanning transmission electron microscopy.

    PubMed

    Krause, Florian F; Schowalter, Marco; Grieb, Tim; Müller-Caspary, Knut; Mehrtens, Thorsten; Rosenauer, Andreas

    2016-02-01

    Several instrumental imperfections of transmission electron microscopes are characterized and their effects on the results of quantitative scanning electron microscopy (STEM) are investigated and quantified using simulations. Methods to either avoid influences of these imperfections during acquisition or to include them in reference calculations are proposed. Particularly, distortions inflicted on the diffraction pattern by an image-aberration corrector can cause severe errors of more than 20% if not accounted for. A procedure for their measurement is proposed here. Furthermore, afterglow phenomena and nonlinear behavior of the detector itself can lead to incorrect normalization of measured intensities. Single electrons accidentally impinging on the detector are another source of error but can also be exploited for threshold-less calibration of STEM images to absolute dose, incident beam current determination and measurement of the detector sensitivity. Copyright © 2015 Elsevier B.V. All rights reserved.

  7. Analysis of Electron Beam Damage of Crystalline Pharmaceutical Materials by Transmission Electron Microscopy

    NASA Astrophysics Data System (ADS)

    S'ari, M.; Cattle, J.; Hondow, N.; Blade, H.; Cosgrove, S.; Brydson, R. M.; Brown, A. P.

    2015-10-01

    We have studied the impact of transmission electron microscopy (TEM) and low dose electron diffraction on ten different crystalline pharmaceutical compounds, covering a diverse chemical space and with differing physical properties. The aim was to establish if particular chemical moieties were more susceptible to damage within the electron beam. We have measured crystalline diffraction patterns for each and indexed nine out of ten of them. Characteristic electron dosages are reported for each material, with no apparent correlation between chemical structure and stability within the electron beam. Such low dose electron diffraction protocols are suitable for the study of pharmaceutical compounds.

  8. Electron microscopy imaging of proteins on gallium phosphide semiconductor nanowires

    NASA Astrophysics Data System (ADS)

    Hjort, Martin; Bauer, Mikael; Gunnarsson, Stefan; Mårsell, Erik; Zakharov, Alexei A.; Karlsson, Gunnel; Sanfins, Elodie; Prinz, Christelle N.; Wallenberg, Reine; Cedervall, Tommy; Mikkelsen, Anders

    2016-02-01

    We have imaged GaP nanowires (NWs) incubated with human laminin, serum albumin (HSA), and blood plasma using both cryo-transmission electron microscopy and synchrotron based X-ray photoemission electron microscopy. This extensive imaging methodology simultaneously reveals structural, chemical and morphological details of individual nanowires and the adsorbed proteins. We found that the proteins bind to NWs, forming coronas with thicknesses close to the proteins' hydrodynamic diameters. We could directly image how laminin is extending from the NWs, maximizing the number of proteins bound to the NWs. NWs incubated with both laminin and HSA show protein coronas with a similar appearance to NWs incubated with laminin alone, indicating that the presence of HSA does not affect the laminin conformation on the NWs. In blood plasma, an intermediate sized corona around the NWs indicates a corona with a mixture of plasma proteins. The ability to directly visualize proteins on nanostructures in situ holds great promise for assessing the conformation and thickness of the protein corona, which is key to understanding and predicting the properties of engineered nanomaterials in a biological environment.We have imaged GaP nanowires (NWs) incubated with human laminin, serum albumin (HSA), and blood plasma using both cryo-transmission electron microscopy and synchrotron based X-ray photoemission electron microscopy. This extensive imaging methodology simultaneously reveals structural, chemical and morphological details of individual nanowires and the adsorbed proteins. We found that the proteins bind to NWs, forming coronas with thicknesses close to the proteins' hydrodynamic diameters. We could directly image how laminin is extending from the NWs, maximizing the number of proteins bound to the NWs. NWs incubated with both laminin and HSA show protein coronas with a similar appearance to NWs incubated with laminin alone, indicating that the presence of HSA does not affect the

  9. SEGMENTATION OF MITOCHONDRIA IN ELECTRON MICROSCOPY IMAGES USING ALGEBRAIC CURVES

    PubMed Central

    Seyedhosseini, Mojtaba; Ellisman, Mark H.; Tasdizen, Tolga

    2014-01-01

    High-resolution microscopy techniques have been used to generate large volumes of data with enough details for understanding the complex structure of the nervous system. However, automatic techniques are required to segment cells and intracellular structures in these multi-terabyte datasets and make anatomical analysis possible on a large scale. We propose a fully automated method that exploits both shape information and regional statistics to segment irregularly shaped intracellular structures such as mitochondria in electron microscopy (EM) images. The main idea is to use algebraic curves to extract shape features together with texture features from image patches. Then, these powerful features are used to learn a random forest classifier, which can predict mitochondria locations precisely. Finally, the algebraic curves together with regional information are used to segment the mitochondria at the predicted locations. We demonstrate that our method outperforms the state-of-the-art algorithms in segmentation of mitochondria in EM images. PMID:25132915

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

    PubMed

    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. Copyright © 2010 Elsevier Inc. All rights reserved.

  11. High-resolution electron microscopy of advanced materials

    SciTech Connect

    Mitchell, T.E.; Kung, H.H.; Sickafus, K.E.; Gray, G.T. III; Field, R.D.; Smith, J.F.

    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.

  12. New developments in electron microscopy for serial image acquisition of neuronal profiles.

    PubMed

    Kubota, Yoshiyuki

    2015-02-01

    Recent developments in electron microscopy largely automate the continuous acquisition of serial electron micrographs (EMGs), previously achieved by laborious manual serial ultrathin sectioning using an ultramicrotome and ultrastructural image capture process with transmission electron microscopy. The new systems cut thin sections and capture serial EMGs automatically, allowing for acquisition of large data sets in a reasonably short time. The new methods are focused ion beam/scanning electron microscopy, ultramicrotome/serial block-face scanning electron microscopy, automated tape-collection ultramicrotome/scanning electron microscopy and transmission electron microscope camera array. In this review, their positive and negative aspects are discussed.

  13. Consecutive light microscopy, scanning-transmission electron microscopy and transmission electron microscopy of traumatic human brain oedema and ischaemic brain damage.

    PubMed

    Castejon, O J; Castejon, H V; Diaz, M; Castellano, A

    2001-10-01

    Cortical biopsies of 11 patients with traumatic brain oedema were consecutively studied by light microscopy (LM) using thick plastic sections, scanning-transmission electron microscopy ((S)TEM) using semithin plastic sections and transmission electron microscopy (TEM) using ultrathin sections. Samples were glutaraldehyde-osmium fixed and embedded in Araldite or Epon. Thick sections were stained with toluidine-blue for light microscopy. Semithin sections were examined unstained and uncoated for (S)TEM. Ultrathin sections were stained with uranyl and lead. Perivascular haemorrhages and perivascular extravasation of proteinaceous oedema fluid were observed in both moderate and severe oedema. Ischaemic pyramidal and non-pyramidal nerve cells appeared shrunken, electron dense and with enlargement of intracytoplasmic membrane compartment. Notably swollen astrocytes were observed in all samples examined. Glycogen-rich and glycogen-depleted astrocytes were identified in anoxic-ischaemic regions. Dark and hydropic satellite, interfascicular and perivascular oligodendrocytes were also found. The status spongiosus of severely oedematous brain parenchyma observed by LM and (S)TEM was correlated with the enlarged extracellular space and disrupted neuropil observed by TEM. The (S)TEM is recommended as a suitable technique for studying pathological processes in the central nervous system and as an informative adjunct to LM and TEM.

  14. Electron microscopy study of direct laser deposited IN718

    SciTech Connect

    Ding, R.G.; Huang, Z.W.; Li, H.Y.; Mitchell, I.; Baxter, G.; Bowen, P.

    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.

  15. Aberration-Coreected Electron Microscopy at Brookhaven National Laboratory

    SciTech Connect

    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 electron

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

    NASA Astrophysics Data System (ADS)

    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

  17. Transmission Electron Microscopy Characterization of Semiconductor Interfacial Structures

    NASA Astrophysics Data System (ADS)

    Robertson, Michael Dennis

    The epitaxial structure and characterization of semiconductor/semiconductor interfacial systems have been studied using transmission electron microscopy as the primary investigative technique. Geometrical and elastic energy theories of epitaxy, as they relate to interfacial structure, have been reviewed to establish the framework necessary for analyzing experimental semiconductor heterostructures. The diffracted electron intensities for cross-sectional semiconductor single layer and superlattice structures have been derived based on the kinematical theory. The expression for the kinematical intensity for electron diffraction from a superlattice was observed to be analogous to the diffraction of light by a diffraction grating. The effects of surface relaxation, present in all strained-layer specimens prepared for the transmission electron microscope, have been investigated using elasticity theory. Conditions where surface relaxation effects can be ignored have also been presented. In order to quantify elastic strains at the nanometer level using high resolution electron microscopy (HREM) images, a new strain analysis technique, based on the cumulative sum of deviations (CUSUM) in lattice-fringe spacings from a target value, has been developed. This technique accurately reproduced the strain profiles in simulated and experimental HREM images and proved to be robust even in the presence of high levels of experimental noise. The above theory and techniques have been applied to three experimental systems, covering three distinct regimes of lattice mismatch (lattice mismatch ranged from -3.4% to +14.6%). These three systems were In_{1-x}Ga_ {x}Sb (0 <=q x <=q 1) single layers on (001) GaAs, rm In_{1-x}Al_{x}Sb/InSb single layers and superlattices on (001) InSb, and a 20 period AlAs/GaAs superlattice on (001) GaAs.

  18. The origins and evolution of freeze-etch electron microscopy

    PubMed Central

    Heuser, John E.

    2011-01-01

    The introduction of the Balzers freeze-fracture machine by Moor in 1961 had a much greater impact on the advancement of electron microscopy than he could have imagined. Devised originally to circumvent the dangers of classical thin-section techniques, as well as to provide unique en face views of cell membranes, freeze-fracturing proved to be crucial for developing modern concepts of how biological membranes are organized and proved that membranes are bilayers of lipids within which proteins float and self-assemble. Later, when freeze-fracturing was combined with methods for freezing cells that avoided the fixation and cryoprotection steps that Moor still had to use to prepare the samples for his original invention, it became a means for capturing membrane dynamics on the millisecond time-scale, thus allowing a deeper understanding of the functions of biological membranes in living cells as well as their static ultrastructure. Finally, the realization that unfixed, non-cryoprotected samples could be deeply vacuum-etched or even freeze-dried after freeze-fracturing opened up a whole new way to image all the other molecular components of cells besides their membranes and also provided a powerful means to image the interactions of all the cytoplasmic components with the various membranes of the cell. The purpose of this review is to outline the history of these technical developments, to describe how they are being used in electron microscopy today and to suggest how they can be improved in order to further their utility for biological electron microscopy in the future. PMID:21844598

  19. Scanning electron microscopy of adult Gongylonema pulchrum (Nematoda: Spirurida).

    PubMed

    Naem, S; Seifi, H; Simon, G T

    2000-05-01

    Scanning electron microscopy (SEM) was used to study the surface ultrastructure of adult worms of Gongylonema pulchrum. The anterior end in both sexes was covered by numerous cuticular platelets. There was a pair of lateral cervical papillac. The buccal opening was small and extended in the dorsoventral direction. Around the mouth a cuticular elevation enclosed the labia, and eight papillae were located laterodorsally and lateroventrally. Two large lateral amphids were seen. On the lateral sides of the female's tail, phasmidal apertures were observed. The caudal end of the male was asymmetrically alate and bore 10 pairs of papillae and two phasmidal apertures.

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

    NASA Technical Reports Server (NTRS)

    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.

  1. Electron microscopy of phages in serotypes of Actinobacillus actinomycetemcomitans.

    PubMed

    Olsen, I; Namork, E; Myhrvold, V

    1993-12-01

    Actinobacillus actinomycetemcomitans, Actinobacillus ureae, Haemophilus aphrophilus, Haemophilus paraphrophilus, Haemophilus influenzae, Haemophilus parainfluenzae, Pasteurella haemolytica and Pasteurella multocida strains were examined by transmission electron microscopy for the presence of bacteriophages. Phages were detected in serotype a (SUNY 75) and e (UOH 1705) and in the fresh clinical isolates UOH Q1243 and UOH Q1247 of A. actinomycetemcomitans. Phages were not found in serotype b, c and d strains of A. actinomycetemcomitans, in the fresh clinical isolate UOH Q1244 of this species or in old strains (including reference strains) of related species from the Actinobacillus-Haemophilus-Pasteurella group.

  2. Site-specific labeling of proteins for electron microscopy

    PubMed Central

    Dambacher, Corey M.; Lander, Gabriel C.

    2015-01-01

    Electron microscopy is commonly employed to determine the subunit organization of large macromolecular assemblies. However, the field lacks a robust molecular labeling methodology for unambiguous identification of constituent subunits. We present a strategy that exploits the unique properties of an unnatural amino acid in order to enable site-specific attachment of a single, readily identifiable protein label at any solvent-exposed position on the macromolecular surface. Using this method, we show clear labeling of a subunit within the 19S proteasome lid subcomplex that has not been amenable to labeling by traditional approaches. PMID:26409249

  3. Measurement of dihedral angles by scanning electron microscopy.

    NASA Technical Reports Server (NTRS)

    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.

  4. Cryo-Electron Microscopy of Biological Macromolecular Structures

    NASA Astrophysics Data System (ADS)

    Yonekura, Koji

    There are many huge macromolecular complexes in living organisms. They are often hard to crystallize because of their size, complexity and heterogeneity. Cryo-electron microscopy (cryo-EM) is a suitable method to analyze the structures of such biological macromolecules, because it can be applied to various forms of samples, e.g. two-dimensional crystal, helical assembly, spherical virus, dispersed particle, cell organelle and cell, although attainable resolution depends on the system. In this review, I introduce these techniques and examples of the structure analysis, and briefly review the perspective of cryo-EM.

  5. Correlated cryogenic photoactivated localization microscopy and cryo-electron tomography.

    PubMed

    Chang, Yi-Wei; Chen, Songye; Tocheva, Elitza I; Treuner-Lange, Anke; Löbach, Stephanie; Søgaard-Andersen, Lotte; Jensen, Grant J

    2014-07-01

    Cryo-electron tomography (CET) produces three-dimensional images of cells in a near-native state at macromolecular resolution, but identifying structures of interest can be challenging. Here we describe a correlated cryo-PALM (photoactivated localization microscopy)-CET method for localizing objects within cryo-tomograms to beyond the diffraction limit of the light microscope. Using cryo-PALM-CET, we identified multiple and new conformations of the dynamic type VI secretion system in the crowded interior of Myxococcus xanthus.

  6. Segmentation of virus particle candidates in transmission electron microscopy images.

    PubMed

    Kylberg, G; Uppström, M; Hedlund, K-O; Borgefors, G; Sintorn, I-M

    2012-02-01

    In this paper, we present an automatic segmentation method that detects virus particles of various shapes in transmission electron microscopy images. The method is based on a statistical analysis of local neighbourhoods of all the pixels in the image followed by an object width discrimination and finally, for elongated objects, a border refinement step. It requires only one input parameter, the approximate width of the virus particles searched for. The proposed method is evaluated on a large number of viruses. It successfully segments viruses regardless of shape, from polyhedral to highly pleomorphic.

  7. Analytical electron microscopy of a hydrated interplanetary dust particle

    NASA Technical Reports Server (NTRS)

    Blake, David F.; Bunch, T. E.; Mardinly, A. J.; Echer, C. J.

    1988-01-01

    Properties of a hydrated interplanetary dust particle (IDP), Ames-Dec86-11, were investigated using TEM and analytical electron microscopy. The particle was found to have mineralogy and chondritic composition indicating an absence of direct kinship with known carbonaceous chondrites. The available data on the Ames-Dec86-11 suggest that at least one aqueous alteration event took place in this hydrated IDP, during which fine-grained material, possibly glass, was transformed to smectite. This event appears to be unique to hydrated IDPs.

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

    NASA Technical Reports Server (NTRS)

    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.

  9. Measurement of dihedral angles by scanning electron microscopy.

    NASA Technical Reports Server (NTRS)

    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.

  10. Analytical electron microscopy of a hydrated interplanetary dust particle

    NASA Technical Reports Server (NTRS)

    Blake, David F.; Bunch, T. E.; Mardinly, A. J.; Echer, C. J.

    1988-01-01

    Properties of a hydrated interplanetary dust particle (IDP), Ames-Dec86-11, were investigated using TEM and analytical electron microscopy. The particle was found to have mineralogy and chondritic composition indicating an absence of direct kinship with known carbonaceous chondrites. The available data on the Ames-Dec86-11 suggest that at least one aqueous alteration event took place in this hydrated IDP, during which fine-grained material, possibly glass, was transformed to smectite. This event appears to be unique to hydrated IDPs.

  11. Analytical electron microscopy of a hydrated interplanetary dust particle

    NASA Astrophysics Data System (ADS)

    Blake, D. F.; Mardinly, A. J.; Echer, C. J.; Bunch, T. E.

    Properties of a hydrated interplanetary dust particle (IDP), Ames-Dec86-11, were investigated using TEM and analytical electron microscopy. The particle was found to have mineralogy and chondritic composition indicating an absence of direct kinship with known carbonaceous chondrites. The available data on the Ames-Dec86-11 suggest that at least one aqueous alteration event took place in this hydrated IDP, during which fine-grained material, possibly glass, was transformed to smectite. This event appears to be unique to hydrated IDPs.

  12. Confocal Microscopy for Modeling Electron Microbeam Irradiation of Skin

    SciTech Connect

    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.

  13. Transmission electron microscopy of electrospun GaN nanofibers

    NASA Astrophysics Data System (ADS)

    Robles-García, Joshua L.; Meléndez, Anamaris; Yates, Douglas; Santiago-Avilés, Jorge J.; Ramos, Idalia; Campo, Eva M.

    2011-06-01

    We have reported earlier progress in producing polycrystalline wurtzite-polymorph and photo-conductive GaN nanofibers by electrospinning. This paper shows grain stacking during heat treatment and suggests the need to understand nucleation and grain growth following electrospinning. Transmission Electron Microscopy (TEM) analysis of GaN shows brittle fibers, grain stacking, and unfinished grain nucleation. X-Ray Diffraction analysis confirmed dominant hexagonal 101-wurtzite preferential overall orientation and the incipient grains are of high crystalline quality as seen by high resolution TEM.

  14. High resolution electron microscopy study of amorphous calcium phosphate

    NASA Astrophysics Data System (ADS)

    Brès, E. F.; Moebus, G.; Kleebe, H.-J.; Pourroy, G.; Werkmann, J.; Ehret, G.

    1993-03-01

    "Amorphous" calcium phosphate (ACP) from human tooth enamel and different synthetic materials has been analysed by high resolution electron microscopy (HREM). All the materials studied showed, in addition to a "truly" amorphous phase, other calcium phosphate phases such as poorly crystalline hydroxyapatite (OHAP), well crystallized OHAP and poorly crystalline CaO type phase. Such structural heterogeneities have not been observed before in ACP, and are only possible to be detected by HREM as this is the only technique able to analyse nanometre size materials in the real space.

  15. Transmission Electron Microscopy Of Lipid Vesicles For Drug Delivery

    NASA Astrophysics Data System (ADS)

    Bello, Valentina; Mattei, Giovanni; Mazzoldi, Paolo; Vivenza, Nicoletta; Gasco, Paolo; Idee, Jean Marc; Robic, Caroline; Borsella, Elisabetta

    2010-10-01

    Iron oxides nanocrystals are largely used for biomedical applications due to their high magnetization. Furthermore for in vivo applications these nanoparticles must be covered with a non-toxic material. Inside the numerous nano-systems for drug delivery, lipid structures, such as Solid Lipid Nanoparticles (SLNs), have been largely developed for various administration routes. In this work SLNs and iron-oxide nanocrystals covered with a lipid shell are characterized by Transmission Electron Microscopy. This technique has revealed to be essential to investigate the ultrafine compositional and morphological properties of these systems.

  16. Photoemission Electron Microscopy of a Plasmonic Silver Nanoparticle Trimer

    SciTech Connect

    Peppernick, Samuel J.; Joly, Alan G.; Beck, Kenneth M.; Hess, Wayne P.; Wang, Jinyong; Wang, Yi-Chung; Wei, Wei

    2013-07-01

    We present a combined experimental and theoretical study to investigate the spatial distribution of photoelectrons emitted from core-shell silver (Ag) nanoparticles. We use two-photon photoemission microscopy (2P-PEEM) to spatially resolve electron emission from a trimeric core-shell aggregate of triangular symmetry. Finite difference time domain (FDTD) simulations are performed to model the intensity distributions of the electromagnetic near-fields resulting from femtosecond (fs) laser excitation of localized surface plasmon oscillations in the triangular core-shell structure. We demonstrate that the predicted FDTD near-field intensity distribution reproduces the 2P-PEEM photoemission pattern.

  17. Droplet Epitaxy Image Contrast in Mirror Electron Microscopy

    NASA Astrophysics Data System (ADS)

    Kennedy, S. M.; Zheng, C. X.; Jesson, D. E.

    2017-01-01

    Image simulation methods are applied to interpret mirror electron microscopy (MEM) images obtained from a movie of GaAs droplet epitaxy. Cylindrical symmetry of structures grown by droplet epitaxy is assumed in the simulations which reproduce the main features of the experimental MEM image contrast, demonstrating that droplet epitaxy can be studied in real-time. It is therefore confirmed that an inner ring forms at the droplet contact line and an outer ring (or skirt) occurs outside the droplet periphery. We believe that MEM combined with image simulations will be increasingly used to study the formation and growth of quantum structures.

  18. Transmission electron microscopy study of flea lymph cell thin sections

    NASA Astrophysics Data System (ADS)

    Volkov, Uryi P.; Konnov, Nikolai P.; Novikova, Olga V.

    2002-07-01

    Transmission electron microscopy investigation of thin sections remains the major method of cells inner structure study with high resolution. However, the present-day technique of cells preparation make it impossible to study a number of biological samples, such as very small quantity of lymph cells of little insects. A new technique of cells preparation has been developed in our lab, which allows to obtain a thin sections of ultra small quantity of cells. Structure of lymph cells of flea was investigated by the technique.

  19. Simultaneous orientation and thickness mapping in transmission electron microscopy

    SciTech Connect

    Tyutyunnikov, Dmitry; Özdöl, V. Burak; Koch, Christoph T.

    2014-12-04

    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.

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

    PubMed Central

    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

  1. Combined Scanning Transmission Electron Microscopy Tilt- and Focal Series

    SciTech Connect

    Dahmen, Tim; Baudoin, Jean-Pierre G; Lupini, Andrew R; Kubel, Christian; Slusallek, Phillip; De Jonge, Niels

    2014-01-01

    In this study, a combined tilt- and focal series is proposed as a new recording scheme for high-angle annular dark-field scanning transmission electron microscopy (STEM) tomography. Three-dimensional (3D) data were acquired by mechanically tilting the specimen, and recording a through-focal series at each tilt direction. The sample was a whole-mount macrophage cell with embedded gold nanoparticles. The tilt focal algebraic reconstruction technique (TF-ART) is introduced as a new algorithm to reconstruct tomograms from such combined tilt- and focal series. The feasibility of TF-ART was demonstrated by 3D reconstruction of the experimental 3D data. The results were compared with a conventional STEM tilt series of a similar sample. The combined tilt- and focal series led to smaller missing wedge artifacts, and a higher axial resolution than obtained for the STEM tilt series, thus improving on one of the main issues of tilt series-based electron tomography.

  2. Biomechanics of DNA structures visualized by 4D electron microscopy

    PubMed Central

    Lorenz, Ulrich J.; Zewail, Ahmed H.

    2013-01-01

    We present a technique for in situ visualization of the biomechanics of DNA structural networks using 4D electron microscopy. Vibrational oscillations of the DNA structure are excited mechanically through a short burst of substrate vibrations triggered by a laser pulse. Subsequently, the motion is probed with electron pulses to observe the impulse response of the specimen in space and time. From the frequency and amplitude of the observed oscillations, we determine the normal modes and eigenfrequencies of the structures involved. Moreover, by selective “nano-cutting” at a given point in the network, it was possible to obtain Young’s modulus, and hence the stiffness, of the DNA filament at that position. This experimental approach enables nanoscale mechanics studies of macromolecules and should find applications in other domains of biological networks such as origamis. PMID:23382239

  3. Electron microscopy of gold nanoparticles at atomic resolution

    PubMed Central

    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

  4. Nanoparticle imaging. Electron microscopy of gold nanoparticles at atomic resolution.

    PubMed

    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

    2014-08-22

    Structure determination of gold nanoparticles (AuNPs) is necessary for understanding their physical and chemical properties, but only one AuNP larger than 1 nanometer in diameter [a 102-gold atom NP (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 a Au68NP at atomic resolution by aberration-corrected transmission electron microscopy, 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 and by comparison of observed infrared absorption spectra with calculations by density functional theory.

  5. Theory and application of scanning electron acoustic microscopy

    NASA Technical Reports Server (NTRS)

    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.

  6. Modeling atomic-resolution scanning transmission electron microscopy images.

    PubMed

    Findlay, Scott D; Oxley, Mark P; Allen, Leslie J

    2008-02-01

    A real-space description of inelastic scattering in scanning transmission electron microscopy is derived with particular attention given to the implementation of the projected potential approximation. A hierarchy of approximations to expressions for inelastic images is presented. Emphasis is placed on the conditions that must hold in each case. The expressions that justify the most direct, visual interpretation of experimental data are also the most approximate. Therefore, caution must be exercised in selecting experimental parameters that validate the approximations needed for the analysis technique used. To make the most direct, visual interpretation of electron-energy-loss spectroscopic images from core-shell excitations requires detector improvements commensurate with those that aberration correction provides for the probe-forming lens. Such conditions can be relaxed when detailed simulations are performed as part of the analysis of experimental data.

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

    PubMed

    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.

  8. Advanced electron microscopy characterization of nanomaterials for catalysis

    DOE PAGES

    Su, Dong

    2017-02-11

    Transmission electron microscopy (TEM) has become one of the most powerful techniques in the fields of material science, inorganic chemistry and nanotechnology. In terms of resolutions, advanced TEM may reach a high spatial resolution of 0.05 nm, a high energy-resolution of 7 meV. In addition, in situ TEM can help researcher to image the process happened within 1 ms. This paper reviews the recent technical approaches of applying advanced TEM characterization on nanomaterials for catalysis. The text is organized according to the demanded information of nanocrystals from the perspective of application: for example, size, composition, phase, strain, and morphology. Themore » electron beam induced effect and in situ TEM are also introduced. As a result, I hope this review can help the scientists in related fields to take advantage of advanced TEM to their own researches.« less

  9. Theory and application of scanning electron acoustic microscopy

    NASA Technical Reports Server (NTRS)

    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.

  10. Development of a Nanoindenter for In Situ Transmission Electron Microscopy.

    PubMed

    Stach, Eric A.; Freeman, Tony; Minor, Andrew M.; Owen, Doug K.; Cumings, John; Wall, Mark A.; Chraska, Tomas; Hull, Robert; Morris, J.W.; Jr, A.; Zettl, Ulrich

    2001-11-01

    In situ transmission electron microscopy is an established experimental technique that permits direct observation of the dynamics and mechanisms of dislocation motion and deformation behavior. In this article, we detail the development of a novel specimen goniometer that allows real-time observations of the mechanical response of materials to indentation loads. The technology of the scanning tunneling microscope is adopted to allow nanometer-scale positioning of a sharp, conductive diamond tip onto the edge of an electron-transparent sample. This allows application of loads to nanometer-scale material volumes coupled with simultaneous imaging of the material's response. The emphasis in this report is qualitative and technique oriented, with particular attention given to sample geometry and other technical requirements. Examples of the deformation of aluminum and titanium carbide as well as the fracture of silicon will be presented.

  11. Some applications of microanalytical electron microscopy in materials research

    SciTech Connect

    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.

  12. Characterization of mesoporosity in ceria particles using electron microscopy.

    PubMed

    Shih, Shao-Ju; Herrero, Pilar Rodrigo; Li, Guoqiang; Chen, Chin-Yi; Lozano-Perez, Sergio

    2011-02-01

    The geometry and three-dimensional (3D) morphology of the ceria particles synthesized by spray pyrolysis (SP) from two different precursors--cerium acetate hydrate and cerium nitrate hydrate (CeA and CeN ceria particles)--were characterized by transmission electron microscopy and electron tomography. Results were compared with surface area measurements, confirming that the surface area of CeA ceria particles is twice as large as that of CeN ceria particles. This result was supported by 3D microstructural observations, which have revealed that CeA ceria particles contain open pores (connected to surfaces) and closed pores (embedded in particles), while CeN ceria particles only contained closed pores. This experimental result suggests that the type of porosity is controlled by the precursors and could be related to their melting temperature during the heating process in SP.

  13. Atomic resolution electron microscopy of small metal clusters

    NASA Astrophysics Data System (ADS)

    Bovin, J.-O.; Malm, J.-O.

    1991-03-01

    Atomic resolution imaging of cluster structures has been performed with high resolution transmission electron microscopy (HRTEM). Metal particles of the sizes 1 nanometer to tens of nanometers have been surface profile imaged on different supports; like zeolites, cordierite and amorphous carbon. It is shown that organic ligands in Schmid-clusters coordinated to the metal surface are desorbed or destroyed by the electron beam. Dynamic events on the surfaces and in the bulk of small metal particles have been recorded for small crystals of Au, Pt, Rh and Pb and can be classified under three headings; The smaller the crystals are the faster rearrangements of the crystal structure; “clouds” of atoms existing outside some surfaces are involved in extensive structural rearrangements of the surface or crystal surface growth; localized atom hopping on surfaces during crystal growth and desorption also occurs.

  14. Investigation of porous asphalt microstructure using optical and electron microscopy.

    PubMed

    Poulikakos, L D; Partl, M N

    2010-11-01

    Direct observations of porous asphalt concrete samples in their natural state using optical and electron microscopy techniques led to useful information regarding the microstructure of two mixes and indicated a relationship between microstructure and in situ performance. This paper presents evidence that suboptimal microstructure can lead to premature failure thus making a first step in defining well or suboptimal performing pavements with a bottom-up approach (microstructure). Laboratory and field compaction produce different samples in terms of the microstructure. Laboratory compaction using the gyratory method has produced more microcracks in mineral aggregates after the binder had cooled. Well-performing mixes used polymer-modified binders, had a more homogeneous void structure with fewer elongated voids and better interlocking of the aggregates. Furthermore, well-performing mixes showed better distribution of the mastic and better coverage of the aggregates with bitumen. Low vacuum scanning electron microscopy showed that styrene butadiene styrene polymer modification in binder exists in the form of discontinuous globules and not continuous networks. A reduction in the polymer phase was observed as a result of aging and in-service use. © 2010 The Authors Journal compilation © 2010 The Royal Microscopical Society.

  15. Golgi apparatus analyzed by cryo-electron microscopy.

    PubMed

    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.

  16. Volume scanning electron microscopy for imaging biological ultrastructure.

    PubMed

    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.

  17. Frontiers of in situ electron microscopy

    SciTech Connect

    Zheng, Haimei; Zhu, Yimei; Meng, Shirley Ying

    2015-01-01

    In situ transmission electron microscopy (TEM) has become an increasingly important tool for materials characterization. It provides key information on the structural dynamics of a material during transformations and the correlation between structure and properties of materials. With the recent advances in instrumentation, including aberration corrected optics, sample environment control, the sample stage, and fast and sensitive data acquisition, in situ TEM characterization has become more and more powerful. In this article, a brief review of the current status and future opportunities of in situ TEM is included. It also provides an introduction to the six articles covered by in this issue of MRS Bulletin explore the frontiers of in situ electron microscopy, including liquid and gas environmental TEM, dynamic four-dimensional TEM, nanomechanics, ferroelectric domain switching studied by in situ TEM, and state-of-the-art atomic imaging of light elements (i.e., carbon atoms) and individual defects.

  18. Collaborative Computational Project for Electron cryo-Microscopy

    SciTech Connect

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

    2015-01-01

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

  19. Amyloid Structure and Assembly: Insights from Scanning Transmission Electron Microscopy

    SciTech Connect

    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).

  20. A national facility for biological cryo-electron microscopy

    PubMed Central

    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. PMID:25615867

  1. A Technique for In Situ Ballistic Electron Emission Microscopy

    NASA Astrophysics Data System (ADS)

    Balsano, Robert; Garramone, John; Labella, Vincent

    2012-02-01

    Ballistic electron emission microscopy (BEEM) is a scanning tunneling microscopy (STM) technique that can measure transport of hot electrons through materials and interfaces with high spatial and energetic resolution. BEEM requires an additional contact to ground the metal base layer of a metal semiconductor junction. Performing BEEM in situ with the sample fabrication requires a custom built STM or modifying a commercial one to facilitate the extra contact, which leaves the technique to highly trained experts. This poster will describe our work to develop a special silicon substrate that has the extra contact built in to enable in situ BEEM without modifications to the STM. Electrically isolated contact traces are lithographically patterned ex situ onto the silicon substrate and connected to the BEEM sample plate which is then inserted into the ultra-high vacuum chamber. The metal is then deposited through a shadow mask and then mounted in situ onto the STM for BEEM measurements. BEEM measurements comparing both in situ and ex situ deposited films will be presented.

  2. Amyloid Structure and Assembly: Insights from Scanning Transmission Electron Microscopy

    PubMed Central

    Goldsbury, Claire; Baxa, Ulrich; Simon, Martha N.; Steven, Alasdair C.; Engel, Andreas; Wall, Joseph S.; Aebi, Ueli; Müller, Shirley A.

    2010-01-01

    Amyloid fibrils are filamentous protein aggregates implicated in several common diseases like Alzheimer’s disease and type II diabetes. Similar structures are also the molecular principle of the infectious spongiform encephalopathies like 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). PMID:20868754

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

    PubMed

    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.

  4. Amyloid structure and assembly: insights from scanning transmission electron microscopy.

    PubMed

    Goldsbury, Claire; Baxa, Ulrich; Simon, Martha N; Steven, Alasdair C; Engel, Andreas; Wall, Joseph S; Aebi, Ueli; Müller, Shirley 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). Copyright © 2010 Elsevier Inc. All rights reserved.

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

    PubMed

    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.

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

    NASA Astrophysics Data System (ADS)

    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.

  7. Relativistic effects in photon-induced near field electron microscopy.

    PubMed

    Park, Sang Tae; Zewail, Ahmed H

    2012-11-26

    Electrons and photons, when interacting via a nanostructure, produce a new way of imaging in space and time, termed photon-induced near field electron microscopy or PINEM [Barwick et al. Nature 2009, 462, 902]. The phenomenon was described by considering the evanescent field produced by the nanostructure, but quantification of the experimental results was achieved by solving the Schrödinger equation for the interaction of the three bodies. The question remained, is the nonrelativistic formulation sufficient for this description? Here, relativistic and nonrelativistic quantum mechanical formulations are compared for electron-photon interaction mediated by nanostructures, and it is shown that there is an exact equivalence for the two formulations. The nonrelativistic formulation was found to be valid in the relativistic regime when using in the former formulation the relativistically corrected velocity (and the corresponding values of momentum and energy). In the PINEM experiment, 200 keV electrons were utilized, giving the experimental (relativistically corrected) velocity to be 0.7c(v without relativistic correction is 0.885c). When this value (0.7c), together with those of the corresponding momentum (p(c) = mv) and energy (E(c) = (1/2)mv(2)), is used in the first order solution of the Schrödinger formulation, an exact equivalence is obtained.

  8. Contamination mitigation strategies for scanning transmission electron microscopy.

    PubMed

    Mitchell, D R G

    2015-06-01

    Modern scanning transmission electron microscopy (STEM) enables imaging and microanalysis at very high magnification. In the case of aberration-corrected STEM, atomic resolution is readily achieved. However, the electron fluxes used may be up to three orders of magnitude greater than those typically employed in conventional STEM. Since specimen contamination often increases with electron flux, specimen cleanliness is a critical factor in obtaining meaningful data when carrying out high magnification STEM. A range of different specimen cleaning methods have been applied to a variety of specimen types. The contamination rate has been measured quantitatively to assess the effectiveness of cleaning. The methods studied include: baking, cooling, plasma cleaning, beam showering and UV/ozone exposure. Of the methods tested, beam showering is rapid, experimentally convenient and very effective on a wide range of specimens. Oxidative plasma cleaning is also very effective and can be applied to specimens on carbon support films, albeit with some care. For electron beam-sensitive materials, cooling may be the method of choice. In most cases, preliminary removal of the bulk of the contamination by methods such as baking or plasma cleaning, followed by beam showering, where necessary, can result in a contamination-free specimen suitable for extended atomic scale imaging and analysis. Copyright © 2015 Elsevier Ltd. All rights reserved.

  9. Correlative Fluorescence Microscopy and Scanning Transmission Electron Microscopy of Quantum Dot Labeled Proteins in Whole Cells in Liquid

    PubMed Central

    Dukes, Madeline J.; Peckys, Diana B.; de Jonge, Niels

    2010-01-01

    Correlative fluorescence microscopy and transmission electron microscopy (TEM) is a state-of-the-art microscopy methodology to study cellular function, combining the functionality of light microscopy with the high resolution of electron microscopy. However, this technique involves complex sample preparation procedures due to its need for either thin sections or frozen samples for TEM imaging. Here, we introduce a novel correlative approach capable of imaging whole eukaryotic cells in liquid with fluorescence microscopy and with scanning transmission electron microscopy (STEM); there is no additional sample preparation necessary for the electron microscopy. Quantum dots (QDs) were bound to epidermal growth factor (EGF) receptors of COS7 fibroblast cells. Fixed whole cells in saline water were imaged with fluorescence microscopy and subsequently with STEM. The STEM images were correlated with fluorescence images of the same cellular regions. QDs of dimensions 7 × 12 nm were visible in a 5 μm thick layer of saline water, consistent with calculations. A spatial resolution of 3 nm was achieved on the QDs. PMID:20550177

  10. Correlative fluorescence microscopy and scanning transmission electron microscopy of quantum-dot-labeled proteins in whole cells in liquid.

    PubMed

    Dukes, Madeline J; Peckys, Diana B; de Jonge, Niels

    2010-07-27

    Correlative fluorescence microscopy and transmission electron microscopy (TEM) is a state-of-the-art microscopy methodology to study cellular function, combining the functionality of light microscopy with the high resolution of electron microscopy. However, this technique involves complex sample preparation procedures due to its need for either thin sections or frozen samples for TEM imaging. Here, we introduce a novel correlative approach capable of imaging whole eukaryotic cells in liquid with fluorescence microscopy and with scanning transmission electron microscopy (STEM); there is no additional sample preparation necessary for the electron microscopy. Quantum dots (QDs) were bound to epidermal growth factor (EGF) receptors of COS7 fibroblast cells. Fixed whole cells in saline water were imaged with fluorescence microscopy and subsequently with STEM. The STEM images were correlated with fluorescence images of the same cellular regions. QDs of dimensions 7x12 nm were visible in a 5 microm thick layer of saline water, consistent with calculations. A spatial resolution of 3 nm was achieved on the QDs.

  11. Electron Microscopy of Staphylococcus aureus Cell Wall Lysis

    PubMed Central

    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

  12. Optimized Negative-Staining Electron Microscopy for Lipoprotein Studies

    PubMed Central

    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

  13. Optimized negative-staining electron microscopy for lipoprotein studies.

    PubMed

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

    2013-01-01

    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. The most popular NS protocols that have been used to examine lipoprotein morphology and structure were reviewed. 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 1nm, but rarely better than 1nm) method which has been used to discover the mechanics of a small protein, 53kDa 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 14Å-resolution IgG antibody three-dimensional map. 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. Published by Elsevier B.V.

  14. Electron microscopy of Staphylococcus aureus cell wall lysis.

    PubMed

    Virgilio, R; González, C; Muñoz, N; Mendoza, S

    1966-05-01

    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.

  15. Transmission electron microscopy characterisation of 0-D nanomaterials

    NASA Astrophysics Data System (ADS)

    Turner, Stuart Matthew

    When materials are scaled down to the nanometre level, a change in physical behaviour is frequently observed. In so-called 0-D nanomaterials (nanoparticles), these unique nanoscale properties are most abundant and are usually linked to either a change in (electronic) structure of the material or to the dominating influence of the particle surface at the nanometre scale. In this doctoral work the nanoscale properties of several nanoparticle systems have been studied using advanced transmission electron microscopy (TEM). Every material that was studied required for its solution a unique approach and a host of transmission electron microscopy techniques. The title of this doctoral work can be freely translated as "retrieving quantitatively the maximal and most accurate chemical, structural and morphological information from nanoparticles by advanced transmission electron microscopy, to uncover and explain their unique properties". Chapter 1 gives a brief general introduction to the world of nanomaterials and nanotechnology in general and more specifically to 0-D nanomaterials (nanoparticles). The unique properties and potential applications of these materials are described. The production of 0-D nanomaterials is not covered in this chapter, as this is an extremely broad field to cover in only a few pages. Instead, the production method for each of the materials is left to the detailed chapters that follow. In Chapter 2 the main transmission electron microscopy techniques used to characterise the materials in the further chapters are described together with the microscopes used to perform these techniques and their parameters of operation. Again, the sample-specific setups are listed in the detailed chapters that follow. Chapter 3 covers all work carried out on luminescent detonation nanodiamond powder for drug delivery and bio-medical imaging applications. Specific attention is paid to the morphology, surface chemistry and nitrogen incorporation of detonation

  16. Light microscopy and scanning electron microscopy study on microstructure of gallbladder mucosa in pig.

    PubMed

    Prozorowska, Ewelina; Jackowiak, Hanna

    2015-03-01

    The present light microscopy (LM) and scanning electron microscopy (SEM) studies on porcine gallbladder mucosa provide a description of the microstructures of great functional importance such as mucosal folds, the epithelium, glands, and lymphatic nodules. The results showed the regional structural differences of the porcine gallbladder wall. Depending on the part of the gallbladder, three types of mucosal structures were described: simple and branched folds and mucosal crypts. An important structural feature found in the mucosa is connected with the structural variety of type of mucosal folds, which change from simple located in the neck, to most composed, i.e., branched or joined, in the polygonal crypts toward the fundus of the gallbladder. The morphometric analysis showed statistically significantly differences in the form and size of the folds and between the fundus, body, and neck of the gallbladder. Differences in the size of mucosal epithelium are discussed in terms of processes of synthesis and secretion of glycoproteins. Regional, species-specific differences in morphology of mucosal subepithelial glands, i.e., their secretory units and openings, and intensity of mucus secretion were described. Our results on the pig gallbladder show adaptation and/or specialization in particular areas of the mucosa for (1) secretion of mucus in the neck or body of gallbladder and (2) for cyclic volume changes, especially in the fundus of gallbladder. The description of the microstructures of mucosa in the porcine gallbladder could be useful as reference data for numerous experiments on the bile tract in the pig.

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

    NASA Astrophysics Data System (ADS)

    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.

  18. Cryo-electron microscopy of extracellular vesicles in fresh plasma.

    PubMed

    Yuana, Yuana; Koning, Roman I; Kuil, Maxim E; Rensen, Patrick C N; Koster, Abraham J; Bertina, Rogier M; Osanto, Susanne

    2013-12-31

    Extracellular vesicles (EV) are phospholipid bilayer-enclosed vesicles recognized as new mediators in intercellular communication and potential biomarkers of disease. They are found in many body fluids and mainly studied in fractions isolated from blood plasma in view of their potential in medicine. Due to the limitations of available analytical methods, morphological information on EV in fresh plasma is still rather limited. To image EV and determine the morphology, structure and size distribution in fresh plasma by cryo-electron microscopy (cryo-EM). Fresh citrate- and ethylenediaminetetraacetic acid (EDTA)-anticoagulated plasma or EV isolated from these plasmas were rapidly cryo-immobilized by vitrification and visualized by cryo-EM. EV isolated from fresh plasma were highly heterogeneous in morphology and size and mostly contain a discernible lipid bilayer (lipid vesicles). In fresh plasma there were 2 types of particles with a median diameter of 30 nm (25-260 nm). The majority of these particles are electron dense particles which most likely represent lipoproteins. The minority are lipid vesicles, either electron dense or electron lucent, which most likely represent EV. Lipid vesicles were occasionally observed in close proximity of platelets in citrate and EDTA-anticoagulated platelet-rich plasma. Cryo-electron tomography (cryo-ET) was employed to determine the 3D structure of platelet secretory granules. Cryo-EM is a powerful technique that enables the characterization of EV in fresh plasma revealing structural details and considerable morphological heterogeneity. Only a small proportion of the submicron structures in fresh plasma are lipid vesicles representing EV.

  19. Fluctuation Electron Microscopy of Amorphous and Polycrystalline Materials

    NASA Astrophysics Data System (ADS)

    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

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

    PubMed

    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.

  1. Three-Dimensional Scanning Transmission Electron Microscopy of Biological Specimens

    PubMed Central

    de Jonge, Niels; Sougrat, Rachid; Northan, Brian M.; Pennycook, Stephen J.

    2010-01-01

    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. PMID:20082729

  2. Three-dimensional scanning transmission electron microscopy of biological specimens

    SciTech Connect

    De Jonge, Niels; Sougrat, Rachid; Northan, Brian; Pennycook, Stephen J

    2010-01-01

    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 data set. The precision of the height determination was 0.2 nm. The resolution of the reconstruction is comparable to that achieved by tilt-series transmission electron microscopy (TEM). 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 data set.

  3. Arc Welders' pneumoconiosis: application of advanced scanning electron microscopy.

    PubMed

    Guidotti, T L; Abraham, J L; DeNee, P B; Smith, J R

    1978-01-01

    Study of lung tissue from necropsy of a 58-year-old arc welder with arc welders' pneumoconiosis, confirmed by history, chest radiography, and pathology, demonstrates the versatility and usefulness of new techniques in scanning electron microscopy (SEM). Secondary electron imaging, the most familiar SEM mode, showed heavy cellular infiltrates in alveoli, the interstitium, and within the interstices of loose whorled fibrotic nodules. Backscattered electron imaging, in which contrast is proportional to elemental atomic number, revealed intracellular metal particles not otherwise visible. Microprobe analysis, energy-dispersive x-ray spectrometry, mapped elemeental iron over the particle image and identified traces of silicon in the whorled nodules. Arc welders' pneumoconiosis appears to be more than a benign siderosis resulting from particulate iron deposition. Simultaneous exposure to other components of welding fumes may alter the pathologic picture, inducing a more complicated fibrotic reaction. The more recently developed advanced techniques of SEM are well suited to the study of pneumoconioses and other problems of heterogenous tissue and mixed chemical systems.

  4. Imaging electronic states on topological semimetals using scanning tunneling microscopy

    DOE PAGES

    Gyenis, András; Inoue, Hiroyuki; Jeon, Sangjun; ...

    2016-10-18

    Following the intense studies on topological insulators, significant efforts have recently been devoted to the search for gapless topological systems. These materials not only broaden the topological classification of matter but also provide a condensed matter realization of various relativistic particles and phenomena previously discussed mainly in high energy physics. Weyl semimetals host massless, chiral, low-energy excitations in the bulk electronic band structure, whereas a symmetry protected pair of Weyl fermions gives rise to massless Dirac fermions.Weemployed scanning tunneling microscopy/spectroscopy to explore the behavior of electronic states both on the surface and in the bulk of topological semimetal phases. Bymore » mapping the quasiparticle interference (QPI) and emerging Landau levels at high magnetic field in Dirac semimetals Cd3As2 and Na3Bi, we observed extended Dirac-like bulk electronic bands. QPI imaged on Weyl semimetal TaAs demonstrated the predicted momentum dependent delocalization of Fermi arc surface states in the vicinity of the surface projected Weyl nodes.« less

  5. Trends in the Electron Microscopy Data Bank (EMDB).

    PubMed

    Patwardhan, Ardan

    2017-06-01

    Recent technological advances, such as the introduction of the direct electron detector, have transformed the field of cryo-EM and the landscape of molecular and cellular structural biology. This study analyses these trends from the vantage point of the Electron Microscopy Data Bank (EMDB), the public archive for three-dimensional EM reconstructions. Over 1000 entries were released in 2016, representing almost a quarter of the total number of entries (4431). Structures at better than 6 Å resolution now represent one of the fastest-growing categories, while the share of annually released tomography-related structures is approaching 20%. The use of direct electron detectors is growing very rapidly: they were used for 70% of the structures released in 2016, in contrast to none before 2011. Microscopes from FEI have an overwhelming lead in terms of usage, and the use of the RELION software package continues to grow rapidly after having attained a leading position in the field. China is rapidly emerging as a major player in the field, supplementing the US, Germany and the UK as the big four. Similarly, Tsinghua University ranks only second to the MRC Laboratory for Molecular Biology in terms of involvement in publications associated with cryo-EM structures at better than 4 Å resolution. Overall, the numbers point to a rapid democratization of the field, with more countries and institutes becoming involved.

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

    PubMed

    de Jonge, Niels; Sougrat, Rachid; Northan, Brian M; Pennycook, Stephen J

    2010-02-01

    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.

  7. On mapping subangstrom electron clouds with force microscopy.

    PubMed

    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.

  8. Applications of inorganic nanoparticles in biological electron microscopy

    NASA Astrophysics Data System (ADS)

    Ni, Thomas Wentung

    Electron microscopy is an immensely powerful for imaging at the cellular level. However, many of the macromolecules of interest are difficult to image due to low electron density. There has been an immense body of work in order to visualize these macromolecules. In the past, many of the methods of visualization revolved around staining samples with heavy metals, however these stains are non-specific. In order to develop more specific methods of tagging macromolecules, there are two different methods to consider: the first being a top-down approach, in which electron dense tags, in this case inorganic nanoparticles, are given specific ligands to take advantage of different chemistries to attach these nanoparticles to macromolecules of interest. The second method is through a bottom-up approach where biomolecules are given the specific ability to form inorganic nanoparticles. Inorganic nanoparticles have been investigated with various ligands in order to enhance binding capability to macromolecules. The chief method of functionalizing these inorganic nanoparticles comes from ligand exchange; much has been studied regarding ligand exchange, but there are still many unanswered questions. Herein, we endeavor to reveal both the mechanism of exchange and the functional unit of exchange. We also report progress towards understanding an enzyme that is capable of forming inorganic nanoparticles, which could be cloned onto proteins as well. This bottom up style has been studied in several other groups; however, none of the previously reported methods have seen much use. Herein, we report a potential NADPH-dependent enzyme that forms selenium nanoparticles.

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

    PubMed

    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.

  10. Imaging electronic states on topological semimetals using scanning tunneling microscopy

    NASA Astrophysics Data System (ADS)

    Gyenis, András; Inoue, Hiroyuki; Jeon, Sangjun; Zhou, Brian B.; Feldman, Benjamin E.; Wang, Zhijun; Li, Jian; Jiang, Shan; Gibson, Quinn D.; Kushwaha, Satya K.; Krizan, Jason W.; Ni, Ni; Cava, Robert J.; Bernevig, B. Andrei; Yazdani, Ali

    2016-10-01

    Following the intense studies on topological insulators, significant efforts have recently been devoted to the search for gapless topological systems. These materials not only broaden the topological classification of matter but also provide a condensed matter realization of various relativistic particles and phenomena previously discussed mainly in high energy physics. Weyl semimetals host massless, chiral, low-energy excitations in the bulk electronic band structure, whereas a symmetry protected pair of Weyl fermions gives rise to massless Dirac fermions. We employed scanning tunneling microscopy/spectroscopy to explore the behavior of electronic states both on the surface and in the bulk of topological semimetal phases. By mapping the quasiparticle interference (QPI) and emerging Landau levels at high magnetic field in Dirac semimetals Cd3As2 and Na3Bi, we observed extended Dirac-like bulk electronic bands. QPI imaged on Weyl semimetal TaAs demonstrated the predicted momentum dependent delocalization of Fermi arc surface states in the vicinity of the surface-projected Weyl nodes.

  11. Robust image alignment for cryogenic transmission electron microscopy.

    PubMed

    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.

  12. Electron microscopy investigations of nanoparticles for cancer diagnostic applications

    NASA Astrophysics Data System (ADS)

    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

  13. An electron microscopy study of wear in polysilicon microelectromechanical systems.

    SciTech Connect

    Dugger, Michael Thomas; Enachescu, M.; Stach, Eric A.; Alsem, Daan Hein; Ritchie, Robert O.

    2005-02-01

    Wear is a critical factor in determining the durability of microelectromechanical systems (MEMS). While the reliability of polysilicon MEMS has received extensive attention, the mechanisms responsible for this failure mode at the microscale have yet to be conclusively determined. We have used on-chip polycrystalline silicon side-wall friction MEMS specimens to study active mechanisms during sliding wear in ambient air. Worn parts were examined by analytical scanning and transmission electron microscopy, while local temperature changes were monitored using advanced infrared microscopy. Observations show that small amorphous debris particles ({approx}50-100 nm) are removed by fracture through the silicon grains ({approx}500 nm) and are oxidized during this process. Agglomeration of such debris particles into larger clusters also occurs. Some of these debris particles/clusters create plowing tracks on the beam surface. A nano-crystalline surface layer ({approx}20-200 nm), with higher oxygen content, forms during wear at and below regions of the worn surface; its formation is likely aided by high local stresses. No evidence of dislocation plasticity or of extreme local temperature increases was found, ruling out the possibility of high temperature-assisted wear mechanisms.

  14. Automation in single-particle electron microscopy connecting the pieces.

    PubMed

    Lyumkis, Dmitry; Moeller, Arne; Cheng, Anchi; Herold, Amber; Hou, Eric; Irving, Christopher; Jacovetty, Erica L; Lau, Pick-Wei; Mulder, Anke M; Pulokas, James; Quispe, Joel D; Voss, Neil R; Potter, Clinton S; Carragher, Bridget

    2010-01-01

    Throughout the history of single-particle electron microscopy (EM), automated technologies have seen varying degrees of emphasis and development, usually depending upon the contemporary demands of the field. We are currently faced with increasingly sophisticated devices for specimen preparation, vast increases in the size of collected data sets, comprehensive algorithms for image processing, sophisticated tools for quality assessment, and an influx of interested scientists from outside the field who might lack the skills of experienced microscopists. This situation places automated techniques in high demand. In this chapter, we provide a generic definition of and discuss some of the most important advances in automated approaches to specimen preparation, grid handling, robotic screening, microscope calibrations, data acquisition, image processing, and computational infrastructure. Each section describes the general problem and then provides examples of how that problem has been addressed through automation, highlighting available processing packages, and sometimes describing the particular approach at the National Resource for Automated Molecular Microscopy (NRAMM). We contrast the more familiar manual procedures with automated approaches, emphasizing breakthroughs as well as current limitations. Finally, we speculate on future directions and improvements in automated technologies. Our overall goal is to present automation as more than simply a tool to save time. Rather, we aim to illustrate that automation is a comprehensive and versatile strategy that can deliver biological information on an unprecedented scale beyond the scope available with classical manual approaches. Copyright © 2010 Elsevier Inc. All rights reserved.

  15. FtsZ Condensates: An In Vitro Electron Microscopy Study

    PubMed Central

    Popp, David; Iwasa, Mitsusada; Narita, Akihiro; Erickson, Harold P.; Maéda, Yuichiro

    2009-01-01

    In vivo cell division protein FtsZ from E. coli forms rings and spirals which have only been observed by low resolution light microscopy. We show that these suprastructures are likely formed by molecular crowding which is a predominant factor in prokaryotic cells and enhances the weak lateral bonds between proto-filaments. Although FtsZ assembles into single proto-filaments in dilute aqueous buffer, with crowding agents above a critical concentration, it forms polymorphic supramolecular structures including rings and toroids (with multiple protofilaments) about 200 nm in diameter, similar in appearance to DNA toroids, and helices with pitches of several hundred nm as well as long, linear bundles. Helices resemble those observed in vivo, whereas the rings and toroids may represent a novel energy minimized state of FtsZ, at a later stage of Z-ring constriction. We shed light on the molecular arrangement of FtsZ filaments within these suprastructures using high resolution electron microscopy. PMID:19137575

  16. Utility of Transmission Electron Microscopy in Small Round Cell Tumors

    PubMed Central

    Kim, Na Rae; Ha, Seung Yeon; Cho, Hyun Yee

    2015-01-01

    Small round cell tumors (SRCTs) are a heterogeneous group of neoplasms composed of small, primitive, and undifferentiated cells sharing similar histology under light microscopy. SRCTs include Ewing sarcoma/peripheral neuroectodermal tumor family tumors, neuroblastoma, desmoplastic SRCT, rhabdomyosarcoma, poorly differentiated round cell synovial sarcoma, mesenchymal chondrosarcoma, small cell osteosarcoma, small cell malignant peripheral nerve sheath tumor, and small cell schwannoma. Non-Hodgkin’s malignant lymphoma, myeloid sarcoma, malignant melanoma, and gastrointestinal stromal tumor may also present as SRCT. The current shift towards immunohistochemistry and cytogenetic molecular techniques for SRCT may be inappropriate because of antigenic overlapping or inconclusive molecular results due to the lack of differentiation of primitive cells and unavailable genetic service or limited moleculocytogenetic experience. Although usage has declined, electron microscopy (EM) remains very useful and shows salient features for the diagnosis of SRCTs. Although EM is not always required, it provides reliability and validity in the diagnosis of SRCT. Here, the ultrastructural characteristics of SRCTs are reviewed and we suggest that EM would be utilized as one of the reliable modalities for the diagnosis of undifferentiated and poorly differentiated SRCTs. PMID:25812730

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

    NASA Astrophysics Data System (ADS)

    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 .

  18. Variability of Protein Structure Models from Electron Microscopy.

    PubMed

    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.

  19. Watershed Merge Tree Classification for Electron Microscopy Image Segmentation

    SciTech Connect

    Liu, TIng; Jurrus, Elizabeth R.; Seyedhosseini, Mojtaba; Ellisman, Mark; Tasdizen, Tolga

    2012-11-11

    Automated segmentation of electron microscopy (EM) images is a challenging problem. In this paper, we present a novel method that utilizes a hierarchical structure and boundary classification for 2D neuron segmentation. With a membrane detection probability map, a watershed merge tree is built for the representation of hierarchical region merging from the watershed algorithm. A boundary classifier is learned with non-local image features to predict each potential merge in the tree, upon which merge decisions are made with consistency constraints in the sense of optimization to acquire the final segmentation. Independent of classifiers and decision strategies, our approach proposes a general framework for efficient hierarchical segmentation with statistical learning. We demonstrate that our method leads to a substantial improvement in segmentation accuracy.

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

    NASA Astrophysics Data System (ADS)

    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.

  1. Temperature Calibration for In Situ Environmental Transmission Electron Microscopy Experiments

    PubMed Central

    Winterstein, JP; Lin, PA; Sharma, R

    2016-01-01

    In situ environmental transmission electron microscopy (ETEM) experiments require specimen heating holders to study material behavior in gaseous environments at elevated temperatures. In order to extract meaningful kinetic parameters, such as activation energies, it is essential to have a direct and accurate measurement of local sample temperature. This is particularly important if the sample temperature might fluctuate, for example when room temperature gases are introduced to the sample area. Using selected-area diffraction (SAD) in an ETEM, the lattice parameter of Ag nanoparticles was measured as a function of the temperature and pressure of hydrogen gas to provide a calibration of the local sample temperature. SAD permits measurement of temperature to an accuracy of ± 30 °C using Ag lattice expansion. Gas introduction can cause sample cooling of several hundred degrees celsius for gas pressures achievable in the ETEM. PMID:26441334

  2. In situ transmission electron microscopy of cadmium selenide nanorod sublimation

    SciTech Connect

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

    2015-01-23

    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. Furthermore, 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.

  3. Photoacoustic microscopy of electronic acupuncture (EA) effect in small animals.

    PubMed

    Yang, Jinge; Wu, Dan; Tang, Yong; Jiang, Huabei

    2017-02-01

    Acupuncture has been an effective treatment for various pain in China for several thousand years. However, the mechanisms underlying this mysterious ancient healing are still largely unknown. Here we applied photoacoustic microscopy (PAM) to investigate brain hemodynamic changes in response to electronic acupuncture (EA) at ST36 (Zusanli). Due to the high optical absorption of blood at 532 nm, PAM could sensitively probe changes in hemoglobin concentration (HbT, i.e., cerebral blood volume [CBV]) of cortical regions in high resolution. Six healthy mice were stimulated at the acupoint and three healthy mice were stimulated at sham points. Remarkable CBV changes in sensorimotor and retrosplenial agranular cortex were observed. Results showed the potential of PAM as a visualization tool to study the acupuncture effect on brain hemodynamics in animal models. (a) Schematic showing the stimulation points. (b) B-scan images overlaid with mouse atlas. (c) & (d) Statistical results of CBV changes from cortical regions.

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

    NASA Astrophysics Data System (ADS)

    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.

  5. Electron microscopy of gallium nitride growth on polycrystalline diamond

    NASA Astrophysics Data System (ADS)

    Webster, R. F.; Cherns, D.; Kuball, M.; Jiang, Q.; Allsopp, D.

    2015-11-01

    Transmission and scanning electron microscopy were used to examine the growth of gallium nitride (GaN) on polycrystalline diamond substrates grown by metalorganic vapour phase epitaxy with a low-temperature aluminium nitride (AlN) nucleation layer. Growth on unmasked substrates was in the (0001) orientation with threading dislocation densities ≈7 × 109 cm-2. An epitaxial layer overgrowth technique was used to reduce the dislocation densities further, by depositing silicon nitride stripes on the surface and etching the unmasked regions down to the diamond substrate. A re-growth was then performed on the exposed side walls of the original GaN growth, reducing the threading dislocation density in the overgrown regions by two orders of magnitude. The resulting microstructures and the mechanisms of dislocation reduction are discussed.

  6. In situ transmission electron microscopy of cadmium selenide nanorod sublimation

    DOE PAGES

    Hellebusch, Daniel J.; Manthiram, Karthish; Beberwyck, Brandon J.; ...

    2015-01-23

    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. Furthermore, we propose a mechanism that accounts for themore » observed sublimation behavior based on the terrace–ledge–kink (TLK) model and how the nanorod surface chemical environment influences the kinetic barrier of sublimation.« less

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

    PubMed

    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.

  8. Fluctuation electron microscopy studies of complex structured materials

    NASA Astrophysics Data System (ADS)

    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.

  9. Advanced analytical electron microscopy for alkali-ion batteries

    SciTech Connect

    Qian, Danna; Ma, Cheng; Meng, Ying Shirley; More, Karren; Chi, Miaofang

    2015-01-01

    Lithium-ion batteries are a leading candidate for electric vehicle and smart grid applications. However, further optimizations of the energy/power density, coulombic efficiency and cycle life are still needed, and this requires a thorough understanding of the dynamic evolution of each component and their synergistic behaviors during battery operation. With the capability of resolving the structure and chemistry at an atomic resolution, advanced analytical transmission electron microscopy (AEM) is an ideal technique for this task. The present review paper focuses on recent contributions of this important technique to the fundamental understanding of the electrochemical processes of battery materials. A detailed review of both static (ex situ) and real-time (in situ) studies will be given, and issues that still need to be addressed will be discussed.

  10. SILVER IMPREGNATION OF ULTRATHIN SECTIONS FOR ELECTRON MICROSCOPY

    PubMed Central

    Marinozzi, Vittorio

    1961-01-01

    A new procedure is described for silver impregnation of thin sections for electron microscopy. Sections of various tissues, fixed in OsO4 and embedded in methacrylate, were treated with an ammoniacal silver solution, directly or after oxidation with periodic acid or hydrogen peroxide. After OsO4 fixation all cellular membranous systems exhibit a primary argentaffinity probably due to the reduction of ammoniacal silver solution by the reduced osmium bound to unsaturated lipids. Bleaching the sections with hydrogen peroxide removes the argentaffinity of protoplasmic structures. Treatment of the sections with periodic acid results in decreased argentaffinity of protoplasmic components while the argentaffinity of metaplasmic structures is greatly enhanced. The latter procedure appears particularly useful for enhancing the contrast of basement membranes. PMID:13766855

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

    PubMed

    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.

  12. Small minded - The characterization of interplanetary dust by electron microscopy

    NASA Astrophysics Data System (ADS)

    Pillinger, C. T.

    1981-12-01

    The collection and analysis of Brownlee particles, interplanetary dust found in the atmosphere, are discussed. The particles are usually around 10 microns in diameter and have slowed to terminal velocity at altitudes near 100 km, having been heated to 550 C for not more than two seconds. Electron microscopy and neutron activation analysis have been employed, noting that the samples are collected by NASA from regions of concentration of one particle per 1000 cu cm. Chondritic compositions have been observed, although inter- and intra-grain abundance ratios do not display a distinct chondritic grouping. Single and polycrystal diffraction studies have identified magnetite, pyroxene, olivine, pyrohotite, taenite, and cohenite. The possibility that the dust particles are of cometary origin will be examined with data from the ESA Halley mission.

  13. Interfacing Microfluidics with Negative Stain Transmission Electron Microscopy

    PubMed Central

    Mukhitov, Nikita; Spear, John M.; Stagg, Scott M.; Roper, Michael G.

    2016-01-01

    A microfluidic platform is presented for preparing negatively stained grids for use in transmission electron microscopy (EM). The microfluidic device is composed of glass etched with readily fabricated features that facilitate the extraction of the grid post-staining and maintains the integrity of the sample. Utilization of this device simultaneously reduced environmental contamination on the grids and improved the homogeneity of the heavy metal stain needed to enhance visualization of biological specimens as compared to conventionally prepared EM grids. This easy-to-use EM grid preparation device provides the basis for future developments of systems with more integrated features, which will allow for high throughput and dynamic structural biology studies. PMID:26642355

  14. Field-electron emission microscopy of carbon-saturated rhenium

    NASA Astrophysics Data System (ADS)

    Bernatskii, D. P.; Pavlov, V. G.

    2017-06-01

    The growth of carbon structures on the surface of a rhenium point field emitter has been studied by field electron microscopy (FEM). It is established that graphene formation takes place on close-packed crystal faces of rhenium and leads to decrease in their work function. For rhenium exposed in benzene vapors, the formation of graphene islands requires a much longer time than that for iridium. Heating of carbon-saturated rhenium point field emitter up to temperatures close to its melting point with subsequent cooling does not lead to changes in the work function and FEM image of the emitter surface. The observed phenomena are explained by high solubility of carbon in rhenium.

  15. Scanning SQUID microscopy with single electron spin sensitivity

    NASA Astrophysics Data System (ADS)

    Vasyukov, Denis

    2014-03-01

    Superconducting interference devices (SQUIDs) have been traditionally used for studying fundamental properties of magnetic materials and superconductors. Although widely used in scanning magnetic microscopy, their progress towards detection of small magnetic moments was stagnating of late due to limitations imposed by conventional designs of planar SQUIDs and contemporary lithography techniques, restricting sample-to-sensor distance smaller than ~ 0.5 micron and SQUIDs diameters smaller than ~ 200 nm. These limitations were overcome by the invention of a SQUID-on-tip device, subsequent realization of a SQUID-on-tip microscope, and by creation of an ultra-small sensor with spatial resolution of 20 nm and sensitivity to a single electron spin per 1 Hz bandwidth. In this talk I will describe the principles of scanning SQUID magnetometry, its applications to study superconductors and its potential for magnetic nano-scale imaging of novel materials.

  16. Transmission electron microscopy analysis of hydroxyapatite nanocrystals from cattle bones

    SciTech Connect

    Patel, Sangeeta; Wei, Shanghai; Han, Jie; Gao, Wei

    2015-11-15

    In this present study, hydroxyapatite which was obtained from cattle bones has been heat treated at temperature 400 °C and 600 °C. The microstructure after the treatment has been studied in detail using Transmission electron microscopy (TEM) and X-ray diffraction techniques. The TEM results indicate that natural bone consists of collagen and hydroxyapatite nano-crystals which are needle shaped. The heat treatment influences the crystallinity and growth of these hydroxyapatite nano-crystals known as ‘crystal maturation’ or ‘crystal ageing’. - Highlights: • Hydroxyapatite is obtained from cattle bones. • Material has been characterised using XRD and TEM. • Crystal growth and orientation has been studied in detail.

  17. Electron microscopy of Nd-Fe-B based magnets

    NASA Astrophysics Data System (ADS)

    Fidler, J.; Knoch, K. G.

    1989-08-01

    Transmission electron microscopy has been used widely to characterize the complex multiphase microstructure of Nd 2Fe 14B based permanent magnets. The grain size of the magnets strongly depends on the processing technique. Our study of sintered magnets revealed two types of grain boundaries: one containing no intergranular phase between hard magnetic grains and one composed of nonmagnetic Nd-rich phases. In doped sintered magnets the dopant is dissolved in the hard magnetic φ-phase. In the case where the solubility of the dopant is low at the sintering temperature (Nb, Mo, Zr), precipitates are formed within the φ-phase. Dopants also form new intergranular phases and influence the wetting of the liquid phase and the smoothness of the surface of the φ-grains during sintering and therefore affect the coercivity.

  18. Simultaneous orientation and thickness mapping in transmission electron microscopy

    DOE PAGES

    Tyutyunnikov, Dmitry; Özdöl, V. Burak; Koch, Christoph T.

    2014-12-04

    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 comparedmore » to those of other techniques available.« less

  19. Annular dark field transmission electron microscopy for protein structure determination.

    PubMed

    Koeck, Philip J B

    2016-02-01

    Recently annular dark field (ADF) transmission electron microscopy (TEM) has been advocated as a means of recording images of biological specimens with better signal to noise ratio (SNR) than regular bright field images. I investigate whether and how such images could be used to determine the three-dimensional structure of proteins given that an ADF aperture with a suitable pass-band can be manufactured and used in practice. I develop an approximate theory of ADF-TEM image formation for weak amplitude and phase objects and test this theory using computer simulations. I also test whether these simulated images can be used to calculate a three-dimensional model of the protein using standard software and discuss problems and possible ways to overcome these.

  20. A Primer to Single-Particle Cryo-Electron Microscopy

    PubMed Central

    Cheng, Yifan; Grigorieff, Nikolaus; Penczek, Pawel A.; Walz, Thomas

    2015-01-01

    Summary Cryo-electron microscopy (cryo-EM) of single-particle specimens is used to determine the structure of proteins and macromolecular complexes without the need for crystals. Recent advances in detector technology and software algorithms now allow images of unprecedented quality to be recorded and structures to be determined at near-atomic resolution. However, compared with X-ray crystallography, cryo-EM is a young technique with distinct challenges. This primer explains the different steps and considerations involved in structure determination by single-particle cryo-EM to provide an overview for scientists wishing to understand more about this technique and the interpretation of data obtained with it, as well as a starting guide for new practitioners. PMID:25910204

  1. Advanced analytical electron microscopy for alkali-ion batteries

    DOE PAGES

    Qian, Danna; Ma, Cheng; Meng, Ying Shirley; ...

    2015-01-01

    Lithium-ion batteries are a leading candidate for electric vehicle and smart grid applications. However, further optimizations of the energy/power density, coulombic efficiency and cycle life are still needed, and this requires a thorough understanding of the dynamic evolution of each component and their synergistic behaviors during battery operation. With the capability of resolving the structure and chemistry at an atomic resolution, advanced analytical transmission electron microscopy (AEM) is an ideal technique for this task. The present review paper focuses on recent contributions of this important technique to the fundamental understanding of the electrochemical processes of battery materials. A detailed reviewmore » of both static (ex situ) and real-time (in situ) studies will be given, and issues that still need to be addressed will be discussed.« less

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

    PubMed Central

    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 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. PMID:27896013

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

    PubMed

    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.

  4. Electron Microscopy Analysis of the Nucleolus of Trypanosoma cruzi

    NASA Astrophysics Data System (ADS)

    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 experimentation of nanostructured materials

    NASA Astrophysics Data System (ADS)

    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.

  6. Characterization of protein immobilization on nanoporous gold using atomic force microscopy and scanning electron microscopy

    NASA Astrophysics Data System (ADS)

    Tan, Yih Horng; Schallom, John R.; Ganesh, N. Vijaya; Fujikawa, Kohki; Demchenko, Alexei V.; Stine, Keith J.

    2011-08-01

    Nanoporous gold (NPG), made by dealloying low carat gold alloys, is a relatively new nanomaterial finding application in catalysis, sensing, and as a support for biomolecules. NPG has attracted considerable interest due to its open bicontinuous structure, high surface-to-volume ratio, tunable porosity, chemical stability and biocompatibility. NPG also has the attractive feature of being able to be modified by self-assembled monolayers. Here we use scanning electron microscopy (SEM) and atomic force microscopy (AFM) to characterize a highly efficient approach for protein immobilization on NPG using N-hydroxysuccinimide (NHS) ester functionalized self-assembled monolayers on NPG with pore sizes in the range of tens of nanometres. Comparison of coupling under static versus flow conditions suggests that BSA (Bovine Serum Albumin) and IgG (Immunoglobulin G) can only be immobilized onto the interior surfaces of free standing NPG monoliths with good coverage under flow conditions. AFM is used to examine protein coverage on both the exterior and interior of protein modified NPG. Access to the interior surface of NPG for AFM imaging is achieved using a special procedure for cleaving NPG. AFM is also used to examine BSA immobilized on rough gold surfaces as a comparative study. In principle, the general approach described should be applicable to many enzymes, proteins and protein complexes since both pore sizes and functional groups present on the NPG surfaces are controllable.Nanoporous gold (NPG), made by dealloying low carat gold alloys, is a relatively new nanomaterial finding application in catalysis, sensing, and as a support for biomolecules. NPG has attracted considerable interest due to its open bicontinuous structure, high surface-to-volume ratio, tunable porosity, chemical stability and biocompatibility. NPG also has the attractive feature of being able to be modified by self-assembled monolayers. Here we use scanning electron microscopy (SEM) and atomic force

  7. Electron Diffraction and High-Resolution Electron Microscopy of Mineral Structures

    NASA Astrophysics Data System (ADS)

    Nord, Gordon L., Jr.

    This book is a well-written English translation of the original 1981 Russian edition, Strukturnoye issledovaniye mineralov metodami mikrodifraktsii i elechtronnoi mikroskopii vysokogo razresheniya. The 1987 English version has been extensively updated and includes references up to 1986. The book is essentially a text on the theoretical and experimental aspects of transmission electron microscopy and has chapters on the reciprocal lattice, electron diffraction (both kinematic and dynamic), and high-resolution electron microscopy.Electron diffraction is emphasized, especially its use for structure analysis of poorly crystalline and fine-grained phases not readily determined by the more exact X ray diffraction method. Two methods of electron diffraction are discussed: selected area electron diffraction (SAED) and oblique-texture electron diffraction (OTED); the latter technique is rarely used in the west and is never discussed in western electron microscopy texts. A SAED pattern is formed by isolating a small micrometer-size area with an aperture and obtaining single-crystal patterns from the diffracted beams. By tilting the sample and obtaining many patterns, a complete picture of the reciprocal lattice can be taken. An OTED pattern is formed when the incident electron beam passes through an inclined preparation consisting of a great number of thin platy crystals lying normal to the texture axis (axis normal to the support grid). To form an OTED pattern, the plates must all lie on a common face, such as a basal plane in phyllosilicates. Upon tilting the plates, an elliptical powder diffraction pattern is formed. Intensities measured from these patterns are used for a structural analysis of the platy minerals.

  8. Microstructure of Mixed Surfactant Solutions by Electron Microscopy

    NASA Astrophysics Data System (ADS)

    Naranjo, Edward

    1995-01-01

    Surfactant mixtures add a new dimension to the design of complex fluid microstructure. By combining different surfactants it is not only possible to modify aggregate morphology and control the macrascopic properties of colloidal dispersions but also to produce a variety of novel synergistic phases. Mixed systems produce new microstructures by altering the intermolecular and interaggregate forces in ways impossible for single component systems. In this dissertation, we report on the phase behavior and microstructure of several synthetic and biological surfactant mixtures as elucidated by freeze-fracture and cryo-transmission electron microscopy. We have discovered that stable, spontaneous unilamellar vesicles can be prepared from aqueous mixtures of commercially available single-tailed cationic and anionic surfactants. Vesicle stability is determined by the length and volume of the hydrocarbon chains of the "catanionic" pairs. Mixtures containing bulky or branched surfactant pairs (C _{16}/C_{12 -14}) in water produce defect-free fairly monodisperse equilibrium vesicles at high dilution. In contrast, mixtures of catanionic surfactants with highly asymmetric tails (C_{16}/C_8 ) form phases of porous vesicles, dilute lamellar L_{alpha}, and anomalous isotropic L_3 phases. Images of the microstructure by freeze-fracture microscopy show that the L_3 phase consists of multiconnected self-avoiding bilayers with saddle shaped curvature. The forces between bilayers of vesicle-forming cationic and anionic surfactant mixtures were also measured using the Surface Force Apparatus (SFA). We find that the vesicles are stabilized by a long range electrostatic repulsion at large separations (>20 A) and an additional salt-independent repulsive force below 20 A. The measured forces correlate very well with the ternary phase diagram and the vesicle microstructures observed by electron microscopy. In addition to studying ionic surfactants, we have also done original work with

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

    SciTech Connect

    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.

  10. Thin dielectric film thickness determination by advanced transmission electron microscopy

    SciTech Connect

    Diebold, A.C.; Foran, B.; Kisielowski, C.; Muller, D.; Pennycook, S.; Principe, E.; Stemmer, S.

    2003-09-01

    High Resolution Transmission Electron Microscopy (HR-TEM) has been used as the ultimate method of thickness measurement for thin films. The appearance of phase contrast interference patterns in HR-TEM images has long been confused as the appearance of a crystal lattice by non-specialists. Relatively easy to interpret crystal lattice images are now directly observed with the introduction of annular dark field detectors for scanning TEM (STEM). With the recent development of reliable lattice image processing software that creates crystal structure images from phase contrast data, HR-TEM can also provide crystal lattice images. The resolution of both methods was steadily improved reaching now into the sub Angstrom region. Improvements in electron lens and image analysis software are increasing the spatial resolution of both methods. Optimum resolution for STEM requires that the probe beam be highly localized. In STEM, beam localization is enhanced by selection of the correct aperture. When STEM measurement is done using a highly localized probe beam, HR-TEM and STEM measurement of the thickness of silicon oxynitride films agree within experimental error. In this paper, the optimum conditions for HR-TEM and STEM measurement are discussed along with a method for repeatable film thickness determination. The impact of sample thickness is also discussed. The key result in this paper is the proposal of a reproducible method for film thickness determination.

  11. High Resolution Transmission Electron Microscopy (HRTEM) of nanophase ferric oxides

    NASA Technical Reports Server (NTRS)

    Golden, D. C.; Morris, R. V.; Ming, D. W.; Lauer, H. V., Jr.

    1994-01-01

    Iron oxide minerals are the prime candidates for Fe(III) signatures in remotely sensed Martian surface spectra. Magnetic, Mossbauer, and reflectance spectroscopy have been carried out in the laboratory in order to understand the mineralogical nature of Martian analog ferric oxide minerals of submicron or nanometer size range. Out of the iron oxide minerals studied, nanometer sized ferric oxides are promising candidates for possible Martian spectral analogs. 'Nanophase ferric oxide (np-Ox)' is a generic term for ferric oxide/oxihydroxide particles having nanoscale (less than 10 nm) particle dimensions. Ferrihydrite, superparamagnetic particles of hematite, maghemite and goethite, and nanometer sized particles of inherently paramagnetic lepidocrocite are all examples of nanophase ferric oxides. np-Ox particles in general do not give X-ray diffraction (XRD) patterns with well defined peaks and would often be classified as X-ray amorphous. Therefore, different np-Oxs preparations should be characterized using a more sensitive technique e.g., high resolution transmission electron microscopy (HRTEM). The purpose of this study is to report the particle size, morphology and crystalline order, of five np-Ox samples by HRTEM imaging and electron diffraction (ED).

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

    SciTech Connect

    Tamer Vestlund, A.; Al-Ashaab, R.; Tyrrel, S.F.; Longhurst, P.J.; Pollard, S.J.T.; Drew, G.H.

    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.

  13. ATOMIC RESOLUTION CRYO ELECTRON MICROSCOPY OF MACROMOLECULAR COMPLEXES

    PubMed Central

    ZHOU, Z. HONG

    2013-01-01

    Single-particle cryo electron microscopy (cryoEM) is a technique for determining three-dimensional (3D) structures from projection images of molecular complexes preserved in their “native,” noncrystalline state. Recently, atomic or near-atomic resolution structures of several viruses and protein assemblies have been determined by single-particle cryoEM, allowing ab initio atomic model building by following the amino acid side chains or nucleic acid bases identifiable in their cryoEM density maps. In particular, these cryoEM structures have revealed extended arms contributing to molecular interactions that are otherwise not resolved by the conventional structural method of X-ray crystallography at similar resolutions. High-resolution cryoEM requires careful consideration of a number of factors, including proper sample preparation to ensure structural homogeneity, optimal configuration of electron imaging conditions to record high-resolution cryoEM images, accurate determination of image parameters to correct image distortions, efficient refinement and computation to reconstruct a 3D density map, and finally appropriate choice of modeling tools to construct atomic models for functional interpretation. This progress illustrates the power of cryoEM and ushers it into the arsenal of structural biology, alongside conventional techniques of X-ray crystallography and NMR, as a major tool (and sometimes the preferred one) for the studies of molecular interactions in supramolecular assemblies or machines. PMID:21501817

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

    PubMed Central

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

    2016-01-01

    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. PMID:27849001

  15. ELECTRON MICROSCOPY OF STRUCTURAL DETAIL IN FROZEN BIOLOGICAL SPECIMENS

    PubMed Central

    Steere, Russell L.

    1957-01-01

    A procedure is described whereby preshadowed replicas can be obtained from frozen biological specimens which have been cut and then etched by sublimation of the ice from their surfaces. Electron micrographs showing details of the internal structure of plant virus crystals are presented to demonstrate the values of the procedure. Crystals of purified tobacco ringspot virus and squash mosaic virus and some portions of turnip yellow mosaic virus crystals have been shown to exhibit hexagonal packing. Sections through in situ crystals of tobacco mosaic virus show the rods to be parallel within each layer and arranged in a square net as viewed end on. Individual rods in each layer of the latter measure 300 mµ in length and are somewhat tilted with respect to the rods of adjacent layers. This results in the formation of a herring-bone appearance when a crystal is cut perpendicular to its hexagonal face. It is suggested that the procedure outlined here might well serve to supplement other procedures for the preparation of many cytological specimens for electron microscopy. PMID:13416310

  16. Analytical electron microscopy in mineralogy; exsolved phases in pyroxenes

    USGS Publications Warehouse

    Nord, G.L.

    1982-01-01

    Analytical scanning transmission electron microscopy has been successfully used to characterize the structure and composition of lamellar exsolution products in pyroxenes. At operating voltages of 100 and 200 keV, microanalytical techniques of x-ray energy analysis, convergent-beam electron diffraction, and lattice imaging have been used to chemically and structurally characterize exsolution lamellae only a few unit cells wide. Quantitative X-ray energy analysis using ratios of peak intensities has been adopted for the U.S. Geological Survey AEM in order to study the compositions of exsolved phases and changes in compositional profiles as a function of time and temperature. The quantitative analysis procedure involves 1) removal of instrument-induced background, 2) reduction of contamination, and 3) measurement of correction factors obtained from a wide range of standard compositions. The peak-ratio technique requires that the specimen thickness at the point of analysis be thin enough to make absorption corrections unnecessary (i.e., to satisfy the "thin-foil criteria"). In pyroxenes, the calculated "maximum thicknesses" range from 130 to 1400 nm for the ratios Mg/Si, Fe/Si, and Ca/Si; these "maximum thicknesses" have been contoured in pyroxene composition space as a guide during analysis. Analytical spatial resolutions of 50-100 nm have been achieved in AEM at 200 keV from the composition-profile studies, and analytical reproducibility in AEM from homogeneous pyroxene standards is ?? 1.5 mol% endmember. ?? 1982.

  17. Combined scanning transmission electron microscopy tilt- and focal series.

    PubMed

    Dahmen, Tim; Baudoin, Jean-Pierre; Lupini, Andrew R; Kübel, Christian; Slusallek, Philipp; de Jonge, Niels

    2014-04-01

    In this study, a combined tilt- and focal series is proposed as a new recording scheme for high-angle annular dark-field scanning transmission electron microscopy (STEM) tomography. Three-dimensional (3D) data were acquired by mechanically tilting the specimen, and recording a through-focal series at each tilt direction. The sample was a whole-mount macrophage cell with embedded gold nanoparticles. The tilt-focal algebraic reconstruction technique (TF-ART) is introduced as a new algorithm to reconstruct tomograms from such combined tilt- and focal series. The feasibility of TF-ART was demonstrated by 3D reconstruction of the experimental 3D data. The results were compared with a conventional STEM tilt series of a similar sample. The combined tilt- and focal series led to smaller "missing wedge" artifacts, and a higher axial resolution than obtained for the STEM tilt series, thus improving on one of the main issues of tilt series-based electron tomography.

  18. The characterization of nanoparticles using analytical electron microscopy

    NASA Astrophysics Data System (ADS)

    Hill, Whitney B.

    2011-06-01

    Nanoparticles are often overlooked during routine trace evidence analyses because of their small size and the degree of difficulty needed to efficiently characterize them. However, analytical electron microscopy (AEM) enables the characterization and/or identification of nanoparticles because of its high magnification capability, the ability to gather elemental data and also the ability to determine the internal structure of a single nanoparticles(1). There is a wide variety of natural and manufactured nanoparticles that are prominent within the environment and their presence becomes very valuable in the absence of larger particles. The combustion of materials produces by-products such as nano-sized carbon soot, fumes, fly ash and gun-shot residue (GSR). Using AEM, nano-sized carbon soot, fumes, fly ash and GSR can not only be distinguished from other nanoparticles within the environment but can also be distinguished from each other because of differences in morphology, elemental composition, and internal structure. The elemental information gathered from combustion by-products during AEM analysis can also give an indication of the original source material. Other nanoparticles such as paint pigments and fillers can also be characterized by AEM using morphology, electron diffraction and elemental composition.

  19. Atomic-scale electron microscopy at ambient pressure.

    PubMed

    Creemer, J F; Helveg, S; Hoveling, G H; Ullmann, S; Molenbroek, A M; Sarro, P M; Zandbergen, H W

    2008-08-01

    We demonstrate a novel nanoreactor for performing atomic-resolution environmental transmission electron microscopy (ETEM) of nanostructured materials during exposure to gases at ambient pressures and elevated temperatures. The nanoreactor is a microelectromechanical system (MEMS) and is functionalized with a micrometer-sized gas-flow channel, electron-transparent windows and a heating device. It fits into the tip of a dedicated sample holder that can be used in a normal CM microscope of Philips/FEI Company. The nanoreactor performance was demonstrated by ETEM imaging of a Cu/ZnO catalyst for methanol synthesis during exposure to hydrogen. Specifically, the nanoreactor facilitated the direct observation of Cu nanocrystal growth and mobility on a sub-second time scale during heating to 500 degrees C and exposure to 1.2 bar of H(2). For the same gas reaction environment, ETEM images show atomic lattice fringes in the Cu nanocrystals with spacing of 0.18 nm, attesting the spatial resolution limit of the system. The nanoreactor concept opens up new possibilities for in situ studies of nanomaterials and the ways they interact with their ambient working environment in diverse areas, such as heterogeneous catalysis, electrochemistry, nanofabrication, materials science and biology.

  20. Preparation of macromolecular complexes for cryo-electron microscopy.

    PubMed

    Grassucci, Robert A; Taylor, Derek J; Frank, Joachim

    2007-01-01

    This protocol describes the preparation of frozen-hydrated single-particle specimens of macromolecular complexes. First, it describes how to create a grid surface coated with holey carbon by first inducing holes in a Formvar film to act as a template for the holey carbon that is stable under cryo-electron microscopy (cryo-EM) conditions and is sample-friendly. The protocol then describes the steps required to deposit the homogeneous sample on the grid and to plunge-freeze the grid into liquid ethane at the temperature of liquid nitrogen, so that it is suitable for cryo-EM visualization. It takes 4-5 h to make several hundred holey carbon grids and about 1 h to make the frozen-hydrated grids. The time required for sample purification varies from hours to days, depending on the sample and the specific procedure required. A companion protocol details how to collect cryo-EM data using an FEI Tecnai transmission electron microscope that can subsequently be processed to obtain a three-dimensional reconstruction of the macromolecular complex.

  1. Disclosing dark mode of femtosecond plasmon with photoemission electron microscopy

    NASA Astrophysics Data System (ADS)

    Ji, Boyu; Wang, Qian; Song, Xiaowei; Tao, Haiyan; Dou, Yinping; Gao, Xun; Hao, Zuoqiang; Lin, Jingquan

    2017-10-01

    The plasmonics dark mode of metal nanorings has potential in the field of e.g. sensing and surface-enhanced Raman spectroscopy (SERS). Most of the investigations on the plasmonics dark mode in a nanoring have been on far-field spectroscopy or near field simulations so far. In this paper, the near-field distribution of the femtosecond dark mode plasmon on an individual gold nanoring is mapped non-invasively using photoemission electron microscopy (PEEM). The experimental results show that strong electron emission distributions in the PEEM images under different polarization directions and wavelengths of femtosecond light are in qualitative agreement with the pattern of quadruple plasmon mode calculated using finite-difference time-domain simulation. In the meantime, it is found that there is a discrepancy in hot spot distribution between the observed PEEM image and the simulated photoelectron emission pattern, which is attributed to some possible factors, such as band structure near the Fermi level of the nanoring material and the temporal profile of femtosecond laser pulse. Real-space near-field imaging of the plasmonics dark mode provides a fundamental understanding of the near field and paves the way for further advancing the applications of dark mode in the field of e.g. sensing and SERS.

  2. Electron microscopy analysis of mineral fibers in human lung tissue

    SciTech Connect

    Friedrichs, K.H.; Brockmann, M.; Fischer, M.; Wick, G. )

    1992-01-01

    In the present study, lung samples from 126 autopsied cases were examined to determine the content of mineral fibers using analytical transmission electron microscopy (ATEM). The cases were divided into four groups (22 lungs of persons exposed to ambient environmental pollution, 32 cases of mesothelioma, 38 cases of primary lung cancer, and 34 asbestosis cases, 13 of these with additional pleural plaques). Fibers were counted, measured, and mineralogically identified using a combination of X-ray microanalysis and electron diffraction of the non-oriented fiber. Concentration of fibrous particles (defined as particles above 1 micron in length with roughly parallel long sides and an aspect ratio of 5:1 and greater) was calculated as fibers 10(6)/g dry lung weight. The concentration of chrysotile was found to be similar throughout the groups except for two cases in the asbestosis group with comparably high numbers of chrysotile. However, a remarkable difference for amphiboles could be observed between the groups. Asbestos bodies were mostly found in the asbestosis group. There was a rather good correlation between numbers of amphibole fibers and asbestos bodies, with an average ratio of 10:1. For comparison purposes between occupationally exposed/non-exposed individuals, a transition was found in the concentration range of 3-10(7) asbestos fibers/g dried lung weight.

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

    NASA Astrophysics Data System (ADS)

    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.

  4. Araldite as an Embedding Medium for Electron Microscopy

    PubMed Central

    Glauert, Audrey M.; Glauert, R. H.

    1958-01-01

    Epoxy resins are suitable media for embedding for electron microscopy, as they set uniformly with virtually no shrinkage. A mixture of araldite epoxy resins has been developed which is soluble in ethanol, and which yields a block of the required hardness for thin sectioning. The critical modifications to the conventional mixtures are the choice of a plasticized resin in conjunction with an aliphatic anhydride as the hardener. The hardness of the final block can be varied by incorporating additional plasticizer, and the rate of setting can be controlled by the use of an amine accelerator. The properties of the araldite mixture can be varied quite widely by adjusting the proportions of the various constituents. The procedure for embedding biological specimens is similar to that employed with methacrylates, although longer soaking times are recommended to ensure the complete penetration of the more viscous epoxy resin. An improvement in the preservation of the fine structure of a variety of specimens has already been reported, and a typical electron microgram illustrates the present paper. PMID:13525433

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

    PubMed

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

    2016-11-16

    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.

  6. Study of dynamic grain growth by electron microscopy and EBSD.

    PubMed

    Rofman, O V; Bate, P S; Brough, I; Humphreys, F J

    2009-03-01

    The effect of hot deformation on fully recrystallized aluminium-copper alloys (Al-4wt%Cu and Al-33wt%Cu) with different volume fractions of CuAl(2) has been studied. The alloys are Zener pinned systems with different superplastic properties. Strain-induced grain growth, observed in both alloys, was quantitatively estimated by means of electron microscopy and EBSD and compared with the rate of static grain growth. Surface marker observations and in situ hot-deformation experiments combined with EBSD were aimed at clarifying the mechanisms responsible for the changes in the deformed microstructures. A sequence of secondary and backscattered electron images and EBSD maps was obtained during in situ SEM deformation with different testing conditions. Overlaying EBSD maps for the Al-4wt%Cu with channelling contrast images showed that grain boundary motion occurred during deformation, creating a layered structure and leading to an increase in size of some grains and shrinkage of others. Of a particular interest are results related to behaviour of CuAl(2) in superplastic Al-33wt%Cu during deformation, including several problems with the use of EBSD in this alloy.

  7. Bone–titanium oxide interface in humans revealed by transmission electron microscopy and electron tomography

    PubMed Central

    Palmquist, Anders; Grandfield, Kathryn; Norlindh, Birgitta; Mattsson, Torsten; Brånemark, Rickard; Thomsen, Peter

    2012-01-01

    Osseointegration, the direct contact between an implant surface and bone tissue, plays a critical role in interfacial stability and implant success. Analysis of interfacial zones at the micro- and nano-levels is essential to determine the extent of osseointegration. In this paper, a series of state-of-the-art microscopy techniques are used on laser-modified implants retrieved from humans. Partially laser-modified implants were retrieved after two and a half months' healing and processed for light and electron microscopy. Light microscopy showed osseointegration, with bone tissue growing both towards and away from the implant surface. Transmission electron microscopy revealed an intimate contact between mineralized bone and the laser-modified surface, including bone growth into the nano-structured oxide. This novel observation was verified by three-dimensional Z-contrast electron tomography, enabling visualization of an apatite layer, with different crystal direction compared with the apatite in the bone tissue, encompassing the nano-structured oxide. In conclusion, the present study demonstrates the nano-scale osseointegration and bonding between apatite and surface-textured titanium oxide. These observations provide novel data in human specimens on the ultrastructure of the titanium–bone interface. PMID:21849383

  8. Atomic-scale APFIM and TEM investigation of grain boundary microchemistry in Astroloy nickel base superalloys

    SciTech Connect

    Blavette, D.; Duval, P.; Letellier, L.; Guttmann, M. |

    1996-12-01

    Electron microscopy and atom-probes techniques, including 3D (three-dimensional) atom-probe, were applied to the investigation of grain-boundary (GB) microchemistry and interfacial segregation phenomena in nickel base superalloys, Astroloy. Diffraction patterns and EDX microanalyses exhibit the presence of intergranular M{sub 23}C{sub 6} chromium enriched carbides as well as intragranular TiC precipitates. Complex phases containing Zr, C and S were also observed. Three-dimensional images as provided by the tomographic atom-probe revealed the presence of a strong segregation of both boron and molybdenum at grain boundaries. Considerable chromium enrichment at {gamma}{prime}{minus}{gamma}{prime} grain boundaries and slight carbon segregation to GBs, whatever their chemical nature, were also detected. All these segregants are distributed in a continuous manner along the boundary over a width close to 0.5 nm. Experiments show that segregation occurs during cooling and more probably between 1,200 and 800 C. Boron, chromium and molybdenum GB enrichments are thought to be mainly controlled by an equilibrium segregation process. No segregation of zirconium was detected.

  9. Interfacial ultramorphology evaluation of resin luting cements to dentin: a correlative scanning electron microscopy and transmission electron microscopy analysis.

    PubMed

    Aguiar, Thaiane Rodrigues; Vermelho, Paulo Moreira; André, Carolina Bosso; Giannini, Marcelo

    2013-12-01

    The objective of this study was to analyze the dentin-resin cements interfacial ultramorphologies using two different methods: scanning (SEM) and transmission electron microscopy (TEM). Four commercial products were evaluated: two conventional cementing system (RelyX ARC/Adper™ Scotchbond™ Multi-Purpose Plus, 3M ESPE and Clearfil Esthetic Cement/DC Bond, Kuraray) and two self-adhesive resin cements (RelyX Unicem, 3M ESPE and Clearfil SA Cement, Kuraray). Prepolymerized resin disks (Sinfony, 3M ESPE) were cemented on oclusal dentin surfaces of 24 third human molars, simulating the indirect restorations. After 24 h, teeth were sectioned into 0.9-mm thick slabs and processed for microscopy analyses (SEM or TEM/ n = 3). Qualitative characterization of dentin-resin cement interface was performed. Hybrid layer formation with long and dense resin tags was observed only for RelyX ARC cementing system. Clearfil Esthetic Cement/DC Bond system revealed few and short resin tags formation, whereas no hybridization and resin tags were detected for self-adhesive resin cements. Some interfacial regions exhibited that the self-adhesive resin cements were not bonded to dentin, presenting bubbles or voids at the interfaces. In conclusion, TEM and SEM bonding interface analyses showed ultramorphological variations among resin cements, which are directly related to dental bonding strategies used for each resin cement tested.

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

    PubMed

    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.

  11. Neurite density from magnetic resonance diffusion measurements at ultrahigh field: comparison with light microscopy and electron microscopy.

    PubMed

    Jespersen, Sune N; Bjarkam, Carsten R; Nyengaard, Jens R; Chakravarty, M Mallar; Hansen, Brian; Vosegaard, Thomas; Østergaard, Leif; Yablonskiy, Dmitriy; Nielsen, Niels Chr; Vestergaard-Poulsen, Peter

    2010-01-01

    Due to its unique sensitivity to tissue microstructure, diffusion-weighted magnetic resonance imaging (MRI) has found many applications in clinical and fundamental science. With few exceptions, a more precise correspondence between physiological or biophysical properties and the obtained diffusion parameters remain uncertain due to lack of specificity. In this work, we address this problem by comparing diffusion parameters of a recently introduced model for water diffusion in brain matter to light microscopy and quantitative electron microscopy. Specifically, we compare diffusion model predictions of neurite density in rats to optical myelin staining intensity and stereological estimation of neurite volume fraction using electron microscopy. We find that the diffusion model describes data better and that its parameters show stronger correlation with optical and electron microscopy, and thus reflect myelinated neurite density better than the more frequently used diffusion tensor imaging (DTI) and cumulant expansion methods. Furthermore, the estimated neurite orientations capture dendritic architecture more faithfully than DTI diffusion ellipsoids.

  12. Effects of ultramorphological changes on adhesion to lased dentin-Scanning electron microscopy and transmission electron microscopy analysis.

    PubMed

    Moretto, Simone G; Azambuja, Nilton; Arana-Chavez, Victor E; Reis, Andre F; Giannini, Marcelo; Eduardo, Carlos de P; De Freitas, Patricia M

    2011-08-01

    Dentin irradiation with erbium lasers has been reported to alter the composite resin bond to this treated surface. There is still a lack of studies reporting the effect of erbium lasers on dentin organic content and elucidating how laser treatment could interfere in the quality of the resin-dentin interface. This study aimed to evaluate the effect of erbium laser irradiation on dentin morphology and microtensile bond strength (μTBS) of an adhesive to dentin. Seventy-two dentin disks were divided into nine groups (n = 8): G1-Control (600-grit SiC paper); Er:YAG groups: G2- 250 mJ/4 Hz; G3- 200 mJ/4 Hz; G4- 180 mJ/10 Hz; G5- 160 mJ/10 Hz; Er,Cr:YSGG groups: G6- 2 W/20 Hz; G7- 2.5 W/20 Hz; G8- 3 W/20 Hz; G9- 4 W/20 Hz. Specimens were processed for cross-sectional analysis by scanning electron microscopy (SEM) (n = 3), transmission electron microscopy (TEM) (n = 2), and adhesive interface (n = 3). Forty-five dentin samples (n = 5) were restored and submitted to μTBS testing. ANOVA (α = 5%) revealed that G1 presented the highest μTBS values and irradiated groups did not differ from each other. TEM micrographs showed a superficial layer of denatured collagen fibrils. For SEM micrographs, it was possible to verify the laser effects extending to dentin subsurface presenting a rough aspect. Cross-sectional dentin micrographs of this hybridized surface revealed a pattern of modified tags with ringlike structures around it. This in vitro study showed that erbium laser irradiation interacts with the dental hard tissue resulting in a specific morphological pattern of dentin and collagen fibrils that negatively affected the bond strength to composite resin. Copyright © 2010 Wiley-Liss, Inc.

  13. Scanning transmission electron microscopy strain measurement from millisecond frames of a direct electron charge coupled device

    NASA Astrophysics Data System (ADS)

    Müller, Knut; Ryll, Henning; Ordavo, Ivan; Ihle, Sebastian; Strüder, Lothar; Volz, Kerstin; Zweck, Josef; Soltau, Heike; Rosenauer, Andreas

    2012-11-01

    A high-speed direct electron detection system is introduced to the field of transmission electron microscopy and applied to strain measurements in semiconductor nanostructures. In particular, a focused electron probe with a diameter of 0.5 nm was scanned over a fourfold quantum layer stack with alternating compressive and tensile strain and diffracted discs have been recorded on a scintillator-free direct electron detector with a frame time of 1 ms. We show that the applied algorithms can accurately detect Bragg beam positions despite a significant point spread each 300 kV electron causes during detection on the scintillator-free camera. For millisecond exposures, we find that strain can be measured with a precision of 1.3 × 10-3, enabling, e.g., strain mapping in a 100×100 nm2 region with 0.5 nm resolution in 40 s.

  14. Scanning transmission electron microscopy strain measurement from millisecond frames of a direct electron charge coupled device

    SciTech Connect

    Mueller, Knut; Rosenauer, Andreas; Ryll, Henning; Ordavo, Ivan; Ihle, Sebastian; Soltau, Heike; Strueder, Lothar; Volz, Kerstin; Zweck, Josef

    2012-11-19

    A high-speed direct electron detection system is introduced to the field of transmission electron microscopy and applied to strain measurements in semiconductor nanostructures. In particular, a focused electron probe with a diameter of 0.5 nm was scanned over a fourfold quantum layer stack with alternating compressive and tensile strain and diffracted discs have been recorded on a scintillator-free direct electron detector with a frame time of 1 ms. We show that the applied algorithms can accurately detect Bragg beam positions despite a significant point spread each 300 kV electron causes during detection on the scintillator-free camera. For millisecond exposures, we find that strain can be measured with a precision of 1.3 Multiplication-Sign 10{sup -3}, enabling, e.g., strain mapping in a 100 Multiplication-Sign 100 nm{sup 2} region with 0.5 nm resolution in 40 s.

  15. Advanced fertility diagnosis in stallion semen using transmission electron microscopy.

    PubMed

    Pesch, Sandra; Bostedt, Hartwig; Failing, Klaus; Bergmann, Martin

    2006-02-01

    Routine semen analysis of stallions is based on light microscopy (LM). However, there are still a number of animals that are subfertile or even infertile not being identified with conventional semen analysis. The objective of this study was to investigate the suitability of transmission electron microscopy (TEM) for advanced fertility diagnosis in stallion. We examined ejaculates of 46 stallions with known fertility. Animals were divided into three different groups: group 1, fertile stallions (pregnant mares> or =70%, n=29); group 2, subfertile stallions (pregnant mares 10-69%, n=14); group 3, infertile stallions (pregnant mares<10%, n=3). Ejaculates were collected in spring 2002. Conventional semen analysis (volume, sperm concentration, motility, live:dead ratio and percentage of morphologically normal sperm) was immediately performed after semen collection. Ultrastructural analysis included the evaluation of 200 acrosomes, heads, midpieces and cross-sections of tails as well as 100 longitudinal sections of tails from every ejaculate. Using LM, we found a significant increase of morphological deviations from 24.5% (x ) in group 1 to 34.5% in group 2 and 73.5% in group 3. Using TEM, we found a significant increase of detached acrosomes from 6.1% in group 1 to 7.6% in group 2 and 21.4% in group 3. Deviations in tubule pattern were also increased (but not significant) from 2.7% in fertile and 2.8% in subfertile to 11.4% in infertile stallions as well as multiple tails from 1.9% in fertile to 2.0% in subfertile and 8.9% in infertile. Our data indicate that TEM is suitable for advanced fertility diagnostic in stallions, giving a connection between fertility and morphology. It suggests that the most likely reason for sub- and infertility in stallion in case of increased LM pathomorphology of semen are acrosomal alterations, especially detached acrosomes.

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

    NASA Astrophysics Data System (ADS)

    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.

  17. Transmission electron microscopy of polymer blends and block copolymers

    NASA Astrophysics Data System (ADS)

    Gomez, Enrique Daniel

    Transmission electron microscopy (TEM) of soft matter is a field that warrants further investigation. Developments in sample preparation, imaging and spectroscopic techniques could lead to novel experiments that may further our understanding of the structure and the role structure plays in the functionality of various organic materials. Unlike most hard materials, TEM of organic molecules is limited by the amount of radiation damage the material can withstand without changing its structure. Despite this limitation, TEM has been and will be a powerful tool to study polymeric materials and other soft matter. In this dissertation, an introduction of TEM for polymer scientists is presented. The fundamentals of interactions of electrons with matter are described using the Schrodinger wave equation and scattering cross-sections to fully encompass coherent and incoherent scattering. The intensity, which is the product of the wave function and its complex conjugate, shows no perceptible change due to the sample. Instead, contrast is generated through the optical system of the microscope by removing scattered electrons or by generating interference due to material-induced phase changes. Perhaps the most challenging aspect of taking TEM images, however, is sample preparation, because TEM experiments require materials with approximately 50 nm thickness. Although ultramicrotomy is a well-established powerful tool for preparing biological and polymeric sections for TEM, the development of cryogenic Focused Ion Beam may enable unprecedented cross-sectional TEM studies of polymer thin films on arbitrary substrates with nanometer precision. Two examples of TEM experiments of polymeric materials are presented. The first involves quantifying the composition profile across a lamellar phase obtained in a multicomponent blend of saturated poly(butadiene) and poly(isobutylene), stabilized by a saturated poly(butadiene) copolymer serving as a surfactant, using TEM and self

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

    PubMed

    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.

  19. Observation of degradation processes of Al electrodes in organic electroluminescence devices by electroluminescence microscopy, atomic force microscopy, scanning electron microscopy, and Auger electron spectroscopy

    NASA Astrophysics Data System (ADS)

    Do, L. M.; Han, E. M.; Niidome, Y.; Fujihira, M.; Kanno, T.; Yoshida, S.; Maeda, A.; Ikushima, A. J.

    1994-11-01

    Degradation of top electrodes is one of the most important factors to determine the lifetimes of organic electroluminescence (EL) devices. An organic EL device (indium thin oxide (ITO/N,N'-diphenyl-N,N'-bis(3-methylphenyl)-(1,1'-biphenyl)-4,4'-diamine (TPD)/tris(8-hydroxy-quinoline)aluminum (Al q(sub 3))/Al) was prepared and a morphological change of the Al top electrode was observed during and/or after applying voltage by atomic force microscopy and scanning electron microscopy (SEM). The change in the electrode surface, i.e., the increase in surface roughness was observed during the current flow. The degradation process started from faint dark core parts and propagated into disks with different rates depending on the magnitude of applied voltage. Degraded sites of the Al electrode, which were analyzed as aluminum oxide by Auger electron spectroscopy, protruded into the air on the organic layers. In SEM images of a life-end electrode, discontinuities due to crevasse formation in the organic layers sandwiched by the ITO base and the metal top electrodes were observed in many places. These results confirm that one of the most crucial factors of the degradation process was deformation of metal and organic layers due to heat, gas evolution, and oxidation caused by applied voltage.

  20. Correlating Intravital Multi-Photon Microscopy to 3D Electron Microscopy of Invading Tumor Cells Using Anatomical Reference Points

    PubMed Central

    Karreman, Matthia A.; Mercier, Luc; Schieber, Nicole L.; Shibue, Tsukasa; Schwab, Yannick; Goetz, Jacky G.

    2014-01-01

    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. PMID:25479106

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

    NASA Astrophysics Data System (ADS)

    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

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

    USDA-ARS?s Scientific Manuscript database

    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...

  3. Imaging doped silicon test structures using low energy electron microscopy.

    SciTech Connect

    Nakakura, Craig Yoshimi; Anderson, Meredith Lynn; Kellogg, Gary Lee

    2010-01-01

    This document is the final SAND Report for the LDRD Project 105877 - 'Novel Diagnostic for Advanced Measurements of Semiconductor Devices Exposed to Adverse Environments' - funded through the Nanoscience to Microsystems investment area. Along with the continuous decrease in the feature size of semiconductor device structures comes a growing need for inspection tools with high spatial resolution and high sample throughput. Ideally, such tools should be able to characterize both the surface morphology and local conductivity associated with the structures. The imaging capabilities and wide availability of scanning electron microscopes (SEMs) make them an obvious choice for imaging device structures. Dopant contrast from pn junctions using secondary electrons in the SEM was first reported in 1967 and more recently starting in the mid-1990s. However, the serial acquisition process associated with scanning techniques places limits on the sample throughput. Significantly improved throughput is possible with the use of a parallel imaging scheme such as that found in photoelectron emission microscopy (PEEM) and low energy electron microscopy (LEEM). The application of PEEM and LEEM to device structures relies on contrast mechanisms that distinguish differences in dopant type and concentration. Interestingly, one of the first applications of PEEM was a study of the doping of semiconductors, which showed that the PEEM contrast was very sensitive to the doping level and that dopant concentrations as low as 10{sup 16} cm{sup -3} could be detected. More recent PEEM investigations of Schottky contacts were reported in the late 1990s by Giesen et al., followed by a series of papers in the early 2000s addressing doping contrast in PEEM by Ballarotto and co-workers and Frank and co-workers. In contrast to PEEM, comparatively little has been done to identify contrast mechanisms and assess the capabilities of LEEM for imaging semiconductor device strictures. The one exception is the

  4. Correlative Fluorescence and Electron Microscopy in 3D-Scanning Electron Microscope Perspective.

    PubMed

    Franks, Jonathan; Wallace, Callen T; Shibata, Masateru; Suga, Mitsuo; Erdman, Natasha; Stolz, Donna B; Watkins, Simon C

    2017-04-03

    The ability to correlate fluorescence microscopy (FM) and electron microscopy (EM) data obtained on biological (cell and tissue) specimens is essential to bridge the resolution gap between the data obtained by these different imaging techniques. In the past such correlations were limited to either EM navigation in two dimensions to the locations previously highlighted by fluorescence markers, or subsequent high-resolution acquisition of tomographic information using a TEM. We present a novel approach whereby a sample previously investigated by FM is embedded and subjected to sequential mechanical polishing and backscatter imaging by scanning electron microscope. The resulting three dimensional EM tomogram of the sample can be directly correlated to the FM data. © 2017 by John Wiley & Sons, Inc. Copyright © 2017 John Wiley & Sons, Inc.

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

    NASA Astrophysics Data System (ADS)

    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

  6. Scanning transmission electron microscopy methods for the analysis of nanoparticles.

    PubMed

    Ponce, Arturo; Mejía-Rosales, Sergio; José-Yacamán, Miguel

    2012-01-01

    Here we review the scanning transmission electron microscopy (STEM) characterization technique and STEM imaging methods. We describe applications of STEM for studying inorganic nanoparticles, and other uses of STEM in biological and health sciences and discuss how to interpret STEM results. The STEM imaging mode has certain benefits compared with the broad-beam illumination mode; the main advantage is the collection of the information about the specimen using a high angular annular dark field (HAADF) detector, in which the images registered have different levels of contrast related to the chemical composition of the sample. Another advantage of its use in the analysis of biological samples is its contrast for thick stained sections, since HAADF images of samples with thickness of 100-120 nm have notoriously better contrast than those obtained by other techniques. Combining the HAADF-STEM imaging with the new aberration correction era, the STEM technique reaches a direct way to imaging the atomistic structure and composition of nanostructures at a sub-angstrom resolution. Thus, alloying in metallic nanoparticles is directly resolved at atomic scale by the HAADF-STEM imaging, and the comparison of the STEM images with results from simulations gives a very powerful way of analysis of structure and composition. The use of X-ray energy dispersive spectroscopy attached to the electron microscope for STEM mode is also described. In issues where characterization at the atomic scale of the interaction between metallic nanoparticles and biological systems is needed, all the associated techniques to STEM become powerful tools for the best understanding on how to use these particles in biomedical applications.

  7. Electron microscopy of iron chalcogenide FeTe(Se) films

    NASA Astrophysics Data System (ADS)

    Shchichko, I. O.; Presnyakov, M. Yu.; Stepantsov, E. A.; Kazakov, S. M.; Antipov, E. V.; Makarova, I. P.; Vasil'ev, A. L.

    2015-05-01

    The structure of Fe1 + δTe1 - x Se x films ( x = 0; 0.05) grown on single-crystal MgO and LaAlO3 substrates has been investigated by transmission and scanning transmission electron microscopy. The study of Fe1.11Te/MgO structures has revealed two crystallographic orientation relationships between the film and substrate. It is shown that the lattice mismatch between the film and substrate is compensated for by the formation of misfit dislocations. The Burgers vector projection is determined. The stresses in the film can partially be compensated for due to the formation of an intermediate disordered layer. It is shown that a FeTe0.5Se0.5 film grown on a LaAlO3 substrate is single-crystal and that the FeTe0.5Se0.5/LaAlO3 interface in a selected region is coherent. The orientation relationships between the film and substrate are also determined for this case.

  8. Low Voltage Transmission Electron Microscopy in Cell Biology.

    PubMed

    Bendayan, Moise; Paransky, Eugene

    2015-07-01

    Low voltage transmission electron microscopy (LVTEM) was employed to examine biological tissues with accelerating voltages as low as 5kV. Tissue preparation was modified to take advantage of the low-voltage techniques. Treatments with heavy metals, such as post-fixation with osmium tetroxide, on block and counterstaining were omitted. Sections (40nm) were thinner than usual and generated highly contrasted images. General appearance of the cells remains similar to that of conventional TEM. New features were however revealed. The matrix of the pancreatic granules displays heterogeneity with partitions that may correspond to the inner-segregation of their secretory proteins. Mitochondria revealed the presence of the ATP synthase granules along their cristea. The nuclear dense chromatin displayed a honeycomb organization while distinct beads, nucleosomes, aligned along thin threads were seen in the dispersed chromatin. Nuclear pore protein complexes revealed their globular nature. The intercalated disks in cardiac muscle displayed their fine structural organization. These features correlate well with data described or predicted by cell and molecular biology. These new aspects are not revealed when thicker and conventionally osmicated tissue sections were examined by LVTEM, indicating that major masking effects are associated with standard TEM techniques. Immunogold was adapted to LVTEM further enhancing its potential in cell biology.

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

    NASA Astrophysics Data System (ADS)

    Lunov, O.; Churpita, O.; Zablotskii, V.; Deyneka, I. G.; Meshkovskii, I. K.; Jäger, A.; Syková, E.; Kubinová, Š.; Dejneka, A.

    2015-02-01

    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. Analytical electron microscopy of biogenic and inorganic carbonates

    NASA Technical Reports Server (NTRS)

    Blake, David F.

    1989-01-01

    In the terrestrial sedimentary environment, the mineralogically predominant carbonates are calcite-type minerals (rhombohedral carbonates) and aragonite-type minerals (orthorhombic carbonates). Most common minerals precipitating either inorganically or biogenically are high magnesium calcite and aragonite. High magnesium calcite (with magnesium carbonate substituting for more than 7 mole percent of the calcium carbonate) is stable only at temperatures greater than 700 C or thereabouts, and aragonite is stable only at pressures exceeding several kilobars of confining pressure. Therefore, these carbonates are expected to undergo chemical stabilization in the diagenetic environment to ultimately form stable calcite and dolomite. Because of the strong organic control of carbonate deposition in organisms during biomineralization, the microchemistry and microstructure of invertebrate skeletal material is much different than that present in inorganic carbonate cements. The style of preservation of microstructural features in skeletal material is therefore often quite distinctive when compared to that of inorganic carbonate even though wholesale recrystallization of the sediment has taken place. Microstructural and microchemical comparisons are made between high magnesium calcite echinoderm skeletal material and modern inorganic high magnesium calcite inorganic cements, using analytical electron microscopy and related techniques. Similar comparisons are made between analogous materials which have undergone stabilization in the diagenetic environment. Similar analysis schemes may prove useful in distinguishing between biogenic and inorganic carbonates in returned Martian carbonate samples.

  11. Electron microscopy of iron chalcogenide FeTe(Se) films

    SciTech Connect

    Shchichko, I. O.; Presnyakov, M. Yu.; Stepantsov, E. A.; Kazakov, S. M.; Antipov, E. V.; Makarova, I. P.; Vasil’ev, A. L.

    2015-05-15

    The structure of Fe{sub 1+δ}Te{sub 1−x}Se{sub x} films (x = 0; 0.05) grown on single-crystal MgO and LaAlO{sub 3} substrates has been investigated by transmission and scanning transmission electron microscopy. The study of Fe{sub 1.11}Te/MgO structures has revealed two crystallographic orientation relationships between the film and substrate. It is shown that the lattice mismatch between the film and substrate is compensated for by the formation of misfit dislocations. The Burgers vector projection is determined. The stresses in the film can partially be compensated for due to the formation of an intermediate disordered layer. It is shown that a FeTe{sub 0.5}Se{sub 0.5} film grown on a LaAlO{sub 3} substrate is single-crystal and that the FeTe{sub 0.5}Se{sub 0.5}/LaAlO{sub 3} interface in a selected region is coherent. The orientation relationships between the film and substrate are also determined for this case.

  12. High-performance probes for light and electron microscopy

    PubMed Central

    Viswanathan, Sarada; Williams, Megan E.; Bloss, Erik B.; Stasevich, Timothy J.; Speer, Colenso M.; Nern, Aljoscha; Pfeiffer, Barret D.; Hooks, Bryan M.; Li, Wei-Ping; English, Brian P.; Tian, Teresa; Henry, Gilbert L.; Macklin, John J.; Patel, Ronak; Gerfen, Charles R.; Zhuang, Xiaowei; Wang, Yalin; Rubin, Gerald M.

    2015-01-01

    We describe an engineered family of highly antigenic molecules based on GFP-like fluorescent proteins. These molecules contain numerous copies of peptide epitopes and simultaneously bind IgG antibodies at each location. These “spaghetti monster” fluorescent proteins (smFPs) distribute well in neurons, notably into small dendrites, spines and axons. smFP immunolabeling localizes weakly expressed proteins not well resolved with traditional epitope tags. By varying epitope and scaffold, we generated a diverse family of mutually orthogonal antigens. In cultured neurons and mouse and fly brains, smFP probes allow robust, orthogonal multi-color visualization of proteins, cell populations and neuropil. smFP variants complement existing tracers, greatly increase the number of simultaneous imaging channels, and perform well in advanced preparations such as array tomography, super-resolution fluorescence imaging and electron microscopy. In living cells, the probes improve single-molecule image tracking and increase yield for RNA-Seq. These probes facilitate new experiments in connectomics, transcriptomics and protein localization. PMID:25915120

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

    SciTech Connect

    Lunov, O. Churpita, O.; Zablotskii, V.; Jäger, A.; Dejneka, A.; Deyneka, I. G.; Meshkovskii, I. K.; Syková, E.; Kubinová, Š.

    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.

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

    PubMed

    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.

  15. Single particle conformations of human serum albumin by electron microscopy.

    PubMed

    Ueno, Yutaka; Mio, Muneyo; Sato, Chikara; Mio, Kazuhiro

    2007-06-01

    Using single particle images taken by electron microscopy, pH-dependent conformational changes of human serum albumin were investigated. Despite the noisy particle images of negatively stained serum albumin (67 kDa), our novel algorithm for automated particle picking and reference-free classification resulted in the appropriate grouping of the particle images. Iteratively aligned particle images in the same group provided recognizable image features for individual groups. In a pH 7.0 study, monomer images were consistent with an available crystal structure model; the dimer images were separated into different classes. At pH 3.5, the monomer images were similar to those at pH 7.0; slight differences included a small number of elongated conformations and increased population of larger multimers. Our images were also compared with projection images of an atomic model from crystallography, and demonstrated consistency of the molecular conformation both at pH 7.0 and pH 3.5. Our classification method was effective in discriminating monomers from a mixture of different conformations of the protein, enabling the study of the conformational dynamics of small proteins, using the atomic model as a reference.

  16. TRANSMISSION ELECTRON MICROSCOPY STUDY OF HELIUM BEARING FUSION WELDS

    SciTech Connect

    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.

  17. Scanning electron microscopy of the endometrium during the secretory phase.

    PubMed Central

    Motta, P M; Andrews, P M

    1976-01-01

    Scanning electron microscopy was used to study the surface morphology of the rabbit endometrium during the secretory phase of the oestrous cycle. The free surfaces of ciliated and of inactive active secretory cells are described. Changes in secretory cell surface morphology resulting from accumulation and secretion of material involve the apparent retraction of microvilli and the formation of one or more bulbous protrusions of the cell's apical surface. These protrusions may be relatively smooth surfaced or exhibit long slender micro-extensions. The protrusions grow in size and are eventually pinched off. Loss of the bulbous protrusions often leaves behind crater-like invaginations of the cell's surface. Secretory cells adjacent to the endometrial glands are the first to exhibit signs of mucin accumulation and secretion. The single cilium of a secretory cell is not apparently affected by the secretory process. Signs of ciliated and secretory cell degeneration, and possible sloughing, are also described. Images Fig. 1 Fig. 2 Fig. 3 Fig. 4 Fig. 5 Fig. 6 Fig. 7 PMID:1033932

  18. Spatial resolution and information transfer in scanning transmission electron microscopy.

    PubMed

    Peng, Yiping; Oxley, Mark P; Lupini, Andrew R; Chisholm, Matthew F; Pennycook, Stephen J

    2008-02-01

    The relation between image resolution and information transfer is explored. It is shown that the existence of higher frequency transfer in the image is just a necessary but not sufficient condition for the achievement of higher resolution. Adopting a two-point resolution criterion, we suggest that a 10% contrast level between two features in an image should be used as a practical definition of resolution. In the context of scanning transmission electron microscopy, it is shown that the channeling effect does not have a direct connection with image resolution because sharp channeling peaks do not move with the scanning probe. Through a quantitative comparison between experimental image and simulation, a Fourier-space approach is proposed to estimate defocus and sample thickness. The effective atom size in Z-contrast imaging depends on the annular detector's inner angle. Therefore, an optimum angle exists for the highest resolution as a trade-off between reduced atom size and reduced signal with limited information transfer due to noise.

  19. Characterization of paired helical filaments by scanning transmission electron microscopy.

    PubMed

    Ksiezak-Reding, Hanna; Wall, Joseph S

    2005-07-01

    Paired helical filaments (PHFs) are abnormal twisted filaments composed of hyperphosphorylated tau protein. They are found in Alzheimer's disease and other neurodegenerative disorders designated as tauopathies. They are a major component of intracellular inclusions known as neurofibrillary tangles (NFTs). The objective of this review is to summarize various structural studies of PHFs in which using scanning transmission electron microscopy (STEM) has been particularly informative. STEM provides shape and mass per unit length measurements important for studying ultrastructural aspects of filaments. These include quantitative comparisons between dispersed and aggregated populations of PHFs as well as comparative studies of PHFs in Alzheimer's disease and other neurodegenerative disorders. Other approaches are also discussed if relevant or complementary to studies using STEM, e.g., application of a novel staining reagent, Nanovan. Our understanding of the PHF structure and the development of PHFs into NFTs is presented from a historical perspective. Others goals are to describe the biochemical and ultrastructural complexity of authentic PHFs, to assess similarities between authentic and synthetic PHFs, and to discuss recent advances in PHF modeling.

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

    NASA Technical Reports Server (NTRS)

    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.

  1. Surface treatment of feldspathic porcelain: scanning electron microscopy analysis

    PubMed Central

    Valian, Azam

    2014-01-01

    PURPOSE Topographic analysis of treated ceramics provides qualitative information regarding the surface texture affecting the micromechanical retention and locking of resin-ceramics. This study aims to compare the surface microstructure following different surface treatments of feldspathic porcelain. MATERIALS AND METHODS This in-vitro study was conducted on 72 porcelain discs randomly divided into 12 groups (n=6). In 9 groups, feldspathic surfaces were subjected to sandblasting at 2, 3 or 4 bar pressure for 5, 10 or 15 seconds with 50 µm alumina particles at a 5 mm distance. In group 10, 9.5% hydrofluoric acid (HF) gel was applied for 120 seconds. In group 11, specimens were sandblasted at 3 bar pressure for 10 seconds and then conditioned with HF. In group 12, specimens were first treated with HF and then sandblasted at 3 bar pressure for 10 seconds. All specimens were then evaluated under scanning electron microscopy (SEM) at different magnifications. RESULTS SEM images of HF treated specimens revealed deep porosities of variable sizes; whereas, the sandblasted surfaces were more homogenous and had sharper peaks. Increasing the pressure and duration of sandblasting increased the surface roughness. SEM images of the two combined techniques showed that in group 11 (sandblasted first), HF caused deeper porosities; whereas in group 12 (treated with HF first) sandblasting caused irregularities with less homogeneity. CONCLUSION All surface treatments increased the surface area and caused porous surfaces. In groups subjected to HF, the porosities were deeper than those in sandblasted only groups. PMID:25352961

  2. Rapid diagnosis of plant virus diseases by transmission electron microscopy.

    PubMed

    Zechmann, Bernd; Zellnig, Günther

    2009-12-01

    A clear and rapid diagnosis of plant virus diseases is of great importance for agriculture and scientific experiments in plant phytopathology. Even though negative staining and transmission electron microscopy (TEM) are often used for detection and identification of viral particles and provide rapid and reliable results, it is necessary to examine ultrastructural changes induced by viruses for clear identification of the disease. With conventional sample preparation for TEM it can take several days to obtain ultrastructural results and it is therefore not suitable for rapid diagnosis of virus diseases of plants. The use of microwave irradiation can reduce the time for sample preparation for TEM investigations. Two model virus-plant systems [Nicotiana tabacum plants infected with Tobacco mosaic virus (TMV), Cucurbita pepo plants infected with Zucchini yellow mosaic virus (ZYMV)] demonstrate that it is possible to diagnose ultrastructural alterations induced by viruses in less than half a day by using microwave irradiation for preparation of samples. Negative staining of the sap of plants infected with TMV and ZYMV and the examination of ultrastructure and size were also carried out during sample preparation thus permitting diagnosis of the viral agent by TEM in a few hours. These methods will contribute towards a rapid and clear identification of virus diseases of plants and will be useful for diagnostic purposes in agriculture and in plant phytopathology.

  3. Residual Deconvolutional Networks for Brain Electron Microscopy Image Segmentation.

    PubMed

    Fakhry, Ahmed; Zeng, Tao; Ji, Shuiwang

    2017-02-01

    Accurate reconstruction of anatomical connections between neurons in the brain using electron microscopy (EM) images is considered to be the gold standard for circuit mapping. A key step in obtaining the reconstruction is the ability to automatically segment neurons with a precision close to human-level performance. Despite the recent technical advances in EM image segmentation, most of them rely on hand-crafted features to some extent that are specific to the data, limiting their ability to generalize. Here, we propose a simple yet powerful technique for EM image segmentation that is trained end-to-end and does not rely on prior knowledge of the data. Our proposed residual deconvolutional network consists of two information pathways that capture full-resolution features and contextual information, respectively. We showed that the proposed model is very effective in achieving the conflicting goals in dense output prediction; namely preserving full-resolution predictions and including sufficient contextual information. We applied our method to the ongoing open challenge of 3D neurite segmentation in EM images. Our method achieved one of the top results on this open challenge. We demonstrated the generality of our technique by evaluating it on the 2D neurite segmentation challenge dataset where consistently high performance was obtained. We thus expect our method to generalize well to other dense output prediction problems.

  4. Cryomesh™: A new substrate for cryo-electron microscopy

    PubMed Central

    Yoshioka, Craig; Carragher, Bridget; Potter, Clint

    2010-01-01

    Here we evaluate a new grid substrate developed by ProtoChips Inc. for cryo-transmission electron microscopy. The new grids are fabricated from doped silicon carbide using processes adapted from the semi-conductor industry. A major motivating purpose in the development of these grids was to increase the low-temperature conductivity of the substrate, a characteristic that is thought to affect the appearance of beam-induced movement (BIM) in TEM images of biological specimens. BIM degrades the quality of data, and is especially severe when frozen biological specimens are tilted in the microscope. Our results show that this new substrate does indeed have a significant impact on reducing the appearance and severity of beam-induced movement in TEM images of tilted cryo-preserved samples. Furthermore, while we have not been able to ascertain the exact causes underlying the BIM phenomenon, we have evidence that the rigidity and flatness of these grids may play a major role in its reduction. This improvement in the reliability of imaging at tilt has a significant impact on using data collection methods such as random conical tilt or orthogonal tilt reconstruction with cryo-preserved samples. Reduction in BIM also has the potential for improving the resolution of 3D cryo-reconstructions in general. PMID:20082728

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

    NASA Technical Reports Server (NTRS)

    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.

  6. Optical and electron microscopy of WC-Co alloys

    SciTech Connect

    Yust, C S; Long, Jr, E L

    1982-02-01

    The microstructures of three commercial cobalt-bonded tungsten carbide alloys have been characterized by optical and electron microscopy and compared with a specially formulated reference alloy composed of tungsten carbide bonded by 6 wt % Co. The first alloy contained additions of chromium as chromium carbides, was similar in microstructure to the reference alloy, and contained secondary carbide grains retained from the chromium addition. An alloy containing metallic chromium also contained grains of the ternary carbide Co/sub 3/W/sub 3/C, or eta phase, which can be rationalized as having formed by reaction of the molten cobalt-chromium binder phase with the tungsten carbide matrix at the processing temperature. The third commercial alloy examined contained a coarse dendritic structure identified as a mixture of eta (Co/sub 3/W/sub 3/C) and chi (Co/sub 3/W/sub 9/C/sub 4/) phases. The reactions responsible for formation of the eta and chi phases in this alloy have not yet been determined.

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

    PubMed

    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.

  8. Ultrastructure of exogenous surfactants using cryogenic scanning electron microscopy.

    PubMed

    Banerjee, R; Bellare, J R

    2001-01-01

    Therapy with specialised biomaterials, exogenous surfactants, is known to significantly decrease the mortality rates in Respiratory Distress Syndrome (RDS). Surfactants available commercially vary widely in composition and biophysical properties. The present paper studies the ultrastructure of three exogenous surfactants used for the treatment of Respiratory Distress Syndrome, namely, Survanta, ALEC and Exosurf Neonatal with respect to their ability to form liposomes using cryogenic scanning electron microscopy. Liposomal organisation is more obvious in Exosurf than in Survanta and is most pronounced in ALEC. ALEC forms closed regular liposomes with an onion-ring-like internal bilayer arrangement. Survanta forms open membranous structures with wavy ribbon-like membranes. The complex membrane-like structures seen with Survanta may be due to the interaction of lipids with surfactant-specific proteins present in this surfactant which is derived from natural lung extracts and might indicate superior spreading at the lipid-water interface. Artificial protein-free surfactants (ALEC and Exosurf) did not appear to form these open membranous structures. Further study of the ultrastructure of possible biomaterials as surfactants could help in the development of new, improved artificial protein-free surfactants with open membranous structures that might facilitate spreading at the air-liquid interface of lungs.

  9. Scanning electron microscopy of lung following alpha irradiation

    SciTech Connect

    Sanders, C.L.; Lauhala, K.E.; McDonald, K.E. )

    1989-09-01

    Pulmonary aggregation of inhaled {sup 239}PuO{sub 2} particles leads to a cellular evolution of focal inflammation, fibrosis, epithelial dysplasia and lung tumor formation. Female Wistar rats were exposed to an aerosol of high-fired {sup 239}PuO{sub 2} (initial lung burden, 3.9 kBq) and the lungs examined at intervals from 1 day to 700 days after exposure by light and scanning electron microscopy and autoradiography. Peribronchiolar Pu particle aggregation increased with time, resulting in well-defined focal inflammatory lesions after 120 days and fibrotic lesions after 180 days. A generalized hypertrophy and hyperplasia of nonciliated bronchiolar cells was seen at 15 days and type II cell hyperplasia by 30 days after exposure. Focal dysplastic changes in type II alveolar epithelium and terminal nonciliated bronchiolar epithelium preceded carcinoma formation. Alveolar bronchiolarization was first noted at 120 days, squamous metaplasia at 210 days, squamous carcinoma at 270 days and adenocarcinoma at 600 days after exposure.

  10. In situ transmission electron microscopy of electron-beam induced damage process in nuclear grade graphite

    NASA Astrophysics Data System (ADS)

    Karthik, C.; Kane, J.; Butt, D. P.; Windes, W. E.; Ubic, R.

    2011-05-01

    Atomic level processes involved in the swelling and crack-closing in nuclear grade graphite under electron irradiation have been observed in real-time using transmission electron microscopy. Noise-filtered lattice images show the formation of vacancy loops, interstitial loops and resulting dislocations with unprecedented clarity. The dislocation dipoles formed via vacancy loops were found to undergo climb resulting in extra basal planes. Concurrent EELS studies showed a reduction in the atomic density because of the breakage of hexagonal carbon rings. The formation of new basal planes via dislocation climb in addition to the bending/breaking of basal planes leads to swelling and closing of micro-cracks.

  11. High-Resolution of Electron Microscopy of Montmorillonite and Montmorillonite/Epoxy Nanocomposites

    DTIC Science & Technology

    2005-01-01

    AFRL-ML-WP-TP-2006-464 HIGH-RESOLUTION OF ELECTRON MICROSCOPY OF MONTMORILLONITE AND MONTMORILLONITE /EPOXY NANOCOMPOSITES Lawrence F...HIGH-RESOLUTION OF ELECTRON MICROSCOPY OF MONTMORILLONITE AND MONTMORILLONITE /EPOXY NANOCOMPOSITES 5c. PROGRAM ELEMENT NUMBER 62102F 5d...transmission electron microscopy the structure and morphology of montmorillonite (MMT), a material of current interest for use in polymer nanocomposites, was

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

    NASA Astrophysics Data System (ADS)

    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.

  13. A Study of Light and Electron Microscopy of the Neural Basis for Diurnal and Nocturnal Vision.

    DTIC Science & Technology

    RETINA, *PHOTORECEPTORS, LIPIDS, MORPHOLOGY, ELECTRON MICROSCOPY, COLORS, NERVE CELLS, VISION, NIGHT VISION, SPAIN, BIRDS, CELLS(BIOLOGY), DAYLIGHT, CELL STRUCTURE, ELECTROPHYSIOLOGY, NOCTURNAL ANIMALS .

  14. New electron microscopy techniques of the study of meteoritic metal.

    SciTech Connect

    Michael, Joseph Richard; Goldstein, Joseph I.; Kotula, Paul Gabriel; Jones, R. H.

    2005-02-01

    Metallic Phases in extraterrestrial materials are composed of Fe-Ni with minor amounts of Co, P, Si, Cr, etc. Electron microscopy techniques (SEM, TEM, EPMA, AEM) have been used for almost 50 years to study micron and submicron microscopic features in the metal phases (Fig. 1) such as clear taenite, cloudy zone, plessite, etc [1,2]. However lack of instrumentation to prepare TEM thin foils in specific sample locations and to obtain micro-scale crystallographic data have limited these investigations. New techniques such as the focused ion beam (FIB) and the electron backscatter electron diffraction (EBSD) techniques have overcome these limitations. The application of the FIB instrument has allowed us to prepare {approx}10 um long by {approx} 5um deep TEM thin sections of metal phases from specific regions of metal particles, in chondrites, irons and stony iron meteorites, identified by optical and SEM observation. Using a FEI dual beam FIB we were able to study very small metal particles in samples of CH chondrites [3] and zoneless plessite (ZP) in ordinary chondrites. Fig. 2 shows a SEM photomicrograph of a {approx}40 um ZP particle in Kernouve, a H6 chondrite. Fig. 3a,b shows a TEM photograph of a section of the FIB prepared TEM foil of the ZP particle and a Ni trace through a tetrataenite/kamacite region of the particle. It has been proposed that the Widmanstatten pattern in low P iron meteorites forms by martensite decomposition, via the reaction {gamma} {yields} {alpha}{sub 2} + {gamma} {yields} {alpha} + {gamma} in which {alpha}{sub 2}, martensite, decomposes to the equilibrium {alpha} and {gamma} phases during the cooling process [4]. In order to show if this mechanism for Widmanstatten pattern formation is correct, crystallographic information is needed from the {gamma} or taenite phases throughout a given meteorite. The EBSD technique was employed in this study to obtain the orientation of the taenite surrounding the initial martensite phase and the

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

    NASA Astrophysics Data System (ADS)

    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

  16. Imaging electron emission from diamond and III V nitride surfaces with photo-electron emission microscopy

    NASA Astrophysics Data System (ADS)

    Nemanich, R. J.; English, S. L.; Hartman, J. D.; Sowers, A. T.; Ward, B. L.; Ade, H.; Davis, R. F.

    1999-05-01

    Wide bandgap semiconductors such as diamond and the III-V nitrides (GaN, AlN, and AlGaN alloys) exhibit small or even negative electron affinities. Results have shown that different surface treatments will modify the electron affinity of diamond to cause a positive or negative electron affinity (NEA). This study describes the characterization of these surfaces with photo-electron emission microscopy (PEEM). The PEEM technique is unique in that it combines aspects of UV photoemission and field emission. In this study, PEEM images are obtained with either a traditional Hg lamp or with tunable UV excitation from a free electron laser. The UV-free electron laser at Duke University provides tunable emission from 3.5 to greater than 7 eV. PEEM images of boron or nitrogen (N)-doped diamond are similar to SEM of the same surface indicating relatively uniform emission. For the N-doped samples, PEEM images were obtained for different photon energies ranging from 5.0 to 6.0 eV. In these experiments, the hydrogen terminated surface showed more intense PEEM images at lower photon energy indicating a lower photothreshold than annealed surfaces which are presumed to be adsorbate free. For the nitrides, the emission properties of an array of GaN emitter structures is imaged. Emission is observed from the peaks, and relatively uniform emission is observed from the array. The field at the sample surface is approximately 10 V/μm which is sufficient to obtain an image without UV light. This process is termed field emission electron microscopy (FEEM).

  17. Sample thickness determination by scanning transmission electron microscopy at low electron energies.

    PubMed

    Volkenandt, Tobias; Müller, Erich; Gerthsen, Dagmar

    2014-02-01

    Sample thickness is a decisive parameter for any quantification of image information and composition in transmission electron microscopy. In this context, we present a method to determine the local sample thickness by scanning transmission electron microscopy at primary energies below 30 keV. The image intensity is measured with respect to the intensity of the incident electron beam and can be directly compared with Monte Carlo simulations. Screened Rutherford and Mott scattering cross-sections are evaluated with respect to fitting experimental data with simulated image intensities as a function of the atomic number of the sample material and primary electron energy. The presented method is tested for sample materials covering a wide range of atomic numbers Z, that is, fluorenyl hexa-peri-hexabenzocoronene (Z = 3.5), carbon (Z = 6), silicon (Z = 14), gallium nitride (Z = 19), and tungsten (Z = 74). Investigations were conducted for two primary energies (15 and 30 keV) and a sample thickness range between 50 and 400 nm.

  18. PREFACE: Electron Microscopy and Analysis Group Conference 2009

    NASA Astrophysics Data System (ADS)

    Baker, Richard

    2010-04-01

    The latest biennial conference of the Electron Microscopy and Analysis Group (EMAG) of the Institute of Physics was held at the University of Sheffield on 9-11 September, 2009. In addition, the Advanced School associated with the conference was run at the University of Sheffield on 8 September. It was particularly pleasing to return to Sheffield after ten years, the successful and memorable EMAG 99 having been held here too. The subject areas covered at EMAG 2009 were advanced electron microscopy techniques; investigating structure-property relationships in advanced materials; nanophysics and nanotechnology. The EMAG 2009 conference attracted 172 delegates while the Advanced School had a full complement of eighteen attendees. Three plenary lectures were given to the whole conference and invited contributions were presented within the theme of each of nine parallel sessions. There were 54 contributed oral presentations within these parallel Sessions and a further 89 poster presentations. All authors were invited to contribute a paper to this Proceedings volume and 108 papers are presented here. I thank all who presented at EMAG 2009 and those who provided a paper for this Proceedings. Each paper was peer reviewed by two reviewers and I also want to thank those colleagues who helped with this essential task. In this volume, the plenary papers are presented first followed by all papers presented in each themed session. These sessions are ordered alphabetically. Within each Session, the invited presentations are presented first, followed by oral and poster contributions together. Another activity of EMAG which is directed primarily at less experienced scientists is the Advanced School. This year, this was on Nanofabrication and Nanomanipulation and I want to thank Guenter Moebus and his colleagues at th University of Sheffield for putting on such an excellent Advanced School. The EMAG series of conferences are well-known not only for the academic conference but also

  19. High resolution electron microscopy of interfaces in fcc materials

    SciTech Connect

    Merkle, K.L.

    1990-08-01

    Modern high-resolution electron microscopy (HREM) instruments, which are capable of a point-to-point resolution of better than 0.2 nm, have allowed atomic-scale observations of a variety of internal interfaces. The application of the HREM technique to fcc model systems for the purpose of addressing a number of interface issues will be examined in this paper. Atomic structure observations for heterophase interfaces of metal/metal and metal/metal-oxide systems as well as HREM studies of grain boundaries in NiO and Au will be discussed with emphasis on generic structural features and the role of the interface plane. Comparisons between observed interface structures and atomistic computer modeling results have shown agreements for some interfaces, as well as certain differences in others. A number of structural features are common to both metal and oxide grain boundaries, as well as certain heterophase boundaries. Of particular importance in close-packed solids appears to be the tendency to preserve, as much as possible, local atomic coordination, giving rise to atomically well-matched regions that alternate along the interface with regions of misfit. It is commonly observed that heterophase interfaces are being preferentially formed on dense-packed planes. Low-index planes are also frequently observed in asymmetric grain boundaries. In addition to the observation of misfit dislocations in heterophase boundaries, misfit-dislocation-like defects have also been found in asymmetric, incommensurate grain boundaries. The tendency for maintaining coherence between dense-packed planes across the interface has resulted in the formation of novel three-dimensional GB structures. HREM observations have brought new insights into the correlations between macroscopic geometry, interfacial energy, and microscopic atomic relaxations.

  20. EDITORIAL: Electron Microscopy and Analysis Group Conference 2013 (EMAG2013)

    NASA Astrophysics Data System (ADS)

    Nellist, Pete

    2014-06-01

    It has once again been my pleasure to act as editor for these proceedings, and I must thank all those who have acted as reviewers. 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, like me, you will be struck by the scientific quality of the 80 papers that follow, and that you will find them interesting and informative. I must also personally thank all the organisers of EMAG2013 for arranging such an excellent meeting. Ian MacLaren, as Chair of the EMAG Group and of the meeting itself, has contributed a foreword to these proceedings describing the meeting in more detail. A particular highlight of the conference was the special symposium in honour of Professor Archie Howie. We all enjoyed a wonderful speech from Archie at the conference dinner, along with some of his electron microscopy-related poetry. I have great pleasure in publishing the conference dinner poems in this proceedings. I hope you will find these proceedings to be an interesting read and an invaluable resource. Pete Nellist Conference committee Conference chair: Dr I MacLaren Programme organiser: Dr C Ducati Proceedings editor: Prof P D Nellist Trade exhibition organiser: C Hockey (CEM Group) Local organisers: Professor E Boyes, Professor P Gai, Dr R Kröger, Dr V Lazarov, Dr P O'Toole, Dr S Tear and Professor J Yuan Advanced school organisers: Dr S Haigh, Dr A Brown Other committee members: Mr K Meade, Mr O Heyning, Dr M Crawford, Mr M Dixon and Dr Z Li

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

    USGS Publications Warehouse

    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.

  2. Characterization of humic substances by environmental scanning electron microscopy.

    PubMed

    Redwood, Paul S; Lead, Jamie R; Harrison, Roy M; Jones, Ian P; Stoll, Serge

    2005-04-01

    Environmental scanning electron microscopy (ESEM) is a new technique capable of imaging micron and submicron particles. Here, we have applied it to image and quantify natural aquatic organic matter (standard Suwannee River humic acid, SRHA). Uniquely, we have observed the humic aggregate structures as a function of humidity and pH. Large aggregates of tens of micrometers were observed as the dominant material under all conditions, although much smaller material was also observed. Fractal dimensions (D) were calculated between 1.48 and 1.70, although these values were not statistically different under conditions of low humidity. However, D values calculated at high humidities (85%) during the rehydration phase were significantly lower (1.48+/-0.01) than in the initial dehydration phase (1.69+/-0.01). This hysteresis indicated that full rehydration of the HS was either kinetically slow or irreversible after dehydration. Fractal analysis of ESEM images was also performed to probe the change in aggregate structure as a function of pH. Minimum values were calculated at neutral pHs, rising by 0.1-0.2 at both high and low pHs because of a combination of the physical chemistry of HS and the impacts of the drying regime within the ESEM. Thus, ESEM was an important complementary technique to other analytical methods. At present, ESEM cannot be used to image nonperturbed natural samples. However, the method is an ideal method for probing the changes in colloid structure as function of hydration state and has the potential to perform fully quantitative and nonperturbing analysis of colloidal structure.

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

    NASA Technical Reports Server (NTRS)

    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.

  4. Transmission electron microscopy investigation of auto catalyst and cobalt germanide

    NASA Astrophysics Data System (ADS)

    Sun, Haiping

    The modern ceria-zirconia based catalysts are used in automobiles to reduce exhaust pollutants. Cobalt germanides have potential applications as electrical contacts in the future Ge-based semiconductor devices. In this thesis, transmission electron microscopy (TEM) techniques were used to study the atomic scale interactions between metallic nanostructures and crystalline substrates in the two material systems mentioned above. The model catalyst samples consisted of precious metal nano-particles (Pd, Rh) supported on the surface of (Ce,Zr)O2 thin films. The response of the microstructure of the metal-oxide interface to the reduction and oxidation treatments was investigated by cross-sectional high resolution TEM. Atomic detail of the metal-oxide interface was obtained. It was found that Pd and Rh showed different sintering and interaction behaviors on the oxide surface. The preferred orientation of Pd particles in this study was Pd(111)//CZO(111). Partial encapsulation of Pd particles by reduced (Ce,Zr)O 2 surface was observed and possible mechanisms of the encapsulation were discussed. The characteristics of the metal-oxide interaction depend on the properties of the oxide, as well as their relative orientation. The results provide experimental evidence for understanding the thermodynamics of the equilibrium morphology of a solid particle supported on a solid surface that is not considered as inert. The reaction of Co with Ge to form epitaxial Co5Ge7 was studied by in situ ultra-high vacuum (UHV) TEM using two methods. One was reactive deposition of Co on Ge, in which the Ge substrate was maintained at 350°C during deposition. The other method was solid state reaction, in which the deposition of Co on Ge was carried out at room temperature followed by annealing to higher temperatures. During reactive deposition, the deposited Co reacted with Ge to form nanosized 3D Co 5Ge7 islands. During solid state reaction, a continuous epitaxial Co5Ge7 film on the (001) Ge

  5. Visualization of Microbial Biomarkers by Scanning Electron Microscopy

    NASA Technical Reports Server (NTRS)

    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.

  6. Visualization of Microbial Biomarkers by Scanning Electron Microscopy

    NASA Technical Reports Server (NTRS)

    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.

  7. A CCD Camera with Electron Decelerator for Intermediate Voltage Electron Microscopy

    SciTech Connect

    Downing, Kenneth H; Downing, Kenneth H.; Mooney, Paul E.

    2008-03-17

    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.

  8. Direct electron imaging in electron microscopy with monolithic active pixel sensors.

    PubMed

    Deptuch, G; Besson, A; Rehak, P; Szelezniak, M; Wall, J; Winter, M; Zhu, Y

    2007-08-01

    A new imaging device for dynamic electron microscopy is in great demand. The detector should provide the experimenter with images having sufficient spatial resolution at high speed. Immunity to radiation damage, accumulated during exposures, is critical. Photographic film, a traditional medium, is not adequate for studies that require large volumes of data or rapid recording and charge coupled device (CCD) cameras have limited resolution, due to phosphor screen coupling. CCD chips are not suitable for direct recording due to their extreme sensitivity to radiation damage. This paper discusses characterization of monolithic active pixel sensors (MAPS) in a scanning electron microscope (SEM) as well as in a transmission electron microscope (TEM). The tested devices were two versions of the MIMOSA V (MV) chip. This 1M pixel device features pixel size of 17 x 17 microm(2) and was designed in a 0.6 microm CMOS process. The active layer for detection is a thin (less than 20 microm) epitaxial layer, limiting the broadening of the electron beam. The first version of the detector was a standard imager with electronics, passivation and interconnection layers on top of the active region; the second one was bottom-thinned, reaching the epitaxial layer from the bottom. The electron energies used range from a few keV to 30 keV for SEM and from 40 to 400 keV for TEM. Deterioration of the image resolution due to backscattering was quantified for different energies and both detector versions.

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

    PubMed Central

    Downing, Kenneth H.; Mooney, Paul E.

    2008-01-01

    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. PMID:18447528

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

    PubMed

    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.

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

    PubMed

    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.

  12. Atomic and electronic structure of polar oxide interfaces: Electron microscopy and density functional theory study

    NASA Astrophysics Data System (ADS)

    Lazarov, Vlado

    Polar oxide interfaces are formed when two polar oxide surfaces join. The apparent presence of an electric dipole moment in the repeat unit parallel to the surface/interface closely relate the polar oxide interfaces instability to that of the of polar oxide surfaces. In this thesis, we combined Electron Microscopy and Density Functional Theory to study how the interface polarity affects the atomic and electronic structure of polar oxide interfaces, by using Fe3O4(111)/MgO(111) as a model system. The formation of Fe nanoinclusions found at the interface and within the polar Fe3 O4(111) film is proposed to be new stabilization mechanism for the magnetite film. High-resolution electron microscopy imaging of the interface together with first principle calculations suggest an atomically abrupt substrate-film interface determined with Fe monolayer in octahedral position (FeB). This interface stacking (O/Mg/O/3FeB/O) provides lowest total interface (system) energy and the most effectively screening of the MgO(111) substrate surface polarity. The results of our study suggest that surface polarity could be used as an additional growth parameter in creating novel material structures, such as metals in oxide matrices.

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

    PubMed

    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.

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

    Cancer.gov

    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...

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

    Cancer.gov

    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. Atomic force microscopy and scanning electron microscopy analysis of daily disposable limbal ring contact lenses

    PubMed Central

    Lorenz, Kathrine Osborn; Kakkassery, Joseph; Boree, Danielle; Pinto, David

    2014-01-01

    Background Limbal ring (also known as ‘circle’) contact lenses are becoming increasingly popular, especially in Asian markets because of their eye-enhancing effects. The pigment particles that give the eye-enhancing effects of these lenses can be found on the front or back surface of the contact lens or ‘enclosed’ within the lens matrix. The purpose of this research was to evaluate the pigment location and surface roughness of seven types of ‘circle’ contact lenses. Methods Scanning electron microscopic (SEM) analysis was performed using a variable pressure Hitachi S3400N instrument to discern the placement of lens pigments. Atomic force microscopy (Dimension Icon AFM from Bruker Nano) was used to determine the surface roughness of the pigmented regions of the contact lenses. Atomic force microscopic analysis was performed in fluid phase under contact mode using a Sharp Nitride Lever probe (SNL-10) with a spring constant of 0.06 N/m. Root mean square (RMS) roughness values were analysed using a generalised linear mixed model with a log-normal distribution. Least square means and their corresponding 95% confidence intervals were estimated for each brand, location and pigment combination. Results SEM cross-sectional images at 500× and 2,000× magnification showed pigment on the surface of six of the seven lens types tested. The mean depth of pigment for 1-DAY ACUVUE DEFINE (1DAD) lenses was 8.1 μm below the surface of the lens, while the remaining lens types tested had pigment particles on the front or back surface. Results of the atomic force microscopic analysis indicated that 1DAD lenses had significantly lower root mean square roughness values in the pigmented area of the lens than the other lens types tested. Conclusions SEM and AFM analysis revealed pigment on the surface of the lens for all types tested with the exception of 1DAD. Further research is required to determine if the difference in pigment location influences on-eye performance. PMID

  17. Atomic force microscopy and scanning electron microscopy analysis of daily disposable limbal ring contact lenses.

    PubMed

    Lorenz, Kathrine Osborn; Kakkassery, Joseph; Boree, Danielle; Pinto, David

    2014-09-01

    Limbal ring (also known as 'circle') contact lenses are becoming increasingly popular, especially in Asian markets because of their eye-enhancing effects. The pigment particles that give the eye-enhancing effects of these lenses can be found on the front or back surface of the contact lens or 'enclosed' within the lens matrix. The purpose of this research was to evaluate the pigment location and surface roughness of seven types of 'circle' contact lenses. Scanning electron microscopic (SEM) analysis was performed using a variable pressure Hitachi S3400N instrument to discern the placement of lens pigments. Atomic force microscopy (Dimension Icon AFM from Bruker Nano) was used to determine the surface roughness of the pigmented regions of the contact lenses. Atomic force microscopic analysis was performed in fluid phase under contact mode using a Sharp Nitride Lever probe (SNL-10) with a spring constant of 0.06 N/m. Root mean square (RMS) roughness values were analysed using a generalised linear mixed model with a log-normal distribution. Least square means and their corresponding 95% confidence intervals were estimated for each brand, location and pigment combination. SEM cross-sectional images at 500× and 2,000× magnification showed pigment on the surface of six of the seven lens types tested. The mean depth of pigment for 1-DAY ACUVUE DEFINE (1DAD) lenses was 8.1 μm below the surface of the lens, while the remaining lens types tested had pigment particles on the front or back surface. Results of the atomic force microscopic analysis indicated that 1DAD lenses had significantly lower root mean square roughness values in the pigmented area of the lens than the other lens types tested. SEM and AFM analysis revealed pigment on the surface of the lens for all types tested with the exception of 1DAD. Further research is required to determine if the difference in pigment location influences on-eye performance. © 2014 The Authors. Clinical and Experimental

  18. Comparison of Scheimpflug-photography, specular microscopy and scanning electron microscopy to detect corneal changes in toxicity studies in rats

    SciTech Connect

    Boeker, T.W.; Wegener, A.; Koch, F.; Hockwin, O. )

    1990-01-01

    With an increasing number of in-vivo methods to examine the eyes of laboratory animals, the rat has become an important animal model in experimental eye research. Specular microscopy is a clinical tool to examine the corneal endothelium in-vivo. To evaluate the versatility of this method for small animal eyes, we studied both corneal endothelial cell-count and corneal thickness in normal rats as well as those with diabetic, naphthalene and UV-B cataract. As a reference scanning electron microscopy (SEM) of the corneal endothelium was performed. For cell-counts the correlation coefficient between both methods was found to be sufficient. The comparison of corneal thickness measurement (SEM-values) with specular microscopy and with Scheimpflugbiometry failed to show a satisfactory correlation. The study proves that specular microscopy is a useful tool to document changes also in the endothelium of the rat-cornea.

  19. High-resolution electron microscopy and electron energy-loss spectroscopy of giant palladium clusters

    NASA Astrophysics Data System (ADS)

    Oleshko, V.; Volkov, V.; Gijbels, R.; Jacob, W.; Vargaftik, M.; Moiseev, I.; van Tendeloo, G.

    1995-12-01

    Combined structural and chemical characterization of cationic polynuclear palladium coordination compounds Pd561L60(OAc)180, where L=1,10-phenantroline or 2,2'-bipyridine has been carried out by high-resolution electron microscopy (HREM) and analytical electron microscopy methods including electron energy-loss spectroscopy (EELS), zero-loss electron spectroscopic imaging, and energy-dispersive X-ray spectroscopy (EDX). The cell structure of the cluster matter with almost completely uniform metal core size distributions centered around 2.3 ±0.5 nm was observed. Zero-loss energy filtering allowed to improve the image contrast and resolution. HREM images showed that most of the palladium clusters had a cubo-octahedral shape. Some of them had a distorted icosahedron structure exhibiting multiple twinning. The selected-area electron diffraction patterns confirmed the face centered cubic structure with lattice parameter close to that of metallic palladium. The energy-loss spectra of the populations of clusters contained several bands, which could be assigned to the delayed Pd M4, 5-edge at 362 eV, the Pd M3-edge at 533 eV and the Pd M2-edge at 561 eV, the NK-edge at about 400 eV, the O K-edge at 532 eV overlapping with the Pd M3-edge and the carbon C K-edge at 284 eV. Background subtraction was applied to reveal the exact positions and fine structure of low intensity elemental peaks. EELS evaluations have been confirmed by EDX. The recorded series of the Pd M-edges and the N K-edge in the spectra of the giant palladium clusters obviously were related to Pd-Pd- and Pd-ligand bonding.

  20. Correlated Cryo-fluorescence and Cryo-electron Microscopy with High Spatial Precision and Improved Sensitivity

    PubMed Central

    Schorb, Martin; Briggs, John A. G.

    2017-01-01

    Performing fluorescence microscopy and electron microscopy on the same sample allows fluorescent signals to be used to identify and locate features of interest for subsequent imaging by electron microscopy. To carry out such correlative microscopy on vitrified samples appropriate for structural cryo-electron microscopy it is necessary to perform fluorescence microscopy at liquid-nitrogen temperatures. Here we describe an adaptation of a cryo-light microscopy stage to permit use of high-numerical aperture objectives. This allows high-sensitivity and high-resolution fluorescence microscopy of vitrified samples. We describe and apply a correlative cryo-fluorescence and cryo-electron microscopy workflow together with a fiducial bead-based image correlation procedure. This procedure allows us to locate fluorescent bacteriophages in cryo-electron microscopy images with an accuracy on the order of 50 nm, based on their fluorescent signal. It will allow the user to precisely and unambiguously identify and locate objects and events for subsequent high-resolution structural study, based on fluorescent signals. PMID:24275379