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Quantitative analytical electron microscopy of multiphase alloys.  


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. PMID:2709131

Prybylowski, J; Ballinger, R; Elliott, C



Quantitative characterization of electron detectors for transmission electron microscopy  

PubMed Central

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

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



Quantitative Transmission Electron Microscopy of Magnetic Minerals T. Kasama1,*  

E-print Network

in rocks on the Earth and on other planets [1]. Magnetite is also found in magnetotactic bacteria and many mineral to magnetite, is less common in nature but is sometimes found in magnetotactic bacteria transmission electron microscopy (TEM) to study (1) greigite-containing magnetotactic bacteria and (2

Dunin-Borkowski, Rafal E.


Quantitative light and scanning electron microscopy of ferret sperm.  


Sperm were obtained via electroejaculation from Domestic ferret, (Mustela putorius furo), Siberian ferret (M. eversmanni), Black-footed ferret (M. nigripes), and a hybrid between Siberian and Domestic, called the Fitch ferret (M. sp.). Comparisons of sperm were made by four different microscopy techniques to determine whether differences exist among species. First, Nomarski differential interference microscopy could be used to distinguish domestic ferret sperm from the others on the basis of the structure of the posterior part of the acrosome. Second, both silver staining, which demonstrates argentophilic protein distribution, and scanning electron microscopy (SEM), revealed differences among the morphology of sperm for each species; variation in the unique appearance of the acrosome in ferret sperm was detected especially well by SEM. To quantify differences in morphology, five sperm head parameters were measured using image analysis; light microscopy produced significantly larger values than did SEM (all parameters and all species but Fitch), and there were significant differences owing to species for all parameters but one. Generally, our data demonstrate the value of complementary techniques to distinguish among sperm of closely related species and more specifically may help establish evolutionary relationships among the ferret species studied. In addition, they provide baseline data important for the captive breeding of the endangered Black-footed ferret. PMID:1793602

Van der Horst, G; Curry, P T; Kitchin, R M; Burgess, W; Thorne, E T; Kwiatkowski, D; Parker, M; Atherton, R W



A technique for quantitative and qualitative viewing of aquatic bacteria using scanning electron microscopy  

E-print Network


Dreier, Thomas Michael



Electron Microscopy.  

ERIC Educational Resources Information Center

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)

Beer, Michael



Quantitative investigation of surface and subsurface fatigue cracks near rivets in riveted joints using acoustic, electron and optical microscopy  

Microsoft Academic Search

Using scanning acoustic microscopy, optical microscopy and scanning electron microscopy, in conjunction with fractography of fractured surfaces, the crack formation and growth kinetics of subsurface fatigue cracks and surface breaking fatigue cracks near rivets have been characterized in detail in this research. The scanning acoustic microscope was used to quantitatively investigate subsurface fatigue cracks (even when they were very small)

Zayna Marie Connor



Compositional analysis of GaAs/AlGaAs heterostructures using quantitative scanning transmission electron microscopy  

SciTech Connect

We demonstrate a method for compositional mapping of Al{sub x}Ga{sub 1–x}As heterostructures with high accuracy and unit cell spatial resolution using quantitative high angle annular dark field scanning transmission electron microscopy. The method is low dose relative to spectroscopic methods and insensitive to the effective source size and higher order lens aberrations. We apply the method to study the spatial variation in Al concentration in cross-sectioned GaAs/AlGaAs core-shell nanowires and quantify the concentration in the Al-rich radial band and the AlGaAs shell segments.

Kauko, H.; Helvoort, A. T. J. van [Department of Physics, Norwegian University of Science and Technology (NTNU), Trondheim (Norway); Zheng, C. L.; Glanvill, S. [Monash Centre for Electron Microscopy, Monash University, VIC 3800 (Australia)] [Monash Centre for Electron Microscopy, Monash University, VIC 3800 (Australia); Zhu, Y.; Etheridge, J., E-mail: [Monash Centre for Electron Microscopy, Monash University, VIC 3800 (Australia); Department of Materials Engineering, Monash University, VIC 3800 (Australia); Dwyer, C. [Monash Centre for Electron Microscopy, Monash University, VIC 3800 (Australia) [Monash Centre for Electron Microscopy, Monash University, VIC 3800 (Australia); Ernst Ruska-Centre for Microscopy and Spectroscopy with Electrons, and Peter Grünberg Institute, Forschungszentrum Jülich, D-52425 Jülich (Germany); Munshi, A. M.; Fimland, B. O. [Department of Electronics and Telecommunications, Norwegian University of Science and Technology (NTNU), Trondheim (Norway)] [Department of Electronics and Telecommunications, Norwegian University of Science and Technology (NTNU), Trondheim (Norway)



A methodology for the extraction of quantitative information from electron microscopy images at the atomic level  

NASA Astrophysics Data System (ADS)

In this paper we describe a methodology developed at the University of Cadiz (Spain) in the past few years for the extraction of quantitative information from electron microscopy images at the atomic level. This work is based on a coordinated and synergic activity of several research groups that have been working together over the last decade in two different and complementary fields: Materials Science and Computer Science. The aim of our joint research has been to develop innovative high-performance computing techniques and simulation methods in order to address computationally challenging problems in the analysis, modelling and simulation of materials at the atomic scale, providing significant advances with respect to existing techniques. The methodology involves several fundamental areas of research including the analysis of high resolution electron microscopy images, materials modelling, image simulation and 3D reconstruction using quantitative information from experimental images. These techniques for the analysis, modelling and simulation allow optimizing the control and functionality of devices developed using materials under study, and have been tested using data obtained from experimental samples.

Galindo, P. L.; Pizarro, J.; Guerrero, E.; Guerrero-Lebrero, M. P.; Scavello, G.; Yáñez, A.; Núñez-Moraleda, B. M.; Maestre, J. M.; Sales, D. L.; Herrera, M.; Molina, S. I.



Quantitative magnetic imaging at the nanometer scale by ballistic electron magnetic microscopy  

SciTech Connect

We demonstrate quantitative ballistic electron magnetic microscopy (BEMM) imaging of simple model Fe(001) nanostructures. We use in situ nanostencil shadow mask resistless patterning combined with molecular beam epitaxy deposition to prepare under ultra-high vacuum conditions nanostructured epitaxial Fe/Au/Fe/GaAs(001) spin-valves. In this epitaxial system, the magnetization of the bottom Fe/GaAs(001) electrode is parallel to the [110] direction, defining accurately the analysis direction for the BEMM experiments. The large hot-electron magnetoresistance of the Fe/Au/Fe/GaAs(001) epitaxial spin-valve allows us to image various stable magnetic configurations on the as-grown Fe(001) microstructures with a high sensitivity, even for small misalignments of both magnetic electrodes. The angular dependence of the hot-electron magnetocurrent is used to convert magnetization maps calculated by micromagnetic simulations into simulated BEMM images. The calculated BEMM images and magnetization rotation profiles show quantitative agreement with experiments and allow us to investigate the magnetic phase diagram of these model Fe(001) microstructures. Finally, magnetic domain reversals are observed under high current density pulses. This opens the way for further BEMM investigations of current-induced magnetization dynamics.

Herve, M.; Tricot, S.; Guezo, S.; Delhaye, G.; Lepine, B.; Schieffer, P.; Turban, P. [Departement Materiaux et Nanosciences, Institut de Physique de Rennes, UMR 6251, CNRS-Universite de Rennes 1, Campus de Beaulieu, Bat 11E, 35042 Rennes cedex (France)] [Departement Materiaux et Nanosciences, Institut de Physique de Rennes, UMR 6251, CNRS-Universite de Rennes 1, Campus de Beaulieu, Bat 11E, 35042 Rennes cedex (France)



Quantitative magnetic imaging at the nanometer scale by ballistic electron magnetic microscopy  

NASA Astrophysics Data System (ADS)

We demonstrate quantitative ballistic electron magnetic microscopy (BEMM) imaging of simple model Fe(001) nanostructures. We use in situ nanostencil shadow mask resistless patterning combined with molecular beam epitaxy deposition to prepare under ultra-high vacuum conditions nanostructured epitaxial Fe/Au/Fe/GaAs(001) spin-valves. In this epitaxial system, the magnetization of the bottom Fe/GaAs(001) electrode is parallel to the [110] direction, defining accurately the analysis direction for the BEMM experiments. The large hot-electron magnetoresistance of the Fe/Au/Fe/GaAs(001) epitaxial spin-valve allows us to image various stable magnetic configurations on the as-grown Fe(001) microstructures with a high sensitivity, even for small misalignments of both magnetic electrodes. The angular dependence of the hot-electron magnetocurrent is used to convert magnetization maps calculated by micromagnetic simulations into simulated BEMM images. The calculated BEMM images and magnetization rotation profiles show quantitative agreement with experiments and allow us to investigate the magnetic phase diagram of these model Fe(001) microstructures. Finally, magnetic domain reversals are observed under high current density pulses. This opens the way for further BEMM investigations of current-induced magnetization dynamics.

Hervé, M.; Tricot, S.; Guézo, S.; Delhaye, G.; Lépine, B.; Schieffer, P.; Turban, P.



Microfibrous quartz varieties: characterization by quantitative X-ray texture analysis and transmission electron microscopy  

NASA Astrophysics Data System (ADS)

Quantitative X-ray texture analysis was used to identify the fiber elongation direction of microfibrous quartz varieties in materials that contain narrow bands of closely spaced, radially divergent spherulitic fiber bundles. The statistical approach of X-ray texture analysis makes it possible to determine the fiber axis direction even when the maximum divergence angle of fibers within spherulitic fiber bundles diverge +/-20° from a common growth direction. Representative samples of chalcedony <11&2macr0>, quartzine [0001], and pseudochalcedony were selected for additional study with transmis-sion electron microscopy. All varieties of microfibrous quartz were found to contain high concentrations of polysynthetic Brazil twins and domains characterized by a modulated microstructure. Trace element profiles of aluminum and iron concentration were measured by electron microprobe analysis for an agate sample that contains alternating bands of chalcedony <11&2macr0> and quartzine fibers. The elemental concentration profiles were measured in a direction parallel to the growth direction of microfibrous quartz. Although fibers within the chalcedony bands initially displayed cooperative helical twisting, the fibers became untwisted during the later stages of growth within any one band. The trace element profiles revealed that higher concentrations of aluminum are associated with quartzine and non-twisted <11&2macr0> fibers, rather than twisted chalcedony <11&2macr0> fibers. Except for the presence of iron-rich inclusions between quartzine and chalcedony bands, trace concentrations of iron were below the detection limit for the different varieties of microfibrous quartz analyzed.

Cady, S. L.; Wenk, H.-R.; Sintubin, M.


Quantitative high-angle annular dark field scanning transmission electron microscopy for materials science  

Microsoft Academic Search

Scanning transmission electron microscopy (STEM) has been widely used for characterization of materials; to identify micro- and nano-structures within a sample and to analyze crystal and defect structures. High-angle annular dark field (HAADF) STEM imaging using atomic number (Z) contrast has proven capable of resolving atomic structures with better than 2 A lateral resolution. In this work, the HAADF STEM

Rumyana V. Petrova



Quantitative scanning electron microscopy of solitary chemoreceptor cells in cyprinids and other teleosts  

Microsoft Academic Search

Synopsis Solitary chemosensory cells (SCC) occur in the epidermis of many lower, aquatic vertebrates. By scanning electron microscopy, SCC apices were counted and density distributions estimated along various transects at the head and body of 12 species of teleost fishes, 7 cyprinids, 2 perciforms, 2 catfish and 1 characinid. In contrast to taste buds (TB), the distribution of SCCs is

Kurt Kotrschal; Konrad Lorenz



Quantitative measurements in in situ straining experiments in transmission electron microscopy.  


Several examples of recent studies by in situ straining experiments in a transmission electron microscope performed in the Toulouse group (France) are presented. In particular, quantitative measurements of the features of the dislocation motion are described. These examples deal with individual or collective propagation of dislocations, which are submitted to various types of obstacle. Different metallic materials are investigated: magnesium, intermetallics, aluminium alloys and gamma phase of a superalloy. PMID:11454154

Pettinari, F; Couret, A; Caillard, D; Molénat, G; Clément, N; Coujou, A



Sub-0.1 nm-resolution quantitative scanning transmission electron microscopy without adjustable parameters  

SciTech Connect

Atomic-resolution imaging in the scanning transmission electron microscope (STEM) constitutes a powerful tool for nanostructure characterization. Here, we demonstrate the quantitative interpretation of atomic-resolution high-angle annular dark-field (ADF) STEM images using an approach that does not rely on adjustable parameters. We measure independently the instrumental parameters that affect sub-0.1 nm-resolution ADF images, quantify their individual and collective contributions to the image intensity, and show that knowledge of these parameters enables a quantitative interpretation of the absolute intensity and contrast across all accessible spatial frequencies. The analysis also provides a method for the in-situ measurement of the STEM's effective source distribution.

Dwyer, C. [Monash Centre for Electron Microscopy, Monash University, Victoria 3800 (Australia); Department of Materials Engineering, Monash University, Victoria 3800 (Australia); ARC Centre of Excellence for Design in Light Metals, Monash University, Victoria 3800 (Australia); Maunders, C. [Department of Materials Engineering, Monash University, Victoria 3800 (Australia); Zheng, C. L. [Monash Centre for Electron Microscopy, Monash University, Victoria 3800 (Australia); Weyland, M.; Etheridge, J. [Monash Centre for Electron Microscopy, Monash University, Victoria 3800 (Australia); Department of Materials Engineering, Monash University, Victoria 3800 (Australia); Tiemeijer, P. C. [FEI Electron Optics, P.O. Box 80066, 5600 KA Eindhoven (Netherlands)



Dose limited reliability of quantitative annular dark field scanning transmission electron microscopy for nano-particle atom-counting.  


Quantitative annular dark field scanning transmission electron microscopy (ADF STEM) has become a powerful technique to characterise nano-particles on an atomic scale. Because of their limited size and beam sensitivity, the atomic structure of such particles may become extremely challenging to determine. Therefore keeping the incoming electron dose to a minimum is important. However, this may reduce the reliability of quantitative ADF STEM which will here be demonstrated for nano-particle atom-counting. Based on experimental ADF STEM images of a real industrial catalyst, we discuss the limits for counting the number of atoms in a projected atomic column with single atom sensitivity. We diagnose these limits by combining a thorough statistical method and detailed image simulations. PMID:25511931

De Backer, A; Martinez, G T; MacArthur, K E; Jones, L; Béché, A; Nellist, P D; Van Aert, S



Calibration-free quantitative surface topography reconstruction in scanning electron microscopy.  


This work presents a new approach to obtain reliable surface topography reconstructions from 2D Scanning Electron Microscopy (SEM) images. In this method a set of images taken at different tilt angles are compared by means of digital image correlation (DIC). It is argued that the strength of the method lies in the fact that precise knowledge about the nature of the rotation (vector and/or magnitude) is not needed. Therefore, the great advantage is that complex calibrations of the measuring equipment are avoided. The paper presents the necessary equations involved in the methods, including derivations and solutions. The method is illustrated with examples of 3D reconstructions followed by a discussion on the relevant experimental parameters. PMID:25255373

Faber, E T; Martinez-Martinez, D; Mansilla, C; Ocelík, V; Hosson, J Th M De



Development of a new quantitative X-ray microanalysis method for electron microscopy.  


Quantitative X-ray microanalysis of thick samples is usually performed by measuring the characteristic X-ray intensities of each element in a sample and in corresponding standards. The ratio of the measured intensities from the unknown material to that from the standard is related to the concentration using the ZAF or ?(?z) equations. Under optimal conditions, accuracies approaching 1% are possible. However, all the experimental conditions must remain the same during the sample and standard measurements. This is not possible with cold field emission scanning electron microscopes (FE-SEMs) where beam current can fluctuate around 5% in its stable regime. Very little work has been done on variable beam current conditions (Griffin, B.J. & Nockolds, C.E., Scanning 13, 307-312, 1991), and none relating to cold FE-SEM applications. To address this issue, a new method was developed using a single spectral measurement. It is similar in approach to the Cliff-Lorimer method developed for the analytical transmission electron microscope. However, corrections are made for X rays generated from thick specimens using the ratio of the characteristic X-ray intensities of two elements in the same material. The proposed method utilizes the ratio of the intensity of a characteristic X-ray normalized by the sum of X-ray intensities of all the elements measured for the sample, which should also reduce the amplitude of error propagation. Uncertainties in the physical parameters of X-ray generation are corrected using a calibration factor that must be previously acquired or calculated. As an example, when this method was applied to the calculation of the composition of Au-Cu National Institute of Standards and Technology standards measured with a cold field emission source SEM, relative accuracies better than 5% were obtained. PMID:20961482

Horny, Paula; Lifshin, Eric; Campbell, Helen; Gauvin, Raynald



Quantitative magnetic imaging at the nanometer scale by Ballistic Electron Magnetic Microscopy  

E-print Network

(BEMM) imaging of simple model Fe(001) nanostructures. We use in situ nanostencil shadow mask resistless direction for the BEMM experiments. The large hot- electron magnetoresistance of the Fe/Au/Fe/GaAs(001 by micromagnetic simulations into simulated BEMM images. The calculated BEMM images and magnetization rotation

Paris-Sud XI, Université de


Application of Quantitative Analytical Electron Microscopy to the Mineral Content of Insect Cuticle  

NASA Astrophysics Data System (ADS)

Quantification of calcium in the cuticle of the fly larva Exeretonevra angustifrons was undertaken at the micron scale using wavelength dispersive X-ray microanalysis, analytical standards, and a full matrix correction. Calcium and phosphorus were found to be present in the exoskeleton in a ratio that indicates amorphous calcium phosphate. This was confirmed through electron diffraction of the calcium-containing tissue. Due to the pragmatic difficulties of measuring light elements, it is not uncommon in the field of entomology to neglect the use of matrix corrections when performing microanalysis of bulk insect specimens. To determine, firstly, whether such a strategy affects the outcome and secondly, which matrix correction is preferable, phi-rho (z) and ZAF matrix corrections were contrasted with each other and without matrix correction. The best estimate of the mineral phase was found to be given by using the phi-rho (z) correction. When no correction was made, the ratio of Ca to P fell outside the range for amorphous calcium phosphate, possibly leading to flawed interpretation of the mineral form when used on its own.

Rasch, Ron; Cribb, Bronwen W.; Barry, John; Palmer, Christopher M.



A combined quantitative mass spectrometry and electron microscopy analysis of ribosomal 30S subunit assembly in E. coli  

PubMed Central

Ribosome assembly is a complex process involving the folding and processing of ribosomal RNAs (rRNAs), concomitant binding of ribosomal proteins (r-proteins), and participation of numerous accessory cofactors. Here, we use a quantitative mass spectrometry/electron microscopy hybrid approach to determine the r-protein composition and conformation of 30S ribosome assembly intermediates in Escherichia coli. The relative timing of assembly of the 3? domain and the formation of the central pseudoknot (PK) structure depends on the presence of the assembly factor RimP. The central PK is unstable in the absence of RimP, resulting in the accumulation of intermediates in which the 3?-domain is unanchored and the 5?-domain is depleted for r-proteins S5 and S12 that contact the central PK. Our results reveal the importance of the cofactor RimP in central PK formation, and introduce a broadly applicable method for characterizing macromolecular assembly in cells. DOI: PMID:25313868

Sashital, Dipali G; Greeman, Candacia A; Lyumkis, Dmitry; Potter, Clinton S; Carragher, Bridget; Williamson, James R



Mitochondrial morphology, topology, and membrane interactions in skeletal muscle: a quantitative three-dimensional electron microscopy study.  


Dynamic remodeling of mitochondrial morphology through membrane dynamics are linked to changes in mitochondrial and cellular function. Although mitochondrial membrane fusion/fission events are frequent in cell culture models, whether mitochondrial membranes dynamically interact in postmitotic muscle fibers in vivo remains unclear. Furthermore, a quantitative assessment of mitochondrial morphology in intact muscle is lacking. Here, using electron microscopy (EM), we provide evidence of interacting membranes from adjacent mitochondria in intact mouse skeletal muscle. Electron-dense mitochondrial contact sites consistent with events of outer mitochondrial membrane tethering are also described. These data suggest that mitochondrial membranes interact in vivo among mitochondria, possibly to induce morphology transitions, for kiss-and-run behavior, or other processes involving contact between mitochondrial membranes. Furthermore, a combination of freeze-fracture scanning EM and transmission EM in orthogonal planes was used to characterize and quantify mitochondrial morphology. Two subpopulations of mitochondria were studied: subsarcolemmal (SS) and intermyofibrillar (IMF), which exhibited significant differences in morphological descriptors, including form factor (means ± SD for SS: 1.41 ± 0.45 vs. IMF: 2.89 ± 1.76, P < 0.01) and aspect ratio (1.97 ± 0.83 vs. 3.63 ± 2.13, P < 0.01) and circularity (0.75 ± 0.16 vs. 0.45 ± 0.22, P < 0.01) but not size (0.28 ± 0.31 vs. 0.27 ± 0.20 ?m(2)). Frequency distributions for mitochondrial size and morphological parameters were highly skewed, suggesting the presence of mechanisms to influence mitochondrial size and shape. In addition, physical continuities between SS and IMF mitochondria indicated mixing of both subpopulations. These data provide evidence that mitochondrial membranes interact in vivo in mouse skeletal muscle and that factors may be involved in regulating skeletal muscle mitochondrial morphology. PMID:23104694

Picard, Martin; White, Kathryn; Turnbull, Douglass M



Transmission Electron Microscopy Basics  

NSDL National Science Digital Library

This extensive site from the University of Liverpool is a set of resources based on the textbook Transmission Electron Microscopy - Basics by D.B.Williams and C.B.Carter. The tutorial is designed to accompany an introductory course on transmission electron microscopy for students with an understanding of elementary physics. Topics include electron scattering, electron atom interactions, the electron gun, probe size, lenses, depth of field and depth of focus, and others. Each chapter includes interactive Java applets that facilitate understanding of the concepts presented.

Peter Goodhew


QEMSCAN° (Quantitative Evaluation of Minerals by Scanning Electron Microscopy): capability and application to fracture characterization in geothermal systems  

NASA Astrophysics Data System (ADS)

Fractures are important conduits for fluids in geothermal systems, and the creation and maintenance of fracture permeability is a fundamental aspect of EGS (Engineered Geothermal System) development. Hydraulic or chemical stimulation techniques are often employed to achieve this. In the case of chemical stimulation, an understanding of the minerals present in the fractures themselves is desirable to better design a stimulation effort (i.e. which chemical to use and how much). Borehole televiewer surveys provide important information about regional and local stress regimes and fracture characteristics (e.g. fracture aperture), and XRD is useful for examining bulk rock mineralogy, but neither technique is able to quantify the distribution of these minerals in fractures. QEMSCAN° is a fully-automated micro-analysis system that enables quantitative chemical analysis of materials and generation of high-resolution mineral maps and images as well as porosity structure. It uses a scanning electron microscopy platform (SEM) with an electron beam source in combination with four energy-dispersive X-ray spectrometers (EDS). The measured backscattered electron and electron-induced secondary X-ray emission spectra are used to classify sample mineralogy. Initial applications of QEMSCAN° technology were predominantly in the minerals industry and application to geothermal problems has remained limited to date. In this pilot study, the potential application of QEMSCAN° technology to fracture characterization was evaluated using samples of representative mineralized fractures in two geothermal systems (Newberry Volcano, Oregon and Brady's geothermal field, Nevada). QEMSCAN° results were compared with XRD and petrographic techniques. Nine samples were analyzed from each field, collected from the drill core in the 1000-1500 m depth range in two shallow wells (GEO-N2 at Newberry Volcano and BCH-3 at Brady's). The samples were prepared as polished thin sections for QEMSCAN° analysis. Results indicate that a sampling resolution of 10 ?m is sufficient to resolve fracture morphology and mineral zonation (where multiple episodes of mineralization occurred), and enables relatively fast data acquisition (3 cm2 can be analyzed in approximately 3 hours). Finer resolutions (down to 2.5 ?m) take significantly longer, but can be used to provide additional spatial detail in areas of interest after a low resolution (10 ?m) scan. Use of XRD data in conjunction with QEMSCAN° data is sometimes needed to distinguish geothermal alteration minerals with similar chemical compositions (clay minerals, micas and chlorite), however overall the technique appears to have excellent potential for geothermal applications.

Ayling, B.; Rose, P. E.; Zemach, E.; Drakos, P. S.; Petty, S.



Scanning ultrafast electron microscopy  

PubMed Central

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

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




Technology Transfer Automated Retrieval System (TEKTRAN)

Transmission electron microscopy (TEM) has long been an important tool in the expanding field of microbiology. A major virtue of TEM is the increased resolution afforded by the use of accelerated electron beams at wavelengths significantly shorter than visible light. When coupled with suitable pre...


Characterization of individual submicrometer aerosol particles collected in Incheon, Korea, by quantitative transmission electron microscopy energy-dispersive X-ray spectrometry  

NASA Astrophysics Data System (ADS)

For the last decade the Monte Carlo calculation method has been proven to be an excellent tool for accurately simulating electron-solid interactions in atmospheric individual particles of micrometer size. Although it was designed for application to scanning electron microscopy, in the present study it is demonstrated that the Monte Carlo calculation can also be applied in a quantitative single particle analysis using transmission electron microscopy (TEM) with an ultrathin window energy-dispersive X-ray (EDX) spectrometer with a high accelerating voltage (200 kV). By utilizing an iterative reverse Monte Carlo simulation combined with successive approximation, atomic elemental concentrations (including low-Z elements) of submicrometer standard particles were determined with high accuracy for electron beam refractory particles such as NaCl, KCl, SiO2, Fe2O3, Na2SO4, K2SO4, CaCO3, and CaSO4. On the basis of quantitative X-ray analysis together with morphological information from TEM images, overall 1638 submicrometer individual particles from 10 sets of aerosol samples collected in Incheon, Korea, were identified. The most frequently encountered particle types are carbonaceous and (NH4)2SO4/NH4HSO4-containing particles, followed by mineral (e.g., aluminosilicate, SiO2, CaCO3), sea salt, K-rich (e.g., K2SO4 and KCl), Fe-rich, fly ash, and transition or heavy-metal-containing (e.g., ZnSO4, ZnCl2, PbSO4) particles. The relative abundances of the submicrometer particle types vary among samples collected in different seasons and also depend on different air mass transport routes. This study demonstrates that the quantitative TEM-EDX individual particle analysis is a useful and reliable technique in characterizing urban submicrometer aerosol particles.

Geng, Hong; Kang, Sujin; Jung, Hae-Jin; ChoëL, Marie; Kim, Hyekyeong; Ro, Chul-Un



Nanoscale analysis of clustered DNA damage after high-LET irradiation by quantitative electron microscopy - The heavy burden to repair.  


Low- and high-linear energy transfer (LET) ionising radiation are effective cancer therapies, but produce structurally different forms of DNA damage. Isolated DNA damage is repaired efficiently; however, clustered lesions may be more difficult to repair, and are considered as significant biological endpoints. We investigated the formation and repair of DNA double-strand breaks (DSBs) and clustered lesions in human fibroblasts after exposure to sparsely (low-LET; delivered by photons) and densely (high-LET; delivered by carbon ions) ionising radiation. DNA repair factors (pKu70, 53BP1, ?H2AX, and pXRCC1) were detected using immunogold-labelling and electron microscopy, and spatiotemporal DNA damage patterns were analysed within the nuclear ultrastructure at the nanoscale level. By labelling activated Ku-heterodimers (pKu70) the number of DSBs was determined in electron-lucent euchromatin and electron-dense heterochromatin. Directly after low-LET exposure (5min post-irradiation), single pKu70 dimers, which reflect isolated DSBs, were randomly distributed throughout the entire nucleus with a linear dose correlation up to 30Gy. Most euchromatic DSBs were sensed and repaired within 40min, whereas heterochromatic DSBs were processed with slower kinetics. Essentially all DNA lesions induced by low-LET irradiation were efficiently rejoined within 24h post-irradiation. High-LET irradiation caused localised energy deposition within the particle tracks, and generated highly clustered DNA lesions with multiple DSBs in close proximity. The dimensions of these clustered lesions along the particle trajectories depended on the chromatin packing density, with huge DSB clusters predominantly localised in condensed heterochromatin. High-LET irradiation-induced clearly higher DSB yields than low-LET irradiation, with up to ?500 DSBs per ?m(3) track volume, and large fractions of these heterochromatic DSBs remained unrepaired. Hence, the spacing and quantity of DSBs in clustered lesions influence DNA repair efficiency, and may determine the radiobiological outcome. PMID:25659339

Lorat, Yvonne; Brunner, Christina U; Schanz, Stefanie; Jakob, Burkhard; Taucher-Scholz, Gisela; Rübe, Claudia E



Silver stain for electron microscopy  

NASA Technical Reports Server (NTRS)

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.

Corbett, R. L.



A quantitative estimation of the exhaust, abrasion and resuspension components of particulate traffic emissions using electron microscopy  

NASA Astrophysics Data System (ADS)

The contribution of the three traffic-related components exhaust, abrasion, and resuspension to kerbside and urban background PM10 and PM1 levels was quantified based on the analysis of individual particles by scanning electron microscopy. A total of 160 samples was collected on 38 days between February and September 2009 at a kerbside and an urban background station in the urban/industrial Ruhr area (Germany). Based on size, morphology, chemical composition and stability under electron bombardment, the 111,003 particles studied in detail were classified into the following 14 particle classes: traffic/exhaust, traffic/abrasion, traffic/resuspension, carbonaceous/organic, industry/metallurgy, industry/power plants, secondary particles, (aged) sea salt, silicates, Ca sulfates, carbonates, Fe oxides/hydroxides, biological particles, and other particles. The traffic/exhaust component consists predominantly of externally mixed soot particles and soot internally mixed with secondary particles. The traffic/abrasion component contains all particles with characteristic tracer elements (Fe, Cu, Ba, Sb, Zn) for brake and tire abrasion. The traffic/resuspension component is defined by the mixing state and comprises all internally mixed particles with a high proportion of silicates or Fe oxides/hydroxides which contain soot or abrasion particles as minor constituent. In addition, silicates and Fe oxides/hydroxides internally mixed with chlorine and sulphur containing particles were also assigned to the traffic/resuspension component. The total contribution of traffic to PM10 was found to be 27% at the urban background station and 48% at the kerbside station, the corresponding values for PM1 are 15% and 39%. These values lie within the range reported in previous literature. The relative share of the different traffic components for PM10 at the kerbside station was 27% exhaust, 15% abrasion, and 58% resuspension (38%, 8%, 54% for PM1). For the urban background, the following relative shares were obtained for PM10: 22% exhaust, 22% abrasion and 56% resuspension (40%, 27%, 33% for PM1). Compared to previous publications we have observed a significantly lower portion of exhaust particles and a significantly higher portion of resuspension particles. The high abundance of resuspension particles underlines their significance for the observed adverse health effects of traffic emissions and for mitigation measures.

Weinbruch, Stephan; Worringen, Annette; Ebert, Martin; Scheuvens, Dirk; Kandler, Konrad; Pfeffer, Ulrich; Bruckmann, Peter



High speed quantitative digital microscopy  

NASA Technical Reports Server (NTRS)

Modern digital image processing hardware makes possible quantitative analysis of microscope images at high speed. This paper describes an application to automatic screening for cervical cancer. The system uses twelve MC6809 microprocessors arranged in a pipeline multiprocessor configuration. Each processor executes one part of the algorithm on each cell image as it passes through the pipeline. Each processor communicates with its upstream and downstream neighbors via shared two-port memory. Thus no time is devoted to input-output operations as such. This configuration is expected to be at least ten times faster than previous systems.

Castleman, K. R.; Price, K. H.; Eskenazi, R.; Ovadya, M. M.; Navon, M. A.



Rigorous quantitative elemental microanalysis by scanning electron microscopy/energy dispersive x-ray spectrometry (SEM/EDS) with spectrum processing by NIST DTSA-II  

NASA Astrophysics Data System (ADS)

Quantitative electron-excited x-ray microanalysis by scanning electron microscopy/silicon drift detector energy dispersive x-ray spectrometry (SEM/SDD-EDS) is capable of achieving high accuracy and high precision equivalent to that of the high spectral resolution wavelength dispersive x-ray spectrometer even when severe peak interference occurs. The throughput of the SDD-EDS enables high count spectra to be measured that are stable in calibration and resolution (peak shape) across the full deadtime range. With this high spectral stability, multiple linear least squares peak fitting is successful for separating overlapping peaks and spectral background. Careful specimen preparation is necessary to remove topography on unknowns and standards. The standards-based matrix correction procedure embedded in the NIST DTSA-II software engine returns quantitative results supported by a complete error budget, including estimates of the uncertainties from measurement statistics and from the physical basis of the matrix corrections. NIST DTSA-II is available free for Java-platforms at:

Newbury, Dale E.; Ritchie, Nicholas W. M.



Electron microscopy and forensic practice  

NASA Astrophysics Data System (ADS)

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).

Kotrlý, Marek; Turková, Ivana



Quantitative analysis on volcanic ash surfaces: Application of extended depth-of-field (focus) algorithm for light and scanning electron microscopy and 3D reconstruction  

Microsoft Academic Search

The depth-of-field mainly affects the image quality either in scanning electron microscopy (SEM) or conventional light microscopy. The limited depth-of-field handicap of microscopy imaging can be used for obtaining “optically sectioned” specimens by moving the object along the optical axis. In this study, multiple images corresponding to different object planes were taken in order to overcome limited depth-of-field on conventional

Orkun Ersoy; Erkan Aydar; Alain Gourgaud; Hasan Bayhan



Electronic Blending in Virtual Microscopy  

ERIC Educational Resources Information Center

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…

Maybury, Terrence S.; Farah, Camile S.



Quantitative imaging of bilirubin by photoacoustic microscopy  

NASA Astrophysics Data System (ADS)

Noninvasive detection of both bilirubin concentration and its distribution is important for disease diagnosis. Here we implemented photoacoustic microscopy (PAM) to detect bilirubin distribution. We first demonstrate that our PAM system can measure the absorption spectra of bilirubin and blood. We also image bilirubin distributions in tissuemimicking samples, both without and with blood mixed. Our results show that PAM has the potential to quantitatively image bilirubin in vivo for clinical applications.

Zhou, Yong; Zhang, Chi; Yao, Da-Kang; Wang, Lihong V.



In Situ Transmission Electron Microscopy  

Microsoft Academic Search

Since the earliest days of transmission electron microscopy, microscopists have realized the potential of microscopy for studying\\u000a dynamic processes. Images recorded sequentially can be used to track the changes caused by deliberate actions, such as heating\\u000a or straining, or uncontrolled processes, such as beam damage. The class of experiments where a specimen is changed or acted\\u000a on while it remains

Frances M. Ross


Aberration corrected Lorentz scanning transmission electron microscopy.  


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. PMID:25677688

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



Application of Electron Diffraction to Biological Electron Microscopy  

PubMed Central

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

Glaeser, Robert M.; Thomas, Gareth



Quantitative phase imaging via Fourier ptychographic microscopy.  


Fourier ptychographic microscopy (FPM) is a recently developed imaging modality that uses angularly varying illumination to extend a system's performance beyond the limit defined by its optical components. The FPM technique applies a novel phase-retrieval procedure to achieve resolution enhancement and complex image recovery. In this Letter, we compare FPM data to theoretical prediction and phase-shifting digital holography measurement to show that its acquired phase maps are quantitative and artifact-free. We additionally explore the relationship between the achievable spatial and optical thickness resolution offered by a reconstructed FPM phase image. We conclude by demonstrating enhanced visualization and the collection of otherwise unobservable sample information using FPM's quantitative phase. PMID:24322147

Ou, Xiaoze; Horstmeyer, Roarke; Yang, Changhuei; Zheng, Guoan



Dynamic imaging with electron microscopy  


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.

Campbell, Geoffrey; McKeown, Joe; Santala, Melissa



Prototype cantilevers for quantitative lateral force microscopy  

SciTech Connect

Prototype cantilevers are presented that enable quantitative surface force measurements using contact-mode atomic force microscopy (AFM). The ''hammerhead'' cantilevers facilitate precise optical lever system calibrations for cantilever flexure and torsion, enabling quantifiable adhesion measurements and friction measurements by lateral force microscopy (LFM). Critically, a single hammerhead cantilever of known flexural stiffness and probe length dimension can be used to perform both a system calibration as well as surface force measurements in situ, which greatly increases force measurement precision and accuracy. During LFM calibration mode, a hammerhead cantilever allows an optical lever ''torque sensitivity'' to be generated for the quantification of LFM friction forces. Precise calibrations were performed on two different AFM instruments, in which torque sensitivity values were specified with sub-percent relative uncertainty. To examine the potential for accurate lateral force measurements using the prototype cantilevers, finite element analysis predicted measurement errors of a few percent or less, which could be reduced via refinement of calibration methodology or cantilever design. The cantilevers are compatible with commercial AFM instrumentation and can be used for other AFM techniques such as contact imaging and dynamic mode measurements.

Reitsma, Mark G.; Gates, Richard S.; Friedman, Lawrence H.; Cook, Robert F. [Nanomechanical Properties Group, National Institute of Standards and Technology, Gaithersburg, Maryland 20899 (United States)




PubMed Central

Thyagarajan, T. R. (Dartmouth Medical School, Hanover, N. H.), S. F. Conti, and H. B. Naylor. Electron microscopy of Rhodotorula glutinis. J. Bacteriol. 83:381–394. 1962.—The structure and manner of nuclear division in Rhodotorula glutinis was studied by electron microscopy of ultrathin sections. Parallel studies with the light microscope, employing conventional staining techniques and phase-contrast microscope observations on nuclei in living cells, were carried out. The nucleus is spherical to oval and is bounded by a nuclear membrane. Intranuclear structures, identified as nucleoli, and electron-transparent areas were observed. The nuclear membrane persists throughout the various stages of cell division. Observations of the nucleus with the electron microscope revealed that nuclear division occurs by a process of elongation and constriction similar to that seen in both living and stained cells. The fine structure of mitochondria and other components of the yeast cell and their behavior during cell division are described. The absence of vacuoles in actively dividing cells of Rhodotorula glutinis lends further support to the view that the vacuole is not an integral part of the nucleus. The results with the electron microscope generally support and considerably extend those obtained with living and stained cells. Images PMID:13921132

Thyagarajan, T. R.; Conti, S. F.; Naylor, H. B.



Spectroscopic imaging in electron microscopy  

SciTech Connect

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.

Pennycook, Stephen J [ORNL; Colliex, C. [Universite Paris Sud, Orsay, France



Direct Detectors for Electron Microscopy  

NASA Astrophysics Data System (ADS)

There is interest in improving the detectors used to capture images in transmission electron microscopy. Detectors with an improved modulation transfer function at high spatial frequencies allow for higher resolution in images at lower magnification, which leads to an increased effective field of view. Detectors with improved detective quantum efficiency are important for low dose applications. One way in which these performance enhancements can be achieved is through direct detection, where primary electrons are converted directly into suitable electrical signals by the detector rather than relying on an indirect electron to photon conversion before detection. In this paper we present the characterisation of detector performance for a number of different direct detection technologies, and compare these technologies to traditional indirect detectors. Overall our results show that direct detection enables a significant improvement in all aspects of detector performance.

Clough, R. N.; Moldovan, G.; Kirkland, A. I.



Cryo-electron microscopy of viruses  

Microsoft Academic Search

Thin vitrified layers of unfixed, unstained and unsupported virus suspensions can be prepared for observation by cryo-electron microscopy in easily controlled conditions. The viral particles appear free from the kind of damage caused by dehydration, freezing or adsorption to a support that is encountered in preparing biological samples for conventional electron microscopy. Cryo-electron microscopy of vitrified specimens offers possibilities for

Marc Adrian; Jacques Dubochet; Jean Lepault; Alasdair W. McDowall



Correlative Stochastic Optical Reconstruction Microscopy and Electron Microscopy  

PubMed Central

Correlative fluorescence light microscopy and electron microscopy allows the imaging of spatial distributions of specific biomolecules in the context of cellular ultrastructure. Recent development of super-resolution fluorescence microscopy allows the location of molecules to be determined with nanometer-scale spatial resolution. However, correlative super-resolution fluorescence microscopy and electron microscopy (EM) still remains challenging because the optimal specimen preparation and imaging conditions for super-resolution fluorescence microscopy and EM are often not compatible. Here, we have developed several experiment protocols for correlative stochastic optical reconstruction microscopy (STORM) and EM methods, both for un-embedded samples by applying EM-specific sample preparations after STORM imaging and for embedded and sectioned samples by optimizing the fluorescence under EM fixation, staining and embedding conditions. We demonstrated these methods using a variety of cellular targets. PMID:25874453

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



Quantitative determination of low-Z elements in single atmospheric particles on boron substrates by automated scanning electron microscopy-energy-dispersive X-ray spectrometry.  


Atmospheric aerosols consist of a complex heterogeneous mixture of particles. Single-particle analysis techniques are known to provide unique information on the size-resolved chemical composition of aerosols. A scanning electron microscope (SEM) combined with a thin-window energy-dispersive X-ray (EDX) detector enables the morphological and elemental analysis of single particles down to 0.1 microm with a detection limit of 1-10 wt %, low-Z elements included. To obtain data statistically representative of the air masses sampled, a computer-controlled procedure can be implemented in order to run hundreds of single-particle analyses (typically 1000-2000) automatically in a relatively short period of time (generally 4-8 h, depending on the setup and on the particle loading). However, automated particle analysis by SEM-EDX raises two practical challenges: the accuracy of the particle recognition and the reliability of the quantitative analysis, especially for micrometer-sized particles with low atomic number contents. Since low-Z analysis is hampered by the use of traditional polycarbonate membranes, an alternate choice of substrate is a prerequisite. In this work, boron is being studied as a promising material for particle microanalysis. As EDX is generally said to probe a volume of approximately 1 microm3, geometry effects arise from the finite size of microparticles. These particle geometry effects must be corrected by means of a robust concentration calculation procedure. Conventional quantitative methods developed for bulk samples generate elemental concentrations considerably in error when applied to microparticles. A new methodology for particle microanalysis, combining the use of boron as the substrate material and a reverse Monte Carlo quantitative program, was tested on standard particles ranging from 0.25 to 10 microm. We demonstrate that the quantitative determination of low-Z elements in microparticles is achievable and that highly accurate results can be obtained using the automatic data processing described here compared to conventional methods. PMID:16131082

Choël, Marie; Deboudt, Karine; Osán, János; Flament, Pascal; Van Grieken, René



Characterization of municipal solid waste incineration (MSWI) bottom ash by scanning electron microscopy and quantitative energy dispersive X-ray microanalysis (SEM/EDX).  


Scanning electron microscopy (SEM) with energy-dispersive X-ray microanalysis (EDX) is frequently used for morphological and qualitative chemical characterization of different materials. The applicability of this method for phase identification, is, however, often underestimated. The application of SEM/EDX for the characterization of different phases in fresh and altered municipal-waste incinerator bottom-ash samples with high lateral resolution is presented. Polished thin sections were prepared from the samples, but fresh fracture surfaces were also used. The EDX analyses were performed by using the correction procedures of a conventional standardless ZAF correction, a peak-to-background ZAF correction, and a correction method for light-element analysis. Because of their highly reactive properties the bottom-ash SEM samples require a special method of preparation. The method facilitates nondestructive preparation of the sensitive bottom-ash alteration phases (e.g. cement phases, hydroxides, salts) and their microstructures. PMID:11508465

Speiser, C; Baumann, T; Niessner, R



Four-dimensional ultrafast electron microscopy  

PubMed Central

Electron microscopy is arguably the most powerful tool for spatial imaging of structures. As such, 2D and 3D microscopies provide static structures with subnanometer and increasingly with ångstrom-scale spatial resolution. Here we report the development of 4D ultrafast electron microscopy, whose capability imparts another dimension to imaging in general and to dynamics in particular. We demonstrate its versatility by recording images and diffraction patterns of crystalline and amorphous materials and images of biological cells. The electron packets, which were generated with femtosecond laser pulses, have a de Broglie wavelength of 0.0335 Å at 120 keV and have as low as one electron per pulse. With such few particles, doses of few electrons per square ångstrom, and ultrafast temporal duration, the long sought after but hitherto unrealized quest for ultrafast electron microscopy has been realized. Ultrafast electron microscopy should have an impact on all areas of microscopy, including biological imaging. PMID:15883380

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



Faculty Position in Materials Electron Microscopy  

E-print Network

Faculty Position in Materials Electron Microscopy at the Ecole Polytechnique Fédérale de Lausanne in electron microscopy of materials within its Institute of Materials. We seek exceptional individuals who community. Top-level applications are invited from candidates at the cutting edge of electron microscopic

Candea, George


Thermal diffuse scattering in transmission electron microscopy.  


In conventional transmission electron microscopy, thermal scattering significantly affects the image contrast. It has been suggested that not accounting for this correctly is the main cause of the Stobbs factor, the ubiquitous, large contrast mismatch found between theory and experiment. In the case where a hard aperture is applied, we show that previous conclusions drawn from work using bright field scanning transmission electron microscopy and invoking the principle of reciprocity are reliable in the presence of thermal scattering. In the aperture-free case it has been suggested that even the most sophisticated mathematical models for thermal diffuse scattering lack in their numerical implementation, specifically that there may be issues in sampling, including that of the contrast transfer function of the objective lens. We show that these concerns can be satisfactorily overcome with modest computing resources; thermal scattering can be modelled accurately enough for the purpose of making quantitative comparison between simulation and experiment. Spatial incoherence of the source is also investigated. Neglect or inadequate handling of thermal scattering in simulation can have an appreciable effect on the predicted contrast and can be a significant contribution to the Stobbs factor problem. PMID:22088442

Forbes, B D; D'Alfonso, A J; Findlay, S D; Van Dyck, D; Lebeau, J M; Stemmer, S; Allen, L J



Quantitative determination of tip parameters in piezoresponse force microscopy  

E-print Network

Quantitative determination of tip parameters in piezoresponse force microscopy Sergei V. Kalinin and Gopalan13 and further developed by Kalinin et al.11 Briefly, a the electrostatic field is determined from

Gopalan, Venkatraman



EPA Science Inventory

Confocal Microscopy System Performance: QA tests, Quantitation and Spectroscopy. Robert M. Zucker 1 and Jeremy M. Lerner 2, 1Reproductive Toxicology Division, National Health and Environmental Effects Research Laboratory, Office of Research Development, U.S. Environmen...


Lock-in thermography, penetrant inspection, and scanning electron microscopy for quantitative evaluation of open micro-cracks at the tooth–restoration interface  

NASA Astrophysics Data System (ADS)

The evaluation of a dental restoration in a non-invasive way is of paramount importance in clinical practice. The aim of this study was to assess the minimum detectable open crack at the cavity–restorative material interface by the lock-in thermography technique, at laser intensities which are safe for living teeth. For the analysis of the interface, 18 box-type class V standardized cavities were prepared on the facial and oral surfaces of each tooth, with coronal margins in enamel and apical margins in dentine. The preparations were restored with the Giomer Beautifil (Shofu) in combination with three different adhesive systems. Three specimens were randomly selected from each experimental group and each slice has been analysed by visible, infrared (IR), and scanning electron microscopy (SEM). Lock-in thermography showed the most promising results in detecting both marginal and internal defects. The proposed procedure leads to a diagnosis of micro-leakages having openings of 1?µm, which is close to the diffraction limit of the IR camera. Clinical use of a thermographic camera in assessing the marginal integrity of a restoration becomes possible. The method overcomes some drawbacks of standard SEM or dye penetration testing. The results support the use of an IR camera in dentistry, for the diagnosis of micro-gaps at bio-interfaces.

Streza, M.; Hodisan, I.; Prejmerean, C.; Boue, C.; Tessier, Gilles



Quantitative analysis of pyrolytic carbon films by polarized light microscopy  

Microsoft Academic Search

The optical properties of differently textured pyrolytic carbon films were quantitatively analyzed by polarized light microscopy. The light intensity for the investigation of optically anisotropic and birefringent materials by polarized light microscopy was calculated as a function of the analyzer angle and the orientation of the material. From these calculations the dependence of the extinction angle on the optical properties

Andreas Pfrang; Thomas Schimmel



The study of epithelial function by quantitative light microscopy  

Microsoft Academic Search

Quantitative light microscopy can be used to analyze the mechanisms of salt and water movement across epithelial cells. Methods for light microscopic visualization and image aquisition are reviewed. Video image recording and processing are shown to be essential for the study of epithelial cell function by light microscopy.

Kenneth R. Spring



Analytical transmission electron microscopy in materials science  

SciTech Connect

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.

Fraser, H.L.



Paul Edwards's tutorial on scanning electron microscopy  

NSDL National Science Digital Library

This site from the University of Durham features descriptions of various scanning electron microscopy methods such as primary and secondary electron imaging, energy-dispersive analysis of X-rays, cathodoluminescence, and electron-beam induced current. Links to other scanning electron microscopic sites are also included.

Paul Edwards


Fast electron microscopy via compressive sensing  


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.

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



Probing the Proton with Electron Microscopy  

NASA Astrophysics Data System (ADS)

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.

Friedman, Jerome I.



Quantitative Time-Lapse Fluorescence Microscopy in Single Cells  

PubMed Central

The cloning of GFP 15 years ago revolutionized cell biology by permitting visualization of a wide range of molecular mechanisms within living cells. Though initially used to make largely qualitative assessments of protein levels and localizations, fluorescence microscopy has since evolved to become highly quantitative and high-throughput. Computational image analysis has catalyzed this evolution, enabling rapid and automated processing of large datasets. Here we review studies that combine time-lapse fluorescence microscopy and automated image analysis to investigate dynamic events at the single-cell level. We highlight examples where single-cell analysis provides unique mechanistic insights into cellular processes that cannot be otherwise resolved in bulk assays. Additionally, we discuss studies where quantitative microscopy facilitates the assembly of detailed 4D lineages in developing organisms. Finally, we describe recent advances in imaging technology, focusing especially on platforms that allow the simultaneous perturbation and quantitative monitoring of biological systems. PMID:19575655

Muzzey, Dale; van Oudenaarden, Alexander



Quantitative analysis of angiogenesis using confocal laser scanning microscopy  

Microsoft Academic Search

Angiogenesis is essential for tumor growth and metastasis. Angiogenesis is commonly quantified by measuring microvessel density\\u000a (MVD) within tumors. In this report, we compared light microscopy with confocal laser scanning microscopy (CLSM) in the qualitative\\u000a and quantitative analysis of angiogenesis. MVDs were determined manually in a lung tumor xenograft and a normal skeletal muscle\\u000a using CD31 immunohistochemical staining and light

Linlang Guo; Patricia Burke; Su-Hao Lo; Regina Gandour-Edwards; Derick Lau



Low voltage transmission electron microscopy of graphene.  


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. PMID:25408379

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



Dinosaur eggshell study using scanning electron microscopy.  


Visualization and analysis of structural features in fossil dinosaur eggs by scanning electron microscopy augment information from traditional petrographic light microscopy. Comparison of characteristics in fossil and modern eggshells allows inferences to be made regarding dinosaur reproductive biology, physiology, and evolutionary relationships. Assessment of diagenetic alteration of primary eggshell calcite structure that occurs during fossilization provides important information necessary for taxonomic identification and paleoenvironmental interpretations. PMID:12392352

Jackson, Frankie D; Schweitzer, Mary H; Schmitt, James G



Simultaneous Correlative Scanning Electron and High-NA Fluorescence Microscopy  

PubMed Central

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

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.



High-speed quantitative interferometric microscopy based phase imaging cytometer  

NASA Astrophysics Data System (ADS)

The paper proposed a simple large scale bio-sample phase detecting equipment called gravity driven phase detecting cytometer, which is based on quantitative interferometric microscopy to realize flowing red blood cells phase distribution detection. The method has advantages on high throughput phase detecting and statistical analysis with high detecting speed and in real-time. The statistical characteristics of red blood cells are useful for biological analysis and disease detection. We believe this method is shedding more light on quantitatively measurement of the phase distribution of bio-samples.

Xue, Liang; Sun, Nan; Yan, Keding; Liu, Fei; Wang, Shouyu



Quantitative sectioning and noise analysis for structured illumination microscopy  

PubMed Central

Structured illumination (SI) has long been regarded as a nonquantitative technique for obtaining sectioned microscopic images. Its lack of quantitative results has restricted the use of SI sectioning to qualitative imaging experiments, and has also limited researchers’ ability to compare SI against competing sectioning methods such as confocal microscopy. We show how to modify the standard SI sectioning algorithm to make the technique quantitative, and provide formulas for calculating the noise in the sectioned images. The results indicate that, for an illumination source providing the same spatially-integrated photon flux at the object plane, and for the same effective slice thicknesses, SI sectioning can provide higher SNR images than confocal microscopy for an equivalent setup when the modulation contrast exceeds about 0.09. PMID:22274364

Hagen, Nathan; Gao, Liang; Tkaczyk, Tomasz S.



Experiences with remote electron microscopy  

SciTech Connect

With the advent of a rapidly proliferating international computer network, it became feasible to consider remote operation of instrumentation normally operated locally. For modern electron microscopes, the growing automation and computer control of many instrumental operations facilitated the task of providing remote operation. In order to provide use of NCEM TEMs by distant users, a project was instituted in 1995 to place a unique instrument, a Kratos EM-1500 operating at 1.5MeV, on-line for remote use. In 1996, the Materials Microcharacterization Collaboratory (MMC) was created as a pilot project within the US Department of Energy's DOE2000 program to establish national collaboratories to provide access via the Internet to unique or expensive DOE research facilities as well as to expertise for remote collaboration, experimentation, production, software development, modeling, and measurement. A major LBNL contribution to the MMC was construction of DeepView, a microscope-independent computer-control system that could be ported to other MMC members to provide a common graphical user-interface (GUI) for control of any MMC instrument over the wide area network.

O'Keefe, Michael A.; Parvin, Bahram



Electron Microscopy of the Cell  

PubMed Central

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

Leeson, T. S.



Subcellular quantitative optical diffraction tomography with digital holographic microscopy  

Microsoft Academic Search

Digital holographic microscopy (DHM) is an interferometric technique, providing quantitative mapping of the phase shift induced by semi-transparent microscopic specimens, such as cells, with sub-wavelength resolution along the optical axis. Thanks to actual PCs and CCDs, DHM provides nowadays cost-effective instruments for real-time measurements at very high acquisition rates, with sub-micron transverse resolution. However, DHM phase images do not reveal

Florian Charrière; Jonas Kühn; Tristan Colomb; Etienne Cuche; Pierre Marquet; Christian Depeursinge



Fast pixel shifting phase unwrapping algorithm in quantitative interferometric microscopy  

NASA Astrophysics Data System (ADS)

Quantitative interferometric microscopy is an important method for observing biological samples such as cells and tissues. In order to obtain continuous phase distribution of the sample from the interferogram, phase extracting and phase unwrapping are both needed in quantitative interferometric microscopy. Phase extracting includes fast Fourier transform method and Hilbert transform method, etc., almost all of them are rapid methods. However, traditional unwrapping methods such as least squares algorithm, minimum network flow method, etc. are time-consuming to locate the phase discontinuities which lead to low processing efficiency. Other proposed high-speed phase unwrapping methods always need at least two interferograms to recover final phase distributions which cannot realize real time processing. Therefore, high-speed phase unwrapping algorithm for single interferogram is required to improve the calculation efficiency. Here, we propose a fast phase unwrapping algorithm to realize high-speed quantitative interferometric microscopy, by shifting mod 2? wrapped phase map for one pixel, then multiplying the original phase map and the shifted one, then the phase discontinuities location can be easily determined. Both numerical simulation and experiments confirm that the algorithm features fast, precise and reliable.

Xu, Mingfei; Shan, Yanke; Yan, Keding; Xue, Liang; Wang, Shouyu; Liu, Fei



D Ultrafast Electron Diffraction, Crystallography, and Microscopy  

Microsoft Academic Search

In this review, we highlight the progress made in the development of 4D ultrafast electron diffraction (UED), crystallography (UEC), and microscopy (UEM) with a focus on concepts, methodologies, and prototypical applications. The joint atomic-scale resolutions in space and time, and sensitivity reached, make it possible to determine complex transient structures and assemblies in different phases. These applications include studies of

Ahmed H. Zewail



Pigment Dispersion Methods for Electron Microscopy  

Microsoft Academic Search

A method of preparing a pigment dispersion for electron microscopy is described, wherein the pigment, such as zinc oxide, is first wet with water and is then dispersed in a solution of cellulose acetate. Another method is described in which mechanical or chemical damage of the particles is avoided by the use of an electrostatic dispersion apparatus.

H. C. O'Brien



Organization of the Primate Retina: Electron Microscopy  

Microsoft Academic Search

The retinae of monkey and man have been studied by electron microscopy to identify cell types, their processes and synaptic contacts. In the inner plexiform layer, the morphological characteristics of the three types of cells (bipolar, ganglion and amacrine) are described and seven synaptic relationships are identified. The bipolar terminals contain ribbons at points of synaptic contact, and, at these

J. E. Dowling; B. B. Boycott



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


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. PMID:25229674

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



Photon-induced near field electron microscopy  

NASA Astrophysics Data System (ADS)

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.

Park, Sang Tae; Zewail, Ahmed H.



Single beam Fourier transform digital holographic quantitative phase microscopy  

NASA Astrophysics Data System (ADS)

Quantitative phase contrast microscopy reveals thickness or height information of a biological or technical micro-object under investigation. The information obtained from this process provides a means to study their dynamics. Digital holographic (DH) microscopy is one of the most used, state of the art single-shot quantitative techniques for three dimensional imaging of living cells. Conventional off axis DH microscopy directly provides phase contrast images of the objects. However, this process requires two separate beams and their ratio adjustment for high contrast interference fringes. Also the use of two separate beams may make the system more vulnerable to vibrations. Single beam techniques can overcome these hurdles while remaining compact as well. Here, we describe the development of a single beam DH microscope providing whole field imaging of micro-objects. A hologram of the magnified object projected on to a diffuser co-located with a pinhole is recorded with the use of a commercially available diode laser and an arrayed sensor. A Fourier transform of the recorded hologram directly yields the complex amplitude at the image plane. The method proposed was investigated using various phase objects. It was also used to image the dynamics of human red blood cells in which sub-micrometer level thickness variation were measurable.

Anand, A.; Faridian, A.; Chhaniwal, V. K.; Mahajan, S.; Trivedi, V.; Dubey, S. K.; Pedrini, G.; Osten, W.; Javidi, B.



Single beam Fourier transform digital holographic quantitative phase microscopy  

SciTech Connect

Quantitative phase contrast microscopy reveals thickness or height information of a biological or technical micro-object under investigation. The information obtained from this process provides a means to study their dynamics. Digital holographic (DH) microscopy is one of the most used, state of the art single-shot quantitative techniques for three dimensional imaging of living cells. Conventional off axis DH microscopy directly provides phase contrast images of the objects. However, this process requires two separate beams and their ratio adjustment for high contrast interference fringes. Also the use of two separate beams may make the system more vulnerable to vibrations. Single beam techniques can overcome these hurdles while remaining compact as well. Here, we describe the development of a single beam DH microscope providing whole field imaging of micro-objects. A hologram of the magnified object projected on to a diffuser co-located with a pinhole is recorded with the use of a commercially available diode laser and an arrayed sensor. A Fourier transform of the recorded hologram directly yields the complex amplitude at the image plane. The method proposed was investigated using various phase objects. It was also used to image the dynamics of human red blood cells in which sub-micrometer level thickness variation were measurable.

Anand, A., E-mail:; Chhaniwal, V. K.; Mahajan, S.; Trivedi, V. [Optics Laboratory, Applied Physics Department, Faculty of Technology and Engineering, M.S. University of Baroda, Vadodara 390001 (India)] [Optics Laboratory, Applied Physics Department, Faculty of Technology and Engineering, M.S. University of Baroda, Vadodara 390001 (India); Faridian, A.; Pedrini, G.; Osten, W. [Institut für Technische Optik, Universität Stuttgart, Pfaffenwaldring 9, 70569 Stuttgart (Germany)] [Institut für Technische Optik, Universität Stuttgart, Pfaffenwaldring 9, 70569 Stuttgart (Germany); Dubey, S. K. [Siemens Technology and Services Pvt. Ltd, Corporate Technology—Research and Technology Centre, Bangalore 560100 (India)] [Siemens Technology and Services Pvt. Ltd, Corporate Technology—Research and Technology Centre, Bangalore 560100 (India); Javidi, B. [Department of Electrical and Computer Engineering, U-4157, University of Connecticut, Storrs, Connecticut 06269-2157 (United States)] [Department of Electrical and Computer Engineering, U-4157, University of Connecticut, Storrs, Connecticut 06269-2157 (United States)



Image Restoration in Cryo-electron Microscopy  

PubMed Central

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

Penczek, Pawel A.



Quantitative phase microscopy and synthetic aperture tomography of live cells  

NASA Astrophysics Data System (ADS)

For more than a decade MIT's George R. Harrison Spectroscopy Laboratory has been developing quantitative phase microscopy (QPM) for biological study. Measurements of a point field were made in the mid 90s, then extended to the full 2D field, and recently, to 3D by using tomography. In the first part of this thesis improvements in the techniques of Fourier Phase Microscopy (FPM) and Hilbert Phase Microscopy (HPM) and their applications to characterize cells and tissues are reported. Tomographic phase microscopy (TPM) provides quantitative information and highly detailed structural information about a live cell, but in its current form it can only examine one cell at a time. Many biological applications including statistical analysis of a large collection of cells such as flow cytometry need a tomography technique that can measure many cells at a time. For the second part of this thesis we have developed a new tomography technique that can measure many cells continuously. In this study we demonstrate the new technique by translating a live cell across a focused beam. This beam is composed of many angular plane waves, and by applying a so-called synthetic aperture algorithm we retrieve individual wave components of the focused beam. We demonstrate for the first time that we can retrieve the field of the focused beam and synthesize any arbitrary angular plane wave. We then construct a 3D map of the variations of the refractive index in a live cell from a series of these synthesized angular plane waves. This new technique is the first step needed to analyze cells flowing through a beam to provide a high-throughput 3D refractive index tomograms that can be used as a new kind of statistical optical assay of living cells.

Lue, Niyom


Rapid quantitative phase imaging for partially coherent light microscopy.  


Partially coherent light provides promising advantages for imaging applications. In contrast to its completely coherent counterpart, it prevents image degradation due to speckle noise and decreases cross-talk among the imaged objects. These facts make attractive the partially coherent illumination for accurate quantitative imaging in microscopy. In this work, we present a non-interferometric technique and system for quantitative phase imaging with simultaneous determination of the spatial coherence properties of the sample illumination. Its performance is experimentally demonstrated in several examples underlining the benefits of partial coherence for practical imagining applications. The programmable optical setup comprises an electrically tunable lens and sCMOS camera that allows for high-speed measurement in the millisecond range. PMID:24921541

Rodrigo, José A; Alieva, Tatiana



Quantitative Phase Contrast Digital Holographic Microscopy in Biophotonics  

NASA Astrophysics Data System (ADS)

Label-free, non-contact, non-destructive, on-line (video repetition rate), high resolution, full field (no scanning), quantitative analysis of morphology and dynamic processes in living cells are required features in life science research and medical diagnostics. Digital Holography combined with microscopic imaging provides these features simultaneously. The modular integration of digital holographic microscopy (DHM) into commercial microscopes yields an axial resolution with interferometric resolution while the lateral resolution is diffraction limited. As amplitude and phase are available by numerical reconstruction from a single digital hologram subsequent automated focus correction is enabled. The evaluation of quantitative digital holographic phase contrast images permits also an effective detection of lateral object movements. Thus, 3D tracking is achieved. The applicability of DHM techniques for dynamic live cell analysis is demonstrated by results from tumor cells and human erythrocytes.

Kemper, Björn; Langehanenberg, Patrik; von Bally, Gert



Quantitative polarized light microscopy of human cochlear sections  

PubMed Central

Dysfunction of the inner ear is the most common cause of sensorineural hearing loss, which is the most common sensory deficit worldwide. Conventional imaging modalities are unable to depict the microanatomy of the human inner ear, hence the need to explore novel imaging modalities. We provide the first characterization of the polarization dependent optical properties of human cochlear sections using quantitative polarized light microscopy (qPLM). Eight pediatric cadaveric cochlear sections, aged 0 (term) to 24 months, were selected from the US National Temporal Bone Registry, imaged with qPLM and analyzed using Image J. Retardance of the bony otic capsule and basilar membrane were substantially higher than that of the stria vascularis, spiral ganglion neurons, organ of Corti and spiral ligament across the half turns of the spiraling cochlea. qPLM provides quantitative information about the human inner ear, and awaits future exploration in vivo. PMID:25780749

Low, Jacob C. M.; Ober, Thomas J.; McKinley, Gareth H.; Stankovic, Konstantina M.



Correlative Photoactivated Localization and Scanning Electron Microscopy  

PubMed Central

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. PMID:24204771

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



Scanning electron microscopy of cold gases  

E-print Network

Ultracold quantum gases offer unique possibilities to study interacting many-body quantum systems. Probing and manipulating such systems with ever increasing degree of control requires novel experimental techniques. Scanning electron microscopy is a high resolution technique which can be used for in situ imaging, single site addressing in optical lattices and precision density engineering. Here, we review recent advances and achievements obtained with this technique and discuss future perspectives.

Santra, Bodhaditya



Fixation of Ejaculated Spermatozoa for Electron Microscopy  

Microsoft Academic Search

EJACULATED spermatozoa cannot be preserved satisfactorily by conventional fixation procedures for electron microscopy. Osmium tetroxide (OsO4) fixation of crude ejaculate consistently produces a variety of artefacts such as separation of the plasma membrane from the acrosome, widening of nuclear vacuoles, erosion of the acrosome, and swelling of mitochondria1-3. These alterations could be the consequence of the rapid destruction of the

Mario Stefanini; Cesare De Martino; Luciano Zamboni



Comparative scanning electron microscopy of bullous diseases*  

PubMed Central

The purpose of this study is to compare scanning electron microscopy findings of the blister roof in three distinct bullous diseases: one intraepidermal acantholytic (pemphigus foliaceus); one due to hemidesmosomal dysfunction (bullous pemphigoid); and one secondary to anchoring fibril dysfunction - type VII collagen (dystrophic epidermolysis bullosa). In pemphigus foliaceus, acantholytic phenomena were readily demonstrated. In bullous pemphigoid, the epidermis had a solid aspect. In dystrophic epidermolysis bullosa a net was seen in the blister roof. PMID:24770520

de Almeida, Hiram Larangeira; Rossi, Gabriela; Karam, Octavio Ruschel; Rocha, Nara Moreira; Silva, Ricardo Marques e



4D electron microscopy: principles and applications.  


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 achievable with short intense pulses containing a large number of electrons, however, are limited to tens of nanometers and nanoseconds, respectively. This is because Coulomb repulsion is significant in such a pulse, and the electrons spread in space and time, thus limiting the beam coherence. It is therefore not possible to image the ultrafast elementary dynamics of complex transformations. The challenge was to retain the high spatial resolution of a conventional TEM while simultaneously enabling the temporal resolution required to visualize atomic-scale motions. In this Account, we discuss the development of four-dimensional ultrafast electron microscopy (4D UEM) and summarize techniques and applications that illustrate the power of the approach. In UEM, images are obtained either stroboscopically with coherent single-electron packets or with a single electron bunch. Coulomb repulsion is absent under the single-electron condition, thus permitting imaging, diffraction, and spectroscopy, all with high spatiotemporal resolution, the atomic scale (sub-nanometer and femtosecond). The time resolution is limited only by the laser pulse duration and energy carried by the electron packets; the CCD camera has no bearing on the temporal resolution. In the regime of single pulses of electrons, the temporal resolution of picoseconds can be attained when hundreds of electrons are in the bunch. The applications given here are selected to highlight phenomena of different length and time scales, from atomic motions during structural dynamics to phase transitions and nanomechanical oscillations. We conclude with a brief discussion of emerging methods, which include scanning ultrafast electron microscopy (S-UEM), scanning transmission ultrafast electron microscopy (ST-UEM) with convergent beams, and time-resolved imaging of biological structures at ambient conditions with environmental cells. PMID:22967215

Flannigan, David J; Zewail, Ahmed H



Electron Microscopy Study of Tin Whisker Growth  

SciTech Connect

The growth of tin whiskers formed on sputtered tin layers deposited on brass was studied using electron microscopy. The occurrence of whiskers appeared to be largely independent of the macroscopic stress state in the film; rather it was microscopic compressive stresses arising from the formation of an intermetallic phase that appeared to be the necessary precursor. Whisker morphology was a result of whether nucleation had occurred on single grains or on multiple grains. In the latter case, the whiskers had a fluted or striated surface. The formation of whiskers on electron transparent samples was demonstrated. These samples showed the whiskers were monocrystalline and defect free, and that the growth direction could be determined.

Norton, Murray G. (Washington State University); Lebret, Joel (8392)



The continuing value of electron microscopy in surgical pathology.  


For decades, transmission electron microscopy has played a valuable diagnostic role in surgical pathology. The continuing importance of electron microscopy, however, can be debated, given the major advances that have occurred in immunohistochemistry and other techniques. Electron microscopy retains excellent educational potential and broad research applicability, and it continues to be a necessity for the evaluation of a small subset of surgical pathology cases, such as renal biopsies and cilia specimens. The real controversy, then, centers on the contribution of electron microscopy in the evaluation of neoplasms. The opinion of many experts indicates that electron microscopy is still vital in the diagnostic assessment of some neoplasms, and that both electron microscopy and immunohistochemistry are more powerful when viewed as complementary rather than competitive techniques. For electron microscopy to be used to its potential, however, electron microscopists must function effectively as consultants. When optimally applied, electron microscopy remains an essential diagnostic tool. PMID:11206335

Tucker, J A



Quantitative Phase Microscopy: how to make phase data meaningful  

PubMed Central

The continued development of hardware and associated image processing techniques for quantitative phase microscopy has allowed superior phase data to be acquired that readily shows dynamic optical volume changes and enables particle tracking. Recent efforts have focused on tying phase data and associated metrics to cell morphology. One challenge in measuring biological objects using interferometrically obtained phase information is achieving consistent phase unwrapping and -dimensions and correct for temporal discrepanices using a temporal unwrapping procedure. The residual background shape due to mean value fluctuations and residual tilts can be removed automatically using a simple object characterization algorithm. Once the phase data are processed consistently, it is then possible to characterize biological samples such as myocytes and myoblasts in terms of their size, texture and optical volume and track those features dynamically. By observing optical volume dynamically it is possible to determine the presence of objects such as vesicles within myoblasts even when they are co-located with other objects. Quantitative phase microscopy provides a label-free mechanism to characterize living cells and their morphology in dynamic environments, however it is critical to connect the measured phase to important biological function for this measurement modality to prove useful to a broader scientific community. In order to do so, results must be highly consistent and require little to no user manipulation to achieve high quality nynerical results that can be combined with other imaging modalities. PMID:25309099

Goldstein, Goldie; Creath, Katherine



Quantitative analysis of live cells using digital holographic microscopy  

NASA Astrophysics Data System (ADS)

During the life time of a cell, it goes through changes to the plasma membrane as well as its internal structures especially distinctive during processes like cell division and death. Different types of microscope are used to fulfill the observation of the cell's variation. In our experiment, Vero cells have been investigated by using phase contrast microscopy and digital holographic microscopy (DHM). A comparison of the images obtained for cell division is presented here. The conventional phase contrast microscope provided a good imaging method in the real time analysis of cell division. The off-axis digital hologram recorded by the DHM system can be reconstructed to obtain both the intensity image and phase contrast image of the test object. These can be used for live cell imaging to provide multiple results from a single equipment setup. The DHM system, besides being a qualitative tool, is able to provide quantitative results and 3D images of the cell division process. The ability of DHM to provide quantitative analysis makes it an ideal tool for life science applications.

Lewis, Tan Rongwei; Qu, Weijuan; Chee, Oi Choo; Singh, Vijay Raj; Asundi, Anand



Quantitative analysis of live cells using digital holographic microscopy  

NASA Astrophysics Data System (ADS)

During the life time of a cell, it goes through changes to the plasma membrane as well as its internal structures especially distinctive during processes like cell division and death. Different types of microscope are used to fulfill the observation of the cell's variation. In our experiment, Vero cells have been investigated by using phase contrast microscopy and digital holographic microscopy (DHM). A comparison of the images obtained for cell division is presented here. The conventional phase contrast microscope provided a good imaging method in the real time analysis of cell division. The off-axis digital hologram recorded by the DHM system can be reconstructed to obtain both the intensity image and phase contrast image of the test object. These can be used for live cell imaging to provide multiple results from a single equipment setup. The DHM system, besides being a qualitative tool, is able to provide quantitative results and 3D images of the cell division process. The ability of DHM to provide quantitative analysis makes it an ideal tool for life science applications.

Lewis, Tan Rongwei; Qu, Weijuan; Chee, Oi Choo; Singh, Vijay Raj; Asundi, Anand



New Developments in Transmission Electron Microscopy for Nanotechnology**  

E-print Network

New Developments in Transmission Electron Microscopy for Nanotechnology** By Zhong Lin Wang* 1. Electron Microscopy and Nanotechnology Nanotechnology, as an international initiative for science manufacturing are the foundation of nanotechnology. Tracking the historical background of why nanotechnology

Wang, Zhong L.


All-Union Conference on Electron Microscopy, 14th, Suzdal, USSR, Oct.-Nov. 1990, Proceedings  

NASA Astrophysics Data System (ADS)

The papers presented in this volume provide an overview of recent developments in electron microscopy, with emphasis on the use of electron microscopy for the study of materials used in electronics and optoelectronics. Topics discussed include quantitative analysis of semiconductor optoelectronics materials by methods of scanning electron microscopy; morphology and structure of epitaxial layers in the multilayer structure InAs/GaAs; reconstruction of Bravais lattices from electronograms; and low-temperature scanning electron microscopy of superconducting YBaCuO films. Papers are also presented on qualitative measurements of the characteristics of SEM detectors; laser scanning microscopy and tomography in scattered light, and a correct method for calculating electron-optic systems.

Petrov, V. I.



Microfluidic system for transmission electron microscopy  

SciTech Connect

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

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



Immunogold labelling for scanning electron microscopy.  


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. PMID:20602226

Goldberg, Martin W; Fiserova, Jindriska



Direct imaging of crystal structure and defects in metastable Ge{sub 2}Sb{sub 2}Te{sub 5} by quantitative aberration-corrected scanning transmission electron microscopy  

SciTech Connect

Knowledge about the atomic structure and vacancy distribution in phase change materials is of foremost importance in order to understand the underlying mechanism of fast reversible phase transformation. In this Letter, by combining state-of-the-art aberration-corrected scanning transmission electron microscopy with image simulations, we are able to map the local atomic structure and composition of a textured metastable Ge{sub 2}Sb{sub 2}Te{sub 5} thin film deposited by pulsed laser deposition with excellent spatial resolution. The atomic-resolution scanning transmission electron microscopy investigations display the heterogeneous defect structure of the Ge{sub 2}Sb{sub 2}Te{sub 5} phase. The obtained results are discussed. Highly oriented Ge{sub 2}Sb{sub 2}Te{sub 5} thin films appear to be a promising approach for further atomic-resolution investigations of the phase change behavior of this material class.

Ross, Ulrich; Lotnyk, Andriy, E-mail:; Thelander, Erik; Rauschenbach, Bernd [Leibniz Institute of Surface Modification, Permoserstr. 15, D-04318 Leipzig (Germany)



Determination of elemental distribution in green micro-algae using synchrotron radiation nano X-ray fluorescence (SR-nXRF) and electron microscopy techniques--subcellular localization and quantitative imaging of silver and cobalt uptake by Coccomyxa actinabiotis.  


The newly discovered unicellular micro-alga Coccomyxa actinabiotis proves to be highly radio-tolerant and strongly concentrates radionuclides, as well as large amounts of toxic metals. This study helps in the understanding of the mechanisms involved in the accumulation and detoxification of silver and cobalt. Elemental distribution inside Coccomyxa actinabiotis cells was determined using synchrotron nano X-ray fluorescence spectroscopy at the ID22 nano fluorescence imaging beamline of the European Synchrotron Radiation Facility. The high resolution and high sensitivity of this technique enabled the assessment of elemental associations and exclusions in subcellular micro-algae compartments. A quantitative treatment of the scans was implemented to yield absolute concentrations of each endogenous and exogenous element with a spatial resolution of 100 nm and compared to the macroscopic content in cobalt and silver determined using inductively coupled plasma-mass spectrometry. The nano X-ray fluorescence imaging was complemented by transmission electron microscopy coupled to X-ray microanalysis (TEM-EDS), yielding differential silver distribution in the cell wall, cytosol, nucleus, chloroplast and mitochondria with unique resolution. The analysis of endogenous elements in control cells revealed that iron had a unique distribution; zinc, potassium, manganese, molybdenum, and phosphate had their maxima co-localized in the same area; and sulfur, copper and chlorine were almost homogeneously distributed among the whole cell. The subcellular distribution and quantification of cobalt and silver in micro-alga, assessed after controlled exposure to various concentrations, revealed that exogenous metals were mainly sequestered inside the cell rather than on mucilage or the cell wall, with preferential compartmentalization. Cobalt was homogeneously distributed outside of the chloroplast. Silver was localized in the cytosol at low concentration and in the whole cell excluding the nucleus at high concentration. Exposure to low concentrations of cobalt or silver did not alter the localization nor the concentration of endogenous elements within the cells. To our knowledge, this is the first report on element co-localization and segregation at the sub-cellular level in micro-algae by means of synchrotron nano X-ray fluorescence spectroscopy. PMID:24394991

Leonardo, T; Farhi, E; Boisson, A-M; Vial, J; Cloetens, P; Bohic, S; Rivasseau, C



Electron microscopy of Giardia lamblia cysts.  

PubMed Central

The flagellated protozoan Giardia lamblia is a recognized public health problem. Intestinal infection can result in acute or chronic diarrhea with associated symptoms in humans. As part of a study to evaluate removal of G. lamblia cysts from drinking water by the processes of coagulation and dual-media filtration, we developed a methodology by using 5.0-microns-porosity membrane filters to evaluate the filtration efficiency. We found that recovery rates of G. lamblia cysts by membrane filtration varied depending upon the type and diameter of the membrane filter. Examination of membrane-filtered samples by scanning electron microscopy revealed flexible and flattened G. lamblia cysts on the filter surface. This feature may be responsible for the low recovery rates with certain filters and, moreover, may have implications in water treatment technology. Formation of the cyst wall is discussed. Electron micrographs of cysts apparently undergoing binary fission and cysts exhibiting a possible bacterial association are shown. Images PMID:7425627

Luchtel, D L; Lawrence, W P; DeWalle, F B



Scanning electron microscopy studies of bacterial cultures  

NASA Astrophysics Data System (ADS)

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.

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



High resolution scanning electron microscopy of plasmodesmata.  


Symplastic transport occurs between neighbouring plant cells through functionally and structurally dynamic channels called plasmodesmata (PD). Relatively little is known about the composition of PD or the mechanisms that facilitate molecular transport into neighbouring cells. While transmission electron microscopy (TEM) provides 2-dimensional information about the structural components of PD, 3-dimensional information is difficult to extract from ultrathin sections. This study has exploited high-resolution scanning electron microscopy (HRSEM) to reveal the 3-dimensional morphology of PD in the cell walls of algae, ferns and higher plants. Varied patterns of PD were observed in the walls, ranging from uniformly distributed individual PD to discrete clusters. Occasionally the thick walls of the giant alga Chara were fractured, revealing the surface morphology of PD within. External structures such as spokes, spirals and mesh were observed surrounding the PD. Enzymatic digestions of cell wall components indicate that cellulose or pectin either compose or stabilise the extracellular spokes. Occasionally, the PD were fractured open and desmotubule-like structures and other particles were observed in their central regions. Our observations add weight to the argument that Chara PD contain desmotubules and are morphologically similar to higher plant PD. PMID:21626150

Brecknock, Sarah; Dibbayawan, Teresa P; Vesk, Maret; Vesk, Peter A; Faulkner, Christine; Barton, Deborah A; Overall, Robyn L



Development of Quantitative electron nano-diffraction  

Microsoft Academic Search

This thesis is a step towards development of quantitative parallel beam electron nano-diffraction (PBED). It is focused on the superstructure determination of zig-zag and zig-zig NaxCoO2 and analysis of charge distribution in the two polymorphs Nb12O29 using PBED. It has been shown that quantitative electron nano-diffraction (parallel beam) has the potential of solving superstructures as well as charge distribution by

V. Kumar



High voltage electron microscopy of lunar samples  

NASA Technical Reports Server (NTRS)

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.

Fernandez-Moran, H.



Improved methods for high resolution electron microscopy  

SciTech Connect

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.

Taylor, J.R.



Scanning electron microscopy of tinea nigra*  

PubMed Central

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. PMID:24770516

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



Scanning electron microscopy of molluscum contagiosum*  

PubMed Central

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

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



Scanning electron microscopy of molluscum contagiosum.  


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

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



Silver nanoparticle-induced degranulation observed with quantitative phase microscopy  

NASA Astrophysics Data System (ADS)

Monitoring a degranulation process in a live mast cell is a quite important issue in immunology and pharmacology. Because the size of a granule is normally much smaller than the resolution limit of an optical microscope system, there is no direct real-time live cell imaging technique for observing degranulation processes except for fluorescence imaging techniques. In this research, we propose optical quantitative phase microscopy (QPM) as a new observation tool to study degranulation processes in a live mast cell without any fluorescence labeling. We measure the cell volumes and the cross sectional profiles (x-z plane) of an RBL-2H3 cell and a HeLa cell, before and after they are exposed to calcium ionophore A23187 and silver nanoparticles (AgNPs). We verify that the volume and the cross sectional line profile of the RBL-2H3 cell were changed significantly when it was exposed to A23187. When 50 ?g/mL of AgNP is used instead of A23187, the measurements of cell volume and cross sectional profiles indicate that RBL-2H3 cells also follow degranulation processes. Degranulation processes for these cells are verified by monitoring the increase of intracellular calcium ([Ca2+]i) and histamine with fluorescent methods.

Yang, Wenzhong; Lee, Seungrag; Lee, Jiyong; Bae, Yoonsung; Kim, Dugyoung



Quantitative high dynamic range beam profiling for fluorescence microscopy  

NASA Astrophysics Data System (ADS)

Modern developmental biology relies on optically sectioning fluorescence microscope techniques to produce non-destructive in vivo images of developing specimens at high resolution in three dimensions. As optimal performance of these techniques is reliant on the three-dimensional (3D) intensity profile of the illumination employed, the ability to directly record and analyze these profiles is of great use to the fluorescence microscopist or instrument builder. Though excitation beam profiles can be measured indirectly using a sample of fluorescent beads and recording the emission along the microscope detection path, we demonstrate an alternative approach where a miniature camera sensor is used directly within the illumination beam. Measurements taken using our approach are solely concerned with the illumination optics as the detection optics are not involved. We present a miniature beam profiling device and high dynamic range flux reconstruction algorithm that together are capable of accurately reproducing quantitative 3D flux maps over a large focal volume. Performance of this beam profiling system is verified within an optical test bench and demonstrated for fluorescence microscopy by profiling the low NA illumination beam of a single plane illumination microscope. The generality and success of this approach showcases a widely flexible beam amplitude diagnostic tool for use within the life sciences.

Mitchell, T. J.; Saunter, C. D.; O'Nions, W.; Girkin, J. M.; Love, G. D.



Web Course Notes and References, Transmission Electron Microscopy  

NSDL National Science Digital Library

This site from the Electron Microscopy Outreach Program features a course devoted to transmission electron microscopy. The course is organized by chapters and covers the principles, design and operation of the electron microscope, contrast and image formation, image analysis, crystals, symmetry and diffraction, and Fourier image processing techniques.

Electron Microscopy Outreach Program


High Resolution Electron Microscopy at the National Cancer Institute

Imaging biological objects with electrons involves principles similar to those used in light microscopy, except that electrons are used for illumination instead of photons and the lenses are magnetic instead of being optical. In the last five decades, electron microscopy (EM) helped to reveal basic cell structures in great detail, allowing researchers to visualize internal structure at resolutions that were about 100 times better than that obtained by optical microscopy.


High Resolution Electron Microscopy at the National Cancer Institute

For many years, electron microscopy has been used to image cells and tissues at high resolution. This technology, invented in the early 20th century, provided breakthrough information in the virology and cell biology fields. Over the last 15 to 20 years, however, rapid advances in imaging and computation technologies have expanded the usefulness of electron microscopy into new realms. Electron microscopy is now poised to close a critical "gap" in the structural biology field.


Aberration-corrected electron microscopy of nanoparticles and intermetallic compounds  

E-print Network

Aberration-corrected electron microscopy of nanoparticles and intermetallic compounds M. Heggen for applications in energy-related catalysis, as well as novel intermetallic compounds, will be presented

Dunin-Borkowski, Rafal E.


Quantitative imaging of living cells by deep ultraviolet microscopy  

E-print Network

Developments in light microscopy over the past three centuries have opened new windows into cell structure and function, yet many questions remain unanswered by current imaging approaches. Deep ultraviolet microscopy ...

Zeskind, Benjamin J



Advanced electron microscopy characterization of multimetallic nanoparticles  

NASA Astrophysics Data System (ADS)

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 synthesis and growth mechanism of highly monodispersed Cu-Pt nanoclusters. The advance electron microscopy of microanalysis allowed us to study the distribution of Cu and Pt with atomistic resolution. The microanalysis revealed that Pt is embedded randomly in the Cu lattice. A novel grand canonical - Langevin dynamics simulation showed the formation of alloy structures in good agreement with the experimental evidence. Finally, we demonstrated the synthesis of AgPd-Pt trimetallic nanoparticles with two different morphologies: multiply twinned core-shell, and hollow particles. We also investigated the growth mechanism of the nanoparticles using grand canonical-Monte Carlo simulations. We found that the Pt regions grow at overpotentials on the AgPd nanoalloys, forming 3D islands at the early stages of the deposition process and presenting very good agreement between the simulated structures and those observed experimentally. Similarly, we also investigated AuCu/Pt core-shell trimetallic nanoparticles, presenting new way to control the nanoparticles morphologies due to the presence of third metal (Pt). Where, we observed the Pt layers are overgrowth on the as prepared AuCu core by Frank-van der Merwe (FM) and Stranski-Krastanov (SK) growth modes. In addition, these nanostructure presents high index facet surfaces with {211} and (321} families, that are highly open structure surfaces and interesting for the catalytic applications. The results of these studies will be useful for the future applications and the design of advanced functional nanomaterials.

Khanal, Subarna Raj


Ultrastructure of Candida albicans pleomorphic forms: phase-contrast microscopy, scanning and transmission electron microscopy.  


A modified method of glutaraldeyde-osmium tetroxide fixation was adjusted to characterize the ultrastructure of Candida albicans pleomorphic forms, using phase-contrast microscopy, scanning electron microscopy and transmission electron microscopy. The discovered morphological criteria defining the individual morphotypes are discussed in terms of mycological and histopathological diagnostics of candidiasis. The relations are discussed between fungal pleomorphism, virulence and susceptibility of different morphotypes to fungicides. PMID:23163212

Staniszewska, Monika; Bondaryk, Ma?gorzata; Siennicka, Katarzyna; Kurzatkowski, Wies?aw



Analytical electron microscopy in mineralogy; exsolved phases in pyroxenes  

USGS Publications Warehouse

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.

Nord, G.L., Jr.



Electron microscopy of seed-storage globulins.  


The quaternary structures of a range of seed globulins, including examples of both the so-called 7 S and 11 S types, have been examined by electron microscopy. The legume 7 S proteins, phaseolin (bean), beta-conglycinin (soybean), and vicilin (pea), appear as flat discs of diameter ca. 8.5 nm and thickness ca. 3.5 nm formed by association of three subunit domains. Phaseolin converts to an 18 S tetramer at acid pH, and images recorded under these conditions suggest that four of the 7 S protomer discs associate to form the faces of a regular tetrahedron. The classical 11 S seed globulins, cucurbitin (pumpkin) and legumin (pea), are approximately spherical molecules of diameter ca. 8.8 nm composed of six subunits. In contrast, the hexameric 10 S storage protein from lupin seed, conglutin gamma, appears toroidal in shape with outer diameter ca. 10.3 nm and thickness ca. 2.2 nm. These results indicate that constraints imposed on seed proteins by their role in sustaining the germinating plant may have allowed a variety of different globulin structures to accumulate in the protein-storage bodies of seeds. PMID:4037802

Tulloch, P A; Blagrove, R J



Ballistic-electron-emission Microscopy of Semiconductor Heterostructures  

NASA Technical Reports Server (NTRS)

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.

Bell, L. Douglas; Narayanamurti, Venkatesh



Reliable strain measurement in transistor arrays by robust scanning transmission electron microscopy  

SciTech Connect

Accurate measurement of the strain field in the channels of transistor arrays is critical for strain engineering in modern electronic devices. We applied atomic-resolution high-angle annular dark-field scanning transmission electron microscopy to quantitative measurement of the strain field in transistor arrays. The quantitative strain profile over 20 transistors was obtained with high reliability and a precision of 0.1%. The strain field was found to form homogeneously in the channels of the transistor arrays. Furthermore, strain relaxation due to the thin foil effect was quantitatively investigated for thicknesses of 35 to 275 nm.

Kim, Suhyun; Kim, Joong Jung; Jung, Younheum; Lee, Kyungwoo; Byun, Gwangsun; Hwang, KyoungHwan; Lee, Sunyoung; Lee, Kyupil [Memory Analysis Science and Engineering Group, Samsung Electronics, San 16, Hwasung City, Gyeonggi-Do 445-701 (Korea, Republic of)] [Memory Analysis Science and Engineering Group, Samsung Electronics, San 16, Hwasung City, Gyeonggi-Do 445-701 (Korea, Republic of)



Electron microscopy: Ultrastable gold substrates for electron cryomicroscopy.  


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 ? helices of individual proteins are resolved. With this improvement, we determine the structure of apoferritin, a smooth octahedral shell of ?-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. PMID:25504723

Russo, Christopher J; Passmore, Lori A



Carving: Scalable Interactive Segmentation of Neural Volume Electron Microscopy Images  

E-print Network

Carving: Scalable Interactive Segmentation of Neural Volume Electron Microscopy Images C. N electron microscopy images. We propose a supervoxel-based en- ergy function with a novel background prior that achieves these goals. This is verified by extensive experiments with a robot mimicking human interactions

Hamprecht, Fred A.


Aberration-Coreected Electron Microscopy at Brookhaven National Laboratory  

SciTech Connect

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 microscopy techniques in strongly correlated electron systems and nanostructured materials. As a first step, with the support of Materials Science Division, Office of Basic Energy Science, US Department of Energy, and the New York State Office of Science, Technology, and Academic Research, recently we acquired three aberration-corrected electron microscopes from the three major microscope manufacturers, i.e., JEOL, Hitachi, and FEI. The Hitachi HD2700C is equipped with a probe corrector, the FEI Titan 80-300 has an imaging corrector, while the JEOL2200MCO has both. All the correctors are of the dual-hexapole type, designed and manufactured by CEOS GmbH based on the design due to Rose and Haider [3, 18]. All these three are one-of-a-kind in the US, designed for specialized capabilities in characterizing nanoscale structure. In this chapter, we review the performance of these state-of-the art instruments and the new challenges associated with the improved spatial resolution, including the environment requirements of the laboratory that hosts these instruments. Although each instrument we describe here has its own strengths and drawbacks, it is not our intention to rank them in terms of their performance, especially their spatial resolution in imaging.

Zhu,Y.; Wall, J.



Field Emission Scanning Electron Microscopy (FE-SEM) and Energy Dispersive X-Ray (EDX) Spectroscopy  

E-print Network

system with 30 take-off angle for quantitative analysis, digital imaging, and X-ray mapping. The EDAXField Emission Scanning Electron Microscopy (FE-SEM) and Energy Dispersive X-Ray (EDX) Spectroscopy of objective aperture. Dual SE detectors allow versatile imaging. The FE-SEM is equipped with fully digital

Gelfond, Michael


Value of electron microscopy in the diagnosis of glomerular diseases.  


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. PMID:20192700

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



Transmission Electron Microscopy of Itokawa Regolith Grains  

NASA Technical Reports Server (NTRS)

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 of [1]. The dis-ordered rim is nanocrystalline with minor amorphous material between crystalline domains. Quantitative element maps show the outermost 10 nm of the disordered rim is Si-rich. Discussion and Conclusions: Both particles record the ef-fects of space weathering processes on Itokawa. Noguchi et al. [2] proposed that the disordered rims they observed on Itokawa particles largely result from solar wind radiation damage and we arrive at a similar conclusion for the two particles we analyzed. The microstructure of the S-depleted layer on the pyrrhotite grain in RA-QD02-0125 is similar to that observed in troilite irradiated with 1018 4 kV He+ [3, 4]. Prolonged irradiation has also been shown to disorder pyrrhotite such that the superstructure reflec-tions are lost [5].

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



Ballistic electrons : microscopy, spectroscopy, devices and luminescence  

E-print Network

This thesis describes research to theoretically model and experimentally measure electronic systems which employ ballistic electron emission. First, a Monte-Carlo framework for simulating electron injection from a tunnel ...

Appelbaum, Ian, 1977-



High Resolution Electron Microscopy at the National Cancer Institute

One key area of interest for the lab has been to close the 3D imaging gap, finding ways to image whole cells and tissues at high resolution. Focused ion beam scanning electron microscopy (FIB-SEM, or otherwise known as ion abrasion scanning electron microscopy, IA-SEM) uses a scanning electron beam to image the face of a fixed, resin-embedded sample, and ion beam to remove “slices” of the sample, resulting in a sequential stack of high resolution images.


Modern Uses of Electron Microscopy for Detection of Viruses  

PubMed Central

Summary: Electron microscopy, considered by some to be an old technique, is still on the forefront of both clinical viral diagnoses and viral ultrastructure and pathogenesis studies. In the diagnostic setting, it is particularly valuable in the surveillance of emerging diseases and potential bioterrorism viruses. In the research arena, modalities such as immunoelectron microscopy, cryo-electron microscopy, and electron tomography have demonstrated how viral structural components fit together, attach to cells, assimilate during replication, and associate with the cellular machinery during replication and egression. These studies provide information for treatment and vaccine strategies. PMID:19822888

Goldsmith, Cynthia S.; Miller, Sara E.



Spectral-domain optical coherence phase microscopy for quantitative biological studies  

E-print Network

Conventional phase-contrast and differential interference contrast microscopy produce high contrast images of transparent specimens such as cells. However, they do not provide quantitative information or do not have enough ...

Joo, Chulmin, 1976-



Advanced Correlative Light/Electron Microscopy: Current Methods and New Developments Using Tokuyasu Cryosections  

PubMed Central

Microscopy is an essential tool for analysis of cellular structures and function. With the advent of new fluorescent probes and super-resolution light microscopy techniques, the study of dynamic processes in living cells has been greatly facilitated. Fluorescence light microscopy provides analytical, quantitative, and three-dimensional (3D) data with emphasis on analysis of live cells using fluorescent markers. Sample preparation is easy and relatively inexpensive, and the use of appropriate tags provides the ability to track specific proteins of interest. Of course, only electron microscopy (EM) achieves the highest definition in terms of ultrastructure and protein labeling. To fill the gap between light microscopy and EM, correlative light and electron microscopy (CLEM) strategies have been developed. In particular, hybrid techniques based upon immuno-EM provide sensitive protein detection combined with high-resolution information on cell structures and protein localization. By adding the third dimension to EM with electron tomography (ET) combined with rapid freezing, CLEM techniques now provide additional tools for quantitative 3D analysis. Here, we overview the major methods applied and highlight the latest advances in the field of CLEM. We then focus on two selected techniques that use cryosections as substrate for combined biomolecular imaging. Finally, we provide a perspective of future developments in the field. (J Histochem Cytochem 57:1103–1112, 2009) PMID:19654103

Cortese, Katia; Diaspro, Alberto; Tacchetti, Carlo



Quantitative Electron Diffraction Structure Analysis (EDSA)  

Microsoft Academic Search

The modern state of EDSA in combination with topological analysis of the ESP and electron density allows to obtain reliable\\u000a and quantitative information about chemical bonding and properties.\\u000a \\u000a The electrostatic field in a crystal is good characterized, the determining factor is however, the introduction of cations\\u000a in the ESP of NaCl type structures. Thus precise EDSA data for calculating the

A. S. Avilov


In Situ Analytical Electron Microscopy for Probing Nanoscale Electrochemistry  

SciTech Connect

Oxides and their tailored structures are at the heart of electrochemical energy storage technologies and advances in understanding and controlling the dynamic behaviors in the complex oxides, particularly at the interfaces, during electrochemical processes will catalyze creative design concepts for new materials with enhanced and better-understood properties. Such knowledge is not accessible without new analytical tools. New innovative experimental techniques are needed for understanding the chemistry and structure of the bulk and interfaces, more importantly how they change with electrochemical processes in situ. Analytical Transmission Electron Microscopy (TEM) is used extensively to study electrode materials ex situ and is one of the most powerful tools to obtain structural, morphological, and compositional information at nanometer scale by combining imaging, diffraction and spectroscopy, e.g., EDS (energy dispersive X-ray spectrometry) and Electron Energy Loss Spectrometry (EELS). Determining the composition/structure evolution upon electrochemical cycling at the bulk and interfaces can be addressed by new electron microscopy technique with which one can observe, at the nanometer scale and in situ, the dynamic phenomena in the electrode materials. In electrochemical systems, for instance in a lithium ion battery (LIB), materials operate under conditions that are far from equilibrium, so that the materials studied ex situ may not capture the processes that occur in situ in a working battery. In situ electrochemical operation in the ultra-high vacuum column of a TEM has been pursued by two major strategies. In one strategy, a 'nano-battery' can be fabricated from an all-solid-state thin film battery using a focused ion beam (FIB). The electrolyte is either polymer based or ceramic based without any liquid component. As shown in Fig. 1a, the interfaces between the active electrode material/electrolyte can be clearly observed with TEM imaging, in contrast to the composite electrodes/electrolyte interfaces in conventional lithium ion batteries, depicted in Fig.1b, where quantitative interface characterization is extremely difficult if not impossible. A second strategy involves organic electrolyte, though this approach more closely resembles the actual operation conditions of a LIB, the extreme volatility In Situ Analytical Electron Microscopy for Probing Nanoscale Electrochemistry by Ying Shirley Meng, Thomas McGilvray, Ming-Che Yang, Danijel Gostovic, Feng Wang, Dongli Zeng, Yimei Zhu, and Jason Graetz of the organic electrolytes present significant challenges for designing an in situ cell that is suitable for the vacuum environment of the TEM. Significant progress has been made in the past few years on the development of in situ electron microscopy for probing nanoscale electrochemistry. In 2008, Brazier et al. reported the first cross-section observation of an all solid-state lithium ion nano-battery by TEM. In this study the FIB was used to make a 'nano-battery,' from an all solid-state battery prepared by pulsed laser deposition (PLD). In situ TEM observations were not possible at that time due to several key challenges such as the lack of a suitable biasing sample holder and vacuum transfer of sample. In 2010, Yamamoto et al. successfully observed changes of electric potential in an all-solid-state lithium ion battery in situ with electron holography (EH). The 2D potential distribution resulting from movement of lithium ions near the positive-electrode/electrolyte interface was quantified. More recently Huang et al. and Wang et al. reported the in situ observations of the electrochemical lithiation of a single SnO{sub 2} nanowire electrode in two different in situ setups. In their approach, a vacuum compatible ionic liquid is used as the electrolyte, eliminating the need for complicated membrane sealing to prevent the evaporation of carbonate based organic electrolyte into the TEM column. One main limitation of this approach is that EELS spectral imaging is not possible due to the high plasmon signal of the ionic li

Graetz J.; Meng, Y.S.; McGilvray, T.; Yang, M.-C.; Gostovic, D.; Wang, F.; Zeng, D.; Zhu, Y.



Relationship between the v2PO4/amide III ratio assessed by Raman spectroscopy and the calcium content measured by quantitative backscattered electron microscopy in healthy human osteonal bone  

NASA Astrophysics Data System (ADS)

Raman microspectroscopy and quantitative backscattered electron imaging (qBEI) of bone are powerful tools to investigate bone material properties. Both methods provide information on the degree of bone matrix mineralization. However, a head-to-head comparison of these outcomes from identical bone areas has not been performed to date. In femoral midshaft cross sections of three women, 99 regions (20×20 ?) were selected inside osteons and interstitial bone covering a wide range of matrix mineralization. As the focus of this study was only on regions undergoing secondary mineralization, zones exhibiting a distinct gradient in mineral content close to the mineralization front were excluded. The same regions were measured by both methods. We found a linear correlation (R2=0.75) between mineral/matrix as measured by Raman spectroscopy and the wt. %Mineral/(100-wt. %Mineral) as obtained by qBEI, in good agreement with theoretical estimations. The observed deviations of single values from the linear regression line were determined to reflect biological heterogeneities. The data of this study demonstrate the good correspondence between Raman and qBEI outcomes in describing tissue mineralization. The obtained correlation is likely sensitive to changes in bone tissue composition, providing an approach to detect potential deviations from normal bone.

Roschger, Andreas; Gamsjaeger, Sonja; Hofstetter, Birgit; Masic, Admir; Blouin, Stéphane; Messmer, Phaedra; Berzlanovich, Andrea; Paschalis, Eleftherios P.; Roschger, Paul; Klaushofer, Klaus; Fratzl, Peter



Fibrin architecture in clots: A quantitative polarized light microscopy analysis  

Microsoft Academic Search

article i nfo Article history: Fibrin plays a vital structural role in thrombus integrity. Thus, the ability to assess fibrin architecture has a potential to provide insight into thrombosis and thrombolysis. Fibrin has an anisotropic molecular structure, which enables it to be seen with polarized light. Therefore, we aimed to determine if automated polarized light microscopy methods of quantifying two

Peter Whittaker; Karin Przyklenk



Fibrin architecture in clots: A quantitative polarized light microscopy analysis  

Microsoft Academic Search

Fibrin plays a vital structural role in thrombus integrity. Thus, the ability to assess fibrin architecture has a potential to provide insight into thrombosis and thrombolysis. Fibrin has an anisotropic molecular structure, which enables it to be seen with polarized light. Therefore, we aimed to determine if automated polarized light microscopy methods of quantifying two structural parameters; fibrin fiber bundle

Peter Whittaker; Karin Przyklenk




EPA Science Inventory

The confocal laser-scanning microscopy (CLSM) has enormous potential in many biological fields. The goal of a CLSM is to acquire and quantify fluorescence and in some instruments acquire spectral characterization of the emitted signal. The accuracy of these measurements demands t...


Writing silica structures in liquid with scanning transmission electron microscopy.  


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. PMID:25207936

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



Transmission electron microscopy characterisation of 0-D nanomaterials  

NASA Astrophysics Data System (ADS)

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 nanodiamond particles cleaned by novel routes, and the possibility of production of luminescent N-V centres within the diamond nanoparticles is studied. Chapter 4 deals with self-arranged Co nanoparticle arrays, so-called superlattices. By closely studying the oxidation behaviour of such arrays, a new intrinsic property has been discovered: enhanced stability against oxidation of self-arranged cobalt nanoparticles. This intriguing physical behaviour of arranged cobalt nanoparticles has never been observed before. Chapter 5 describes and discusses all results obtained from TEM investigation of hybrid nanoporous-nanoparticle materials for advanced catalysis applications: first, the possibilities of TEM for the characterisation of the metal MOF material family; and second, the example of Au ZIF. Finally, in Chapter 6, assisted spray-pyrolysis generated ZnO nanoparticles are studied. The ZnO nanomaterial produced by a novel assisted spray pyrolysis method is compared to conventionally spray pyrolysed ZnO nanomaterials. The influence of assisted spray pyrolysis production on the size, morphology and optical properties (UV blocking capabilities) of the ZnO nanoparticles is studied for the case of citric-acid assisted spray pyrolysis.

Turner, Stuart Matthew


Ion-induced electron emission microscopy  


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.

Doyle, Barney L. (Albuquerque, NM); Vizkelethy, Gyorgy (Albuquerque, NM); Weller, Robert A. (Brentwood, TN)



Chromatic Confocal Electron Microscopy with a Finite Pinhole Size  

NASA Astrophysics Data System (ADS)

Scanning confocal electron microscopy (SCEM) is a new imaging mode in electron microscopy. Spherical aberration corrected electron microscope instruments fitted with two aberration correctors can be used in this mode which provides improved depth resolution and selectivity compared to optical sectioning in a conventional scanning transmission geometry. In this article, we consider the depth resolution and energy resolution in the confocal optical configuration for SCEM using inelastically scattered electrons with a finite pinhole size. We experimentally demonstrate energy-filtered optical sectioning in a double aberration-corrected instrument with uncorrected chromatic aberration without using a dedicated energy filter.

Wang, P.; Kirkland, A. I.; Nellist, P. D.



Applications of plasma cleaning for electron microscopy  

Microsoft Academic Search

Summary form only given. Specimen contamination and amorphous irradiation damage severely limit the ability to perform accurate electron microscope analysis of materials, especially as specimen areas of interest decrease in size. To analyze smaller areas of interest, electron probe sizes have decreased, while probe currents have increased. The combination of these two factors results in an increase in the amount

T. C. Isabell; P. E. Fischione; E. A. Fischione



Entanglement-assisted electron microscopy based on a flux qubit  

SciTech Connect

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.

Okamoto, Hiroshi, E-mail: [Department of Electronics and Information Systems, Akita Prefectural University, Yurihonjo 015-0055 (Japan); Nagatani, Yukinori [National Institute for Physiological Sciences, Okazaki 444-8787 (Japan)



Aberration-Coreected Electron Microscopy at Brookhaven National Laboratory  

Microsoft Academic Search

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

Yimei Zhu; Joe Wall



Drift correction for scanning-electron microscopy  

E-print Network

Scanning electron micrographs at high magnification (100,000x and up) are distorted by motion of the sample during image acquisition, a phenomenon called drift. We propose a method for correcting drift distortion in images ...

Snella, Michael T



Noninvasive electron microscopy with interaction-free quantum measurements  

SciTech Connect

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.

Putnam, William P.; Yanik, Mehmet Fatih [Department of Electrical Engineering and Computer Science and Research Laboratory of Electronics, Massachusetts Institute of Technology, Cambridge, Massachusetts 02139 (United States)



Stimulated excitation electron microscopy and spectroscopy.  


Recent advances in instrumentation for electron optics and spectroscopy have prompted exploration of ultra-low excitations such as phonons, bond vibrations and Johnson noise. These can be excited not just with fast electrons but also thermally or by other external sources of radiation. The near-field theory of electron energy loss and gain provides a convenient platform for analysing these processes. Possibilities for selected phonon mapping and imaging are discussed. Effects should certainly be observable in atomic resolution structure imaging but diffraction contrast imaging could perhaps be more informative. Additional exciting prospects to be explored include the transition from phonon excitation to single atom recoil and the boosting of energy loss and gain signals with tuned laser illumination. PMID:25312246

Howie, A



Semi-automated methods for simulation and measurement of amyloid fiber distributions obtained from transmission electron microscopy experiments  

Microsoft Academic Search

Transmission electron microscopy (TEM) is the standard procedure for qualitatively confirming the presence of amyloid fibers in a protein aggregation reaction product. However, extracting quantitative information about the amyloid size distribution from the electron micrographs is a nontrivial problem. Here we describe methods for (i) the simulation of pseudo-TEM images of amyloid fiber distributions having known characteristic properties and (ii)

Damien Hall


Quantitative flaw characterization with scanning laser acoustic microscopy  

NASA Technical Reports Server (NTRS)

Surface roughness and diffraction are two factors that have been observed to affect the accuracy of flaw characterization with scanning laser acoustic microscopy. Inaccuracies can arise when the surface of the test sample is acoustically rough. It is shown that, in this case, Snell's law is no longer valid for determining the direction of sound propagation within the sample. The relationship between the direction of sound propagation within the sample, the apparent flaw depth, and the sample's surface roughness is investigated. Diffraction effects can mask the acoustic images of minute flaws and make it difficult to establish their size, depth, and other characteristics. It is shown that for Fraunhofer diffraction conditions the acoustic image of a subsurface defect corresponds to a two-dimensional Fourier transform. Transforms based on simulated flaws are used to infer the size and shape of the actual flaw.

Generazio, E. R.; Roth, D. J.



Quantitative analysis of scanning force microscopy data using harmonic models  

NASA Astrophysics Data System (ADS)

The separate identification of dissipative and elastic force contributions in Atomic Force Microscopy (AFM) is discussed. We show that within a harmonic approximation the interaction of the AFM tip with the sample surface can be described by average interaction parameters, namely an effective elastic tip-sample interaction kts and an effective dissipation ?ts, which can be extracted in a simple way from measured data. The method is applied to force spectroscopy curves on hard and soft polymeric model surfaces. The approach enables a thorough discussion of the influence of experimental parameters on the measured data. In imaging a clear identification of phases in systems with hard-soft contrast as for instance in semicrystalline polymers is made possible.

Henze, Thomas; Schroeter, Klaus; Petzold, Albrecht; Thurn-Albrecht, Thomas



Non-interferometric transient quantitative phase microscopy for ultrafast engineering  

NASA Astrophysics Data System (ADS)

The accelerating developments in micro- & nanotechnology require faster and more precise tools for application and diagnostics. A new ultra-fast diagnostics is presented as an advancement of a conventional phase microscopy method (Iatia QPmTM). Contrary to the conventional method using one CCD to detect three object planes, three CCDs detect these planes separately. This concept, named TQPm, has been analyzed and validated by measuring the phase of a commercial fiber. This new method has been implemented into a pump&probe set-up with an ultra-long delay line (? t<1.6 ?s) and ablation of metals by ultra-fast laser radiation with large intensities has been investigated measuring the melt geometry.

Horn, Alexander; Mingareev, Ilja; Werth, Alexander; Kachel, Martin; Brenk, Udo



Transmission electron microscopy of the bacterial nucleoid  

Microsoft Academic Search

Water-containing biological material cannot withstand the vacuum of the transmission electron microscope. The classical solution to this problem has been to dehydrate chemically fixed biological samples and then embed them in resin. During such treatment, the bacterial nucleoid is especially prone to aggregation, which affects its global shape and fine structure. Initial attempts to deal with aggregation by optimizing chemical

Mikhail Eltsov; Benoît Zuber



Electron microscopy studies of CNT layers  

NASA Astrophysics Data System (ADS)

SEM and TEM use in an investigation of CNT-Ni layers different properties is shown. We present the possibilities of using different SEM modes (SE - secondary electrons, LABE - low angle backscattered electrons) for studies of C-Ni and CNT-Ni layers topography, morphology and cross-sectional investigations (adhesion, pores size and shape, uniformity). Correlation between concentration of Ni in studied layers and technological parameters as well as in a case of CNT-Ni films correlations of Ni concentration and a diameter of carbon nanotubes are discussed. TEM studies concentrate on structure of Ni nanograins in C-Ni layers and CNT-Ni layers, CNT structure and defects, nanoonion structure. We present methods of determination of graphite plane number in MWCNTs, distance between these planes, role of catalyst position in CNT growth and interaction between catalyst and substrate. EDS method for qualitative analysis of Ni catalyst in these layers was also presented.

Koz?owski, Miros?aw; Radomska, Joanna; Wronka, Halina; Czerwosz, El?bieta; Sobczak, Kamil



Axial ion–electron emission microscopy of IC radiation hardness  

Microsoft Academic Search

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

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



Axial ion-electron emission microscopy of IC radiation hardness  

Microsoft Academic Search

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

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



Scanning electron microscopy of bacteria Tetrasphaera duodecadis.  


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. PMID:25156672

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



Imaging Hydrated Microbial Extracellular Polymers: Comparative Analysis by Electron Microscopy? †  

PubMed Central

Microbe-mineral and -metal interactions represent a major intersection between the biosphere and geosphere but require high-resolution imaging and analytical tools for investigation of 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 by conventional electron microscopy approaches with 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 the nature of interactions between microbial extracellular polymers and their environment. PMID:21169451

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



Imaging hydrated microbial extracellular polymers: Comparative analysis by electron microscopy  

SciTech Connect

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.

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



Biological applications of phase-contrast electron microscopy.  


Here, I review the principles and applications of phase-contrast electron microscopy using phase plates. First, I develop the principle of phase contrast based on a minimal model of microscopy, introducing a double Fourier-transform process to mathematically formulate the image formation. Next, I explain four phase-contrast (PC) schemes, defocus PC, Zernike PC, Hilbert differential contrast, and schlieren optics, as image-filtering processes in the context of the minimal model, with particular emphases on the Zernike PC and corresponding Zernike phase plates. Finally, I review applications of Zernike PC cryo-electron microscopy to biological systems such as protein molecules, virus particles, and cells, including single-particle analysis to delineate three-dimensional (3D) structures of protein and virus particles and cryo-electron tomography to reconstruct 3D images of complex protein systems and cells. PMID:24357373

Nagayama, Kuniaki



CI Slide: calibration slide for quantitative microscopy imaging in absorbance  

NASA Astrophysics Data System (ADS)

New imaging technologies are changing the field of digital pathology. This field faces numerous challenges and there is a pressing need for standardization, calibration protocols, quality control and quantitative assessment. We have designed a new calibration imaging slide (Cancer Imaging Slide), specifically to measure the characteristics of old or new imaging systems or scanners. The layout of the slide consists of 138 boxes with the side length of 1.6 mm, containing objects of known morphologic and photometric characteristics. Among them, 112 boxes contain different permutations of circles, ovals, and squares. The circles have different radii, radius/pitch ratios and step transmissions. The ovals have different sizes and orientations. The squares are consistent in size and orientation but have different step transmission values. Also, 16 boxes contain three resolution test targets: crosses, USAF target and Siemens star. The last 10 boxes are blank boxes with different transmission values. Four slides were scanned and imaged on one commercial whole-slide scanner and one high resolution imaging system. After segmenting the images, about 200 features (photometric, morphologic and architectural) were measured with our in-house image processing software. The objective of the project is to develop a statistical process control using this new slide. In this paper, we describe the characteristics of the slide and present our preliminary results.

Sheikhzadeh, Fahime; Ye, Qian; Zulkafly, Nasir; Carraro, Anita; Korbelic, Jagoda; Chen, Zhaoyang; Harrison, Alan; Follen, Michele; MacAulay, Calum; Ward, Rabab K.; Guillaud, Martial



Quantitative orientation-independent differential interference contrast (DIC) microscopy  

NASA Astrophysics Data System (ADS)

We describe a new DIC technique, which records phase gradients within microscopic specimens independently of their orientation. The proposed system allows the generation of images representing the distribution of dry mass (optical path difference) in the specimen. Unlike in other forms of interference microscopes, this approach does not require a narrow illuminating cone. The orientation-independent differential interference contrast (OI-DIC) system can also be combined with orientation-independent polarization (OI-Pol) measurements to yield two complementary images: one showing dry mass distribution (which is proportional to refractive index) and the other showing distribution of birefringence (due to structural or internal anisotropy). With a model specimen used for this work -- living spermatocytes from the crane fly, Nephrotoma suturalis --- the OI-DIC image clearly reveals the detailed shape of the chromosomes while the polarization image quantitatively depicts the distribution of the birefringent microtubules in the spindle, both without any need for staining or other modifications of the cell. We present examples of a pseudo-color combined image incorporating both orientation-independent DIC and polarization images of a spermatocyte at diakinesis and metaphase of meiosis I. Those images provide clear evidence that the proposed technique can reveal fine architecture and molecular organization in live cells without perturbation associated with staining or fluorescent labeling. The phase image was obtained using optics having a numerical aperture 1.4, thus achieving a level of resolution never before achieved with any interference microscope.

Shribak, Michael; LaFountain, James; Biggs, David; Inoué, Shinya



Evaluations of carbon nanotube field emitters for electron microscopy  

NASA Astrophysics Data System (ADS)

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.

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



Prokaryote viruses studied by electron microscopy.  


This review summarizes the electron microscopical descriptions of prokaryote viruses. Since 1959, nearly 6300 prokaryote viruses have been described morphologically, including 6196 bacterial and 88 archaeal viruses. As in previous counts, the vast majority (96.3 %) are tailed, and only 230 (3.7 %) are polyhedral, filamentous, or pleomorphic. The family Siphoviridae, whose members are characterized by long, noncontractile tails, is by far the largest family (over 3600 descriptions, or 57.3 %). Prokaryote viruses are found in members of 12 bacterial and archaeal phyla. Archaeal viruses belong to 15 families or groups of family level and infect members of 16 archaeal genera, nearly exclusively hyperthermophiles or extreme halophiles. Tailed archaeal viruses are found in the Euryarchaeota only, whereas most filamentous and pleomorphic archaeal viruses occur in the Crenarchaeota. Bacterial viruses belong to 10 families and infect members of 179 bacterial genera, mostly members of the Firmicutes and ?-proteobacteria. PMID:22752841

Ackermann, H-W; Prangishvili, D



Electron microscopy of biomaterials based on hydroxyapatite  

SciTech Connect

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.

Suvorova, E. I., E-mail:; Klechkovskaya, V. V. [Russian Academy of Sciences, Shubnikov Institute of Crystallography (Russian Federation); Komarov, V. F.; Severin, A. V.; Melikhov, I. V. [Moscow State University (Russian Federation); Buffat, P. A. [Ecole Polytechnique Federale de Lausanne, Centre Interdisciplinaire de Microscopie Electronique (Switzerland)



Surface morphology of Trichinella spiralis by scanning electron microscopy  

SciTech Connect

The surface morphology of larval and adult Trichinella spiralis was studied by scanning electron microscopy (SEM) of fixed, dried, and metal-coated specimens. The results are compared with those found earlier by various investigators using light and transmission electron microscopy. Some morphological features reported here are revealed uniquely by SEM. These include the pores of the cephalic sense organs, the character of secondary cuticular folds, variations of the hypodermal gland cell openings or pores, and the presence of particles on the copulatory bell.

Kim, C.W. (State Univ. of New York, Stony Brook); Ledbetter, M.C.



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

Microsoft Academic Search

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

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



Atomic force microscopy and transmission electron microscopy of cellulose from Micrasterias denticulata; evidence for a chiral helical microfibril twist  

Microsoft Academic Search

Atomic force microscopy (AFM), tapping mode atomic force microscopy (TM-AFM) and transmission electron microscopy (TEM) have\\u000a been used to image the cell wall, ultrathin sections of whole cells and cellulose microfibrils prepared from the green alga\\u000a Micrasterias denticulata. Measurements of the microfibril dimensions are in agreement with earlier observations carried out by electron microscopy.\\u000a Images at the molecular level of




Scanning electron microscopy of Dermatobia hominis reveals cutaneous anchoring features.  


We report the case of a 45-year-old Caucasian woman suffering from cutaneous myiasis. With the use of scanning electron microscopy, we placed special focus on the mechanisms by which Dermatobia hominis can fasten securely within the human skin. PMID:17599666

Möhrenschlager, Matthias; Mempel, Martin; Weichenmeier, Ingrid; Engst, Reinhard; Ring, Johannnes; Behrendt, Heidrun




EPA Science Inventory

A recently developed in vitro excystation procedure results in almost total excystation of Giardia muris, an intestinal parasite of mice. The present experiment examines the G. muris cyst morphology by scanning electron microscopy and evaluates the efficacy of the excystation pro...


Collaboration at the Nanoscale: Exploring Viral Genetics with Electron Microscopy  

ERIC Educational Resources Information Center

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…

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




Microsoft Academic Search

ABS>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

M. L. Watson



Electron Microscopy of the Carboxysomes (Polyhedral Bodies) of Thiobacillus neapolitanus  

PubMed Central

The carboxysomes of Thiobacillus neapolitanus are shown, by electron microscopy, to consist of a paracrystalline array of 10-nm particles surrounded by a “membrane.” The 10-nm particles have a center hole or depression and have been previously identified as ribulose diphosphate carboxylase. The membrane is a monolayer approximately 3.5-nm thick. Images PMID:4127632

Shively, J. M.; Ball, Frances L.; Kline, Betty W.



Refining Mitochondria Segmentation in Electron Microscopy Imagery with Active Surfaces  

E-print Network

Refining Mitochondria Segmentation in Electron Microscopy Imagery with Active Surfaces Anne Jorstad for refining the boundary surfaces of mitochondria segmentation data. We exploit the fact that mitochondria imagery. Our resulting surfaces are seen to fit very accurately to the mitochondria membranes, more

Fua, Pascal


Araldite as an Embedding Medium for Electron Microscopy  

Microsoft Academic Search

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 conven- tional mixtures are the choice of a plasticized resin in





Technology Transfer Automated Retrieval System (TEKTRAN)

Low temperature scanning electron microscopy (LTSEM) has been used to observe and characterize fresh and metamorphosed naturally formed snow crystals1. These crystals form in the atmosphere by a process known as vapor deposition, in which molecules of water vapor (gas) bind to form a crystal (solid...


Quantifying Nanoscale Order in Amorphous Materials via Fluctuation Electron Microscopy  

ERIC Educational Resources Information Center

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…

Bogle, Stephanie Nicole



High Resolution Electron Microscopy at the National Cancer Institute

Bennett AE, Narayan K, Shi D, Hartnell L, Gousset K, He H, Lowekamp BC, Yoo TS, Bliss D, Freed EO and Subramaniam S. (2009) Ion-abrasion scanning electron microscopy reveals surface-connected tubular conduits in HIV-infected macrophages. PLoS Pathog.


Electron microscopy of oligodendroglia in severe mental illness  

Microsoft Academic Search

Qualitative electron microscopy was performed to verify whether brain pathology in schizophrenia and bipolar disorder is associated with alterations of oligodendroglial cells and myelinated fibers. Ultrastructural signs of apoptosis and necrosis of oligodendroglial cells were found in the prefrontal area 10 and the caudate nucleus in both schizophrenia and bipolar disorder. Damage of myelin sheath lamellae, with the formation of

Natalya Uranova; Diana Orlovskaya; Olga Vikhreva; Ivetta Zimina; Natalya Kolomeets; Victor Vostrikov; Valentina Rachmanova




EPA Science Inventory

Large sulfate artifacts up to 2 um in diameter were observed by scanning electron microscopy for the fine particle fraction collected in dichotomous samplers. he artifacts were attributed to small liquid particles that piled up on the filter, coalesced, and later dried as larger ...


Scanning electron microscopy imaging of hydraulic cement microstructure  

E-print Network

- ground, homogenizedblend of limestone,shaleand iron ore sintered in a rotary kiln to temperaturesofScanning electron microscopy imaging of hydraulic cement microstructure by Paul Stutzman Building Reprinted from Cement and Concrete Composites, Vol. 26, No. 8, 957-966 pp., November 2004. NOTE: This paper

Bentz, Dale P.


Electron microscopy and diffraction of aluminium oxide whiskers  

Microsoft Academic Search

Whiskers of ?-Al2O3 have been grown by the condensation and oxidation of aluminium on an alumina substrate, and examined by transmission electron microscopy and diffraction. Unbroken whiskers invariably terminate in a small globule of aluminium and have a ‘drumstick’ form. The most perfect whiskers are ribbons with their principal surfaces parallel to the (0001) planes; the majority of these have

D. J. Barber



Detection of parvoviruses in wolf feces by electron microscopy  

USGS Publications Warehouse

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.

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



Rapid characterization of ultrafiltration membranes by scanning electron microscopy  

Microsoft Academic Search

Physicochemical properties of ultrafiltration membranes were studied by scanning electron microscopy. The membrane elemental composition (carbon, oxygen, and sulfur) was determined by energy dispersion analysis. The elements were shown to be homogeneously distributed along the membrane. A homogeneous pore distribution on the membrane surface was found after covering it with a thin gold layer. The pore sizes are ~50 nm.

F. J. Márquez-Rocha; M. Aguilar-Juárez; M. J. Acosta-Ruíz; M. I. Gradilla



Contextual analysis for both light and electronic microscopy applications  

Microsoft Academic Search

The present paper describes the developement of an image analysis system devoted to applications in histology and electronic microscopy. The main characteristic of the system is its capacity to work simultaneously on more than one field of view and to handle different working magnifications. Finally it generates a complete map of analyzed sections.

G. Colliot; O. Lejeune; S. de Bentzmann; M. Tajani; N. Bonnet; E. C. Hirsch



Scanning electron microscopy of Setaria cervi adult male worms.  


Scanning electron microscopy studies of adult Setaria cervi male worms showed the presence of horn-like lateral appendages and characteristic striated bands on the ventral side of the tail. All other features were indistinguishable from other members of the genus Setaria. PMID:2040560

Almeida, A J; Deobhankar, K P; Bhopale, M K; Zaman, V; Renapurkar, D M



Laboratory-based quantitative hard x-ray phase microscopy in one dimension using waveguides  

NASA Astrophysics Data System (ADS)

We report on the quantitative hard x-ray phase microscopy obtained with a laboratory source equipped with an x-ray planar waveguide. The waveguide, acting as a small secondary source with increased coherence, allows for phase contrast microscopy to be measured from a phase-only one-dimensional object. We analyzed different strategies and their performances for the case studied of low absorbing one-dimensional sample. It was found that the phase-only approximation for the sample enables the best performance in phase retrieval. Results obtained from experimental data are supported by phase retrieval performed on simulated data allowing an estimation of the performance of the algorithms. The ability to perform quantitative phase contrast microscopy with waveguides is an important advance for this novel x-ray phase contrast method, well suited to compact laboratory setups.

Roberts, Matthew D.; Pelliccia, Daniele



Molecular and Cellular Quantitative Microscopy: theoretical investigations, technological developments and applications to neurobiology  

NASA Astrophysics Data System (ADS)

This PhD project aims at the development and evaluation of microscopy techniques for the quantitative detection of molecular interactions and cellular features. The primarily investigated techniques are F?rster Resonance Energy Transfer imaging and Fluorescence Lifetime Imaging Microscopy. These techniques have the capability to quantitatively probe the biochemical environment of fluorophores. An automated microscope capable of unsupervised operation has been developed that enables the investigation of molecular and cellular properties at high throughput levels and the analysis of cellular heterogeneity. State-of-the-art Förster Resonance Energy Transfer imaging, Fluorescence Lifetime Imaging Microscopy, Confocal Laser Scanning Microscopy and the newly developed tools have been combined with cellular and molecular biology techniques for the investigation of protein-protein interactions, oligomerization and post-translational modifications of ?-Synuclein and Tau, two proteins involved in Parkinson’s and Alzheimer’s disease, respectively. The high inter-disciplinarity of this project required the merging of the expertise of both the Molecular Biophysics Group at the Debye Institute - Utrecht University and the Cell Biophysics Group at the European Neuroscience Institute - G?ttingen University. This project was conducted also with the support and the collaboration of the Center for the Molecular Physiology of the Brain (Göttingen), particularly with the groups associated with the Molecular Quantitative Microscopy and Parkinson’s Disease and Aggregopathies areas. This work demonstrates that molecular and cellular quantitative microscopy can be used in combination with high-throughput screening as a powerful tool for the investigation of the molecular mechanisms of complex biological phenomena like those occurring in neurodegenerative diseases.

Esposito, Alessandro



Imaging and microanalysis of thin ionomer layers by scanning transmission electron microscopy  

SciTech Connect

Improved conditions for imaging and spectroscopic mapping of thin perfluorosulfonic acid (PFSA) ionomer layers in fuel cell electrodes by scanning transmission electron microscopy (STEM) have been investigated. These conditions are first identified on model systems of Nafion ionomer-coated nanostructured thin films and nanoporous Si. The optimized conditions are then applied in a quantitative study of the ionomer through-layer loading for two typical electrode catalyst coatings using electron energy loss and energy dispersive X-ray spectroscopy in the transmission electron microscope. The e-beam induced damage to the perfluorosulfonic acid (PFSA) ionomer is quantified by following the fluorine mass loss with electron exposure and is then mitigated by a few orders of magnitude using cryogenic specimen cooling and a higher incident electron voltage. Multivariate statistical analysis is also applied to the analysis of spectrum images for data denoising and unbiased separation of independent components related to the catalyst, ionomer, and support.

Cullen, David A [ORNL; Koestner, Roland [General Motors Corporation; Kukreja, Ratan [General Motors Corporation; Minko, Sergiy [Clarkson University, Potsdam, NY; Trotsenko, Oleksandr [Clarkson University, Potsdam, NY; Tokarev, Alexander V [ORNL; Guetaz, Laure [French Atomic Energy Commission (CEA), Grenoble; Meyer III, Harry M [ORNL; Parish, Chad M [ORNL; More, Karren Leslie [ORNL



Correlated Light and Electron Microscopy/Electron Tomography of Mitochondria In Situ  

PubMed Central

Three-dimensional light microscopy and three-dimensional electron microscopy (electron tomography) separately provide very powerful tools to study cellular structure and physiology, including the structure and physiology of mitochondria. Fluorescence microscopy allows one to study processes in live cells with specific labels and stains that follow the movement of labeled proteins and changes within cellular compartments but does not have sufficient resolution to define the ultrastructure of intracellular organelles such as mitochondria. Electron microscopy and electron tomography provide the highest resolution currently available to study mitochondrial ultrastructure but cannot follow processes in living cells. We describe the combination of these two techniques in which fluorescence confocal microscopy is used to study structural and physiologic changes in mitochondria within apoptotic HeLa cells to define the apoptotic timeframe. Cells can then be selected at various stages of the apoptotic timeframe for examination at higher resolution by electron microscopy and electron tomography. This is a form of “virtual” 4-dimensional electron microscopy that has revealed interesting structural changes in the mitochondria of HeLa cells during apoptosis. The same techniques can be applied, with modification, to study other dynamic processes within cells in other experimental contexts. PMID:19348881

Perkins, Guy A.; Sun, Mei G.; Frey, Terrence G.



Microanalysis using secondary electrons in scanning electron microscopy.  


A recent study of secondary electron (SE) spectra in an Auger spectrometer demonstrated unique features indicative of the chemical nature of the tested material. The scanning electron microscope (SEM) naturally generates SEs; therefore, in this paper, we combine the concept of using differential voltage contrast (DVC) with SE spectroscopy to identify the chemical nature of a material. It is demonstrated that this method reveals the uniqueness of electron energy distribution in the conduction band of a solid or, what is the same, the uniqueness of a build-up of the outer electron shell system, and avoids errors due to the changes in the angular distribution or yield of the SE in the SEM. A theory of this new microanalytical method is developed. The experimental limitations of the SEM for this type of study are examined as well. PMID:11587321

Mil'shtein, S; Joy, D C



Robert Feulgen Prize Lecture 1995. Electronic light microscopy: present capabilities and future prospects.  


Electronic light microscopy involves the combination of microscopic techniques with electronic imaging and digital image processing, resulting in dramatic improvements in image quality and ease of quantitative analysis. In this review, after a brief definition of digital images and a discussion of the sampling requirements for the accurate digital recording of optical images, I discuss the three most important imaging modalities in electronic light microscopy--video-enhanced contrast microscopy, digital fluorescence microscopy and confocal scanning microscopy--considering their capabilities, their applications, and recent developments that will increase their potential. Video-enhanced contrast microscopy permits the clear visualisation and real-time dynamic recording of minute objects such as microtubules, vesicles and colloidal gold particles, an order of magnitude smaller than the resolution limit of the light microscope. It has revolutionised the study of cellular motility, and permits the quantitative tracking of organelles and gold-labelled membrane bound proteins. In combination with the technique of optical trapping (optical tweezers), it permits exquisitely sensitive force and distance measurements to be made on motor proteins. Digital fluorescence microscopy enables low-light-level imaging of fluorescently labelled specimens. Recent progress has involved improvements in cameras, fluorescent probes and fluorescent filter sets, particularly multiple bandpass dichroic mirrors, and developments in multiparameter imaging, which is becoming particularly important for in situ hybridisation studies and automated image cytometry, fluorescence ratio imaging, and time-resolved fluorescence. As software improves and small computers become more powerful, computational techniques for out-of-focus blur deconvolution and image restoration are becoming increasingly important. Confocal microscopy permits convenient, high-resolution, non-invasive, blur-free optical sectioning and 3D image acquisition, but suffers from a number of limitations. I discuss advances in confocal techniques that address the problems of temporal resolution, spherical and chromatic aberration, wavelength flexibility and cross-talk between fluorescent channels, and describe new optics to enhance axial resolution and the use of two-photon excitation to reduce photobleaching. Finally, I consider the desirability of establishing a digital image database, the BioImage database, which would permit the archival storage of, and public Internet access to, multidimensional image data from all forms of biological microscopy. Submission of images to the BioImage database would be made in coordination with the scientific publication of research results based upon these data.(ABSTRACT TRUNCATED AT 400 WORDS) PMID:8536077

Shotton, D M



Organization of the Outer Plexiform Layer of the Primate Retina: Electron Microscopy of Golgi-Impregnated Cells  

Microsoft Academic Search

Golgi-impregnated retinae of rhesus monkeys have been examined by serial section electron microscopy to establish in a quantitative manner the neural connexions in the outer plexiform layer. The results have shown that there are two types of midget bipolar cell, here called the invaginating midget bipolar and the flat midget bipolar. Both types of midget bipolar are exclusive to a

Helga Kolb



Cooled CCD camera with tapered fibre optics for electron microscopy  

NASA Astrophysics Data System (ADS)

A CCD camera for use in electron microscopy, with 1286 × 1152, 37 ?m pixels and an input aperture of 60 mm diameter, is described in this paper. An attempt is made to optimise the phosphor resolution for 120 keV electrons using Monte-Carlo simulation methods. Incident electrons are converted to visible light in a polycrystalline phosphor (P43) deposited on tapered fibre optics and imaged on to a cooled slow-scan CCD which is controlled from a Sun sparc-station, running under a Unix platform, through a VME-based drive and read-out electronics system. The camera is attached to a Philips CM12 microscope and is used mainly for recording electron-diffraction patterns from two-dimensiónally ordered protein arrays. Data can be displayed rapidly on the Sun monitor and can also be transferred for further analysis to a Dec Alpha computer via Ethernet for application of various image-processing programs.

Faruqi, A. R.; Andrews, H. N.



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

NASA Astrophysics Data System (ADS)

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.

Stoll, Joshua D.; Kolmakov, Andrei



Color metallography and electron microscopy techniques applied to the characterization of 413.0 aluminum alloys.  


The influence on alloy 413.0 of the refinement and modification of its microstructure was analyzed by means of several microscopy techniques, as well as the effect of the application of high pressure during solidification. For each treatment and solidification pressure condition employed, the most suitable microscopy techniques for identifying and characterizing the phases present were investigated. Color metallography and electron microscopy techniques were applied to the qualitative microstructural analysis. Volume fraction and grain size of the primary ?-Al were characterized by quantitative metallographic techniques. The results show that the effect caused by applying high pressure during solidification of the alloy is more pronounced than that caused by modification and refinement of the microstructure when it solidifies at atmospheric pressure. Furthermore, it has been shown that, for Al-Si alloy characterization, when aiming to characterize the primary ?-Al phase, optical color metallography observed under crossed polarized light plus a sensitive tint filter is the most suitable technique. When the goal is to characterize the eutectic Si, the use of optical color metallography or electron microscopy is equally valid. The characterization of iron-rich intermetallic compounds should preferably be performed by means of backscattered electron imaging. PMID:23701972

Vander Voort, George; Asensio-Lozano, Juan; Suárez-Peña, Beatriz



Detective quantum efficiency of electron area detectors in electron microscopy  

PubMed Central

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

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



Low temperature electron microscopy and electron diffraction of the purple membrane of Halobacterium halobium  

Microsoft Academic Search

The structure of the purple membrane of Halobacterium halobium was studied by high resolution electron microscopy and electron diffraction, primarily at low temperature. The handedness of the purple membrane diffraction pattern with respect to the cell membrane was determined by electron diffraction of purple membranes adsorbed to polylysine. A new method of preparing frozen specimens was used to preserve the



Vibrational and optical spectroscopies integrated with environmental transmission electron microscopy.  


Here, we present a measurement platform for collecting multiple types of spectroscopy data during high-resolution environmental transmission electron microscopy observations of dynamic processes. Such coupled measurements are made possible by a broadband, high-efficiency, free-space optical system. The critical element of the system is a parabolic mirror, inserted using an independent hollow rod and placed below the sample holder which can focus a light on the sample and/or collect the optical response. We demonstrate the versatility of this optical setup by using it to combine in situ atomic-scale electron microscopy observations with Raman spectroscopy. The Raman data is also used to measure the local temperature of the observed sample area. Other applications include, but are not limited to: cathodo- and photoluminescence spectroscopy, and use of the laser as a local, high-rate heating source. PMID:25490533

Picher, Matthieu; Mazzucco, Stefano; Blankenship, Steve; Sharma, Renu



Fracture characteristics of uranium alloys by scanning electron microscopy  

Microsoft Academic Search

The fracture characteristics of uranium alloys were determined by scanning electron microscopy. The fracture mode of stress-corrosion cracking (SCC) of uranium-7.5 weight percent niobium-2.5 weight percent zirconium (Mulberry) alloy, uranium--niobium alloys, and uranium--molybdenum alloys in aqueous chloride solutions is intergranular. The SCC fracture surface of the Mulberry alloy is characterized by very clean and smooth grain facets. The tensile-overload fracture

J. W. Koger; R. K. Jr. Bennett



Visualization of neural cell adhesion molecule by electron microscopy  

Microsoft Academic Search

The 130- and 160-kD polypeptide forms of the neural cell adhesion molecule (NCAM) were ana- lyzed by electron microscopy after low angle rotary shadowing and freeze replication. Individual NCAM molecules appeared as uniformly thick rods, with a distinct bend or hinge region near their middle. Ag- gregates were also present, containing two to six rods in a pinwheel-like configuration without

Alison K. Hall; Urs Rutishauser



[Acquired zinc deficiency mimicking glucagonoma dermatitis. Histology and electron microscopy].  


A 47-year-old man suffering from colitis ulcerosa developed serious anaemia, oedema, stomatitis with burning tongue, dry skin and erosive lesions with peripheral spreading on the ankles. Histology and electron microscopy showed characteristic signs for necrolytic migratory erythema, zinc deficiency acrodermatitis, or pellagra. Decreased serum zinc level and response to oral zinc substitution finally proved the diagnosis of acquired zinc deficiency. PMID:6464530

Haneke, E



Detection and identification of light impurities by electron microscopy  

Microsoft Academic Search

For over 40 years impurities have been believed to stabilize the ceramic -SiN but there is no direct evidence for their identity or lattice location. In bulk materials electron microscopy can generally image heavy impurities. Here we report direct imaging of N columns in -SiN that suggests the presence of excess light elements in specific N columns. First-principles calculations rule

Juan C Idrobo Tapia; Mark P Oxley; Weronika Walkosz; Robert F Klie; Serdar Ogut; B Mikijelj; Stephen J Pennycook; Sokrates T. Pantelides



Interactions of elastic and rigid vesicles with human skin in vitro: electron microscopy and two-photon excitation microscopy  

Microsoft Academic Search

Interactions between vesicle formulations and human skin were studied, in vitro, in relation to their composition and elasticity. The skin ultrastructure was investigated using transmission electron microscopy (TEM), freeze-fracture electron microscopy (FFEM) and two-photon fluorescence microscopy (TPE). The main difference between the vesicle formulations was their elasticity. Elastic vesicle formulations contained bilayer forming surfactants\\/lipids and single-chain surfactant octaoxyethylenelaurate-ester (PEG-8-L), whereas

Benedicte A. I. van den Bergh; Jurrien Vroom; Hans Gerritsen; Hans E. Junginger; Joke A. Bouwstra



Three-dimensional volume imaging with electron microscopy toward connectome.  


Ultrastructural analyses with electron microscopy have provided indispensable information to understand physiology and pathology of the nervous system. Recent advancement in imaging methodology paved the way for complete reconstruction of the neuronal connection map in the central nervous system, which is termed 'connectome' and would provide key insights to understand the functions of the brain. The critical advancement includes serial ultrastructural observation with scanning electron microscopy (SEM) instead of conventional serial sectioning transmission electron microscopy along with specific tissue preparation methods to increase heavy metal deposition for efficient SEM imaging. The advanced imaging methods using SEM have distinct advantages and disadvantages in multiple aspects, such as resolution and imaging speed, and should be selected depending on the observation conditions, such as target tissue sizes, required spatial resolution and necessity for re-observation. Dealing with the huge dataset remained to be a major obstacle, and automation in segmentation and 3D reconstruction would be critical to understand neuronal circuits in a larger volume of the brain. Future improvement in acquisition and analyses of the morphological data obtained with the advanced SEM imaging is awaited to elucidate the significance of whole connectome as the structural basis of the consciousness, intelligence and memory of a subject. PMID:25550364

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



Experiments in electron microscopy: from metals to nerves  

NASA Astrophysics Data System (ADS)

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.

Unwin, Nigel



Fixation methods for electron microscopy of human and other liver  

PubMed Central

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

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



High sensitivity piezomagnetic force microscopy for quantitative probing of magnetic materials at the nanoscale.  


Accurate scanning probing of magnetic materials at the nanoscale is essential for developing and characterizing magnetic nanostructures, yet quantitative analysis is difficult using the state of the art magnetic force microscopy, and has limited spatial resolution and sensitivity. In this communication, we develop a novel piezomagnetic force microscopy (PmFM) technique, with the imaging principle based on the detection of magnetostrictive response excited by an external magnetic field. In combination with the dual AC resonance tracking (DART) technique, the contact stiffness and energy dissipation of the samples can be simultaneously mapped along with the PmFM phase and amplitude, enabling quantitative probing of magnetic materials and structures at the nanoscale with high sensitivity and spatial resolution. PmFM has been applied to probe magnetic soft discs and cobalt ferrite thin films, demonstrating it as a powerful tool for a wide range of magnetic materials. PMID:23720016

Chen, Qian Nataly; Ma, Feiyue; Xie, Shuhong; Liu, Yuanming; Proksch, Roger; Li, Jiangyu



Investigation of Nematode Diversity using Scanning Electron Microscopy and Fluorescent Microscopy  

NASA Astrophysics Data System (ADS)

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.

Seacor, Taylor; Howell, Carina



Quantitative measurement of in-plane cantilever torsion for calibrating lateral piezoresponse force microscopy.  

SciTech Connect

A simple quantitative measurement procedure of in-plane cantilever torsion for calibrating lateral piezoresponse force microscopy is presented. This technique enables one to determine the corresponding lateral inverse optical lever sensitivity (LIOLS) of the cantilever on the given sample. Piezoelectric coefficient, d{sub 31} of BaTiO{sub 3} single crystal (-81.62 {+-} 40.22 pm/V) which was calculated using the estimated LIOLS was in good agreement with the reported value in literature.

Choi, H.; Hong, S.; No, K. (Materials Science Division); (Korea Adv. Inst. Sci. Tech.)



Analysis of mixed cell cultures with quantitative digital holographic phase microscopy  

NASA Astrophysics Data System (ADS)

In order to study, for example, the influence of pharmaceuticals or pathogens on different cell types under identical measurement conditions and to analyze interactions between different cellular specimens a minimally-invasive quantitative observation of mixed cell cultures is of particular interest. Quantitative phase microscopy (QPM) provides high resolution detection of optical path length changes that is suitable for stain-free minimally-invasive live cell analysis. Due to low light intensities for object illumination, QPM minimizes the interaction with the sample and is in particular suitable for long term time-lapse investigations, e.g., for the detection of cell morphology alterations due to drugs and toxins. Furthermore, QPM has been demonstrated to be a versatile tool for the quantification of cellular growth, the extraction morphological parameters and cell motility. We studied the feasibility of QPM for the analysis of mixed cell cultures. It was explored if quantitative phase images provide sufficient information to distinguish between different cell types and to extract cell specific parameters. For the experiments quantitative phase imaging with digital holographic microscopy (DHM) was utilized. Mixed cell cultures with different types of human pancreatic tumor cells were observed with quantitative DHM phase contrast up to 35 h. The obtained series of quantitative phase images were evaluated by adapted algorithms for image segmentation. From the segmented images the cellular dry mass and the mean cell thickness were calculated and used in the further analysis as parameters to quantify the reliability the measurement principle. The obtained results demonstrate that it is possible to characterize the growth of cell types with different morphologies in a mixed cell culture separately by consideration of specimen size and cell thickness in the evaluation of quantitative DHM phase images.

Kemper, Björn; Wibbeling, Jana; Ketelhut, Steffi



Electron microscopy study of antioxidant interaction with bacterial cells  

NASA Astrophysics Data System (ADS)

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.

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



New immunolatex spheres: visual markers of antigens on lymphocytes for scanning electron microscopy  

PubMed Central

New immunochemical reagents consisting of antibodies bound to small latex spheres were used as visual markers for the detection and localization of cell surface antigens by scanning electron microscopy. Cross-linked latex spheres of various sizes from 300 to 3,4000 A in diameter were synthesized by aqueous emulsion copolymerization of methacrylate derivatives containing hydroxyl and carboxyl functional groups. Proteins and other molecules containing primary amino groups were covalently bonded to the acrylic spheres under a variety of mild conditions by the aqueous carbodiimide, cyanogen bromide, and glutaraldehyde methods. For use in the indirect immunochemical-labeling technique, goat antibodies directed against rabbit immunoglobulins were bonded to the spheres. These immunolatex reagents were shown to bind only to cells (red blood and lymphocytes) which had previously been sensitized with rabbit antibodies against cell surface antigens. Mouse spleen lymphocytes with exposed immunoglobulins on their surface (B cells) were labeled with these spheres and distinguished from unlabeled or T lymphocytes by scanning electron microscopy. The distribution of Ig receptors on lymphocytes was also studied using the spheres as visual markers. When lymphocytes were fixed with glutaraldehyde and subsequently labeled with the immunolatex reagents, a random distribution was observed by scanning electron microscopy; a patchy distribution was observed when unfixed lymphocytes were used. These results are consistent with studies using ferritin-labeled antibodies (S. De Petris and M. Raff. 1973. Nature [Lond.]. 241:257.) and support the view that Ig receptors on lymphocytes undergo translational diffusion. In addition to serving as visual markers for scanning electron microscopy, these latex spheres tagged with fluorescent or radioactive molecules have applications as highly sensitive markers for fluorescent microscopy and as reagents for quantitative studies of cell surface antigens and other receptors. PMID:803228



Quantitative imaging of electrospun fibers by PeakForce Quantitative NanoMechanics atomic force microscopy using etched scanning probes.  


Electrospun polymeric submicron and nanofibers can be used as tissue engineering scaffolds in regenerative medicine. In physiological conditions fibers are subjected to stresses and strains from the surrounding biological environment. Such stresses can cause permanent deformation or even failure to their structure. Therefore, there is a growing necessity to characterize their mechanical properties, especially at the nanoscale. Atomic force microscopy is a powerful tool for the visualization and probing of selected mechanical properties of materials in biomedical sciences. Image resolution of atomic force microscopy techniques depends on the equipment quality and shape of the scanning probe. The probe radius and aspect ratio has huge impact on the quality of measurement. In the presented work the nanomechanical properties of four different polymer based electrospun fibers were tested using PeakForce Quantitative NanoMechanics atomic force microscopy, with standard and modified scanning probes. Standard, commercially available probes have been modified by etching using focused ion beam (FIB). Results have shown that modified probes can be used for mechanical properties mapping of biomaterial in the nanoscale, and generate nanomechanical information where conventional tips fail. PMID:25710786

Chlanda, Adrian; Rebis, Janusz; Kije?ska, Ewa; Wozniak, Michal J; Rozniatowski, Krzysztof; Swieszkowski, Wojciech; Kurzydlowski, Krzysztof J



Transmission electron microscopy: isotope substitution extends the lifetime of organic molecules in transmission electron microscopy (small 5/2015).  


Decreasing the energy of the electron beam in transmission electron microscopy (TEM) improves the stability of inorganic molecules, however, organic molecules become surprisingly less stable under these conditions. On page 622, E. Bichoutskaia, U. Kaiser, A. N. Khlobystov, and co-workers analyse the interactions of the e-beam with C-H bonds and are able to highlight the low atomic weight of hydrogen as a key issue. Exchanging protium for deuterium resolves the issue and brings atomic-resolution TEM imaging of individual organic molecules closer to reality. PMID:25644255

Chamberlain, Thomas W; Biskupek, Johannes; Skowron, Stephen T; Bayliss, Peter A; Bichoutskaia, Elena; Kaiser, Ute; Khlobystov, Andrei N



Correlative cryo-electron tomography and optical microscopy of cells.  


The biological processes occurring in a cell are complex and dynamic, and to achieve a comprehensive understanding of the molecular mechanisms underlying these processes, both temporal and spatial information is required. While cryo-electron tomography (cryoET) provides three-dimensional (3D) still pictures of near-native state cells and organelles at molecular resolution, fluorescence light microscopy (fLM) offers movies of dynamic cellular processes in living cells. Combining and integrating these two commonly used imaging modalities (termed correlative microscopy) provides a powerful means to not only expand the imaging scale and resolution but also to complement the dynamic information available from optical microscopy with the molecular-level, 3D ultrastructure detail provided by cryoET. As such, a correlative approach performed on a given specimen can provide high resolution snapshots of dynamic cellular events. In this article, I review recent advances in correlative light microscopy and cryoET and discuss major findings made available by applying this method. PMID:23962486

Zhang, Peijun



4D multiple-cathode ultrafast electron microscopy  

PubMed Central

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

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



Scanning electron microscopy and energy dispersive analysis: applications in the field of cultural heritage.  


Scanning electron microscopy has been extensively used for the material characterization of objects of artistic and archaeological importance, especially in combination with energy dispersive X-ray microanalysis (SEM/EDX). The advantages and limitations of SEM/EDX are presented in a few case studies: analysis of pigments in cross-sections of paint layers, quantitative analysis of archaeological glass from the Roman period excavated in Ephesos/Turkey, and investigations on glasses with medieval composition concerning their weathering stability and degradation phenomena. PMID:17031630

Schreiner, Manfred; Melcher, Michael; Uhlir, Katharina



Advances in quantitative nanoscale subsurface imaging by mode-synthesizing atomic force microscopy  

NASA Astrophysics Data System (ADS)

This paper reports on advances toward quantitative non-destructive nanoscale subsurface investigation of a nanofabricated sample based on mode synthesizing atomic force microscopy with heterodyne detection, addressing the need to correlate the role of actuation frequencies of the probe fp and the sample fs with depth resolution for 3D tomography reconstruction. Here, by developing a simple model and validating the approach experimentally through the study of the nanofabricated calibration depth samples consisting of buried metallic patterns, we demonstrate avenues for quantitative nanoscale subsurface imaging. Our findings enable the reconstruction of the sample depth profile and allow high fidelity resolution of the buried nanostructures. Non-destructive quantitative nanoscale subsurface imaging offers great promise in the study of the structures and properties of complex systems at the nanoscale.

Vitry, P.; Bourillot, E.; Plassard, C.; Lacroute, Y.; Tetard, L.; Lesniewska, E.



Advances in quantitative nanoscale subsurface imaging by mode-synthesizing atomic force microscopy  

SciTech Connect

This paper reports on advances toward quantitative non-destructive nanoscale subsurface investigation of a nanofabricated sample based on mode synthesizing atomic force microscopy with heterodyne detection, addressing the need to correlate the role of actuation frequencies of the probe f{sub p} and the sample f{sub s} with depth resolution for 3D tomography reconstruction. Here, by developing a simple model and validating the approach experimentally through the study of the nanofabricated calibration depth samples consisting of buried metallic patterns, we demonstrate avenues for quantitative nanoscale subsurface imaging. Our findings enable the reconstruction of the sample depth profile and allow high fidelity resolution of the buried nanostructures. Non-destructive quantitative nanoscale subsurface imaging offers great promise in the study of the structures and properties of complex systems at the nanoscale.

Vitry, P.; Bourillot, E.; Plassard, C.; Lacroute, Y.; Lesniewska, E. [ICB, UMR CNRS 6303 CNRS-University of Bourgogne, Dijon F-21078 (France); Tetard, L. [Nanoscience Technology Center, University of Central Florida, Orlando, Florida 32826 (United States)



Time resolved electron microscopy for in situ experiments  

SciTech Connect

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.

Campbell, Geoffrey H., E-mail:; McKeown, Joseph T.; Santala, Melissa K. [Lawrence Livermore National Laboratory, Livermore, California 94550 (United States)



Opportunities for electron microscopy in space radiation biology  

SciTech Connect

Densely ionizing, particulate radiations in outer space are likely to cause to mammalian tissues biological damage that is particularly amenable to examination by the techniques of electron microscopy. This situation arises primarily from the fact that once the density of ionization along the particle track exceeds a certain value, small discrete lesions involving many adjacent cells may be caused in organized tissues. Tissue damage produced by ionization densities below the critical value also afford opportunities for electron microscopic evaluation, as is shown by the damage produced in optic and proximate tissues of the New Zealand white rabbit in terrestrial experiments. Late radiation sequelae in nondividing, or terminally differentiating, tissues, and in stem cell populations, are of special importance in these regards. It is probable that evaluations of the hazards posed to astronauts by galactic particulate radiations during prolonged missions in outer space will not be complete without adequate electron microscopic evaluation of the damage those radiations cause to organized tissues.

Lett, J.T.



The Electron Microscopy Outreach Program: A Web-Based Resource for Research and Education  

E-print Network

The Electron Microscopy Outreach Program: A Web-Based Resource for Research and Education Gina E and expertise for biological elec- tron microscopy. A major focus is molecular electron microscopy, but the site site, called the Electron Micros- copy (EM) Outreach Program (URL: http://em- outreach

Baker, Timothy S.


Application for Visitors to the Boulder Laboratory for 3D Electron Microscopy of Cells  

E-print Network

Application for Visitors to the Boulder Laboratory for 3D Electron Microscopy of Cells The Boulder Laboratory for 3D Electron Microscopy of Cells is supported by the National Center for Research Resources)735-0770 University of Colorado, Dept. MCDB The Boulder Laboratory for 3D Electron Microscopy of Cells 347 UCB Boulder

Stowell, Michael


Fast microstructure and phase analyses of nanopowders using combined analysis of transmission electron microscopy scattering patterns.  


The full quantitative characterization of nanopowders using transmission electron microscopy scattering patterns is shown. This study demonstrates the feasibility of the application of so-called combined analysis, a global approach for phase identification, structure refinement, characterization of anisotropic crystallite sizes and shapes, texture analysis and texture variations with the probed scale, using electron diffraction patterns of TiO2 and Mn3O4 nanocrystal aggregates and platinum films. Electron diffraction pattern misalignments, positioning, and slight changes from pattern to pattern are directly integrated and refined within this approach. The use of a newly developed full-pattern search-match methodology for phase identification of nanopowders and the incorporation of the two-wave dynamical correction for diffraction patterns are also reported and proved to be efficient. PMID:25176993

Boullay, P; Lutterotti, L; Chateigner, D; Sicard, L



High-resolution electron microscopy of advanced materials  

SciTech Connect

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.

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



The origins and evolution of freeze-etch electron microscopy  

PubMed Central

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

Heuser, John E.



Simultaneous orientation and thickness mapping in transmission electron microscopy.  


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 22nm technology transistor structures. The performance and limitations of our method are discussed and compared to those of other techniques available. PMID:25497718

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



Electron Spin Magnetic Resonance Force Microscopy of Nitroxide Spin Labels  

NASA Astrophysics Data System (ADS)

Nitroxide spin labels are widely used in electron spin resonance studies of biological and polymeric systems. Magnetic resonance force microscopy (MRFM) is a magnetic resonance technique that couples the high spatial resolution of a scanning probe microscope with the species selectivity of magnetic resonance. We report on our investigations of 4-amino TEMPO, a nitroxide spin label, by force-gradient MRFM. Our microscope operates at high vacuum in liquid helium, using a custom fabricated ultra-soft silicon cantilever in the magnet-on-cantilever geometry. An 18 GHz gap coupled microstripline resonator supplies the transverse field.

Moore, Eric W.; Lee, Sanggap; Hickman, Steven A.; Wright, Sarah J.; Marohn, John A.



Segmentation of virus particle candidates in transmission electron microscopy images.  


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. PMID:21972793

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



Immunochemistry and electron microscopy of head and neck rhabdomyoma.  

PubMed Central

Rhabdomyomas are rare benign tumours originating in skeletal or cardiac muscle. Extracardiac tumours are usually situated in the head and neck. Four cases are presented, three arising in the larynx and the other in the cervical region. All four cases were studied by light and electron microscopy, and in three immunohistochemical staining for myoglobin, desmin, and vimentin was carried out to study the diagnostic features of the lesions and their histogenesis. Images Fig 1 Fig 2 Fig 3 Fig 4 Fig 5 Fig 6 Fig 7 Fig 8 Fig 9 Fig 10 PMID:3056977

Helliwell, T R; Sissons, M C; Stoney, P J; Ashworth, M T



Ballistic Electron Emission Microscopy of Metal/Group IV Interfaces  

NASA Technical Reports Server (NTRS)

Ballistic electron emission microscopy and spectroscopy, together with related techniques, have been applied with great success to the study of buried interfaces. These probes, known collectively as BEEM, have yielded important information on interface transport, interface band structure, and carrier scattering, with lateral spatial resolution on the nanometer scale. Recent applications of the technique to polycrystalline metal/semiconductor interfaces have demonstrated an ability to spatially map both conduction band and valence band semiconductor structure. BEEM studies of epitaxial silicide/silicon interfaces have been particularly fruitful, as the rich silicide band structure results in complex and often surprising transport behavior...

Hecht, M. H.; Kaiser, W. J.; Bell, L. D.; Fathauer, R.; Manion, S. J.



Analytical electron microscopy of a hydrated interplanetary dust particle  

NASA Astrophysics Data System (ADS)

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.

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


Analytical electron microscopy of a hydrated interplanetary dust particle  

NASA Technical Reports Server (NTRS)

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.

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



Confocal Microscopy for Modeling Electron Microbeam Irradiation of Skin  

SciTech Connect

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.

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



Transmission Electron Microscopy Of Lipid Vesicles For Drug Delivery  

NASA Astrophysics Data System (ADS)

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.

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



Imaging and quantitative data acquisition of biological cell walls with Atomic Force Microscopy and Scanning Acoustic Microscopy  

SciTech Connect

This chapter demonstrates the feasibility of Atomic Force Microscopy (AFM) and High Frequency Scanning Acoustic Microscopy (HF-SAM) as tools to characterize biological tissues. Both the AFM and the SAM have shown to provide imaging (with different resolution) and quantitative elasticity measuring abilities. Plant cell walls with minimal disturbance and under conditions of their native state have been examined with these two kinds of microscopy. After descriptions of both the SAM and AFM, their special features and the typical sample preparation is discussed. The sample preparation is focused here on epidermal peels of onion scales and celery epidermis cells which were sectioned for the AFM to visualize the inner surface (closest to the plasma membrane) of the outer epidermal wall. The nm-wide cellulose microfibrils orientation and multilayer structure were clearly observed. The microfibril orientation and alignment tend to be more organized in older scales compared with younger scales. The onion epidermis cell wall was also used as a test analog to study cell wall elasticity by the AFM nanoindentation and the SAM V(z) feature. The novelty in this work was to demonstrate the capability of these two techniques to analyze isolated, single layered plant cell walls in their natural state. AFM nanoindentation was also used to probe the effects of Ethylenediaminetetraacetic acid (EDTA), and calcium ion treatment to modify pectin networks in cell walls. The results suggest a significant modulus increase in the calcium ion treatment and a slight decrease in EDTA treatment. To complement the AFM measurements, the HF-SAM was used to obtain the V(z) signatures of the onion epidermis. These measurements were focused on documenting the effect of pectinase enzyme treatment. The results indicate a significant change in the V(z) signature curves with time into the enzyme treatment. Thus AFM and HF-SAM open the door to a systematic nondestructive structure and mechanical property study of complex biological cell walls. A unique feature of this approach is that both microscopes allow the biological samples to be examined in their natural fluid (water) environment.

Tittmann, B. R. [Penn State; Xi, X. [Penn State



Quantitative sub-surface and non-contact imaging using scanning microwave microscopy.  


The capability of scanning microwave microscopy for calibrated sub-surface and non-contact capacitance imaging of silicon (Si) samples is quantitatively studied at broadband frequencies ranging from 1 to 20 GHz. Calibrated capacitance images of flat Si test samples with varying dopant density (10(15)-10(19) atoms cm(-3)) and covered with dielectric thin films of SiO2 (100-400 nm thickness) are measured to demonstrate the sensitivity of scanning microwave microscopy (SMM) for sub-surface imaging. Using standard SMM imaging conditions the dopant areas could still be sensed under a 400 nm thick oxide layer. Non-contact SMM imaging in lift-mode and constant height mode is quantitatively demonstrated on a 50 nm thick SiO2 test pad. The differences between non-contact and contact mode capacitances are studied with respect to the main parameters influencing the imaging contrast, namely the probe tip diameter and the tip-sample distance. Finite element modelling was used to further analyse the influence of the tip radius and the tip-sample distance on the SMM sensitivity. The understanding of how the two key parameters determine the SMM sensitivity and quantitative capacitances represents an important step towards its routine application for non-contact and sub-surface imaging. PMID:25751635

Gramse, Georg; Brinciotti, Enrico; Lucibello, Andrea; Patil, Samadhan B; Kasper, Manuel; Rankl, Christian; Giridharagopal, Rajiv; Hinterdorfer, Peter; Marcelli, Romolo; Kienberger, Ferry



Intracellular subsurface imaging using a hybrid shear-force feedback/scanning quantitative phase microscopy technique  

NASA Astrophysics Data System (ADS)

Quantitative phase microscopy (QPM) allows for the imaging of translucent or transparent biological specimens without the need for exogenous contrast agents. This technique is usually applied towards the investigation of simple cells such as red blood cells which are typically enucleated and can be considered to be homogenous. However, most biological cells are nucleated and contain other interesting intracellular organelles. It has been established that the physical characteristics of certain subsurface structures such as the shape and roughness of the nucleus is well correlated with onset and progress of pathological conditions such as cancer. Although the acquired quantitative phase information of biological cells contains surface information as well as coupled subsurface information, the latter has been ignored up until now. A novel scanning quantitative phase imaging system unencumbered by 2pi ambiguities is hereby presented. This system is incorporated into a shear-force feedback scheme which allows for simultaneous phase and topography determination. It will be shown how subsequent image processing of these two data sets allows for the extraction of the subsurface component in the phase data and in vivo cell refractometry studies. Both fabricated samples and biological cells ranging from rat fibroblast cells to malaria infected human erythrocytes were investigated as part of this research. The results correlate quite well with that obtained via other microscopy techniques.

Edward, Kert


Interferometric time-stretch microscopy for ultrafast quantitative cellular and tissue imaging at 1 ?m  

NASA Astrophysics Data System (ADS)

Quantitative phase imaging (QPI) has been proven to be a powerful tool for label-free characterization of biological specimens. However, the imaging speed, largely limited by the image sensor technology, impedes its utility in applications where high-throughput screening and efficient big-data analysis are mandated. We here demonstrate interferometric time-stretch (iTS) microscopy for delivering ultrafast quantitative phase cellular and tissue imaging at an imaging line-scan rate >20 MHz-orders-of-magnitude faster than conventional QPI. Enabling an efficient time-stretch operation in the 1-?m wavelength window, we present an iTS microscope system for practical ultrafast QPI of fixed cells and tissue sections, as well as ultrafast flowing cells (at a flow speed of up to 8 m/s). To the best of our knowledge, this is the first time that time-stretch imaging could reveal quantitative morphological information of cells and tissues with nanometer precision. As many parameters can be further extracted from the phase and can serve as the intrinsic biomarkers for disease diagnosis, iTS microscopy could find its niche in high-throughput and high-content cellular assays (e.g., imaging flow cytometry) as well as tissue refractometric imaging (e.g., whole-slide imaging for digital pathology).

Lau, Andy K. S.; Wong, Terence T. W.; Ho, Kenneth K. Y.; Tang, Matthew T. H.; Chan, Antony C. S.; Wei, Xiaoming; Lam, Edmund Y.; Shum, Ho Cheung; Wong, Kenneth K. Y.; Tsia, Kevin K.



Quantitative sub-surface and non-contact imaging using scanning microwave microscopy  

NASA Astrophysics Data System (ADS)

The capability of scanning microwave microscopy for calibrated sub-surface and non-contact capacitance imaging of silicon (Si) samples is quantitatively studied at broadband frequencies ranging from 1 to 20 GHz. Calibrated capacitance images of flat Si test samples with varying dopant density (1015–1019 atoms cm?3) and covered with dielectric thin films of SiO2 (100–400 nm thickness) are measured to demonstrate the sensitivity of scanning microwave microscopy (SMM) for sub-surface imaging. Using standard SMM imaging conditions the dopant areas could still be sensed under a 400 nm thick oxide layer. Non-contact SMM imaging in lift-mode and constant height mode is quantitatively demonstrated on a 50 nm thick SiO2 test pad. The differences between non-contact and contact mode capacitances are studied with respect to the main parameters influencing the imaging contrast, namely the probe tip diameter and the tip–sample distance. Finite element modelling was used to further analyse the influence of the tip radius and the tip–sample distance on the SMM sensitivity. The understanding of how the two key parameters determine the SMM sensitivity and quantitative capacitances represents an important step towards its routine application for non-contact and sub-surface imaging.

Gramse, Georg; Brinciotti, Enrico; Lucibello, Andrea; Patil, Samadhan B.; Kasper, Manuel; Rankl, Christian; Giridharagopal, Rajiv; Hinterdorfer, Peter; Marcelli, Romolo; Kienberger, Ferry



Correlative light and electron microscopy imaging of autophagy in a zebrafish infection model.  


High-resolution imaging of autophagy has been used intensively in cell culture studies, but so far it has been difficult to visualize this process in detail in whole animal models. In this study we present a versatile method for high-resolution imaging of microbial infection in zebrafish larvae by injecting pathogens into the tail fin. This allows visualization of autophagic compartments by light and electron microscopy, which makes it possible to correlate images acquired by the 2 techniques. Using this method we have studied the autophagy response against Mycobacterium marinum infection. We show that mycobacteria during the progress of infection are frequently associated with GFP-Lc3-positive vesicles, and that 2 types of GFP-Lc3-positive vesicles were observed. The majority of these vesicles were approximately 1 ?m in size and in close vicinity of bacteria, and a smaller number of GFP-Lc3-positive vesicles was larger in size and were observed to contain bacteria. Quantitative data showed that these larger vesicles occurred significantly more in leukocytes than in other cell types, and that approximately 70% of these vesicles were positive for a lysosomal marker. Using electron microscopy, it was found that approximately 5% of intracellular bacteria were present in autophagic vacuoles and that the remaining intracellular bacteria were present in phagosomes, lysosomes, free inside the cytoplasm or occurred as large aggregates. Based on correlation of light and electron microscopy images, it was shown that GFP-Lc3-positive vesicles displayed autophagic morphology. This study provides a new approach for injection of pathogens into the tail fin, which allows combined light and electron microscopy imaging in vivo and opens new research directions for studying autophagy process related to infectious diseases. PMID:25126731

Hosseini, Rohola; Lamers, Gerda Em; Hodzic, Zlatan; Meijer, Annemarie H; Schaaf, Marcel Jm; Spaink, Herman P



Visualizing aquatic bacteria by light and transmission electron microscopy.  


The understanding of the functional role of aquatic bacteria in microbial food webs is largely dependent on methods applied to the direct visualization and enumeration of these organisms. While the ultrastructure of aquatic bacteria is still poorly known, routine observation of aquatic bacteria by light microscopy requires staining with fluorochromes, followed by filtration and direct counting on filter surfaces. Here, we used a new strategy to visualize and enumerate aquatic bacteria by light microscopy. By spinning water samples from varied tropical ecosystems in a cytocentrifuge, we found that bacteria firmly adhere to regular slides, can be stained by fluorochoromes with no background formation and fast enumerated. Significant correlations were found between the cytocentrifugation and filter-based methods. Moreover, preparations through cytocentrifugation were more adequate for bacterial viability evaluation than filter-based preparations. Transmission electron microscopic analyses revealed a morphological diversity of bacteria with different internal and external structures, such as large variation in the cell envelope and capsule thickness, and presence or not of thylakoid membranes. Our results demonstrate that aquatic bacteria represent an ultrastructurally diverse population and open avenues for easy handling/quantification and better visualization of bacteria by light microscopy without the need of filter membranes. PMID:24132727

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



Scanning moiré fringe imaging by scanning transmission electron microscopy.  


A type of artificial contrast found in annular dark-field imaging is generated by spatial interference between the scanning grating of the electron beam and the specimen atomic lattice. The contrast is analogous to moiré fringes observed in conventional transmission electron microscopy. We propose using this scanning interference for retrieving information about the atomic lattice structure at medium magnifications. Compared with the STEM atomic imaging at high magnifications, this approach might have several advantages including easy observation of lattice discontinuities and reduction of image degradation from carbon contamination and beam damage. Application of the technique to reveal the Burgers vector of misfit dislocations at the interface of epitaxial films is demonstrated and its potential for studying strain fields is discussed. PMID:20006440

Su, Dong; Zhu, Yimei



Detection and identification of light impurities by electron microscopy  

SciTech Connect

For over 40 years impurities have been believed to stabilize the ceramic {alpha}-Si{sub 3}N{sub 4} but there is no direct evidence for their identity or lattice location. In bulk materials electron microscopy can generally image heavy impurities. Here we report direct imaging of N columns in {alpha}-Si{sub 3}N{sub 4} that suggests the presence of excess light elements in specific N columns. First-principles calculations rule out Si or N interstitials and suggest O impurities, which are then confirmed by atomically resolved electron-energy-loss spectroscopy. The result provides a possible explanation for the stability of {alpha}-Si{sub 3}N{sub 4} with implications for the design of next-generation structural ceramics.

Idrobo Tapia, Juan C [ORNL; Oxley, Mark P [ORNL; Walkosz, Weronika [University of Illinois, Chicago; Klie, Robert F [University of Illinois, Chicago; Ogut, Serdar [University of Illinois, Chicago; Mikijelj, B [Ceradyne Inc., Costa Mesa, CA; Pennycook, Stephen J [ORNL; Pantelides, Sokrates T. [Vanderbilt University



Combined Scanning Transmission Electron Microscopy Tilt- and Focal Series  

SciTech Connect

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.

Dahmen, Tim [German Research Center for Artificial Intelligence (DFKI), Germany] [German Research Center for Artificial Intelligence (DFKI), Germany; Baudoin, Jean-Pierre G [ORNL] [ORNL; Lupini, Andrew R [ORNL] [ORNL; Kubel, Christian [Karlsruhe Institute of Technology, Leopoldshafen, Germany] [Karlsruhe Institute of Technology, Leopoldshafen, Germany; Slusallek, Phillip [German Research Center for Artificial Intelligence (DFKI), Germany] [German Research Center for Artificial Intelligence (DFKI), Germany; De Jonge, Niels [ORNL] [ORNL



Quantitative tracking of tumor cells in phase-contrast microscopy exploiting halo artifact pattern  

NASA Astrophysics Data System (ADS)

Tumor cell morphology is closely related to its invasiveness characteristics and migratory behaviors. An invasive tumor cell has a highly irregular shape, whereas a spherical cell is non-metastatic. Thus, quantitative analysis of cell features is crucial to determine tumor malignancy or to test the efficacy of anticancer treatment. We use phase-contrast microscopy to analyze single cell morphology and to monitor its change because it enables observation of long-term activity of living cells without photobleaching and phototoxicity, which is common in other fluorescence-labeled microscopy. Despite this advantage, there are image-level drawbacks to phase-contrast microscopy, such as local light effect and contrast interference ring, among others. Thus, we first applied a local filter to compensate for non-uniform illumination. Then, we used intensity distribution information to detect the cell boundary. In phase-contrast microscopy images, the cell normally appears as a dark region surrounded by a bright halo. As the halo artifact around the cell body is minimal and has an asymmetric diffusion pattern, we calculated the cross-sectional plane that intersected the center of each cell and was orthogonal to the first principal axis. Then, we extracted the dark cell region by level set. However, a dense population of cultured cells still rendered single-cell analysis difficult. Finally, we measured roundness and size to classify tumor cells into malignant and benign groups. We validated segmentation accuracy by comparing our findings with manually obtained results.

Kang, Mi-Sun; Song, Soo-Min; Lee, Hana; Kim, Myoung-Hee



Quantitative 3D molecular cutaneous absorption in human skin using label free nonlinear microscopy.  


Understanding the penetration mechanisms of drugs into human skin is a key issue in pharmaceutical and cosmetics research. To date, the techniques available for percutaneous penetration of compounds fail to provide a quantitative 3D map of molecular concentration distribution in complex tissues as the detected microscopy images are an intricate combination of concentration distribution and laser beam attenuation upon deep penetration. Here we introduce and validate a novel framework for imaging and reconstructing molecular concentration within the depth of artificial and human skin samples. Our approach combines the use of deuterated molecular compounds together with coherent anti-Stokes Raman scattering spectroscopy and microscopy that permits targeted molecules to be unambiguously discriminated within skin layers. We demonstrate both intercellular and transcellular pathways for different active compounds, together with in-depth concentration profiles reflecting the detailed skin barrier architecture. This method provides an enabling platform for establishing functional activity of topically applied products. PMID:25550155

Chen, Xueqin; Grégoire, Sébastien; Formanek, Florian; Galey, Jean-Baptiste; Rigneault, Hervé



Transmission electron microscopy of liquid phase densified SiC  

SciTech Connect

Transmission electron microscopy was used to characterize microstructures of SiC densified using a transient liquid phase (resulting from the reaction of Al{sub 2}O{sub 3} with Al{sub 4}C{sub 3}) by hot pressing at 1875 {degree}C for 10 min in N{sub 2}. High resolution electron microscopy showed that the SiC grain boundaries were free of glassy phases, suggesting that all liquid phases crystallized upon cooling. Phases that might be expected due to reactive sintering (i.e., AlN, Al{sub 2}OC, Al{sub 2}O{sub 3}, Al{sub 4}O{sub 4}C, Al{sub 3}O{sub 3}N, or solid solutions of SiC, AlN, and Al{sub 2}OC) were not observed. However, significant Al, Si, O, and C concentrations were found at all triple junctions of these rapidly densified ceramics.

Carpenter, R.W.; Braue, W. (Center for Solid State Science, the Arizona State University, Tempe, Arizona (USA)); Cutler, R.A. (Ceramatec, Inc., 2425 South 900 West, Salt Lake City, Utah (USA))



An electron microscopy study of tritium decay in vanadium  

NASA Astrophysics Data System (ADS)

Pre-thinned electron microscopy specimens of vanadium have been tritium-charged to produce ?-phase tritide precipitates (VT 0.5) embedded in a dilute V-T ?-phase. After aging for only 2-3 months, both at room temperature and at -196°C, significant effects due to the decay of tritium to 3He were observed by transmission electron microscopy: (1) the formation of small 3He bubbles (diameters 1.0 - 1.5 nm) and high densities of interstitial dislocation loops in the ?-phase areas (2) the decoration of dislocations in the ?-phase with 3He filled tubes and/or bubbles (3) isolated 3He bubbles in the ?-phase (which were not attached to dislocations) which had relieved their high internal pressure by prismatic loop punching. The observed clustering indicates that helium is mobile in both phases at room temperature. Moreover, the formation of helium bubbles in the absence of irradiation-induced defects strongly suggests precipitation via a self-trapping mechanism. This process can result in a microstructure similar to that found after He implantation and may be an important factor in material degradation.

Schober, T.; Lässer, R.; Jäger, W.; Thomas, G. J.



Scanning electron microscopy of mycoplasmas adhering to erythrocytes.  

PubMed Central

The interaction of Mycoplasma pneumoniae and Mycoplasma gallisepticum with human erythrocytes (RBC) was studied by scanning electron microscopy. The tight nature of the attachment of the microorganisms to the RBC was indicated by the indentation of the RBC surface at the site of attachment of M. gallisepticum cells and by traction and resulting distortion in the shape of the RBC at the point of its attachment to M. pneumoniae filaments growing on glass or plastic. In many cases attachment took place via the tip of the filaments, the membrane of the parasite appearing to be fused with that of the RBC. The morphology of the mycoplasmas growing on cover slips conformed in general with previous descriptions obtained by scanning electron microscopy. Growth of M. pneumoniae on glass or plastic consisted of branching filaments spread on the inert surface and microcolonies made up of intertwining filaments projecting into the medium. The filaments had a bulbous swelling adjacent to a tapered tip end. A few filaments were shown to have a ropelike helical twist. M. gallisepticum grown on the cover slips of Leighton tubes had a peculiar fusiform or teardrop shape with blebs at one or both poles of the cells. Elongated filamentous forms and chains of coccobacillary bodies were observed as well. Images Fig. 1 Fig. 2 Fig. 3 Fig. 4 Fig. 5 Fig. 4 Fig. 5 Fig. 6 Fig. 7a Fig. 7b-7e Fig. 8 Fig. 9 PMID:6777306

Razin, S; Banai, M; Gamliel, H; Polliack, A; Bredt, W; Kahane, I



A national facility for biological cryo-electron microscopy  

PubMed Central

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

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



Use of electron microscopy to classify canine perivascular wall tumors.  


The histologic classification of canine perivascular wall tumors (PWTs) is controversial. Many PWTs are still classified as hemangiopericytomas (HEPs), and the distinction from peripheral nerve sheath tumors (PNSTs) is still under debate. A recent histologic classification of canine soft tissue sarcomas included most histologic types of PWT but omitted those that were termed undifferentiated. Twelve cases of undifferentiated canine PWTs were evaluated by transmission electron microscopy. The ultrastructural findings supported a perivascular wall origin for all cases with 4 categories of differentiation: myopericytic (n = 4), myofibroblastic (n = 1), fibroblastic (n = 2), and undifferentiated (n = 5). A PNST was considered unlikely in each case based on immunohistochemical expression of desmin and/or the lack of typical ultrastructural features, such as basal lamina. Electron microscopy was pivotal for the subclassification of canine PWTs, and the results support the hypothesis that canine PWTs represent a continuum paralleling the phenotypic plasticity of vascular mural cells. The hypothesis that a subgroup of PWTs could arise from a pluripotent mesenchymal perivascular wall cell was also considered and may explain the diverse differentiation of canine PWTs. PMID:22865645

Palmieri, C; Avallone, G; Cimini, M; Roccabianca, P; Stefanello, D; Della Salda, L



Amyloid Structure and Assembly: Insights from Scanning Transmission Electron Microscopy  

SciTech Connect

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).

Goldsbury, C.; Wall, J.; Baxa, U.; Simon, M. N.; Steven, A. C.; Engel, A.; Aebi, U.; Muller, S. A.



Low-Cost Cryo-Light Microscopy Stage Fabrication for Correlated Light/Electron Microscopy  

PubMed Central

The coupling of cryo-light microscopy (cryo-LM) and cryo-electron microscopy (cryo-EM) poses a number of advantages for understanding cellular dynamics and ultrastructure. First, cells can be imaged in a near native environment for both techniques. Second, due to the vitrification process, samples are preserved by rapid physical immobilization rather than slow chemical fixation. Third, imaging the same sample with both cryo-LM and cryo-EM provides correlation of data from a single cell, rather than a comparison of "representative samples". While these benefits are well known from prior studies, the widespread use of correlative cryo-LM and cryo-EM remains limited due to the expense and complexity of buying or building a suitable cryogenic light microscopy stage. Here we demonstrate the assembly, and use of an inexpensive cryogenic stage that can be fabricated in any lab for less than $40 with parts found at local hardware and grocery stores. This cryo-LM stage is designed for use with reflected light microscopes that are fitted with long working distance air objectives. For correlative cryo-LM and cryo-EM studies, we adapt the use of carbon coated standard 3-mm cryo-EM grids as specimen supports. After adsorbing the sample to the grid, previously established protocols for vitrifying the sample and transferring/handling the grid are followed to permit multi-technique imaging. As a result, this setup allows any laboratory with a reflected light microscope to have access to direct correlative imaging of frozen hydrated samples. PMID:21673645

Carlson, David B.; Evans, James E.



Electron microscopy investigations of nanoparticles for cancer diagnostic applications  

NASA Astrophysics Data System (ADS)

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 preserved after a chemical etch. Magnetic measurements show a slight decrease in magnetic moment after ion milling. From TEM characterization, the introduction of oxygen into the copper release layer, prior the film deposition process, can effectively control the topography of the oxidized-copper grains and, consequently, lead to the production of SAF nanoparticles with flatter layers. Size distribution studies performed on SAFs fabricated using self-assembled stamps show that it is possible to produce monodisperse nanoparticles with diameters from 70 nm up. Part Two of the dissertation describes structural characterization experiments performed on Composite Organic-Inorganic Nanoparticles (COINs), which are a novel type of SERS nanoclusters formed by aggregating silver nanoparticles with Raman molecules, and then encapsulating them with an organic coating that stabilizes the aggregates and promotes subsequent functionalization with antibodies. Part Three of this dissertation focuses on the development and application of electron microscopy-based techniques to characterize the nanomaterial-biology interactions, to assess how, or indeed whether, nanoparticles are attaching to the cancer cells. The technique of negative staining was applied to simultaneously visualize inorganic nanoparticles and their biofunctionalized entities under the TEM and to verify the successful functionalization of nanoparticles with antibodies. The interpretation of the negatively-stained COINs was consistent with the EFTEM data. Next, the localization and characterization of CD54-functionalized COINs on the apicolateral portions of U937 leukemia cell lines was determined using TEM, SEM and Scanning Auger Microscopy. The analyses show that CD54 antigens are localized at a specific region on U937 leukemia cell surfaces. SEM imaging and SER spectroscopy correlation studies of different antibody-conjugated COINs attached onto different cancer cell lines show a direct correlation between the number of COINs binding to cells and the corresponding SER intensity. Finally, TEM was used to l

Koh, Ai Leen


An open-source deconvolution software package for 3-D quantitative fluorescence microscopy imaging  

PubMed Central

Summary Deconvolution techniques have been widely used for restoring the 3-D quantitative information of an unknown specimen observed using a wide-field fluorescence microscope. Deconv, an open-source deconvolution software package, was developed for 3-D quantitative fluorescence microscopy imaging and was released under the GNU Public License. Deconv provides numerical routines for simulation of a 3-D point spread function and deconvolution routines implemented three constrained iterative deconvolution algorithms: one based on a Poisson noise model and two others based on a Gaussian noise model. These algorithms are presented and evaluated using synthetic images and experimentally obtained microscope images, and the use of the library is explained. Deconv allows users to assess the utility of these deconvolution algorithms and to determine which are suited for a particular imaging application. The design of Deconv makes it easy for deconvolution capabilities to be incorporated into existing imaging applications. PMID:19941558




Accurate single-shot quantitative phase imaging of biological specimens with telecentric digital holographic microscopy.  


The advantages of using a telecentric imaging system in digital holographic microscopy (DHM) to study biological specimens are highlighted. To this end, the performances of nontelecentric DHM and telecentric DHM are evaluated from the quantitative phase imaging (QPI) point of view. The evaluated stability of the microscope allows single-shot QPI in DHM by using telecentric imaging systems. Quantitative phase maps of a section of the head of the drosophila melanogaster fly and of red blood cells are obtained via single-shot DHM with no numerical postprocessing. With these maps we show that the use of telecentric DHM provides larger field of view for a given magnification and permits more accurate QPI measurements with less number of computational operations. PMID:24781590

Doblas, Ana; Sánchez-Ortiga, Emilio; Martínez-Corral, Manuel; Saavedra, Genaro; Garcia-Sucerquia, Jorge



Gravity driven high throughput phase detecting cytometer based on quantitative interferometric microscopy  

NASA Astrophysics Data System (ADS)

Phase distribution detection of cells and tissues is concerned since it is an important auxiliary method for observing biological samples. High speed and large amount cell detection is needed for its high detecting efficiency. In this paper, we have proposed a simple large scale biological sample phase detection device called gravity driven high throughput phase detecting cytometer based on quantitative interferometric microscopy to obtain flowing red blood cells phase. The system could realize high throughput phase detecting and statistical analysis with high detecting speed and in real time. The statistical characteristics of red blood cells could be obtained which might be helpful for biological analysis and disease detection. We believe this method is a powerful tool to quantitatively measure the phase distribution of biological samples.

Xue, Liang; Wang, Shouyu; Yan, Keding; Sun, Nan; Ferraro, Pietro; Li, Zhenhua; Liu, Fei



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

PubMed Central

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. PMID:21750834

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



Electron microscopy and atomic force microscopy studies of chromatin and metaphase chromosome structure.  


The folding of the chromatin filament and, in particular, the organization of genomic DNA within metaphase chromosomes has attracted the interest of many laboratories during the last five decades. This review discusses our current understanding of chromatin higher-order structure based on results obtained with transmission electron microscopy (TEM), cryo-electron microscopy (cryo-EM), and different atomic force microscopy (AFM) techniques. Chromatin isolated from different cell types in buffers without cations form extended filaments with nucleosomes visible as separated units. In presence of low concentrations of Mg(2+), chromatin filaments are folded into fibers having a diameter of ? 30 nm. Highly compact fibers were obtained with isolated chromatin fragments in solutions containing 1-2mM Mg(2+). The high density of these fibers suggested that the successive turns of the chromatin filament are interdigitated. Similar results were obtained with reconstituted nucleosome arrays under the same ionic conditions. This led to the proposal of compact interdigitated solenoid models having a helical pitch of 4-5 nm. These findings, together with the observation of columns of stacked nucleosomes in different liquid crystal phases formed by aggregation of nucleosome core particles at high concentration, and different experimental evidences obtained using other approaches, indicate that face-to-face interactions between nucleosomes are very important for the formation of dense chromatin structures. Chromatin fibers were observed in metaphase chromosome preparations in deionized water and in buffers containing EDTA, but chromosomes in presence of the Mg(2+) concentrations found in metaphase (5-22 mM) are very compact, without visible fibers. Moreover, a recent cryo-electron microscopy analysis of vitreous sections of mitotic cells indicated that chromatin has a disordered organization, which does not support the existence of 30-nm fibers in condensed chromosomes. TEM images of partially denatured chromosomes obtained using different procedures that maintain the ionic conditions of metaphase showed that bulk chromatin in chromosomes is organized forming multilayered plate-like structures. The structure and mechanical properties of these plates were studied using cryo-EM, electron tomography, AFM imaging in aqueous media, and AFM-based nanotribology and force spectroscopy. The results obtained indicated that the chromatin filament forms a flexible two-dimensional network, in which DNA is the main component responsible for the mechanical strength observed in friction force measurements. The discovery of this unexpected structure based on a planar geometry has opened completely new possibilities for the understanding of chromatin folding in metaphase chromosomes. It was proposed that chromatids are formed by many stacked thin chromatin plates oriented perpendicular to the chromatid axis. Different experimental evidences indicated that nucleosomes in the plates are irregularly oriented, and that the successive layers are interdigitated (the apparent layer thickness is 5-6 nm), allowing face-to-face interactions between nucleosomes of adjacent layers. The high density of this structure is in agreement with the high concentration of DNA observed in metaphase chromosomes of different species, and the irregular orientation of nucleosomes within the plates make these results compatible with those obtained with mitotic cell cryo-sections. The multilaminar chromatin structure proposed for chromosomes allows an easy explanation of chromosome banding and of the band splitting observed in stretched chromosomes. PMID:21703860

Daban, Joan-Ramon



Trace metal and mineral speciation of remediated wastes using electron microscopy  

Microsoft Academic Search

Electron microscopic techniques, including scanning electron microscopy (SEM), transmission electron microscopy (TEM), and electron probe microanalyses (EPMA), were used to evaluate metal species and mineralogical phases associated with metal-bearing contaminated soil and industrial wastes that have been solidified and stabilized with Portland cement. Metals present in the wastes included arsenic, barium, cadmium, chromium, copper, lead, nickel, and zinc. In addition,

I. Klich; L. P. Wilding; L. R. Drees



Optimized Negative-Staining Electron Microscopy for Lipoprotein Studies  

PubMed Central

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

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




PubMed Central

A study has been made of the fine structure of hepatic parenchymal cells of human biopsy material in a case of pancreatic tumor with obstructive jaundice. Dense particles about 60 A in diameter have been found in the cytoplasm, which are considered to be ferritin molecules by electron microscopy. They are encountered throughout the cytoplasmic matrix and are often aggregated in electron-transparent areas, most of which are enclosed by an apparently single-layered membrane. Identification of the elemental iron has been pursued by the application of the x-ray scanning microanalyser which reveals a quantitative value within 1.0 per cent of the pure iron sample. The use of x-ray scanning microanalysis enables one to obtain accurate data from extremely small and precisely defined volumes of biological specimens. PMID:14002530

Yasuzumi, G.



Direct mapping of Li distribution in electrochemically lithiated graphite anodes using scanning Auger electron microscopy  

NASA Astrophysics Data System (ADS)

The spatial distribution of Li ions in electrochemically lithiated graphite anodes for Li-ion battery is characterized by scanning Auger electron microscopy. We show that direct mapping of Li KVV peak intensity reveal the spatial distribution of intercalated Li and its chemical state in a quantitative manner. Furthermore, we demonstrate that mapping using a C KVV peak also reflects the spatial distribution of Li due to the change in the electronic properties of C atoms induced by the electrode reaction (Li intercalation). Mapping measurements on three samples with different charging states (20%, 50%, and 100%) show that at the early stage of charging Li ions do not intercalate homogenously into all the graphite particles but selectively into some specific ones with higher rates. Our method provides the criteria to evaluate structure-correlated Li intercalation from nanometer- to micrometer-scale, such as conductivity network in the electrodes due to a non-uniform morphology of binder and conductive additives.

Ishida, Nobuyuki; Fukumitsu, Hitoshi; Kimura, Hiroshi; Fujita, Daisuke



Clinical applications of scanning electron microscopy and X-ray microanalysis in dermatology  

SciTech Connect

Scanning electron microscopy is frequently applied to dermatological problems, as is evident from a review of the recent literature. In this paper, preparation methods and new techniques allowing experimental studies on the integumentary system are emphasized. Quantitative analysis in the electron microscope by use of energy-dispersive X-ray microanalysis (EDX) has become an important accessory technique. EDX can, for instance, be used to study problems involving physiological changes induced in skin by agents causing contact reactions. Recently, it has been shown that treatment with DNCB, chromate and nickel causes changes in elemental distribution in guinea-pig skin. In addition, elemental uptake in the integumentary system and in pathological inclusions in skin can be analyzed.

Forslind, B.



Quantitative analysis of zinc in rat hippocampal mossy fibers by nuclear microscopy.  


Zinc (Zn) is involved in regulating mental and motor functions of the brain. Previous approaches have determined Zn content in the brain using semi-quantitative histological methods. We present here an alternative approach to map and quantify Zn levels in the synapses from mossy fibers to CA3 region of the hippocampus. Based on the use of nuclear microscopy, which is a combination of imaging and analysis techniques encompassing scanning transmission ion microscopy (STIM), Rutherford backscattering spectrometry (RBS), and particle induced X-ray emission (PIXE), it enables quantitative elemental mapping down to the parts per million (?g/g dry weight) levels of zinc in rat hippocampal mossy fibers. Our results indicate a laminar-specific Zn concentration of 240±9?M in wet weight level (135±5?g/g dry weight) in the stratum lucidum (SL) compared to 144±6?M in wet weight level (81±3?g/g dry weight) in the stratum pyramidale (SP) and 78±10?M in wet weight level (44±5?g/g dry weight) in the stratum oriens (SO) of the hippocampus. The mossy fibers terminals in CA3 are mainly located in the SL. Hence the Zn concentration is suggested to be within this axonal presynaptic terminal system. PMID:22766378

Zhang, Binbin; Ren, Minqin; Sheu, Fwu-Shan; Watt, Frank; Routtenberg, Aryeh



High-resolution microscopy of plasmon field distributions by scanning tunneling luminescence and photoemission electron microscopies  

NASA Astrophysics Data System (ADS)

The exploitation of plasmon resonances to promote the interaction between conjugated molecules and optical fields motivates intensive research. The objectives are to understand the mechanisms of plasmon-mediated interactions, and to realize molecularly- or atomically-precise metal nanostructures, combining field enhancements and optical antenna effects. In this review paper, we present examples of plasmonic-field mappings based on scanning tunneling microscope (STM)-induced light emission or multiphoton photoemission (PEEM), two techniques among those which offer today's best spatial resolutions for plasmon microscopy. An unfamiliar property of the junction of an STM is its ability to behave as a highly localized source of light. It can be exploited to probe optoelectronic properties, in particular plasmonic fields, with ultimate subnanometer spatial resolution, an advantage balanced by a sometimes delicate deconvolution of local-probe influence. Alternatively, local-probe disadvantages can be overcome by imaging the photoemitted electrons, using well-established electron optics. This allows obtaining two-dimensional intensity maps reflecting the unperturbed distribution of the optical near field. This approach provides full field spectroscopic images with a routine spatial resolution of the order of 20 nm (down to 5 nm with recent aberration corrected instruments).

Douillard, Ludovic; Charra, Fabrice



Utility of Transmission Electron Microscopy in Small Round Cell Tumors  

PubMed Central

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.

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



Nanoscale mechanical drumming visualized by 4D electron microscopy.  


With four-dimensional (4D) electron microscopy, we report in situ imaging of the mechanical drumming of a nanoscale material. The single crystal graphite film is found to exhibit global resonance motion that is fully reversible and follows the same evolution after each initiating stress pulse. At early times, the motion appears "chaotic" showing the different mechanical modes present over the micron scale. At longer time, the motion of the thin film collapses into a well-defined fundamental frequency of 1.08 MHz, a behavior reminiscent of mode locking; the mechanical motion damps out after approximately 200 micros and the oscillation has a "cavity" quality factor of 150. The resonance time is determined by the stiffness of the material, and for the 75 nm thick and 40 microm square specimen used here we determined Young's modulus to be 1.0 TPa for the in-plane stress-strain profile. Because of its real-time dimension, this 4D microscopy should have applications in the study of these and other types of materials structures. PMID:19006405

Kwon, Oh-Hoon; Barwick, Brett; Park, Hyun Soon; Baskin, J Spencer; Zewail, Ahmed H



Quantitative visualization of colloidal and intracellular gold nanoparticles by confocal microscopy.  


Gold nanoparticles (AuNPs) have the potential to become a versatile biomarker. For further use of AuNPs labeled with functionalized molecules, their visualization in biological systems by routine laboratory tools such as light microscopy is crucial. However, the size far below the diffraction limit affords specialized parameters for microscopical detection, which stimulated the current study, aimed to determine from which size onward AuNPs, either in dispersion or cell-associated, can be reliably detected by standard confocal microscopy. First, gold colloids of size-restricted fractions are examined in dispersion. At a minimum particle size of 60 nm, detection appears to be reliable. Particle counts in dilution series confirm these results by revealing single particle detection of 60-nm colloids. Second, AuNPs are visualized and quantified in cells, which interestingly cause a phase shift in the reflection of AuNPs. Gold mass spectroscopy confirms the number of AuNPs counted microscopically inside cells. Furthermore, it demonstrates for the first time a very high diffusion rate of 15-nm particles into the cells. In conclusion, the results back the suitability of confocal microscopy for the quantitative tracking of colloidal and intracellular gold nanoparticles sized 60 nm. PMID:20615017

Klein, Sabine; Petersen, Svea; Taylor, Ulrike; Rath, Detlef; Barcikowski, Stephan



Morphological classification of bioaerosols from composting using scanning electron microscopy.  


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 ?m) single, spherical cells, followed by larger (>1 ?m) single cells, with aggregates occurring in smaller proportions. Most aggregates consisted of clusters of 2-3 particles as opposed to chains, and were <10 ?m 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. PMID:24565805

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



Electron microscopy in kidney research: seeing is believing.  


Electron microscopy (EM) has been an indispensable tool for kidney research since its inception more than half a century ago. Much of the substantial advances were propelled by the need to find methods to best visualize and analyze the kidney's structure deduced from the fundamental principle that has structure and function intimately related. The result of 3 decades of experimental kidney work between 1950 and 1980 coincided with remarkable advances in nephrology that marked a renaissance era for renal pathology and resulted in the morphologic classification of medical kidney diseases. In the era of genetics and molecular medicine TEM continues to contribute significant clinical and pathogenetic insights in kidney disease. The basic principles as applied to kidney disease experimental models are discussed with emphasis on crescent formation in Col4A3-deficient mice and a mouse model of experimental oxalosis (CaOx). PMID:23876007

Liapis, Helen



Watershed Merge Tree Classification for Electron Microscopy Image Segmentation  

SciTech Connect

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.

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



Studies on human lacteal rotavirus antibodies by immune electron microscopy.  


In vitro studies carried out by immune electron microscopy (IEM) indicate that rotavirus aggregation detected in the stools of newborn breast-fed infants with rotavirus infection is antibody-induced. Aggregation of rotavirus particles occurred with the IgA-containing fraction of expressed breast milk (EBM) obtained five days postpartum and with the IgA- and IgG-containing fractions of a pool of EBMs containing samples collected 2-3 days postpartum. Bovine milk fractions also demonstrated this activity in the IgG- and IgA-containing fraction. Studies on unfractionated EBMs from a mother who experienced a rotavirus infection during the 43rd week of lactation showed that following rotavirus infection all three major classes of rotavirus-specific antibodies were present in breast milk, this being confirmed by enzyme immunoassay. PMID:6302221

Totterdell, B M; Banatvala, J E; Chrystie, I L



Collaborative computational project for electron cryo-microscopy.  


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. PMID:25615866

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



Pneumocystis carinii in vitro: A study by scanning electron microscopy.  

PubMed Central

Pneumocystis carinii is a parasitic microorganism which induces an often fatal pneumonitis in a variety of compromised patients (e.g., premature infants, those with congenital immune deficiency disorders, those therapeutically immune suppressed, etc.). Organisms derived from murine sources were cultivated in vitro on monolayers of primary embryonic chick epithelial lung cells. The infected cultures were then examined by scanning electron microscopy. An examination of the surface ultrastructure revealed pleomorphic organisms which were not only attached to host cells with anchoring fibers but also connected to other parasites via fine fibrils. Another type of fine fibril was observed which may play a role in the organism's nutrition. Images Figure 7 Figure 2 Figure 8 Figure 3 Figure 4 Figure 6 Figure 5 Figure 1 PMID:299987

Murphy, M. J.; Pifer, L. L.; Hughes, W. T.



Neuron Segmentation in Electron Microscopy Images Using Partial Differential Equations  

PubMed Central

In connectomics, neuroscientists seek to identify the synaptic connections between neurons. Segmentation of cell membranes using supervised learning algorithms on electron microscopy images of brain tissue is often done to assist in this effort. Here we present a partial differential equation with a novel growth term to improve the results of a supervised learning algorithm. We also introduce a new method for representing the resulting image that allows for a more dynamic thresholding to further improve the result. Using these two processes we are able to close small to medium sized gaps in the cell membrane detection and improve the Rand error by as much as 9% over the initial supervised segmentation. PMID:25143802

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



Near field and exit wave computations for electron microscopy.  


The partial wave phase shift formalism of atomic scattering is applied to compute exit wave functions for isolated Au and Si atoms under both plane wave and focused probe illumination. Connections between the far field and near field (exit) waves are clarified. This approach treats the Coulomb singularity properly though at 100 keV large numbers of phase shifts are required. In principle any form of incident wave can be handled so it may provide a means for testing traditional scattering theories used in electron microscopy. By applying the analysis to an atom embedded in a constant potential rather than free space, exit spheres of radius half the interatomic spacing can be used. PMID:23726769

Howie, A



High Resolution Scanning Electron Microscopy of Cells Using Dielectrophoresis  

PubMed Central

Ultrastructural analysis of cells can reveal valuable information about their morphological, physiological, and biochemical characteristics. Scanning electron microscopy (SEM) has been widely used to provide high-resolution images from the surface of biological samples. However, samples need to be dehydrated and coated with conductive materials for SEM imaging. Besides, immobilizing non-adherent cells during processing and analysis is challenging and requires complex fixation protocols. In this work, we developed a novel dielectrophoresis based microfluidic platform for interfacing non-adherent cells with high-resolution SEM at low vacuum mode. The system enables rapid immobilization and dehydration of samples without deposition of chemical residues over the cell surface. Moreover, it enables the on-chip chemical stimulation and fixation of immobilized cells with minimum dislodgement. These advantages were demonstrated for comparing the morphological changes of non-budding and budding yeast cells following Lyticase treatment. PMID:25089528

Tang, Shi-Yang; Zhang, Wei; Soffe, Rebecca; Nahavandi, Sofia; Shukla, Ravi; Khoshmanesh, Khashayar



Identification and quantitive analysis of calcium phosphate microparticles in intestinal tissue by nuclear microscopy  

NASA Astrophysics Data System (ADS)

Microscopic particles (0.5-2 ?m diameter), rich in calcium and phosphorus, are found in the lumen of the mid-distal gut of all mammals investigated, including humans, and these may play a role in immuno-surveillance and immune regulation of antigens from food and symbiotic bacteria that are contained in the gut. Whether these particles can cross in to tissue of the intestinal mucosa is unclear. If so, characterising their morphology and chemical composition is an important task in elucidating their function. The analysis of calcium phosphate in biological tissues has been approached in several ways including optical microscopy, scanning electron microscopy and, most recently in this work, with nuclear microscopy. In this paper, we describe the use of microPIXE and microRBS to locate these particles and to determine, accurately, the ratio of phosphorus to calcium using the information on sample thickness obtained from RBS to allow the PIXE ratios to be corrected. A commercial sample of hydroxy apatite was used to demonstrate accuracy and precision of the technique. Then, in a pilot study on intestinal tissue of mice, we demonstrated the presence of calcium phosphate microparticles, consistent with confocal microscopy observations, and we identified the average molar P:Ca molar ratio as 1.0. Further work will confirm the exact chemical speciation of these particles and will examine the influence of differing calcium containing diets on the formation of these microparticles.

Gomez-Morilla, Inmaculada; Thoree, Vinay; Powell, Jonathan J.; Kirkby, Karen J.; Grime, Geoffrey W.



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

PubMed Central

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

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



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


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. Microsc. Res. Tech. 78:220-229, 2015. © 2015 Wiley Periodicals, Inc. PMID:25604381

Prozorowska, Ewelina; Jackowiak, Hanna



Measuring porosity at the nanoscale by quantitative electron tomography.  


Quantitative electron tomography is proposed to characterize porous materials at a nanoscale. To achieve reliable three-dimensional (3D) quantitative information, the influence of missing wedge artifacts and segmentation methods is investigated. We are presenting the "Discrete Algebraic Reconstruction Algorithm" as the most adequate tomography method to measure porosity at the nanoscale. It provides accurate 3D quantitative information, regardless the presence of a missing wedge. As an example, we applied our approach to nanovoids in La2Zr2O7 thin films. PMID:21090668

Biermans, E; Molina, L; Batenburg, K J; Bals, S; Van Tendeloo, G



Electronic environment for a field emission gun in electron microscopy H. Pinna, K. Liang (*), M. Denizart and B. Jouffrey  

E-print Network

659 Electronic environment for a field emission gun in electron microscopy H. Pinna, K. Liang of the source given by a field emis- sion gun is particularly interesting in electron microscopy. This paper ofthe numerous disruptive breakdowns which can occur in that type of gun. The fluctuations

Paris-Sud XI, Université de


Versatile Silicon Photodiode Detector Technology for Scanning Electron Microscopy with High-Efficiency Sub-5 keV Electron Detection  

E-print Network

Versatile Silicon Photodiode Detector Technology for Scanning Electron Microscopy with High: +31 (0)15 26 22163, Email: Abstract A new silicon electron detector technology for Scanning Electron Microscopy, based on ultrashallow p+ n boron-layer photodiodes, features nm-thin anodes

Technische Universiteit Delft


Progress towards critical dimension low vacuum scanning electron microscopy  

NASA Astrophysics Data System (ADS)

Low vacuum scanning electron microscopy (LVSEM) is proposed and evaluated for next generation Critical Dimension (CD) metrology. Its ability to control charging artifacts and hydrocarbon contamination in order to obtain high signal-to-noise ratio, high resolution image data from insulating materials make the technology an excellent match for the increased use of high-k dielectrics and shrinking feature size in the semiconductor industry. The presence of a gas in the LVSEM chamber means that the probe characteristics and secondary electron amplification, detection, and signal-to-noise ratio differ significantly from conventional high vacuum tools. In order for low vacuum CD approaches to be viable, all of the processes must be understood and described to the degree of accuracy currently available on high vacuum systems. Consequently, the focus of this thesis is to determine an analytic form of the signal-to-noise ratio for two detector configurations: the simplified steady-state cascade system operating in the well defined Townsend's discharge region, and the high resolution, low vacuum immersion lens secondary electron detector, for which the physical amplification process has not been studied in the past. A physically realistic and ultimately predictive model, which could potentially be incorporated in CD simulation codes such as NIST's MONSEL, is developed. Its effectiveness is verified with experimental data acquired as a function of gas pressure for all important operating parameters, such as electron beam energy and current, detector bias, cascade distance, and gas type, and its capability for optimization of the imaging conditions is discussed. Noise characteristics are also analyzed using Monte Carlo gain histograms and pure statistical methods.

Tileli, Vasiliki


Absence of elastic clamping in quantitative piezoelectric force microscopy measurements of nanostructures  

NASA Astrophysics Data System (ADS)

We establish that clamping effects, which limit accurate determination of piezoelectric responses in bulk materials and films using piezoelectric force microscopy (PFM), are not present when measuring discrete nanostructures with radii less than five times the tip radius. This conclusion is established by comparing the piezoelectric response in ZnO rods using two electrode configurations: one with the conducting atomic force microscopy tip acting as the top electrode and the other using a uniform metal top electrode. The distributions of piezoelectric coefficients measured with these two types of electrode configurations are the same. Hence, clamping issues do not play a role in the piezoelectric property measurement of nanomaterials using PFM. The role of conduction electrons on the piezoelectric measurement in both cases is also discussed.

Scrymgeour, David A.; Hsu, Julia W. P.



Partial liquid ventilation with perfluorocarbon in acute lung injury: light and transmission electron microscopy studies.  


Liquid ventilation using perfluorocarbon has been shown to improve gas exchange in animal models of acute lung injury as well as in children with acute respiratory distress syndrome. This study was designed to define structural features of lung injury following partial liquid ventilation (PLV) using light and transmission electron microscopy in a rabbit model of acute respiratory distress. Animals were treated with either conventional mechanical ventilation (CMV-gas) (n = 6) or PLV (n = 5) for 4 h after the induction of acute lung injury with saline lavage. Control animals were killed after the lung injury. PLV significantly improved alveolar-arterial oxygen tension and the oxygen index compared with CMV (P < 0.05). Morphometric studies using light microscopy show less alveolar hemorrhage, less edema, and fewer hyaline membranes in the PLV group (P < 0.05). Polymorphonuclear leukocyte sequestration in lung capillaries (11.4 +/- 1.5 versus 19.2 +/- 3 x 10(8)/ml, P < 0.05, PLV versus CMV) and migration into airspaces (3.1 +/- 1.2 versus 4.5 +/- 1.1 x 10(8)/ml, P < 0.05, PLV versus CMV) were lower in the gravity-dependent lung regions. There were fewer alveolar macrophages in the PLV group compared with other groups (P < 0.05). Fluorescence microscopy analysis shows fewer type II alveolar epithelial cells in the CMV group and brighter type II cells in the PLV group. Transmission electron microscopy studies show more alveolar wall damage in the CMV group, with type II cells detached from their basement membrane with fewer surfactant-containing lamellar bodies. We conclude that quantitative histologic analysis shows less lung damage and inflammation when perfluorocarbon is combined with CMV in the management of acute respiratory distress syndrome. PMID:10745025

van Eeden, S F; Klut, M E; Leal, M A; Alexander, J; Zonis, Z; Skippen, P



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

Technology Transfer Automated Retrieval System (TEKTRAN)

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...


Thin dielectric film thickness determination by advanced transmission electron microscopy  

SciTech Connect

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.

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



Transmission Electron Microscopy Investigations of Domain Wall and Dislocation Interactions  

NASA Astrophysics Data System (ADS)

The effects of dislocations on domain wall motion in magnetite are important to a variety of paleomagnetic applications. Here we directly image these effects using a transmission electron microscope. Four samples were cut from a single, fully oriented crystal of magnetite. These magnetites were deformed in the dislocation glide regime using a one-atmosphere rig at varying temperatures and pressures to create dislocations. In one dislocation-rich sample, traditional and Lorentz-mode transmission electron microscopy was used to investigate the crystallographic nature of the dislocations and to observe their interactions with domain walls. Dislocations were primarily, though not exclusively, found in the {111} planes with Burgers vectors in the expected <110> directions. Domain wall pinning at dislocations was recorded as the sample was tilted within the microscope in the presence of a 16.8 mT axial field. This allowed us to experimentally determine the microcoercivity, which was 0.5 mT, on average. FORC diagram measurements from all samples showed typical multidomain coercivity distributions, with a central higher coercivity ridge extending out to between 5 and 20 mT. The microcoercivity measurements are in good agreement with theoretical models for domain wall pinning at a dislocation, and the bulk coercivity measurements are in good agreement with models for domain wall pinning at larger dislocation populations.

Lindquist, A. K.; Feinberg, J. M.; Harrison, R. J.; Newell, A. J.



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

NASA Technical Reports Server (NTRS)

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).

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



Scanning tunneling and scanning transmission electron microscopy of biological membranes  

NASA Astrophysics Data System (ADS)

The feasibility of imaging porin membrane, which is a reconstituted biological membrane consisting of phospholipid and protein, was studied by scanning tunneling microscopy (STM). Due to detailed knowledge of its composition from biochemical and its three-dimensional (3D) structure from electron microscopical analysis, porin vesicles seem to be a suitable model specimen for exploring the application of STM in biology. Unstained vesicles adsorbed onto a thin amorphous carbon film supported by a finder grid were localized using a scanning transmission electron microscope (STEM) at low irradiation doses ( < 100 {e -}/{nm 2}). Suitable areas of the sample were then positioned in the STM by a light optical telescope. STM images taken under ambient pressure from empty amorphous carbon films exhibited corrugations in the range of ? 1 nm, whereas steps having a height of 5 nm were reproducibly observed on grids with porin vesicles. Since this value is in good agreement with that obtained from air-dried metal shadowed vesicles, we interpret these steps as the edges of porin membranes.

Stemmer, A.; Reichelt, R.; Engel, A.; Rosenbusch, J. P.; Ringger, M.; Hidber, H. R.; Güntherodt, H. J.



Electron microscopy analysis of mineral fibers in human lung tissue  

SciTech Connect

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.

Friedrichs, K.H.; Brockmann, M.; Fischer, M.; Wick, G. (Medical Institute of Environmental Hygiene, Heinrich Heine University, Duesseldorf (Germany))



Using advanced electron microscopy for the characterization of catalytic materials  

NASA Astrophysics Data System (ADS)

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-corrected electron microscopy was used to systematically examine, atomic column by atomic column, the effect of elemental substitution on the long-range crystalline order, atomic coordinates, and site occupancies of the various formulations such that trends could be developed linking these properties to catalytic yields. To accomplish this task, an algorithm was developed that enabled the direct extraction of atomic coordinates and site occupancies from high-angle annular dark-field (HAADF) images to within 1% and 15% uncertainty, respectively. Furthermore, this general method could be applied to various crystalline systems and may dramatically improve the quality of initial structural models used in Rietveld refinements. Improvement in the quality of starting models may increase the structural and chemical complexity of inorganic structures that can be solved by using "powder methods" alone. In addition to the development of these trends, HAADF analyses also revealed the presence of coherent compositional miscibility gaps, rotational twin domains, and structural intergrowths in the complex Mo-V-M-O oxide system. Other catalytic systems that are addressed in this dissertation include Pd, Ag, and bimetallic Pd-Ag catalysts for the selective hydrogenation of acetylene in excess ethylene, alkali and alkaline earth promoted Ru catalysts for the production of clean hydrogen through the decomposition of ammonia, the production of Pt nanoparticles using dendrimer templates, and Pt-Re bimetallic catalysts for the conversion of glycerol to hydrocarbons and syn gas. In each of these studies, electron microscopy was used as a complimentary tool to synthetic and reaction studies to better understand interactions between the nanoparticles and the support/template, to determine the effect of adding various promoters, or to understand the nanoscale structural and chemical changes associated with the formation of bimetallic nanoparticles. A final area addressed in this dissertation is the interaction between the electron beam and the specimen. In one particular study direct

Pyrz, William D.


Customized patterned substrates for highly versatile correlative light-scanning electron microscopy  

PubMed Central

Correlative light electron microscopy (CLEM) combines the advantages of light and electron microscopy, thus making it possible to follow dynamic events in living cells at nanometre resolution. Various CLEM approaches and devices have been developed, each of which has its own advantages and technical challenges. We here describe our customized patterned glass substrates, which improve the feasibility of correlative fluorescence/confocal and scanning electron microscopy. PMID:25391455

Benedetti, Lorena; Sogne, Elisa; Rodighiero, Simona; Marchesi, Davide; Milani, Paolo; Francolini, Maura



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

PubMed Central

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

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



Combined use of confocal laser scanning microscopy and transmission electron microscopy for visualisation of identical cells processed by cryotechniques  

Microsoft Academic Search

Summary.   Successive visualisation of identical plant cells by light and electron microscopy is reported. For this purpose segments\\u000a of pea and barley leaves were prepared by high-pressure freezing, freeze-substitution, and low-temperature embedding. The\\u000a use of Safranin O during low-temperature dehydration allowed, on one hand, staining of all cellular components as investigated\\u000a by confocal laser scanning microscopy and, on the other

S. Pfeiffer; M. Beese; M. Boettcher; K. Kawaschinski; K. Krupinska



Aerosols over the Amazon Rainforest Investigated by Scanning Transmission X-ray Microscopy and Scanning Electron Microscopy  

Microsoft Academic Search

We collected atmospheric aerosols over the central Amazon Basin near Manaus, Brazil, during the wet season in February 2008, as part of the AMAZE-08 (Amazonian Aerosol Characterization Experiment) campaign. Aerosol samples were analyzed by Scanning Transmission X-ray Microscopy (STXM) coupled with Near-Edge X-ray Absorption Fine Structure Spectroscopy (NEXAFS) and Scanning Electron Microscopy (SEM) coupled with Energy-Dispersive X-ray Analysis (EDX). A

M. O. Andreae; P. P. Artaxo; M. K. Gilles; S. T. Martin; E. Mikhailov; R. Moffet; U. Poeschl; B. Sinha; H. Yang



Towards quantitative electrochemical measurements on the nanoscale by scanning probe microscopy: environmental and current spreading effects  

SciTech Connect

The application of electric bias across tip-surface junctions in scanning probe microscopy can readily induce surface and bulk electrochemical processes that can be further detected though changes in surface topography, Faradaic or conductive currents, or electromechanical strain responses. However, the basic factors controlling tip-induced electrochemical processes, including the relationship between applied tip bias and the thermodynamics of local processes remains largely unexplored. Using the model Li-ion reduction reaction on the surface in Li-ion conducting glass ceramic, we explore the factors controlling Li-metal formation and find surprisingly strong effects of atmosphere and back electrode composition on the process. These studies suggest the feasibility of SPM-based quantitative electrochemical studies under proper environmental controls, extending the concepts of ultramicroelectrodes to the single-digit nanometer scale.

Arruda, Thomas M [ORNL] [ORNL; Kumar, Amit [Oak Ridge National Laboratory (ORNL)] [Oak Ridge National Laboratory (ORNL); Veith, Gabriel M [ORNL] [ORNL; Jesse, Stephen [ORNL] [ORNL; Tselev, Alexander [ORNL] [ORNL; Baddorf, Arthur P [ORNL] [ORNL; Balke, Nina [ORNL] [ORNL; Kalinin, Sergei V [ORNL] [ORNL



Quantitative interferometric microscopy with improved full-field phase aberration compensation  

NASA Astrophysics Data System (ADS)

Single-shot quantitative interferometric microscopy (QIM) needs a high-accuracy and rapid phase retrieval algorithm. Retrieved phase distributions are often influenced by phase aberration background caused by both imaging system and phase retrieval algorithms. Here, we propose an improved phase aberration compensation (PAC) approach in order to eliminate the phase aberrations inherent in the data. With this method, sample-free parts are identified and used to calculate the background phase, reducing phase errors induced in samples and providing high-quality phase images. We now demonstrate that QIM based on this PAC approach realizes high-quality phase imaging from a single interferogram. This is of great potential for a real-time speedy diagnosis.

Xue, Liang; Wang, Shouyu; Yan, Keding; Sun, Nan; Li, Zhenhua; Liu, Fei



Nanoscale-resolved elasticity: contact mechanics for quantitative contact resonance atomic force microscopy  

NASA Astrophysics Data System (ADS)

Contact resonance atomic force microscopy (CR-AFM) constitutes a powerful approach for nanometer-resolved mechanical characterization of surfaces. Yet, absolute accuracy is frequently impaired by ad hoc assumptions on the dynamic AFM cantilever characteristics as well as contact model. Within the present study, we clarify the detailed interplay of stress fields and geometries for full quantitative understanding, employing combined experimental numerical studies for real AFM probes. Concerning contact description, a two-parameter ansatz is utilized that takes tip geometries and their corresponding indentation moduli into account. Parameter sets obtained upon experimental data fitting for different tip blunting states, are discussed in terms of model-specific artificiality versus real contact physics at the nanoscale. Unveiling the underlying physics in detail, these findings pave the way for accurate characterization of nanomechanical properties with highest resolution.

Jakob, A. M.; Buchwald, J.; Rauschenbach, B.; Mayr, S. G.



Quantitative magnetic force microscopy on permalloy dots using an iron filled carbon nanotube probe.  


An iron filled carbon nanotube (FeCNT), a 10-40 nm ferromagnetic nanowire enclosed in a protective carbon tube, is an attractive candidate for a magnetic force microscopy (MFM) probe as it provides a mechanically and chemically robust, nanoscale probe. We demonstrate the probe's capabilities with images of the magnetic field gradients close to the surface of a Py dot in both the multi-domain and vortex states. We show the FeCNT probe is accurately described by a single magnetic monopole located at its tip. Its effective magnetic charge is determined by the diameter of the iron wire and its saturation magnetization 4?M(s) ? 2.2 × 10(4)G. A magnetic monopole probe is advantageous as it enables quantitative measurements of the magnetic field gradient close to the sample surface. The lateral resolution is defined by the diameter of the iron wire and the probe-sample separation. PMID:21864777

Wolny, F; Obukhov, Y; Mühl, T; Weissker, U; Philippi, S; Leonhardt, A; Banerjee, P; Reed, A; Xiang, G; Adur, R; Lee, I; Hauser, A J; Yang, F Y; Pelekhov, D V; Büchner, B; Hammel, P C



Quantitative Characterization of Biological Liquids for Third-Harmonic Generation Microscopy  

PubMed Central

Third-harmonic generation (THG) microscopy provides images of unstained biological samples based on spatial variations in third-order nonlinear susceptibility, refractive index, and dispersion. In this study, we establish quantitative values for the third-order nonlinear susceptibilities of several solvents (water, ethanol, glycerol), physiological aqueous (ions, amino acids, polypeptides, bovine serum albumin, glucose) and lipid (triglycerides, cholesterol) solutions as a function of solute concentration in the 1.05–1.25 ?m excitation range. We use these data in conjunction with imaging experiments to show that THG imaging with ?1.2 ?m excitation lacks specificity and sensitivity to detect physiological ion concentration changes, and that nonaqueous structures such as lipid bodies provide a more robust source of signal. Finally, we illustrate the impact of index-matching liquids in THG images. These data provide a basis for interpreting biological THG images and for developing additional applications. PMID:17085492

Débarre, Delphine; Beaurepaire, Emmanuel



Quantitative microscopy and nanoscopy of sickle red blood cells performed by wide field digital interferometry  

NASA Astrophysics Data System (ADS)

We have applied wide-field digital interferometry (WFDI) to examine the morphology and dynamics of live red blood cells (RBCs) from individuals who suffer from sickle cell anemia (SCA), a genetic disorder that affects the structure and mechanical properties of RBCs. WFDI is a noncontact, label-free optical microscopy approach that can yield quantitative thickness profiles of RBCs and measurements of their membrane fluctuations at the nanometer scale reflecting their stiffness. We find that RBCs from individuals with SCA are significantly stiffer than those from a healthy control. Moreover, we show that the technique is sensitive enough to distinguish classes of RBCs in SCA, including sickle RBCs with apparently normal morphology, compared to the stiffer crescent-shaped sickle RBCs. We expect that this approach will be useful for diagnosis of SCA and for determining efficacy of therapeutic agents.

Shaked, Natan T.; Satterwhite, Lisa L.; Telen, Marilyn J.; Truskey, George A.; Wax, Adam



Quantitative imaging of microvasculature in deep tissue with a spectrum-based photo-acoustic microscopy.  


We analyze photo-acoustic signals from capillaries and theoretically demonstrate the quantitative relationship between vascular diameter and spectral slope in a low-frequency band. Phantom experiments validate the theoretical analysis. Based on this finding, spectral slope is proposed as the imaging parameter of a photo-acoustic microscopy. This system effectively quantifies the microvasculature with diameters of 60 and 150 ?m, which are smaller than the wavelength 342 ?m at the central frequency 4.39 MHz of ultrasound transducer. The low frequency also guarantees the imaging depth in the order of centimeters. The proposed scheme could be potential for noninvasive diagnosis of diseases related to abnormal vasoconstriction or angiectasis. PMID:25768159

Gao, Xiaoxiang; Tao, Chao; Wang, Xueding; Liu, Xiaojun



Nanoscale-resolved elasticity: contact mechanics for quantitative contact resonance atomic force microscopy.  


Contact resonance atomic force microscopy (CR-AFM) constitutes a powerful approach for nanometer-resolved mechanical characterization of surfaces. Yet, absolute accuracy is frequently impaired by ad hoc assumptions on the dynamic AFM cantilever characteristics as well as contact model. Within the present study, we clarify the detailed interplay of stress fields and geometries for full quantitative understanding, employing combined experimental numerical studies for real AFM probes. Concerning contact description, a two-parameter ansatz is utilized that takes tip geometries and their corresponding indentation moduli into account. Parameter sets obtained upon experimental data fitting for different tip blunting states, are discussed in terms of model-specific artificiality versus real contact physics at the nanoscale. Unveiling the underlying physics in detail, these findings pave the way for accurate characterization of nanomechanical properties with highest resolution. PMID:24838534

Jakob, A M; Buchwald, J; Rauschenbach, B; Mayr, S G



Quantitative optical microscopy: measurement of cellular biophysical features with a standard optical microscope.  


We describe the use of a standard optical microscope to perform quantitative measurements of mass, volume, and density on cellular specimens through a combination of bright field and differential interference contrast imagery. Two primary approaches are presented: noninterferometric quantitative phase microscopy (NIQPM), to perform measurements of total cell mass and subcellular density distribution, and Hilbert transform differential interference contrast microscopy (HTDIC) to determine volume. NIQPM is based on a simplified model of wave propagation, termed the paraxial approximation, with three underlying assumptions: low numerical aperture (NA) illumination, weak scattering, and weak absorption of light by the specimen. Fortunately, unstained cellular specimens satisfy these assumptions and low NA illumination is easily achieved on commercial microscopes. HTDIC is used to obtain volumetric information from through-focus DIC imagery under high NA illumination conditions. High NA illumination enables enhanced sectioning of the specimen along the optical axis. Hilbert transform processing on the DIC image stacks greatly enhances edge detection algorithms for localization of the specimen borders in three dimensions by separating the gray values of the specimen intensity from those of the background. The primary advantages of NIQPM and HTDIC lay in their technological accessibility using "off-the-shelf" microscopes. There are two basic limitations of these methods: slow z-stack acquisition time on commercial scopes currently abrogates the investigation of phenomena faster than 1 frame/minute, and secondly, diffraction effects restrict the utility of NIQPM and HTDIC to objects from 0.2 up to 10 (NIQPM) and 20 (HTDIC) ?m in diameter, respectively. Hence, the specimen and its associated time dynamics of interest must meet certain size and temporal constraints to enable the use of these methods. Excitingly, most fixed cellular specimens are readily investigated with these methods. PMID:24747818

Phillips, Kevin G; Baker-Groberg, Sandra M; McCarty, Owen J T



Probing the Plasma Membrane Structure of Immune Cells Through the Analysis of Membrane Sheets by Electron Microscopy  

PubMed Central

This chapter describes a method to generate plasma membrane sheets that are large enough to visualize the membrane architecture and perform quantitative analyses of protein distributions. This procedure places the sheets on electron microscopy grids, parallel to the imaging plane of the microscope, where they can be characterized by transmission electron microscopy. The basic principle of the technique is that cells are broken open (“ripped”) through mechanical forces applied by the separation of two opposing surfaces sandwiching the cell, with one of the surfaces coated onto an EM grid. The exposed inner membrane surfaces can then be visualized with electron dense stains and specific proteins can be detected with gold conjugated probes. PMID:21701974

Lillemeier, Björn F.; Davis, Mark M.



Rapid estimation of catalyst nanoparticle morphology and atomic-coordination by high-resolution Z-contrast electron microscopy.  


Heterogeneous nanoparticle catalyst development relies on an understanding of their structure-property relationships, ideally at atomic resolution and in three-dimensions. Current transmission electron microscopy techniques such as discrete tomography can provide this but require multiple images of each nanoparticle and are incompatible with samples that change under electron irradiation or with surveying large numbers of particles to gain significant statistics. Here, we make use of recent advances in quantitative dark-field scanning transmission electron microscopy to count the number atoms in each atomic column of a single image from a platinum nanoparticle. These atom-counts, along with the prior knowledge of the face-centered cubic geometry, are used to create atomistic models. An energy minimization is then used to relax the nanoparticle's 3D structure. This rapid approach enables high-throughput statistical studies or the analysis of dynamic processes such as facet-restructuring or particle damage. PMID:25340541

Jones, Lewys; MacArthur, Katherine E; Fauske, Vidar T; van Helvoort, Antonius T J; Nellist, Peter D



Transmission electron microscopy of polymer blends and block copolymers  

NASA Astrophysics Data System (ADS)

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-consistent field theory (SCFT). The liquid-like nature of this system at room temperature makes traditional staining methods for the enhancement of contrast ineffective. As an alternative, we take advantage of the large inelastic scattering cross-section of soft materials to generate contrast in zero-loss TEM images. Independent spatially resolved thickness measurements enable quantification of electron scattering. This enabled a comparison between the TEM data and predictions based on SCFT without any adjustable parameters. The second example involves the utilization of energy-filtered transmission electron microscopy (EFTEM) to compute elemental maps by taking advantage of ionization events. Elemental mapping of lithium is used to determine the distribution of salt in nanostructured poly(styrene-block-ethylene oxide) (SEO) copolymer/lithium salt electrolytes. Surprisingly, the concentration of lithium within a poly(ethylene oxide) (PEO) domain is found to be inhomogeneous; the salt is localized to the middle of the channels. Self-consistent field theory simulations suggest that localization of lithium is due to chain stretching at the interface, which increases with molecular weight. EFTEM and SCFT results show that the segregation of lithium salt to the middle of the PEO lamellae is greater for higher molecular weight polymers. This is correlated with the ionic conductivity of the copolymer electrolyte, which is found to show a higher conductivity for thinner lithium lamellae.

Gomez, Enrique Daniel


Large-scale automatic reconstruction of neuronal processes from electron microscopy images.  


Automated sample preparation and electron microscopy enables acquisition of very large image data sets. These technical advances are of special importance to the field of neuroanatomy, as 3D reconstructions of neuronal processes at the nm scale can provide new insight into the fine grained structure of the brain. Segmentation of large-scale electron microscopy data is the main bottleneck in the analysis of these data sets. In this paper we present a pipeline that provides state-of-the art reconstruction performance while scaling to data sets in the GB-TB range. First, we train a random forest classifier on interactive sparse user annotations. The classifier output is combined with an anisotropic smoothing prior in a Conditional Random Field framework to generate multiple segmentation hypotheses per image. These segmentations are then combined into geometrically consistent 3D objects by segmentation fusion. We provide qualitative and quantitative evaluation of the automatic segmentation and demonstrate large-scale 3D reconstructions of neuronal processes from a 27,000?m(3) volume of brain tissue over a cube of 30?m in each dimension corresponding to 1000 consecutive image sections. We also introduce Mojo, a proofreading tool including semi-automated correction of merge errors based on sparse user scribbles. PMID:25791436

Kaynig, Verena; Vazquez-Reina, Amelio; Knowles-Barley, Seymour; Roberts, Mike; Jones, Thouis R; Kasthuri, Narayanan; Miller, Eric; Lichtman, Jeff; Pfister, Hanspeter



Cryo-Electron Microscopy Structure of Lactococcal Siphophage 1358 Virion  

PubMed Central

ABSTRACT Lactococcus lactis, a Gram+ lactic acid-producing bacterium used for the manufacture of several fermented dairy products, is subject to infection by diverse virulent tailed phages, leading to industrial fermentation failures. This constant viral risk has led to a sustained interest in the study of their biology, diversity, and evolution. Lactococcal phages now constitute a wide ensemble of at least 10 distinct genotypes within the Caudovirales order, many of them belonging to the Siphoviridae family. Lactococcal siphophage 1358, currently the only member of its group, displays a noticeably high genomic similarity to some Listeria phages as well as a host range limited to a few L. lactis strains. These genomic and functional characteristics stimulated our interest in this phage. Here, we report the cryo-electron microscopy structure of the complete 1358 virion. Phage 1358 exhibits noteworthy features, such as a capsid with dextro handedness and protruding decorations on its capsid and tail. Observations of the baseplate of virion particles revealed at least two conformations, a closed and an open, activated form. Functional assays uncovered that the adsorption of phage 1358 to its host is Ca2+ independent, but this cation is necessary to complete its lytic cycle. Taken together, our results provide the complete structural picture of a unique lactococcal phage and expand our knowledge on the complex baseplate of phages of the Siphoviridae family. IMPORTANCE Phages of Lactococcus lactis are investigated mainly because they are sources of milk fermentation failures in the dairy industry. Despite the availability of several antiphage measures, new phages keep emerging in this ecosystem. In this study, we provide the cryo-electron microscopy reconstruction of a unique lactococcal phage that possesses genomic similarity to particular Listeria phages and has a host range restricted to only a minority of L. lactis strains. The capsid of phage 1358 displays the almost unique characteristic of being dextro handed. Its capsid and tail exhibit decorations that we assigned to nonspecific sugar binding modules. We observed the baseplate of 1358 in two conformations, a closed and an open form. We also found that the adsorption to its host, but not infection, is Ca2+ independent. Overall, this study advances our understanding of the adhesion mechanisms of siphophages. PMID:24872584

Spinelli, Silvia; Bebeacua, Cecilia; Orlov, Igor; Tremblay, Denise; Klaholz, Bruno P.



Quantitative analysis of immunofluorescent punctate staining of synaptically localized proteins using confocal microscopy and stereology.  


We established a protocol for the immunofluorescent detection of glutamate receptor subunits at synaptic sites using laser scanning confocal microscopy and stereological procedures. An in vitro model of eyeblink classical conditioning from turtles was used for this study. Triple-labeling of the presynaptic marker synaptophysin, the NR1 subunit of NMDA receptors, and the GluR4 subunit of AMPA receptors was performed on pseudoconditioned (control) and conditioned in vitro brain stem preparations in which punctate staining for each individual protein, as well as for the colocalization of GluR4 and NR1 with synaptophysin, was analyzed. For every tissue section analyzed, images of two consecutive optical planes were taken using confocal microscopy. Protein puncta were counted in one optical section (sample section) if they were not present in the optical section immediately above the sample section (look-up section). We found a significant increase in the colocalization of GluR4-containing AMPA receptors with synaptophysin after conditioning compared with the control group. Colocalization of NR1 subunits with synaptophysin was unchanged after conditioning. The described protocol, therefore, can be used for the quantitative analysis of changes in synaptic localization of different types of proteins. The protocol is designed to provide a more accurate and uniform approach in studying receptor trafficking during various forms of synaptic plasticity. PMID:16740315

Mokin, Maxim; Keifer, Joyce



Quantitative analysis of intrinsic skin aging in dermal papillae by in vivo harmonic generation microscopy  

PubMed Central

Chronological skin aging is associated with flattening of the dermal-epidermal junction (DEJ), but to date no quantitative analysis focusing on the aging changes in the dermal papillae (DP) has been performed. The aim of the study is to determine the architectural changes and the collagen density related to chronological aging in the dermal papilla zone (DPZ) by in vivo harmonic generation microscopy (HGM) with a sub-femtoliter spatial resolution. We recruited 48 Asian subjects and obtained in vivo images on the sun-protected volar forearm. Six parameters were defined to quantify 3D morphological changes of the DPZ, which we analyzed both manually and computationally to study their correlation with age. The depth of DPZ, the average height of isolated DP, and the 3D interdigitation index decreased with age, while DP number density, DP volume, and the collagen density in DP remained constant over time. In vivo high-resolution HGM technology has uncovered chronological aging-related variations in DP, and sheds light on real-time quantitative skin fragility assessment and disease diagnostics based on collagen density and morphology. PMID:25401037

Liao, Yi-Hua; Kuo, Wei-Cheng; Chou, Sin-Yo; Tsai, Cheng-Shiun; Lin, Guan-Liang; Tsai, Ming-Rung; Shih, Yuan-Ta; Lee, Gwo-Giun; Sun, Chi-Kuang



Quantitative chemical imaging and unsupervised analysis using hyperspectral coherent anti-Stokes Raman scattering microscopy.  


In this work, we report a method to acquire and analyze hyperspectral coherent anti-Stokes Raman scattering (CARS) microscopy images of organic materials and biological samples resulting in an unbiased quantitative chemical analysis. The method employs singular value decomposition on the square root of the CARS intensity, providing an automatic determination of the components above noise, which are retained. Complex CARS susceptibility spectra, which are linear in the chemical composition, are retrieved from the CARS intensity spectra using the causality of the susceptibility by two methods, and their performance is evaluated by comparison with Raman spectra. We use non-negative matrix factorization applied to the imaginary part and the nonresonant real part of the susceptibility with an additional concentration constraint to obtain absolute susceptibility spectra of independently varying chemical components and their absolute concentration. We demonstrate the ability of the method to provide quantitative chemical analysis on known lipid mixtures. We then show the relevance of the method by imaging lipid-rich stem-cell-derived mouse adipocytes as well as differentiated embryonic stem cells with a low density of lipids. We retrieve and visualize the most significant chemical components with spectra given by water, lipid, and proteins segmenting the image into the cell surrounding, lipid droplets, cytosol, and the nucleus, and we reveal the chemical structure of the cells, with details visualized by the projection of the chemical contrast into a few relevant channels. PMID:24099603

Masia, Francesco; Glen, Adam; Stephens, Phil; Borri, Paola; Langbein, Wolfgang



Hyperspectral and differential CARS microscopy for quantitative chemical imaging in human adipocytes.  


In this work, we demonstrate the applicability of coherent anti-Stokes Raman scattering (CARS) micro-spectroscopy for quantitative chemical imaging of saturated and unsaturated lipids in human stem-cell derived adipocytes. We compare dual-frequency/differential CARS (D-CARS), which enables rapid imaging and simple data analysis, with broadband hyperspectral CARS microscopy analyzed using an unsupervised phase-retrieval and factorization method recently developed by us for quantitative chemical image analysis. Measurements were taken in the vibrational fingerprint region (1200-2000/cm) and in the CH stretch region (2600-3300/cm) using a home-built CARS set-up which enables hyperspectral imaging with 10/cm resolution via spectral focussing from a single broadband 5 fs Ti:Sa laser source. Through a ratiometric analysis, both D-CARS and phase-retrieved hyperspectral CARS determine the concentration of unsaturated lipids with comparable accuracy in the fingerprint region, while in the CH stretch region D-CARS provides only a qualitative contrast owing to its non-linear behavior. When analyzing hyperspectral CARS images using the blind factorization into susceptibilities and concentrations of chemical components recently demonstrated by us, we are able to determine vol:vol concentrations of different lipid components and spatially resolve inhomogeneities in lipid composition with superior accuracy compared to state-of-the art ratiometric methods. PMID:24877002

Di Napoli, Claudia; Pope, Iestyn; Masia, Francesco; Watson, Peter; Langbein, Wolfgang; Borri, Paola



Quantitative phase imaging of live cells using fast Fourier phase microscopy  

NASA Astrophysics Data System (ADS)

Using the decomposition of an image field in two spatial components that can be controllably shifted in phase with respect to each other, a new quantitative-phase microscope has been developed. The new instrument, referred to as the fast Fourier phase microscope (f-FPM), provides a factor of 100 higher acquisition rate compared with our previously reported Fourier phase microscope. The resulting quantitative-phase images are characterized by diffraction limited transverse resolution and path-length stability better than 2 nm at acquisition rates of 10 frames/s or more. These features make the f-FPM particularly appealing for investigating the structure and dynamics of live cells over a broad range of time scales. In addition, we demonstrate the possibility of examining subcellular structures by digitally processing the amplitude and phase information provided by the instrument. Thus we developed software that can emulate phase contrast and differential interference contrast microscopy images by numerically processing FPM images. This approach adds the flexibility of digitally varying the phase shift between the two interfering beams. The images obtained appear as if they were recorded by variable phase contrast or differential interference contrast microscopes that deliver an enhanced view to the subcellular structure when compared with the typical commercial microscope.

Lue, Niyom; Choi, Wonshik; Popescu, Gabriel; Ikeda, Takahiro; Dasari, Ramachandra R.; Badizadegan, Kamran; Feld, Michael S.



Quantitative analysis of intrinsic skin aging in dermal papillae by in vivo harmonic generation microscopy.  


Chronological skin aging is associated with flattening of the dermal-epidermal junction (DEJ), but to date no quantitative analysis focusing on the aging changes in the dermal papillae (DP) has been performed. The aim of the study is to determine the architectural changes and the collagen density related to chronological aging in the dermal papilla zone (DPZ) by in vivo harmonic generation microscopy (HGM) with a sub-femtoliter spatial resolution. We recruited 48 Asian subjects and obtained in vivo images on the sun-protected volar forearm. Six parameters were defined to quantify 3D morphological changes of the DPZ, which we analyzed both manually and computationally to study their correlation with age. The depth of DPZ, the average height of isolated DP, and the 3D interdigitation index decreased with age, while DP number density, DP volume, and the collagen density in DP remained constant over time. In vivo high-resolution HGM technology has uncovered chronological aging-related variations in DP, and sheds light on real-time quantitative skin fragility assessment and disease diagnostics based on collagen density and morphology. PMID:25401037

Liao, Yi-Hua; Kuo, Wei-Cheng; Chou, Sin-Yo; Tsai, Cheng-Shiun; Lin, Guan-Liang; Tsai, Ming-Rung; Shih, Yuan-Ta; Lee, Gwo-Giun; Sun, Chi-Kuang



Microscopy environment for quantitative spatial and temporal analysis of multicellular interactions  

NASA Astrophysics Data System (ADS)

Quantitative analysis of spatial and temporal concurrent responses of multiple markers in 3-dimensional cell cultures is hampered by the routine mode of sequential image acquisition, measurement and analysis of specific targets. A system was developed for detailed analysis of multi-dimensional, time-sequence responses and in order to relate features in novel and meaningful ways that will further our understanding of basic biology. Optical sectioning of the 3-dimensional structures is achieved with structured light illumination using the Wilson grating as described by Lanni. The automated microscopy system can image multicellular structures and track dynamic events, and is equipped for simultaneous/ sequential imaging of multiple fluorescent markers. Computer-controlled perfusion of external stimuli into the culture system allows (i) real-time observations of multiple cellular responses and (ii) automatic and intelligent adjustment of experimental parameters. This creates a feedback loop in real-time that directs desired responses in a given experiment. On-line image analysis routines provide cell-by-cell measurement results through segmentation and feature extraction (i.e. intensity, localization, etc.), and quantitation of meta-features such as dynamic responses of cells or correlations between different cells. Off-line image and data analysis is used to derive models of the processes involved, which will deepen the understanding of the basic biology.

Sudar, Damir; Parvin, Bahram; Callahan, Daniel E.; Schwarz, Richard I.; Knowles, David W.; Ortiz de Solorzano, Carlos; Barcellos-Hoff, Mary H.



Analytical electron microscopy study on gallium nitride systems doped with manganese and iron  

NASA Astrophysics Data System (ADS)

Modulated structures of gallium nitride (GaN) doped with transition metal ions (here Fe, Mn) are investigated by analytical (scanning) transmission electron microscopy to gain insight into the structural arrangement and chemical composition of the material, known to be critically correlated to the magnetic response and hence the functionality of these technologically relevant systems. Three classes of samples are considered: (i) homogeneous (dilute) (Ga, Mn)N; (ii) ?-Mn-doped (Ga, ?-Mn)N and phase separated (Ga, Fe)N, containing Fe-rich nanocrystals. The combination of various microscopic techniques employed, allows for a quantitative determination of the distribution of the magnetic ions in the samples, providing essential information on the structural and chemical asset of these systems.

Meingast, Arno; Navarro Quezada, Andrea; Devillers, Thibaut; Kovács, András; Albu, Mihela; Fladischer, Stefanie; Bonanni, Alberta; Kothleitner, Gerald



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


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. PMID:21844587

Colliex, Christian



Environmental Scanning Electron Microscopy of Ice Crystal Nucleation and Growth  

NASA Astrophysics Data System (ADS)

Ice crystal nucleation and growth are dual processes that can be studied uniquely through Environmental Scanning Electron Microscopy (ESEM). By utilizing differential pumping systems and a Peltier element to vary the vapor pressure and to achieve temperatures below the freezing point, respectively, it is possible to obtain supersaturated conditions relative to ice in the sample chamber of an Environmental Scanning Electron Microscope. Ice crystals were nucleated on a variety of atmospherically relevant substrates and grown in a pure water vapor environment in the chamber of a FEI-Quanta 200 ESEM. To initiate ice crystal nucleation, the Peltier element was set at a temperature between -10°C and -25°C, while the chamber water vapor pressure was adjusted to just below the frost point. Ice crystal nucleation and growth was then controlled by careful adjustments of chamber pressure and temperature, where high-magnification images of hexagonal ice crystals were acquired at nanoscale resolution. These images display prominent mesoscopic surface topography including linear strands, crevasses, islands, and steps. The surface features are seen to be ubiquitously present at all observed temperatures, at many supersaturated and subsaturated conditions, and on all crystal facets. Additionally, a pre-growth "shadow" resembling a dark spot sometimes appeared on areas of the sample stage immediately preceding ice crystal nucleation and growth. The observations represent the most highly magnified images of ice surfaces yet reported and significantly expand the range of ambient conditions where the features are conspicuous. New knowledge of the presence and characteristics of these features could transform the fundamental understanding of ice crystal growth kinetics and its physical parameterization in the context of atmospheric and cryospheric science. To the extent these observations are applicable to atmospheric ice, the results suggest that the radiative representation of ice and mixed-phase cloud properties in climate models could be markedly affected.

Amaral, M.; Miller, A. L.; Magee, N. B.



Transmission electron microscopy analysis of corroded metal waste forms.  

SciTech Connect

This report documents the results of analyses with transmission electron microscopy (TEM) combined with energy dispersive X-ray spectroscopy (EDS) and selected area electron diffraction (ED) of samples of metallic waste form (MWF) materials that had been subjected to various corrosion tests. The objective of the TEM analyses was to characterize the composition and microstructure of surface alteration products which, when combined with other test results, can be used to determine the matrix corrosion mechanism. The examination of test samples generated over several years has resulted in refinements to the TEM sample preparation methods developed to preserve the orientation of surface alteration layers and the underlying base metal. The preservation of microstructural spatial relationships provides valuable insight for determining the matrix corrosion mechanism and for developing models to calculate radionuclide release in repository performance models. The TEM results presented in this report show that oxide layers are formed over the exposed steel and intermetallic phases of the MWF during corrosion in aqueous solutions and humid air at elevated temperatures. An amorphous non-stoichiometric ZrO{sub 2} layer forms at the exposed surfaces of the intermetallic phases, and several nonstoichiometric Fe-O layers form over the steel phases in the MWF. These oxide layers adhere strongly to the underlying metal, and may be overlain by one or more crystalline Fe-O phases that probably precipitated from solution. The layer compositions are consistent with a corrosion mechanism of oxidative dissolution of the steel and intermetallic phases. The layers formed on the steel and intermetallic phases form a continuous layer over the exposed waste form, although vertical splits in the layer and corrosion in pits and crevices were seen in some samples. Additional tests and analyses are needed to verify that these layers passivate the underlying metals and if passivation can break down as the MWF corrodes. The importance of localized corrosion should also be determined.

Dietz, N. L.



Quantitation of glucocorticoid receptor DNA-binding dynamics by single-molecule microscopy and FRAP.  


Recent advances in live cell imaging have provided a wealth of data on the dynamics of transcription factors. However, a consistent quantitative description of these dynamics, explaining how transcription factors find their target sequences in the vast amount of DNA inside the nucleus, is still lacking. In the present study, we have combined two quantitative imaging methods, single-molecule microscopy and fluorescence recovery after photobleaching, to determine the mobility pattern of the glucocorticoid receptor (GR) and the mineralocorticoid receptor (MR), two ligand-activated transcription factors. For dexamethasone-activated GR, both techniques showed that approximately half of the population is freely diffusing, while the remaining population is bound to DNA. Of this DNA-bound population about half the GRs appeared to be bound for short periods of time (? 0.7 s) and the other half for longer time periods (? 2.3 s). A similar pattern of mobility was seen for the MR activated by aldosterone. Inactive receptors (mutant or antagonist-bound receptors) show a decreased DNA binding frequency and duration, but also a higher mobility for the diffusing population. Likely, very brief (? 1 ms) interactions with DNA induced by the agonists underlie this difference in diffusion behavior. Surprisingly, different agonists also induce different mobilities of both receptors, presumably due to differences in ligand-induced conformational changes and receptor complex formation. In summary, our data provide a consistent quantitative model of the dynamics of GR and MR, indicating three types of interactions with DNA, which fit into a model in which frequent low-affinity DNA binding facilitates the search for high-affinity target sequences. PMID:24632838

Groeneweg, Femke L; van Royen, Martin E; Fenz, Susanne; Keizer, Veer I P; Geverts, Bart; Prins, Jurrien; de Kloet, E Ron; Houtsmuller, Adriaan B; Schmidt, Thomas S; Schaaf, Marcel J M



Quantitation of Glucocorticoid Receptor DNA-Binding Dynamics by Single-Molecule Microscopy and FRAP  

PubMed Central

Recent advances in live cell imaging have provided a wealth of data on the dynamics of transcription factors. However, a consistent quantitative description of these dynamics, explaining how transcription factors find their target sequences in the vast amount of DNA inside the nucleus, is still lacking. In the present study, we have combined two quantitative imaging methods, single-molecule microscopy and fluorescence recovery after photobleaching, to determine the mobility pattern of the glucocorticoid receptor (GR) and the mineralocorticoid receptor (MR), two ligand-activated transcription factors. For dexamethasone-activated GR, both techniques showed that approximately half of the population is freely diffusing, while the remaining population is bound to DNA. Of this DNA-bound population about half the GRs appeared to be bound for short periods of time (?0.7 s) and the other half for longer time periods (?2.3 s). A similar pattern of mobility was seen for the MR activated by aldosterone. Inactive receptors (mutant or antagonist-bound receptors) show a decreased DNA binding frequency and duration, but also a higher mobility for the diffusing population. Likely, very brief (?1 ms) interactions with DNA induced by the agonists underlie this difference in diffusion behavior. Surprisingly, different agonists also induce different mobilities of both receptors, presumably due to differences in ligand-induced conformational changes and receptor complex formation. In summary, our data provide a consistent quantitative model of the dynamics of GR and MR, indicating three types of interactions with DNA, which fit into a model in which frequent low-affinity DNA binding facilitates the search for high-affinity target sequences. PMID:24632838

Groeneweg, Femke L.; van Royen, Martin E.; Fenz, Susanne; Keizer, Veer I. P.; Geverts, Bart; Prins, Jurrien; de Kloet, E. Ron; Houtsmuller, Adriaan B.; Schmidt, Thomas S.; Schaaf, Marcel J. M.



Hybrid Electron Microscopy Normal Mode Analysis graphical interface and protocol.  


This article presents an integral graphical interface to the Hybrid Electron Microscopy Normal Mode Analysis (HEMNMA) approach that was developed for capturing continuous motions of large macromolecular complexes from single-particle EM images. HEMNMA was shown to be a good approach to analyze multiple conformations of a macromolecular complex but it could not be widely used in the EM field due to a lack of an integral interface. In particular, its use required switching among different software sources as well as selecting modes for image analysis was difficult without the graphical interface. The graphical interface was thus developed to simplify the practical use of HEMNMA. It is implemented in the open-source software package Xmipp 3.1 ( and only a small part of it relies on MATLAB that is accessible through the main interface. Such integration provides the user with an easy way to perform the analysis of macromolecular dynamics and forms a direct connection to the single-particle reconstruction process. A step-by-step HEMNMA protocol with the graphical interface is given in full details in Supplementary material. The graphical interface will be useful to experimentalists who are interested in studies of continuous conformational changes of macromolecular complexes beyond the modeling of continuous heterogeneity in single particle reconstruction. PMID:25268657

Sorzano, Carlos Oscar S; de la Rosa-Trevín, José Miguel; Tama, Florence; Joni?, Slavica



Scanning electron microscopy of lung following alpha irradiation  

SciTech Connect

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.

Sanders, C.L.; Lauhala, K.E.; McDonald, K.E. (Pacific Northwest Laboratory, Richland, WA (USA))



Cryogenic Transmission Electron Microscopy Nanostructural Study of Shed Microparticles  

PubMed Central

Microparticles (MPs) are sub-micron membrane vesicles (100–1000 nm) shed from normal and pathologic cells due to stimulation or apoptosis. MPs can be found in the peripheral blood circulation of healthy individuals, whereas elevated concentrations are found in pregnancy and in a variety of diseases. Also, MPs participate in physiological processes, e.g., coagulation, inflammation, and angiogenesis. Since their clinical properties are important, we have developed a new methodology based on nano-imaging that provides significant new data on MPs nanostructure, their composition and function. We are among the first to characterize by direct-imaging cryogenic transmitting electron microscopy (cryo-TEM) the near-to-native nanostructure of MP systems isolated from different cell types and stimulation procedures. We found that there are no major differences between the MP systems we have studied, as most particles were spherical, with diameters from 200 to 400 nm. However, each MP population is very heterogeneous, showing diverse morphologies. We investigated by cryo-TEM the effects of standard techniques used to isolate and store MPs, and found that either high-g centrifugation of MPs for isolation purposes, or slow freezing to –80°C for storage introduce morphological artifacts, which can influence MP nanostructure, and thus affect the efficiency of these particles as future diagnostic tools. PMID:24386253

Issman, Liron; Brenner, Benjamin; Talmon, Yeshayahu; Aharon, Anat



Histological preparation of developing vestibular otoconia for scanning electron microscopy  

NASA Technical Reports Server (NTRS)

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.

Huss, D.; Dickman, J. D.



Transmission electron microscopy (TEM) study of minerals in coal  

SciTech Connect

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

Hsieh, Kuang-Chien



Microanalysis of ambient particles from Lexington, KY, by electron microscopy  

NASA Astrophysics Data System (ADS)

Analytical transmission electron microscopy has been used for in-depth characterization of the individual submicron (physical size<1 ?m) particulate matter (PM) from the urban atmosphere of Lexington, Kentucky, USA. Carbonaceous particles and sulfur-bearing particles dominate the PM samples on a particle count basis. Soot aggregates are the major form of carbonaceous particles. Organic-type and biogenic carbonaceous particles are also observed and exhibit quite different morphologies and microstructures compared with combustion-generated soot and char particles. Ammonium sulfate is the dominant chemical type of sulfur-bearing particles. Other types of sulfur-bearing particles include crystalline sodium, potassium and calcium sulfates. Silicon-bearing particles consist mainly of silicon oxides and aluminosilicates. Iron and titanium oxide particles are the major metal oxides found in this study. Considerable amounts of iron oxide with other transition metal elements (e.g. Mn and Zn) have been observed, typically in the form of spinel nanocrystals. Minor amounts of other species (e.g. phosphates, lead compounds, and carbonates) have also been observed. The combined use of these analytical techniques is capable of revealing much new information on the microcharacteristics of individual submicron particles; such information should be relevant to toxicological, epidemiological, and source apportionment studies.

Chen, Yuanzhi; Shah, Naresh; Huggins, Frank E.; Huffman, Gerald P.


Surface treatment of feldspathic porcelain: scanning electron microscopy analysis  

PubMed Central

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

Valian, Azam



Analytical electron microscopy of biogenic and inorganic carbonates  

NASA Technical Reports Server (NTRS)

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.

Blake, David F.



Scanning electron microscopy and roughness study of dental composite degradation.  


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. PMID:22325725

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



High resolution electron microscopy of nonstoichiometric apatite crystals.  


The application of high resolution electron microscopy, computer image processing, and image simulation techniques to the investigation of synthetic nonstoichiometric apatites has provided new details of apatite crystal growth mechanisms. Under certain precipitation conditions, calcium-deficient apatites with distinct octacalcium phosphate (OCP)-apatite intergrowths have been observed. Apatite crystals with unit-cell thick overgrowths of OCP on their surfaces confirmed the stepwise hydrolysis crystal growth mechanism initially proposed by Brown (Nature 196:1048-1050). However, many crystals also contained a central two-dimensional OCP inclusion one to two unit cells thick, embedded in an apatite matrix. Similar planar defects have been observed in dental enamel, dentin, and bone crystals. We have developed a modified version of Brown's stepwise OCP hydrolysis apatite crystal growth mechanism to explain the formation of crystals with OCP central planar defects. The mechanism involves the nucleation of an OCP seed that grows until it reaches a critical size, rh, before OCP hydrolysis occurs. Apatite subsequently grows epitaxially on the OCP seed, thereby embedding it in the center of an apatite crystal. Apatite growth is facilitated by partial screw dislocations emanating from the planar defect. PMID:2672890

Nelson, D G; Barry, J C



Non-thermal plasma mills bacteria: Scanning electron microscopy observations  

NASA Astrophysics Data System (ADS)

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.

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



Study of the Electron Mean Free Path by Ballistic Electron Emission Microscopy  

NASA Astrophysics Data System (ADS)

Ballistic Electron Emission Microscopy allows buried interfaces to be characterized with a subnanometer resolution. Additionnally, the electron mean free path in a thin metal layer can thus be investigated with a good level of accuracy. This paper presents the latest results in this field for an Au/n-Si Schottky diode. A discussion is also proposed to link these results with the still controversial interpretations of BEEM contrasts. La microscopie par émission d'électrons balistiques permet la caractérisation d'interfaces enfouies, avec une résolution inférieure au nanomètre. De plus, le libre parcours moyen des électrons dans une fine couche métallique peut être étudié avec une excellente précision. Cet article présente les derniers résultats dans ce domaine, pour des diodes Schottky Au/n-Si. Une discussion est également proposée pour relier ces résultats aux interprétations, toujours très controversées, des contrastes fournis par les images BEEM.

Girardin, C.; Coratger, R.; Pechou, R.; Ajustron, F.; Beauvillain, J.



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

PubMed Central

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??m 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:24689948

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



Electron diffraction and microscopy study of nanotubes and nanowires  

NASA Astrophysics Data System (ADS)

Carbon nanotubes have many excellent properties that are strongly influenced by their atomic structure. The realization of the ultimate potential of carbon nanotubes in technological applications necessitates a precise control of the structure of as-grown nanotubes as well as the identification of their atomic structures. Transmission electron microscopy (TEM) is a technique that can deliver this by combining the high resolution imaging and electron diffraction simultaneously. In this study, a new catalyst system (the Co/Si) was investigated in the production of single-walled carbon nanotubes (SWNTs) by laser ablation. It was discovered that the Co/Si mixture as a catalyst was as successful as the Ni/Co in the synthesis of SWNTs. The isolated individual SWNTs were examined by using nanobeam electron diffraction for the structure identification and it was found that carbon nanotubes grown by this catalyst mixture tend to be slightly more metallic. The electron diffraction technique has been refined to establish a new methodology to determine the chirality of each shell in a carbon nanotube and it has been applied to determine the atomic structure of double-walled carbon nanotubes (DWNT), few-walled carbon nanotubes (FWNT) and multi-walled carbon nanotubes (MWNT). We observed that there is no strong correlation in the structure of two adjacent shells in DWNTs. Several FWNTs and MWNTs have been examined by our new electron diffraction method to determine their atomic structures and to test the efficiency and the reliability of this method for structure identification. We now suggest that a carbon nanotube of up to 25 shells can be studied and the chirality of each shell can be identified by this new technique. The guidelines for the automation of such procedure have been laid down and explained in this work. The atomic structure of tungsten disulfide (WS2) nanotubes was studied by using the methods developed for the structure determination of carbon nanotubes. The WS2 nanotubes are another example of the tube forming ability of the layered structures and a member of the family of inorganic fullerene-like structures. These nanotubes are much larger in diameter than carbon nanotubes. The tubes studied here have helicities less than 18° and usually have near zigzag structure. The short-range order (SRO) in the atomic structure of carbon soot produced by laser ablation was investigated using electron diffraction and radial distribution function (RDF) analysis. The effects of the furnace temperature and the metal catalyst on the SRO in the carbon soot were also studied. It was discovered that the SRO structure is the same for all carbon soot samples studied and is very similar to that of amorphous carbon. These techniques were also applied to determine the atomic structure of amorphous boron nanowires. We found out that the atomic structure of these boron nanowires agree well with the previously reported structure of bulk amorphous boron.

Deniz, Hakan


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

SciTech Connect

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.

Boeker, T.W.; Wegener, A.; Koch, F.; Hockwin, O. (University-Eye-Clinic Bonn (Germany))



Investigation of viability of plant tissue in the environmental scanning electron microscopy.  


The advantages of environmental scanning electron microscopy (ESEM) make it a suitable technique for studying plant tissue in its native state. There have been few studies on the effects of ESEM environment and beam damage on the viability of plant tissue. A simple plant tissue, Allium cepa (onion) upper epidermal tissue was taken as the model for study. The change of moisture content of samples was studied at different relative humidities. Working with the electron beam on, viability tests were conducted for samples after exposure in the ESEM under different operating conditions to investigate the effect of electron beam dose on the viability of samples. The results suggested that without the electron beam, the ESEM chamber itself can prevent the loss of initial moisture if its relative humidity is maintained above 90%. With the electron beam on, the viability of Allium cepa (onion) cells depends both on the beam accelerating voltage and the electron dose/unit area hitting the sample. The dose can be controlled by several of the ESEM instrumental parameters. The detailed process of beam damage on cuticle-down and cuticle-up samples was investigated and compared. The results indicate that cuticular adhesion to the cell wall is relatively weak, but highly resistant to electron beam damage. Systematic study on the effect of ESEM operation parameters has been done. Results qualitatively support the intuitive expectations, but demonstrate quantitatively that Allium cepa epidermal cells are able to be kept in a hydrated and viable state under relevant operation condition inside ESEM, providing a basis for further in situ experiments on plant tissues. PMID:20183923

Zheng, Tao; Waldron, K W; Donald, Athene M



Sensitivity and spatial resolution for electron-spin-resonance detection by magnetic resonance force microscopy  

E-print Network

Sensitivity and spatial resolution for electron-spin-resonance detection by magnetic resonance The signal intensity of electron spin resonance in magnetic resonance force microscopy MRFM experiments that magnetic resonance force microscopy MRFM is a new 3D imaging technique8,9 with the potential of achieving

Hammel, P. Chris


3D reconstruction and comparison of shapes of DNA minicircles observed by cryo-electron microscopy  

E-print Network

3D reconstruction and comparison of shapes of DNA minicircles observed by cryo-electron microscopy, 2006 ABSTRACT We use cryo-electron microscopy to compare 3D shapes of 158 bp long DNA minicircles activator protein binding site. We present a sorting algorithm that correlates the reconstructed shapes

Alvarez, Nadir


Zinc speciation in a contaminated aquatic environment: Characterization of environmental particles by analytical electron microscopy  

Microsoft Academic Search

Analytical electron microscopy (AEM) was used to characterize individual aquatic particles in a lake that has been contaminated by zinc smelting operations. Samples were collected from the sediments and the water column of the lake along a gradient of metal contamination. The samples were prepared to preserve their aqueous nature, and thin sections were observed by transmission electron microscopy (TEM).

Samuel M. Webb; Gary G. Leppard; Jean-François Gaillard



FtsZ Condensates: An In Vitro Electron Microscopy Study David Popp,1  

E-print Network

FtsZ Condensates: An In Vitro Electron Microscopy Study David Popp,1 Mitsusada Iwasa,1 Akihiro of these structures are toroids, spheroids, and rod-like, and these condensates were extensively investigated in vitro by fluorescent light microscopy (FLM) techniques,3,4 yet it has never been observed by direct in vivo electron

Erickson, Harold P.


Spermiogenesis and spermatozoon of Echinostoma caproni ( Platyhelminthes, Digenea): transmission and scanning electron microscopy, and tubulin immunocytochemistry  

Microsoft Academic Search

Spermiogenesis and the spermatozoon of Echinostoma caproni (from experimentally infested laboratory mice) were investigated by several methods. Transmission electron microscopy shows that spermiogenesis consists of a proximo-distal fusion of three processes followed by elongation of the spermatid. Scanning electron microscopy shows that the spermatozoon is a filiform cell, 235 ?m in length, with a cylindrical anterior extremity and a broader

C. Iomini; J.-L. Justine



Nonlinear optical microscopy: use of second harmonic generation and two-photon microscopy for automated quantitative liver fibrosis studies  

E-print Network

Liver fibrosis is associated with an abnormal increase in an extracellular matrix in chronic liver diseases. Quantitative characterization of fibrillar collagen in intact tissue is essential for both fibrosis studies and ...

Sun, Wanxin


Quantitative determination of maximal imaging depth in all-NIR multiphoton microscopy images of thick tissues  

NASA Astrophysics Data System (ADS)

We report two methods for quantitatively determining maximal imaging depth from thick tissue images captured using all-near-infrared (NIR) multiphoton microscopy (MPM). All-NIR MPM is performed using 1550 nm laser excitation with NIR detection. This method enables imaging more than five-fold deep in thick tissues in comparison with other NIR excitation microscopy methods. In this study, we show a correlation between the multiphoton signal along the depth of tissue samples and the shape of the corresponding empirical probability density function (pdf) of the photon counts. Histograms from this analysis become increasingly symmetric with the imaging depth. This distribution transitions toward the background distribution at higher imaging depths. Inspired by these observations, we propose two independent methods based on which one can automatically determine maximal imaging depth in the all-NIR MPM images of thick tissues. At this point, the signal strength is expected to be weak and similar to the background. The first method suggests the maximal imaging depth corresponds to the deepest image plane where the ratio between the mean and median of the empirical photon-count pdf is outside the vicinity of 1. The second method suggests the maximal imaging depth corresponds to the deepest image plane where the squared distance between the empirical photon-count mean obtained from the object and the mean obtained from the background is greater than a threshold. We demonstrate the application of these methods in all-NIR MPM images of mouse kidney tissues to study maximal depth penetration in such tissues.

Sarder, Pinaki; Akers, Walter J.; Sudlow, Gail P.; Yazdanfar, Siavash; Achilefu, Samuel



Transmission Electron Microscopy of Iron Metal in Almahata Sitta Ureilite  

NASA Technical Reports Server (NTRS)

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.

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



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

NASA Astrophysics Data System (ADS)

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

Nellist, Pete



Simplified setup for imaging with digital holographic microscopy and enhanced quantitative phase contrast by osmotic stimulation of living cells  

NASA Astrophysics Data System (ADS)

Many interferometry-based quantitative phase contrast imaging techniques require the generation of a coherent reference wave, which results in a phase stability decrease and the demand for a precise adjustment of the intensity ratio between object and reference wave. Thus, investigations on a simplified digital holographic microscopy approach that avoids a separate reference wave were performed. Results from live cell investigations demonstrate the capability of the method for quantitative phase contrast imaging. In further experiments the modification of the intracellular refractive index distribution by osmotic stimulation was analyzed. Data from human pancreas tumor cells show that by choice of suitable buffer solutions live cell imaging with enhanced quantitative phase contrast is achieved.

Kemper, Björn; Przibilla, Sabine; Rommel, Christina E.; Vollmer, Angelika; Ketelhut, Steffi; Schnekenburger, Jürgen; von Bally, Gert



Visualization of Microbial Biomarkers by Scanning Electron Microscopy  

NASA Technical Reports Server (NTRS)

We are developing tools to link the biochemical structure of selected biomarkers with putative biogenic structures observed in mineralized samples. The detection of evidence of life on Mars and other planets will rely on methods that can discriminate compounds formed exclusively by living organisms. While biogenic compounds, such as amino acids and nucleotides have been discovered in extraterrestrial sources, such as meteorites and comets, their formation can be explained by abiotic means. The formation of cellular structures, or more elaborate organic molecules, such as complex lipids, proteins or nucleic acids, however, is strongly correlated to the presence of even the most primitive life processes. Recent evidence lends support to the hypothesis that life may have once existed on Mars. Carbonate globules and ppm concentrations of polycyclic aromatic hydrocarbons (PAHs) have been described in ALH84001, a meteorite originating from Mars ejecta captured by Earth over 13,000 years ago. The localized high concentration of PAHs that follow an increasing gradient from the intact fusion crust towards the interior corresponds to microgram quantities of hydrocarbon. Even though ALH84001 and other similar meteorites have withstood the forces capable of ejecting rock through Mars' escape velocity, upon entering Earth's atmosphere, their core temperatures are likely not to have been raised significantly, as evidenced by the survival of remanent magnetic signatures. Ideal biomarkers of ancient or modern biological life would include molecules that are (or were) pervasive and highly resistant to degradation. Also, requisite methods of detection should be simple, extremely sensitive and broadly inclusive (NASA SP-530). Lipopolysaccharide (LPS), peptidoglycan or pseudopeptidoglycan and beta-glucan are microbial cell wall components which together cover the entire microbial spectrum of eubacteria, archea and fungi. They are all remarkably resistant to thermal degradation. 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.

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



Application of frequency-domain Fluorescence Lifetime Imaging Microscopy as a quantitative analytical tool for microfluidic devices.  


We describe the application of wide-field frequency domain Fluorescence Lifetime Imaging Microscopy (FLIM) to imaging in microfluidic devices. FLIM is performed using low cost, intensity modulated Light Emitting Diodes (LEDs) for illumination. The use of lifetime imaging for quantitative analysis within such devices is demonstrated by mapping the molecular diffusion of iodide ions across a microchannel. PMID:19516711

Elder, A D; Matthews, S M; Swartling, J; Yunus, K; Frank, J H; Brennan, C M; Fisher, A C; Kaminski, C F



Diffusion tensor microscopy in human nervous tissue with quantitative correlation based on direct histological comparison  

PubMed Central

Thanks to its proven utility in both clinical and research applications, diffusion tensor tractography (DTT) is regularly employed as a means of delineating white matter tracts. While successful efforts have been made to validate tractographic predictions, comparative methods which would permit the validation of such predictions at microscopic resolutions in complex biological tissues have remained elusive. In a previous study, we attempted to validate for the first time such predictions at microscopic resolutions in spinal cords of the rat and pig using a semi-quantitative analysis method. In the current study, we report improved quantitative analysis methods that can be used to determine the accuracy of DTT through comparative histology and apply theses techniques for the first time to human tissue (spinal cord) samples. Histological images are downsampled to resolutions equivalent to our magnetic resonance microscopy (MRM) and converted to binary maps using an automated thresholding tool. These maps (n = 3) are coregistered to the MRM allowing us to quantify the agreement based on the number of pixels which contain tracts common to both imaging datasets. In our experiments we find that—on average—89% of imaging pixels predicted by DTT to contain in-plane white matter tract structure correspond to physical tracts identified by histology. In addition, angular analysis comparing the orientation of fiber tracts measured in histology to their corresponding in-plane primary eigenvector components is presented. Thus, as well as demonstrating feasibility in human tissue, we report a robust agreement between imaging datasets taken at microscopic resolution and confirm the primary eigenvector’s role as a fundamental parameter with clear physical correlates in the microscopic regime. PMID:21575730

Hansen, Brian; Flint, Jeremy J.; Heon-Lee, Choong; Fey, Michael; Vincent, Franck; King, Michael A.; Vestergaard-Poulsen, Peter; Blackband, Stephen J.



Vortex Matter Research by Electron Microscopy - Memorial to Dr. Akira Tonomura -  

NASA Astrophysics Data System (ADS)

Electron phase microscopy represented by electron holography and Lorentz microscopy, has opened the way to observing a single quantized magnetic flux-line - "vortex" - and its dynamical behavior inside superconductors. It was realized by Dr. Akira Tonomura and his collaborators by developing a field emission electron microscope and by advancing the technologies for visualizing the vortices, for example, low-temperature specimen stage and magnetic-field application system. Electron phase microscopy was clarified to be a powerful tool for investigating the flux-line features inside materials and their configurations with defects. This paper intends to give a review of the research results on vortex matter physics.

Harada, K.


Method for local temperature measurement in a nanoreactor for in situ high-resolution electron microscopy.  


In situ high-resolution transmission electron microscopy (TEM) of solids under reactive gas conditions can be facilitated by microelectromechanical system devices called nanoreactors. These nanoreactors are windowed cells containing nanoliter volumes of gas at ambient pressures and elevated temperatures. However, due to the high spatial confinement of the reaction environment, traditional methods for measuring process parameters, such as the local temperature, are difficult to apply. To address this issue, we devise an electron energy loss spectroscopy (EELS) method that probes the local temperature of the reaction volume under inspection by the electron beam. The local gas density, as measured using quantitative EELS, is combined with the inherent relation between gas density and temperature, as described by the ideal gas law, to obtain the local temperature. Using this method we determined the temperature gradient in a nanoreactor in situ, while the average, global temperature was monitored by a traditional measurement of the electrical resistivity of the heater. The local gas temperatures had a maximum of 56 °C deviation from the global heater values under the applied conditions. The local temperatures, obtained with the proposed method, are in good agreement with predictions from an analytical model. PMID:23831940

Vendelbo, S B; Kooyman, P J; Creemer, J F; Morana, B; Mele, L; Dona, P; Nelissen, B J; Helveg, S



Nanoscale Electronic Inhomogeneity in In_2Se_3 Nanoribbons Revealed by Microwave Impedance Microscopy  

SciTech Connect

Driven by interactions due to the charge, spin, orbital, and lattice degrees of freedom, nanoscale inhomogeneity has emerged as a new theme for materials with novel properties near multiphase boundaries. As vividly demonstrated in complex metal oxides and chalcogenides, these microscopic phases are of great scientific and technological importance for research in hightemperature superconductors, colossal magnetoresistance effect, phase-change memories, and domain switching operations. Direct imaging on dielectric properties of these local phases,however, presents a big challenge for existing scanning probe techniques. Here, we report the observation of electronic inhomogeneity in indium selenide (In{sub 2}Se{sub 3}) nanoribbons by near-field scanning microwave impedance microscopy. Multiple phases with local resistivity spanning six orders of magnitude are identified as the coexistence of superlattice, simple hexagonal lattice and amorphous structures with {approx}100nm inhomogeneous length scale, consistent with high-resolution transmission electron microscope studies. The atomic-force-microscope-compatible microwave probe is able to perform quantitative sub-surface electronic study in a noninvasive manner. Finally, the phase change memory function in In{sub 2}Se{sub 3} nanoribbon devices can be locally recorded with big signal of opposite signs.

Lai, K.J.



Atomic-Scale Imaging and Spectroscopy for In Situ Liquid Scanning Transmission Electron Microscopy  

SciTech Connect

Observation of growth, synthesis, dynamics and electrochemical reactions in the liquid state is an important yet largely unstudied aspect of nanotechnology. The only techniques that can potentially provide the insights necessary to advance our understanding of these mechanisms is simultaneous atomic-scale imaging and quantitative chemical analysis (through spectroscopy) under environmental conditions in the transmission electron microscope (TEM). In this study we describe the experimental and technical conditions necessary to obtain electron energy loss (EEL) spectra from a nanoparticle in colloidal suspension using aberration corrected scanning transmission electron microscopy (STEM) combined with the environmental liquid stage. At a fluid path length below 400 nm, atomic resolution images can be obtained and simultaneous compositional analysis can be achieved. We show that EEL spectroscopy can be used to quantify the total fluid path length around the nanoparticle, and demonstrate characteristic core-loss signals from the suspended nanoparticles can be resolved and analyzed to provide information on the local interfacial chemistry with the surrounding environment. The combined approach using aberration corrected STEM and EEL spectra with the in situ fluid stage demonstrates a plenary platform for detailed investigations of solution based catalysis and biological research.

Jungjohann, K. L.; Evans, James E.; Aguiar, Jeff; Arslan, Ilke; Browning, Nigel D.



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

SciTech Connect

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

Lansåker, Pia C., E-mail:; Niklasson, Gunnar A.; Granqvist, Claes G. [Department of Engineering Sciences, The Ångström Laboratory, Uppsala University, P. O. Box 534, SE-751 21 Uppsala (Sweden); Hallén, Anders [Royal Institute of Technology, KTH-ICT, Elektrum 229, Kista, SE-164 40 Stockholm (Sweden)



Quantitative Imaging of Rapidly Decaying Evanescent Fields Using Plasmonic Near-Field Scanning Optical Microscopy  

PubMed Central

Non-propagating evanescent fields play an important role in the development of nano-photonic devices. While detecting the evanescent fields in far-field can be accomplished by coupling it to the propagating waves, in practice they are measured in the presence of unwanted propagating background components. It leads to a poor signal-to-noise ratio and thus to errors in quantitative analysis of the local evanescent fields. Here we report on a plasmonic near-field scanning optical microscopy (p-NSOM) technique that incorporates a nanofocusing probe for adiabatic focusing of propagating surface plasmon polaritons at the probe apex, and for enhanced coupling of evanescent waves to the far-field. In addition, a harmonic demodulation technique is employed to suppress the contribution of the background. Our experimental results show strong evidence of background free near-field imaging using the new p-NSOM technique. Furthermore, we present measurements of surface plasmon cavity modes, and quantify their contributing sources using an analytical model. PMID:24076563

Zhang, Zhen; Ahn, Phillip; Dong, Biqin; Balogun, Oluwaseyi; Sun, Cheng



Direct visualization and quantitative analysis of water diffusion in complex biological tissues using CARS microscopy  

PubMed Central

To date, it has not been possible to measure microscopic diffusive water movements in epithelia and in the interstitial space of complex tissues and organs. Diffusive water movements are essential for life because they convey physiologically important small molecules, e.g. nutrients and signaling ligands throughout the extracellular space of complex tissues. Here we report the development of a novel method for the direct observation and quantitative analysis of water diffusion dynamics in a biologically organized tissue using Coherent Anti-Stokes Raman Scattering (CARS) microscopy. Using a computer simulation model to analyze the CARS O-H bond vibration data during H2O/D2O exchange in a 3D epithelial cyst, we succeeded in measuring the diffusive water permeability of the individual luminal and basolateral water pathways and also their response to hormonal stimulation. Our technique will be applicable to the measurement of diffusive water movements in other structurally complex and medically important tissues and organs. PMID:24067894

Yu, Ying-Chun; Sohma, Yoshiro; Takimoto, Shinichi; Miyauchi, Takayuki; Yasui, Masato



Microscopy, Culture, and Quantitative Real-Time PCR Examination Confirm Internalization of Mycobacteria in Plants  

PubMed Central

The environment is a reservoir of nontuberculous mycobacteria and is considered a source of infection for animals and humans. Mycobacteria can persist in different types of environments for a relatively long time. We have studied their possible internalization into plant tissue through intact, as well as damaged, root systems of different types of plants grown in vitro and under field conditions. The substrate into which plants were seeded was previously contaminated with different strains of Mycobacterium avium (108 to 1010 cells/g of soil) and feces from animals with paratuberculosis. We detected M. avium subsp. avium, hominissuis, and paratuberculosis in the stems and leaves of the plants by both culture and real-time quantitative PCR. The presence of mycobacteria in the plant tissues was confirmed by microscopy. The concentration of mycobacteria found inside plant tissue was several orders of magnitude lower (up to 104 cells/g of tissue) than the initial concentration of mycobacteria present in the culture medium or substrate. These findings led us to the hypothesis that plants may play a role in the spread and transmission of mycobacteria to other organisms in the environment. PMID:24747896

Lvoncik, S.; Slana, I.; Kulich, P.; Kralik, P.



Nanoparticle interactions with live cells: Quantitative fluorescence microscopy of nanoparticle size effects  

PubMed Central

Summary Engineered nanomaterials are known to enter human cells, often via active endocytosis. Mechanistic details of the interactions between nanoparticles (NPs) with cells are still not well enough understood. NP size is a key parameter that controls the endocytic mechanism and affects the cellular uptake yield. Therefore, we have systematically analyzed the cellular uptake of fluorescent NPs in the size range of 3.3–100 nm (diameter) by live cells. By using spinning disk confocal microscopy in combination with quantitative image analysis, we studied the time courses of NP association with the cell membrane and subsequent internalization. NPs with diameters of less than 10 nm were observed to accumulate at the plasma membrane before being internalized by the cells. In contrast, larger NPs (100 nm) were directly internalized without prior accumulation at the plasma membrane, regardless of their surface charges. We attribute this distinct size dependence to the requirement of a sufficiently strong local interaction of the NPs with the endocytic machinery in order to trigger the subsequent internalization. PMID:25551067

Shang, Li; Nienhaus, Karin; Jiang, Xiue; Yang, Linxiao; Landfester, Katharina; Mailänder, Volker; Simmet, Thomas



Modulation of hexokinase association with mitochondria analyzed with quantitative three-dimensional confocal microscopy  

PubMed Central

Hexokinase isozyme I is proposed to be associated with mitochondria in vivo. Moreover, it has been suggested that this association is modulated in coordination with changes in cell metabolic state. To test these hypotheses, we analyzed the subcellular distribution of hexokinase relative to mitochondria in paraformaldehyde-fixed astrocytes using immunocytochemistry and quantitative three-dimensional confocal microscopy. Analysis of the extent of colocalization between hexokinase and mitochondria revealed that approximately 70% of cellular hexokinase is associated with mitochondria under basal metabolic conditions. In contrast to the immunocytochemical studies, between 15 to 40% of cellular hexokinase was found to be associated with mitochondria after fractionation of astrocyte cultures depending on the exact fractionation conditions. The discrepancy between fractionation studies and those based on imaging of distributions in fixed cells indicates the usefulness of using techniques that can evaluate the distributions of "cytosolic" enzymes in cells whose subcellular ultrastructure is not severely disrupted. To determine if hexokinase distribution is modulated in concert with changes in cell metabolism, the localization of hexokinase with mitochondria was evaluated after inhibition of glucose metabolism with 2-deoxyglucose. After incubation with 2-deoxyglucose there was an approximate 35% decrease in the amount of hexokinase associated with mitochondria. These findings support the hypothesis that hexokinase is bound to mitochondria in rat brain astrocytes in vivo, and that this association is sensitive to cell metabolic state. PMID:1991789



ANG-2 for quantitative Na(+) determination in living cells by time-resolved fluorescence microscopy.  


Sodium ions (Na(+)) play an important role in a plethora of cellular processes, which are complex and partly still unexplored. For the investigation of these processes and quantification of intracellular Na(+) concentrations ([Na(+)]i), two-photon coupled fluorescence lifetime imaging microscopy (2P-FLIM) was performed in the salivary glands of the cockroach Periplaneta americana. For this, the novel Na(+)-sensitive fluorescent dye Asante NaTRIUM Green-2 (ANG-2) was evaluated, both in vitro and in situ. In this context, absorption coefficients, fluorescence quantum yields and 2P action cross-sections were determined for the first time. ANG-2 was 2P-excitable over a broad spectral range and displayed fluorescence in the visible spectral range. Although the fluorescence decay behaviour of ANG-2 was triexponential in vitro, its analysis indicates a Na(+)-sensitivity appropriate for recordings in living cells. The Na(+)-sensitivity was reduced in situ, but the biexponential fluorescence decay behaviour could be successfully analysed in terms of quantitative [Na(+)]i recordings. Thus, physiological 2P-FLIM measurements revealed a dopamine-induced [Na(+)]i rise in cockroach salivary gland cells, which was dependent on a Na(+)-K(+)-2Cl(-) cotransporter (NKCC) activity. It was concluded that ANG-2 is a promising new sodium indicator applicable for diverse biological systems. PMID:25311309

Roder, Phillip; Hille, Carsten



Backscattered Electron Microscopy as an Advanced Technique in Petrography.  

ERIC Educational Resources Information Center

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

Krinsley, David Henry; Manley, Curtis Robert



Materials characterization using high-resolution scanning-electron microscopy and x-ray microanalysis  

NASA Astrophysics Data System (ADS)

Low-voltage field-emission scanning-electron microscopy offers the possibility to characterize a wide range of materials. Electron optics in an electron beam column have improved in recent decades and now probe diameters of 1-10 nm can be obtained, allowing a wide range of applications to be explored. This article discusses the applications of low-voltage microscopy, including the characterization of nanoparticles, super-lattice structures, and carbon nanotubes.

Gauvin, Raynald; Robertson, Kevin; Horny, Paula; Elwazri, Adbelbaset M.; Yue, Steve



Cryo Electron Microscopy Reconstructions of the Leviviridae Unveil the Densest Icosahedral RNA Packing Possible  

Microsoft Academic Search

We solved the structures of the single-stranded RNA bacteriophages Q?, PP7 and AP205 by cryo-electron microscopy. On the outside, the symmetrized electron density maps resemble the previously described cryo-electron microscopy structure of MS2. RNA density is present inside the capsids, suggesting that the genomic RNA of Q?, PP7 and AP205, analogous to MS2, contains many coat protein-binding sites in addition

Roman I. Koning; Hans J. Warmenhoven; Henk K. Koerten; Jan van Duin



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


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

Downing, Kenneth H; Mooney, Paul E



An image intensifier for electron microscopy of polymers  

Microsoft Academic Search

A unique manipulator for a channel electron multiplier array (CEMA) is described. It allows for easy installation into the viewing chamber of a transmission electron microscope. This device, through a series of motions, positions the CEMA normal to the electron beam, thus amplifying the electron flux while preserving its image. This increase in image brightness permits operation of the microscope

P. W. Bratt; W. R. Even; S. H. Carr



Transmission electron microscopy analysis of a coated conductor produced by chemical deposition methods  

Microsoft Academic Search

A water-based precursor solution was used to deposit a thin La2Zr2O7 layer (70nm) on a NiW substrate. The characterization of the La2Zr2O7 (LZO) consisted of X-ray diffraction, pole figures, reflection high-energy electron diffraction, scanning electron microscopy, atomic force microscopy and transmission electron microscopy (TEM) of the cross-section. The results reveal that high-quality LZO layers can be produced using completely water-based

V. Cloet; T. Thersleff; O. Stadel; S. Hoste; B. Holzapfel; I. Van Driessche



A workflow for the automatic segmentation of organelles in electron microscopy image stacks  

PubMed Central

Electron microscopy (EM) facilitates analysis of the form, distribution, and functional status of key organelle systems in various pathological processes, including those associated with neurodegenerative disease. Such EM data often provide important new insights into the underlying disease mechanisms. The development of more accurate and efficient methods to quantify changes in subcellular microanatomy has already proven key to understanding the pathogenesis of Parkinson's and Alzheimer's diseases, as well as glaucoma. While our ability to acquire large volumes of 3D EM data is progressing rapidly, more advanced analysis tools are needed to assist in measuring precise three-dimensional morphologies of organelles within data sets that can include hundreds to thousands of whole cells. Although new imaging instrument throughputs can exceed teravoxels of data per day, image segmentation and analysis remain significant bottlenecks to achieving quantitative descriptions of whole cell structural organellomes. Here, we present a novel method for the automatic segmentation of organelles in 3D EM image stacks. Segmentations are generated using only 2D image information, making the method suitable for anisotropic imaging techniques such as serial block-face scanning electron microscopy (SBEM). Additionally, no assumptions about 3D organelle morphology are made, ensuring the method can be easily expanded to any number of structurally and functionally diverse organelles. Following the presentation of our algorithm, we validate its performance by assessing the segmentation accuracy of different organelle targets in an example SBEM dataset and demonstrate that it can be efficiently parallelized on supercomputing resources, resulting in a dramatic reduction in runtime. PMID:25426032

Perez, Alex J.; Seyedhosseini, Mojtaba; Deerinck, Thomas J.; Bushong, Eric A.; Panda, Satchidananda; Tasdizen, Tolga; Ellisman, Mark H.



Mass and molecular composition of vesicular stomatitis virus: a scanning transmission electron microscopy analysis.  

PubMed Central

Dark-field scanning transmission electron microscopy was used to perform mass analyses of purified vesicular stomatitis virions, pronase-treated virions, and nucleocapsids, leading to a complete self-consistent account of the molecular composition of vesicular stomatitis virus. The masses obtained were 265.6 +/- 13.3 megadaltons (MDa) for the native virion, 197.5 +/- 8.4 MDa for the pronase-treated virion, and 69.4 +/- 4.9 MDa for the nucleocapsid. The reduction in mass effected by pronase treatment, which corresponds to excision of the external domains (spikes) of G protein, leads to an average of 1,205 molecules of G protein per virion. The nucleocapsid mass, after compensation for the RNA (3.7 MDa) and residual amounts of other proteins, yielded a complement of 1,258 copies of N protein. Calibration of the amounts of M, NS, and L proteins relative to N protein by biochemical quantitation yielded values of 1,826, 466, and 50 molecules, respectively, per virion. Assuming that the remaining virion mass is contributed by lipids in the viral envelope, we obtained a value of 56.1 MDa for its lipid content. In addition, four different electron microscopy procedures were applied to determine the nucleocapsid length, which we conclude to be 3.5 to 3.7 micron. The nucleocapsid comprises a strand of repeating units which have a center-to-center spacing of 3.3 nm as measured along the middle of the strand. We show that these repeating units represent monomers of N protein, each of which is associated with 9 +/- 1 bases of single-stranded RNA. From scanning transmission electron microscopy images of negatively stained nucleocapsids, we inferred that N protein has a wedge-shaped, bilobed structure with dimensions of approximately 9.0 nm (length), approximately 5.0 nm (depth), and approximately 3.3 nm (width, at the midpoint of its long axis). In the coiled configuration of the in situ nucleocapsid, the long axis of N protein is directed radially, and its depth corresponds to the pitch of the nucleocapsid helix. Images PMID:2985822

Thomas, D; Newcomb, W W; Brown, J C; Wall, J S; Hainfeld, J F; Trus, B L; Steven, A C



Enhanced quantitative confocal microscopy and its application for the measurement of tympanic membrane thickness  

NASA Astrophysics Data System (ADS)

This work shows that confocal microscopy allows a quantitative study of delicate 3D-biotissue in fresh condition, thus avoiding histological preparation processes. The developed procedure results in exact and accurate thickness data for mum-sized objects with a measuring error of less than 1mum. It is, however, necessary to take into account the effect of focal shift in the case of refractive index mismatch to obtain such precise data. The use of the proposed method is advised instead of the use of a paraxial approximation for the axial scale correction because the method improves measurement precision by a factor of four. The axial scaling correction factors obtained in this work show that for most practical situations the correction cannot be ignored when one wants to obtain precise quantitative data. The thickness correction method can also be used to determine with high accuracy the index of refraction of biological tissue. The thickness measurement method was applied to fresh, untreated tympanic membranes of the gerbil, the cat and the human. Thickness had to be measured at many points as it differs strongly across the membrane. Similar thickness distributions were found in all pars tensas measured even across the species studied: (1) a very thin, central region with a rather constant thickness, curving as a horse shoe upwards around the manubrium (thickness: gerbil: about 7mum, cat: about 10mum, human: large inter-specimen variation: 40mum-120mum), (2) a thinnest zone at the inferior side, (3) a thicker zone at the supero-anterior side, (4) superior to the umbo, an anterior region thicker than the posterior region, (5) maximal thicknesses in a very small region near the entire manubrium and the entire annular periphery. The pars flaccida is found to be thicker than the pars tensa. It shows no central homogeneous zone: the thickness varies irregularly and very rapidly over short distances. Arbitrarily spaced bumps and notches are present over the entire pars flaccida surface. The thickness results advise against the use of single thickness values in mathematical models. The presented thickness and its spatial distribution can be introduced into middle ear computer models to further improve model realism.

Kuypers, Liesbeth



In Situ Transmission Electron Microscopy of Lead Dendrites and Lead Ions in Aqueous Solution  

PubMed Central

An ideal technique for observing nanoscale assembly would provide atomic-resolution images of both the products and the reactants in real time. Using a transmission electron microscope (TEM) we image in situ the electrochemical deposition of lead from an aqueous solution of lead(II) nitrate. Both the lead deposits and the local Pb2+ concentration can be visualized. Depending on the rate of potential change and the potential history, lead deposits on the cathode in a structurally compact layer or in dendrites. In both cases the deposits can be removed and the process repeated. Asperities that persist through many plating and stripping cycles consistently nucleate larger dendrites. Quantitative digital image analysis reveals excellent correlation between changes in the Pb2+ concentration, the rate of lead deposition, and the current passed by the electrochemical cell. Real-time electron microscopy of dendritic growth dynamics and the associated local ionic concentrations can provide new insight into the functional electrochemistry of batteries and related energy storage technologies. PMID:22702348

White, Edward R.; Singer, Scott B.; Augustyn, Veronica; Hubbard, William A.; Mecklenburg, Matthew; Dunn, Bruce; Regan, Brian C.



Practical factors affecting the performance of a thin-film phase plate for transmission electron microscopy  

PubMed Central

A number of practical issues must be addressed when using thin carbon films as quarter-wave plates for Zernike phase-contrast electron microscopy. We describe, for example, how we meet the more stringent requirements that must be satisfied for beam alignment in this imaging mode. In addition we address the concern that one might have regarding the loss of some of the scattered electrons as they pass through such a phase plate. We show that two easily measured parameters, (1) the low-resolution image contrast produced in cryo-EM images of tobacco mosaic virus particles and (2) the fall-off of the envelope function at high resolution, can be used to quantitatively compare the data quality for Zernike phase-contrast images and for defocused bright-field images. We describe how we prepare carbon-film phase plates that are initially free of charging or other effects that degrade image quality. We emphasize, however, that even though the buildup of hydrocarbon contamination can be avoided by heating the phase plates during use, their performance nevertheless deteriorates over the time scale of days to weeks, thus requiring their frequent replacement in order to maintain optimal performance. PMID:19157711

Danev, Radostin; Glaeser, Robert M.; Nagayama, Kuniaki



Boron concentration profiling by high angle annular dark field-scanning transmission electron microscopy in homoepitaxial ?-doped diamond layers  

SciTech Connect

To develop further diamond related devices, the concentration and spatial location of dopants should be controlled down to the nanometer scale. Scanning transmission electron microscopy using the high angle annular dark field mode is shown to be sensitive to boron doping in diamond epilayers. An analytical procedure is described, whereby local boron concentrations above 10{sup 20} cm{sup ?3} were quantitatively derived down to nanometer resolution from the signal dependence on thickness and boron content. Experimental boron local doping profiles measured on diamond p{sup ?}/p{sup ++}/p{sup ?} multilayers are compared to macroscopic profiles obtained by secondary ion mass spectrometry, avoiding reported artefacts.

Araújo, D.; Alegre, M. P.; Piñero, J. C. [Dpto Ciencia de los Materiales, Facultad de Ciencias, Universidad de Cádiz, 11510 Puerto Real (Cádiz) (Spain)] [Dpto Ciencia de los Materiales, Facultad de Ciencias, Universidad de Cádiz, 11510 Puerto Real (Cádiz) (Spain); Fiori, A.; Bustarret, E. [Institut Néel, CNRS-Université Joseph Fourier, 25 av. des Martyrs, 38042 Grenoble (France)] [Institut Néel, CNRS-Université Joseph Fourier, 25 av. des Martyrs, 38042 Grenoble (France); Jomard, F. [Groupe d'Etude de la Matière Condensée (GEMaC), UMR 8635 du CNRS, UVSQ, 45 av. des Etats-Unis, 78035 Versailles Cedex (France)] [Groupe d'Etude de la Matière Condensée (GEMaC), UMR 8635 du CNRS, UVSQ, 45 av. des Etats-Unis, 78035 Versailles Cedex (France)



Hierarchical super-structure identified by polarized light microscopy, electron microscopy and nanoindentation: Implications for the limits of biological control over the growth mode of abalone sea shells  

PubMed Central

Background Mollusc shells are commonly investigated using high-resolution imaging techniques based on cryo-fixation. Less detailed information is available regarding the light-optical properties. Sea shells of Haliotis pulcherina were embedded for polishing in defined orientations in order to investigate the interface between prismatic calcite and nacreous aragonite by standard materialographic methods. A polished thin section of the interface was prepared with a defined thickness of 60 ?m for quantitative birefringence analysis using polarized light and LC-PolScope microscopy. Scanning electron microscopy images were obtained for comparison. In order to study structural-mechanical relationships, nanoindentation experiments were performed. Results Incident light microscopy revealed a super-structure in semi-transparent regions of the polished cross-section under a defined angle. This super-structure is not visible in transmitted birefringence analysis due to the blurred polarization of small nacre platelets and numerous organic interfaces. The relative orientation and homogeneity of calcite prisms was directly identified, some of them with their optical axes exactly normal to the imaging plane. Co-oriented "prism colonies" were identified by polarized light analyses. The nacreous super-structure was also visualized by secondary electron imaging under defined angles. The domains of the super-structure were interpreted to consist of crystallographically aligned platelet stacks. Nanoindentation experiments showed that mechanical properties changed with the same periodicity as the domain size. Conclusions In this study, we have demonstrated that insights into the growth mechanisms of nacre can be obtained by conventional light-optical methods. For example, we observed super-structures formed by co-oriented nacre platelets as previously identified using X-ray Photo-electron Emission Microscopy (X-PEEM) [Gilbert et al., Journal of the American Chemical Society 2008, 130:17519–17527]. Polarized optical microscopy revealed unprecedented super-structures in the calcitic shell part. This bears, in principle, the potential for in vivo studies, which might be useful for investigating the growth modes of nacre and other shell types. PMID:22967319



Electron Microscopy of Interactions Between Engineered Nanomaterials and Cells  

E-print Network

like free-electron lasers and backward wave oscillators (BWOs). The third is the utilization of optical- rently available THz source technologies, particularly the tunable emitters, such as free-electron lasers

Pala, Nezih


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


Over a hundred years an atomistic point of view has been indispensable to explore fascinating properties of various materials and to develop novel functional materials. High-resolution microscopies, rapidly developed during the period, have taken central roles in promoting materials science and related techniques to observe and analyze the materials. As microscopies with the capability of atom-imaging, field ion microscopy (FIM), scanning tunneling microscopy (STM), atomic force microscopy (AFM) and transmission electron microscopy (TEM) can be cited, which have been highly evaluated as methods to ultimately bring forward the viewpoint of reductionism in materials science. On one hand, there have been difficulties to derive useful and practical information on large (micro) scale unique properties of materials using these excellent microscopies and to directly advance the engineering for practical materials. To make bridges over the gap between an atomic scale and an industrial engineering scale, we have to develop emergence science step-by-step as a discipline having hierarchical structures for future prospects by combining nanoscale and microscale techniques; as promising ways, the combined microscopic instruments covering the scale gap and the extremely sophisticated methods for sample preparation seem to be required. In addition, it is noted that spectroscopic and theoretical methods should implement the emergence science.Fundamentally, the function of microscope is to determine the spatial positions of a finite piece of material, that is, ultimately individual atoms, at an extremely high resolution with a high stability. To define and control the atomic positions, the STM and AFM as scanning probe microscopy (SPM) have successfully demonstrated their power; the technological heart of SPM lies in an atomically sharpened tip, which can be observed by FIM and TEM. For emergence science we would like to set sail using the tip as a base. Meanwhile, it is significant to extend a model sample prepared for the microscopies towards a microscale sample while keeping the intrinsic properties found by the microscopies.In this study we present our trial of developing microscopic combined instruments among FIM, field emission microscopy (FEM), STM, AFM and scanning electron microscopy (SEM), in which we prepared and characterized the tips for the SPM, and in addition, the sample preparation to take a correlation between nanoscale and microscale properties of functional materials. Recently, we developed a simple sample preparation method of a rutile single crystal TiO2 covered with an epitaxially-grown monolayer of SiO2 by annealing the crystals in a furnace at high temperatures in air; the crystal samples were placed into a quartz container in the furnace [1]. The vapor of SiO evaporated from the quartz container were adsorbed on the crystal while the crystal surfaces being fully oxidized in air. The SiO2-TiO2 composite systems are promising to protect catalytic TiO2 performance; the photo-catalytic activity is kept by coating with hard and stable SiO2 layers and to extend the lifetime of water super-hydrophilicity even in dark, though understanding of their properties is insufficient due to the lack of techniques to fabricate a well-characterized system on a nanoscale to conduct control experiments. The SiO2 overlayers were observed by low energy electron diffraction (LEED) in vacuum and frequency-modulation (FM) AFM in water [1,2], and water contact angles (WCA) were measured [2]. Although the WCA measurement seems a classic characterization, this method possesses a high potential to make a bridge by controlling the environmental conditions. We will discuss the details. PMID:25359799

Tomitori, Masahiko; Sasahara, Akira



Stereological characterization of the {gamma}' particles in a nickel base superalloy: Comparison between transmission electron microscopy and atomic force microscopy techniques  

SciTech Connect

Critical comparison of transmission electron microscopy and atomic force microscopy techniques was provided concerning size measurements of {gamma}' precipitates in a nickel-base superalloy. The divergence between results is explained in terms of the resolution limit for atomic force microscopy, linked both to the tip dimension and the diameter of the investigated particles.

Risbet, M. [Universite de Technologie de Compiegne, Laboratoire Roberval, 60205 Compiegne Cedex (France)], E-mail:; Feaugas, X. [Universite de la Rochelle, L.E.M.M.A., 17042 La Rochelle cedex 01 (France); Guillemer-Neel, C. [Universite Picardie Jules Verne, L.T.R.M.C., 80025 Amiens (France); Clavel, M. [Ecole Centrale Paris, M.S.S.Mat., 92295 Chatenay-Malabry (France)



FEATURE ARTICLE Transmission Electron Microscopy of Shape-Controlled Nanocrystals and Their Assemblies  

E-print Network

is the quantum confinement effect in small size metallic and semiconductor quantum dots.4,5 The shift of electron the electronic structure, bonding, and possibly chemical reactivities. The sublimation activation energy of AuFEATURE ARTICLE Transmission Electron Microscopy of Shape-Controlled Nanocrystals

Wang, Zhong L.


Outlook of application of aberration corrected-electron microscopy in the ligand-protected metal clusters  

E-print Network

Outlook of application of aberration corrected-electron microscopy in the ligand- protected metal microscopy(HREM) and high angle annular dark field (HAADF) -scanning transmission EM (STEM) imaging clusters has been observed. For example, both Fresnel fringes around the particles and strong phase

Frenkel, Anatoly


Plasmolysis of Pteridium protoplasts: A study using light and scanning-electron microscopy  

Microsoft Academic Search

A study was undertaken using gametophytes of the fern Pteridium aquilinum to examine the effects of plasmolysis on the topography of protoplasts. Methods are described whereby the surfaces of non-isolated protoplasts can be observed in the plasmolysed condition using scanning electron microscopy. Plasmolysed gametophytes were also examined in the light microscope using differential interference contrast and ultra-violet fluorescence microscopy after

S. M. Attree; E. Sheffield



Domain and Microstructural Characterization of Ferroelectric Materials Using Transmission Electron Microscopy  

Microsoft Academic Search

Ferroelectric materials exhibit many intriguing properties, and their microstructures play very important roles. In this article, domain formation, microstructural features and domain dynamics in ferroelectric materials were investigated using optical microscopy and electron microscopy. Domain arrangements and their characteristics significantly influence their polarization switchability and therefore electrical properties. 90° domain boundaries inclined or perpendicular to applied electric field may dissociate

Chen-Chia Chou; Cheng-Sao Chen; Tsung-Heryeh



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


In the following article an electron/ion microscopy study will be presented which investigates the uptake of silver nanoparticles (AgNPs) by the marine diatom Thalassiosira pseudonana, a primary producer aquatic species. This organism has a characteristic silica exoskeleton that may represent a barrier for the uptake of some chemical pollutants, including nanoparticles (NPs), but that presents a technical challenge when attempting to use electron-microscopy (EM) methods to study NP uptake. Here we present a convenient method to detect the NPs interacting with the diatom cell. It is based on a fixation procedure involving critical point drying which, without prior slicing of the cell, allows its inspection using transmission electron microscopy. Employing a combination of electron and ion microscopy techniques to selectively cut the cell where the NPs were detected, we are able to demonstrate and visualize for the first time the presence of AgNPs inside the cell membrane. PMID:24797958

García, César Pascual; Burchardt, Alina D; Carvalho, Raquel N; Gilliland, Douglas; António, Diana C; Rossi, François; Lettieri, Teresa



Engineered ascorbate peroxidase as a genetically encoded reporter for electron microscopy  

E-print Network

Electron microscopy (EM) is the standard method for imaging cellular structures with nanometer resolution, but existing genetic tags are inactive in most cellular compartments[superscript 1] or require light and can be ...

Deerinck, Thomas J


Development of multiplexing strategies for electron and super-resolution optical microscopy/  

E-print Network

The aim of this work is to increase the multiplexing capabilities of electron and super resolution optical microscopy. This will be done through the development of molecular-scale barcodes that can be resolved in one of ...

Tillberg, Paul W



Surface-enhanced Raman spectroscopy and correlated scanning electron microscopy of individual carbon nanotubes  

E-print Network

Surface-enhanced Raman spectroscopy and correlated scanning electron microscopy of individual, which generally limits its potential uses. Surface-enhanced Raman spectroscopy SERS can be used before and after depositing silver nanoparticles. Local regions exhibiting surface enhanced Raman

Cronin, Steve


Segmentation and registration of molecular components in 3-dimensional density maps from cryo-electron microscopy  

E-print Network

Cryo-electron microscopy is a method that produces 3D density maps of macromolecular complexes. Segmentation and registration methods are heavily used to extract structural information from such density maps. Segmentation ...

Pintilie, Grigore Dimitrie, 1976-



Normal-Incidence Photoemission Electron Microscopy (NI-PEEM) for Imaging Surface Plasmon Polaritons  

E-print Network

Philip Kahl & Simone Wall & Christian Witt & Christian Schneider & Daniela Bayer & Alexander Fischer across a metal surface. Keywords Surface plasmon polariton . Nonlinear photoemission microscopy . Normal of the free electron gas in the metal and an electric and magnetic field

Aeschlimann, Martin



EPA Science Inventory

Concerns about the environmental and public health effects of particulate matter (PM) have stimulated interest in analytical techniques capable of measuring the size and chemical composition of individual aerosol particles. Computer-controlled scanning electron microscopy (CCSE...


Detection of Silver Nanoparticles inside Marine Diatom Thalassiosira pseudonana by Electron Microscopy and Focused Ion Beam  

PubMed Central

In the following article an electron/ion microscopy study will be presented which investigates the uptake of silver nanoparticles (AgNPs) by the marine diatom Thalassiosira pseudonana, a primary producer aquatic species. This organism has a characteristic silica exoskeleton that may represent a barrier for the uptake of some chemical pollutants, including nanoparticles (NPs), but that presents a technical challenge when attempting to use electron-microscopy (EM) methods to study NP uptake. Here we present a convenient method to detect the NPs interacting with the diatom cell. It is based on a fixation procedure involving critical point drying which, without prior slicing of the cell, allows its inspection using transmission electron microscopy. Employing a combination of electron and ion microscopy techniques to selectively cut the cell where the NPs were detected, we are able to demonstrate and visualize for the first time the presence of AgNPs inside the cell membrane. PMID:24797958

Pascual García, César; Burchardt, Alina D.; Carvalho, Raquel N.; Gilliland, Douglas; C. António, Diana; Rossi, François; Lettieri, Teresa



Diffusive and inelastic scattering in ballistic-electron-emission spectroscopy and ballistic-electron-emission microscopy  

SciTech Connect

Ballistic-electron-emission microscopy (BEEM) of Au/Si(001) n type was done to study whether elastic scattering in the Au overlayer is dominant. It was found that there is no dependence of the BEEM current on the relative gradient of the Au surface with respect to the Si interface, and this demonstrates that significant elastic scattering must occur in the Au overlayer. Ballistic-electron-emission spectroscopy (BEES) was also done, and, rather than using the conventional direct-current BEES, alternating-current (ac) BEES was done on Au/Si and also on Au/PtSi/Si(001) n type. The technique of ac BEES was found to give linear threshold for the Schottky barrier, and it also clearly showed the onset of electron-hole pair creation and other inelastic scattering events. The study of device quality PtSi in Au/PtSi/Si(001) yielded an attenuation length of 4 nm for electrons of energy 1 eV above the PtSi Fermi energy. 20 refs., 5 figs.

Lee, E.Y.; Turner, B.R.; Schowalter, L.J. [Rensselaer Polytechnic Institute, Troy, NY (United States)] [and others] [Rensselaer Polytechnic Institute, Troy, NY (United States); and others



Structure and magnetism in ultrathin iron oxides characterized by low energy electron microscopy.  


We have grown epitaxial films a few atomic layers thick of iron oxides on ruthenium. We characterize the growth by low energy electron microscopy. Using selected-area diffraction and intensity-versus-voltage spectroscopy, we detect two distinct phases which are assigned as wüstite and magnetite. Spin-polarized low energy electron microscopy reveals magnetic domain patterns in the magnetite phase at room temperature. PMID:21828572

Santos, B; Loginova, E; Mascaraque, A; Schmid, A K; McCarty, K F; de la Figuera, J



High yield production of long branched Au nanoparticles characterized by atomic resolution transmission electron microscopy.  


Long multi-branched gold nanoparticles have been synthesized in a very high yield through a facile synthesis combining two different capping agents. The stability of these materials with the time has been tested and their characterization have been performed by diverse advanced electron microscopy techniques, paying special attention to aberration corrected transmission electron microscopy in order to unambiguously analyze the surface structure of the branches and provide insights for the formation of stellated gold nanoparticles. PMID:22125420

Mayoral, Alvaro; Magen, Cesar; Jose-Yacaman, Miguel



High Resolution Electron Microscopy at the National Cancer Institute

The long-term mission of our research program is to obtain an integrated, quantitative understanding of cells and viruses at molecular resolution. We take an interdisciplinary approach to this problem by combining novel technologies for 3D imaging with computational and cell biological tools.


Logic upsets in SRAMs using ion electron emission microscopy  

Microsoft Academic Search

A high linear energy transfer (LET) system for performing radiation effects microscopy (REM) has been developed at Sandia National Laboratories in Albuquerque. A radio frequency quadrupole (RFQ) linac is used to boost the energy of ions accelerated by a conventional Tandem Van de Graaff - Pelletron from 0.25 to 1.9 MeV\\/amu. The LET for heavy ions is near a maximum

B. L. Doyle; F. D. McDaniel; G. Vizkelethy; P. E. Dodd; P. Rossi; D. S. Walsh



Logic upsets in SRAMs using ion electron emission microscopy  

Microsoft Academic Search

A high linear energy transfer (LET) system for performing radiation effects microscopy (REM) has been developed at Sandia National Laboratories in Albuquerque. A radio frequency quadrupole (RFQ) linac is used to boost the energy of ions accelerated by a conventional Tandem Van de Graaff – Pelletron from 0.25 to 1.9 MeV\\/amu. The LET for heavy ions is near a maximum

B. L Doyle; F. D McDaniel; G Vizkelethy; P. E Dodd; P Rossi; D. S Walsh



High Resolution Electron Microscopy at the National Cancer Institute

The development of new technology and new methods has been central to our lab’s mission. One key development has been that of a complete framework for alignment, classification, and averaging of volumes derived by electron tomography that is computationally efficient and effectively accounts for the missing wedge that is inherent to limited angle electron tomography.


Transmission electron microscopy, scanning tunneling microscopy, and atomic force microscopy of the cell envelope layers of the archaeobacterium Methanospirillum hungatei GP1.  

PubMed Central

Methanospirillum hungatei GP1 possesses paracrystalline cell envelope components including end plugs and a sheath formed from stacked hoops. Both negative-stain transmission electron microscopy (TEM) and scanning tunneling microscopy (STM) distinguished the 2.8-nm repeat on the outer surface of the sheath, while negative-stain TEM alone demonstrated this repeat around the outer circumference of individual hoops. Thin sections revealed a wave-like outer sheath surface, while STM showed the presence of deep grooves that precisely defined the hoop-to-hoop boundaries at the waveform nodes. Atomic force microscopy of sheath tubes containing entrapped end plugs emphasized the end plug structure, suggesting that the sheath was malleable enough to collapse over the end plugs and deform to mimic the shape of the underlying structure. High-resolution atomic force microscopy has revised the former idea of end plug structure so that we believe each plug consists of at least four discs, each of which is approximately 3.5 nm thick. PT shadow TEM and STM both demonstrated the 14-nm hexagonal, particulate surface of an end plug, and STM showed the constituent particles to be lobed structures with numerous smaller projections, presumably corresponding to the molecular folding of the particle. Images PMID:8458836

Southam, G; Firtel, M; Blackford, B L; Jericho, M H; Xu, W; Mulhern, P J; Beveridge, T J



Extracellular vesicles release by cardiac telocytes: electron microscopy and electron tomography  

PubMed Central

Telocytes have been reported to play an important role in long-distance heterocellular communication in normal and diseased heart, both through direct contact (atypical junctions), as well as by releasing extracellular vesicles (EVs) which may act as paracrine mediators. Exosomes and ectosomes are the two main types of EVs, as classified by size and the mechanism of biogenesis. Using electron microscopy (EM) and electron tomography (ET) we have found that telocytes in culture release at least three types of EVs: exosomes (released from endosomes; 45 ± 8 nm), ectosomes (which bud directly from the plasma membrane; 128 ± 28 nm) and multivesicular cargos (MVC; 1 ± 0.4 ?m), the latter containing tightly packaged endomembrane-bound vesicles (145 ± 35 nm). Electron tomography revealed that endomembrane vesicles are released into the extracellular space as a cargo enclosed by plasma membranes (estimated area of up to 3 ?m2). This new type of EV, also released by telocytes in tissue, likely represents an essential component in the paracrine secretion of telocytes and may consequently be directly involved in heart physiology and regeneration. PMID:25257228

Fertig, Emanuel T; Gherghiceanu, Mihaela; Popescu, Laurentiu M



Three-Dimensional Electron Microscopy Simulation with the CASINO Monte Carlo Software  

PubMed Central

Monte Carlo softwares are widely used to understand the capabilities of electron microscopes. To study more realistic applications with complex samples, 3D Monte Carlo softwares are needed. In this paper, the development of the 3D version of CASINO is presented. The software feature a graphical user interface, an efficient (in relation to simulation time and memory use) 3D simulation model, accurate physic models for electron microscopy applications, and it is available freely to the scientific community at this website: It can be used to model backscattered, secondary, and transmitted electron signals as well as absorbed energy. The software features like scan points and shot noise allow the simulation and study of realistic experimental conditions. This software has an improved energy range for scanning electron microscopy and scanning transmission electron microscopy applications. PMID:21769885

Demers, Hendrix; Poirier-Demers, Nicolas; Couture, Alexandre Réal; Joly, Dany; Guilmain, Marc; de Jonge, Niels; Drouin, Dominique



3D Quantitative Confocal Laser Microscopy of Ilmenite Volume Distribution in Alpe Arami Olivine  

NASA Astrophysics Data System (ADS)

The deep origin of the Alpe Arami garnet lherzolite massif in the Swiss Alps proposed by Dobrzhinetskaya et al. (Science, 1996) has been a focus of heated debate. One of the lines of evidence supporting an exhumation from more than 200 km depth includes the abundance, distribution, and orientation of magnesian ilmenite rods in the oldest generation of olivine. This argument has been disputed in terms of the abundance of ilmenite and consequently the maximum TiO2 content in the discussed olivine. In order to address this issue, we have directly measured the volume fraction of ilmenite of the oldest generation of olivine by applying confocal laser scanning microscopy (CLSM). CLSM is a method which allows for three-dimensional imaging and quantitative volume determination by optical sectioning of the objects. The images for 3D reconstruction and measurements were acquired from petrographic thin sections in reflected laser light with 488 nm wavelength. Measurements of more than 80 olivine grains in six thin sections of our material yielded an average volume fraction of 0.31% ilmenite in the oldest generation of olivine from Alpe Arami. This translates into 0.23 wt.% TiO2 in olivine with error in determination of ±0.097 wt.%, a value significantly different from that of 0.02 to 0.03 wt.% TiO2 determined by Hacker et al. (Science, 1997) by a broad-beam microanalysis technique. During the complex geological history of the Alpe Arami massif, several events of metamorphism are recorded which all could have caused increased mobility of the mineral components. Evidence for loss of TiO2 from olivine is the tendency for high densities of ilmenite to be restricted to cores of old grains, the complete absence of ilmenite inclusions from the younger, recrystallized, generation of olivine, and reduction in ilmenite size and abundance in more serpentinized specimens. These observations suggest that only olivine grains with the highest concentrations of ilmenite are close to the original amount of TiO2 incorporated in the olivine. Our measurements show maximum volume fraction of ilmenite of 1.21%, corresponding to 0.9±0.38 wt.% of TiO2. Even the most conservative scenario reveals concentration of TiO2 in olivine of more than 0.5 wt.%, a value comparable to that reported by Dobrzhinetskaya et al. (1996) and an order of magnitude greater than any previous measurement of TiO2 in olivine. Experiments by Dobrzhinetskaya et al. (Chem. Geol, 2000) found that such high solubility can occur, but only at P>10GPa at mantle temperatures.

Bozhilov, K. N.



A quantum mechanical scheme to reduce radiation damage in electron microscopy  

SciTech Connect

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

Okamoto, Hiroshi; Latychevskaia, Tatiana; Fink, Hans-Werner [Institute of Physics, University of Zurich, Winterthurerstrasse 190, Zurich CH-8057 (Switzerland)



Comparative morphology of zebra (Dreissena polymorpha) and quagga (Dreissena bugensis) mussel sperm: Light and electron microscopy  

USGS Publications Warehouse

Adult zebra (Dreissena polymorpha) and quagga (Dreissena bugensis) mussels were induced to release large quantities of live spermatozoa by the administration of 5-hydroxytryptamine (serotonin). Sperm were photographed alive using phase-contrast microscopy and were fixed subsequently with glutaraldehyde followed by osmium tetroxide for eventual examination by transmission or scanning electron microscopy. The sperm of both genera are of the ect-aquasperm type. Their overall dimensions and shape allow for easy discrimination at the light and scanning electron microscopy level. Transmission electron microscopy of the cells reveals a barrel-shaped nucleus in zebra mussel sperm and an elongated nucleus in quagga mussel sperm. In both species, an acrosome is cradled in a nuclear fossa. The ultrastructure of the acrosome and axial body, however, is distinctive for each species. The structures of the midpiece are shown, including a unique mitochondrial 'skirt' that includes densely packed parallel cristae and extends in a narrow sheet from the mitochondria.

Walker, G.K.; Black, M.G.; Edwards, C.A.



Electron microscopy: a brief history and review of current clinical application.  


This chapter describes the historic development of techniques that has made it possible to use electron microscopy, principally transmission electron microscopy, for diagnostic purposes. It was described how the standard techniques for preparing tissue for light microscopy had been modified to make it possible to view the ultrastructural components of a cell, tissue, or organ that cannot be resolved with a light microscope. There is a discussion of the types of tissues and cells that were and are currently observed by electron microscopy for diagnostic purposes. All of the materials that are used in tissue preparation and the general protocols for processing the tissues are also included. There are also notes which describe steps that can be changed or modified and why depending on conditions and anticipated outcome. PMID:25015145

Gordon, Ronald E



Theoretical study of ferroelectric nanoparticles using phase reconstructed electron microscopy  

NASA Astrophysics Data System (ADS)

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

Phatak, C.; Petford-Long, A. K.; Beleggia, M.; De Graef, M.



Materials Advances through Aberration-Corrected Electron Microscopy  

SciTech Connect

Over the last few years, the performance of electron microscopes has undergone a dramatic improvement, with achievable resolution having more than doubled. It is now possible to probe individual atomic sites in many materials and to determine atomic and electronic structure with single-atom sensitivity. This revolution has been enabled by the successful correction of the dominant aberrations present in electron lenses. In this review, the authors present a brief overview of these instrumental advances, emphasizing the new insights they provide to several areas of materials research.

Pennycook, Stephen J [ORNL; Varela del Arco, Maria [ORNL; Hetherington, Crispin J. D. [University of Oxford; Kirkland, Angus I. [University of Oxford



Nanopaleomagnetism of meteoritic Fe-Ni studied using X-ray photoemission electron microscopy  

NASA Astrophysics Data System (ADS)

X-ray photoemission electron microscopy (XPEEM) enables natural remanent magnetisation to be imaged with ˜30 nm resolution across a field of view of 5-20 ?m. The method is applied to structural features typical of the Widmanstätten microstructure (kamacite - tetrataenite rim - cloudy zone - plessite) in the Tazewell IIICD iron meteorite. Kamacite lamellae and the tetrataenite rim are multidomain, whereas plessite consists of laths of different phases displaying a range of stable magnetisation directions. The cloudy zone (CZ) displays a complex interlocking domain pattern resulting from nanoscale islands of tetrataenite with easy axes distributed along three possible crystallographic directions. Quantitative analysis of the coarse and intermediate CZ was achieved using a combination of image simulations and histogram profile matching. Remanence information was extracted from individual regions of interest ˜400 nm wide, demonstrating for the first time the capability of XPEEM to perform quantitative paleomagnetic analysis at sub-micron length scales. The three tetrataenite easy axis orientations occur with equal probability in the coarse and intermediate CZ, suggesting that spinodal decomposition in these regions was not strongly influenced by internal interaction fields, and that they are suitable candidates for future paleomagnetic studies. The fine CZ shows a strong dominance of one easy axis. This effect is attributed to island-island exchange interactions that render the fine CZ unsuitable for paleomagnetic study. Variations in the relative strength (proportion of dominant easy axis) and direction (direction of dominant easy axis) of a paleomagnetic field can be resolved from different regions of the CZ using XPEEM, raising the prospect of obtaining a time-resolved measurement of the active dynamo period in meteorites originating from the upper unmelted regions of differentiated asteroids (e.g. chondrites, pallasites, mesosiderites).

Bryson, James F. J.; Herrero-Albillos, Julia; Kronast, Florian; Ghidini, Massimo; Redfern, Simon A. T.; van der Laan, Gerrit; Harrison, Richard J.



Developing a denoising filter for electron microscopy and tomography data in the cloud.  


The low radiation conditions and the predominantly phase-object image formation of cryo-electron microscopy (cryo-EM) result in extremely high noise levels and low contrast in the recorded micrographs. The process of single particle or tomographic 3D reconstruction does not completely eliminate this noise and is even capable of introducing new sources of noise during alignment or when correcting for instrument parameters. The recently developed Digital Paths Supervised Variance (DPSV) denoising filter uses local variance information to control regional noise in a robust and adaptive manner. The performance of the DPSV filter was evaluated in this review qualitatively and quantitatively using simulated and experimental data from cryo-EM and tomography in two and three dimensions. We also assessed the benefit of filtering experimental reconstructions for visualization purposes and for enhancing the accuracy of feature detection. The DPSV filter eliminates high-frequency noise artifacts (density gaps), which would normally preclude the accurate segmentation of tomography reconstructions or the detection of alpha-helices in single-particle reconstructions. This collaborative software development project was carried out entirely by virtual interactions among the authors using publicly available development and file sharing tools. PMID:23066432

Starosolski, Zbigniew; Szczepanski, Marek; Wahle, Manuel; Rusu, Mirabela; Wriggers, Willy



Combination of imaging mass spectrometry and electron microscopy for quasi nondestructive surface analysis  

NASA Astrophysics Data System (ADS)

We report on a combination of imaging mass spectrometry (MS) and scanning electron microscopy (SEM) developed in a custom designed time-of-flight (TOF) MS instrument with laser post-ionization of sputtered atoms. Elemental (by MS) and topographical (by SEM) mapping of surfaces of heavily contaminated Si collectors from the NASA Genesis sample return mission enabled obtaining much more accurate and detailed depth distribution of the Solar Wind Mg and Ca implanted in these collectors. This is because the cleanest areas were identified by the SEM/MS mapping, and high resolution sputter depth profiling at these locations revealed near-surface (0-15 nm) depth distribution of Mg and Ca, that were used for more accurate fluence calculations of these Solar Wind species. MS imaging was virtually nondestructive at primary ion fluence 1012 cm-2, causing no effect on accuracy and precision of quantitative depth profiling that followed the imaging. We also demonstrate importance of such an approach by directly comparing high resolution depth profiles measured on clean areas versus arbitrarily selected areas.

Baryshev, S. V.; Zinovev, A. V.; Tripa, C. E.; Veryovkin, I. V.



Analysis of the phiX DNA replication cycle by electron microscopy.  


We have monitored the development of intracellular phiX DNA forms during the course of a virus life cycle that duplicates as closely as possible the normal infection of individual cells by single virions. The viral DNA was isolated in a one-step purification procedure, and quantitative electron microscopy was performed on the samples, resulting in the following conclusions: (i) Early in the life cycle, when the cells accumulate duplex rings, two types of DNA replication intermediates are observed: a rolling circle with a single-stranded tail; and a novel form, a single-stranded circle that is partially duplex. Thus, duplex ring synthesis appears to occur in two asymmetric steps, with positive strand DNA first being processed from the tail of the rolling circle and circularized, before it acts as a template for negative strand synthesis. (ii) Late in the life cycle, as single-stranded circles are synthesized and virus particles are assembled, only one replicating intermediate is observed--the rolling circle with a single-stranded tail. At this stage, the number of rolling circles reaches a level of about 35 per cell. (iii) The net rate of polymerization in the rolling circle intermediates is about 200 nucleotides per sec. PMID:273222

Koths, K; Dressler, D



Quantification of metallic nanoparticle morphology with tilt series imaging by transmission electron microscopy  

NASA Astrophysics Data System (ADS)

We report on the quantitative analysis of electrolessly deposited Au and Ag nanoparticles (NPs) on SU8 polymer with the help of High-Angle Annular Dark-Field Scanning Transmission Electron Microscopy (HAADF-STEM) in tilt series. Au NPs act as nucleating agents for the electroless deposition of silver. Au NPs were prepared by attachingAu^3+cations to amine functionalized SU8 polymeric surfaces and then reducing it with aqueous NaBH4. The nanoscale morphology of the deposited NPs on the surface of polymer has been studied from the dark field TEM cross sectional images. Ag NPs were deposited on the cross-linked polymeric surface from a silver citrate solution reduced by hydroquinone. HAADF-STEM enables us to determine the distances between the NPs and their exact locations at and near the surface. The particle distribution, sizes and densities provide us with the data necessary to control the parameters for the development of the electroless deposition technique for emerging nanoscale technologies.

Dutta, Aniruddha; Yuan, Biao; Clukay, Christopher J.; Grabill, Christopher N.; Heinrich, Helge; Bhattacharya, Aniket; Kuebler, Stephen M.



Developing a denoising filter for electron microscopy and tomography data in the cloud  

PubMed Central

The low radiation conditions and the predominantly phase-object image formation of cryo-electron microscopy (cryo-EM) result in extremely high noise levels and low contrast in the recorded micrographs. The process of single particle or tomographic 3D reconstruction does not completely eliminate this noise and is even capable of introducing new sources of noise during alignment or when correcting for instrument parameters. The recently developed Digital Paths Supervised Variance (DPSV) denoising filter uses local variance information to control regional noise in a robust and adaptive manner. The performance of the DPSV filter was evaluated in this review qualitatively and quantitatively using simulated and experimental data from cryo-EM and tomography in two and three dimensions. We also assessed the benefit of filtering experimental reconstructions for visualization purposes and for enhancing the accuracy of feature detection. The DPSV filter eliminates high-frequency noise artifacts (density gaps), which would normally preclude the accurate segmentation of tomography reconstructions or the detection of alpha-helices in single-particle reconstructions. This collaborative software development project was carried out entirely by virtual interactions among the authors using publicly available development and file sharing tools. PMID:23066432

Starosolski, Zbigniew; Szczepanski, Marek; Wahle, Manuel; Rusu, Mirabela



Transmission electron microscopy characterization of Zircaloy-4 and ZIRLO™ oxide layers  

NASA Astrophysics Data System (ADS)

Waterside corrosion of zirconium alloy nuclear fuel cladding varies markedly from one alloy to another. In addition, for a given alloy, the corrosion rate evolves during the corrosion process, most notably when the oxide loses its stability at the oxide transition. In an effort to understand the mechanism resulting in the variations of corrosion rate observed at the oxide transition, oxide layers formed on Zircaloy-4 and ZIRLO™ in high temperature water autoclave environments, and archived before and after the transition, are characterized using transmission electron microscopy. The study characterizes and compares the oxide morphology in both alloys at different times during the corrosion process, in an effort to understand the oxide growth mechanism for these alloys. Results show that the oxide is mainly composed of monoclinic ZrO2 Hydrides are also observed in that region, with a definite orientation relationship with the matrix. The observations of the oxide/metal interface are qualitatively similar for the two alloys but quantitatively different. The incorporation of intermetallic precipitates into the oxide layer is also studied, and compared between the two alloys. These results are discussed in terms of previous observations and of current models of oxide growth.

de Gabory, Benoit; Motta, Arthur T.; Wang, Ke



Electron microscopy of nano-inclusion formation in (Ga,Mn)As magnetic semiconductors  

E-print Network

corrected high-resolution TEM images and nano-beam diffraction patterns from individual precipitatesElectron microscopy of nano-inclusion formation in (Ga,Mn)As magnetic semiconductors A. Kovács1 , T for Electron Nanoscopy, Technical University of Denmark, Kgs. Lyngby 2800, Denmark 2 MAX-Lab, Lund University

Dunin-Borkowski, Rafal E.