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Sample records for microscopy electron scanning

  1. Scanning ultrafast electron microscopy.

    PubMed

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

    2010-08-24

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

  2. Scanning ultrafast electron microscopy

    PubMed Central

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

    2010-01-01

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

  3. Environmental scanning electron microscopy in cell biology.

    PubMed

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

    2013-01-01

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

  4. Scanning Electron Microscopy Sample Preparation and Imaging.

    PubMed

    Nguyen, Jenny Ngoc Tran; Harbison, Amanda M

    2017-01-01

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

  5. Scanning electron microscopy study of Trichomonas gallinae.

    PubMed

    Tasca, Tiana; De Carli, Geraldo A

    2003-12-01

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

  6. Feature Adaptive Sampling for Scanning Electron Microscopy

    PubMed Central

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

    2016-01-01

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

  7. Scanning electron microscopy of superficial white onychomycosis*

    PubMed Central

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

    2015-01-01

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

  8. Aberration corrected Lorentz scanning transmission electron microscopy.

    PubMed

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

    2015-05-01

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

  9. Immunogold Labeling for Scanning Electron Microscopy.

    PubMed

    Goldberg, Martin W; Fišerová, Jindřiška

    2016-01-01

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

  10. Immunogold labelling for scanning electron microscopy.

    PubMed

    Goldberg, Martin W; Fiserova, Jindriska

    2010-01-01

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

  11. Scanning electron microscopy studies of bacterial cultures

    NASA Astrophysics Data System (ADS)

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

    2012-02-01

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

  12. Scanning Electron Microscopy of the Presbylarynx.

    PubMed

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

    2016-06-01

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

  13. Scanning electron microscopy study of Tritrichomonas augusta.

    PubMed

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

    2004-09-01

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

  14. Phase-contrast scanning transmission electron microscopy.

    PubMed

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

    2015-06-01

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

  15. Scanning electron microscopy of lichen sclerosus*

    PubMed Central

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

    2013-01-01

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

  16. Correlative photoactivated localization and scanning electron microscopy.

    PubMed

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

    2013-01-01

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

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

    SciTech Connect

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

    2016-07-25

    Abstract

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

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

    PubMed

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

    2016-08-01

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

  19. Electric fields in Scanning Electron Microscopy simulations

    NASA Astrophysics Data System (ADS)

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

    2016-03-01

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

  20. Scanning electron microscopy of tinea nigra.

    PubMed

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

    2014-01-01

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

  1. Scanning electron microscopy of molluscum contagiosum*

    PubMed Central

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

    2013-01-01

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

  2. Hexamethyldisilazane for scanning electron microscopy of Gastrotricha.

    PubMed

    Hochberg, R; Litvaitis, M K

    2000-01-01

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

  3. [Scanning electron microscopy of Paragonimus proliferus].

    PubMed

    Zhou, Ben-jiang

    2005-10-30

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

  4. Scanning electron microscopy of softened enamel.

    PubMed

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

    2004-01-01

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

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

    PubMed

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

    2008-01-01

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

  6. Optical microscopy versus scanning electron microscopy in urolithiasis.

    PubMed

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

    2009-10-01

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

  7. Scanning Transmission Electron Microscopy at High Resolution

    PubMed Central

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

    1974-01-01

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

  8. Atmospheric pressure scanning transmission electron microscopy.

    PubMed

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

    2010-03-10

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

  9. Preparation of nematodes for scanning electron microscopy.

    PubMed

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

    1975-01-01

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

  10. Scanning electron microscopy of bacteria Tetrasphaera duodecadis.

    PubMed

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

    2014-01-01

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

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

    PubMed

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

    2014-10-07

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

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

    PubMed

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

    2017-08-01

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

  13. Image Resolution in Scanning Transmission Electron Microscopy

    SciTech Connect

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

    2008-06-26

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

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

    PubMed Central

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

    1969-01-01

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

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

    PubMed

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

    2015-02-04

    Silica nanoparticles are imaged in solution with scanning transmission electron microscopy (STEM) using a liquid cell with silicon nitride (SiN) membrane windows. The STEM images reveal that silica structures are deposited in well-defined patches on the upper SiN membranes upon electron beam irradiation. The thickness of the deposits is linear with the applied electron dose. Scanning electron microscopy (SEM) and atomic force microscopy (AFM) demonstrate that the deposited patches are a result of the merging of the original 20 nm-diameter nanoparticles, and that the related surface roughness depends on the electron dose rate used. Using this approach, sub-micrometer scale structures are written on the SiN in liquid by controlling the electron exposure as function of the lateral position. © 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

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

    PubMed

    Abraham, J L

    1979-08-01

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

  17. System and method for compressive scanning electron microscopy

    DOEpatents

    Reed, Bryan W

    2015-01-13

    A scanning transmission electron microscopy (STEM) system is disclosed. The system may make use of an electron beam scanning system configured to generate a plurality of electron beam scans over substantially an entire sample, with each scan varying in electron-illumination intensity over a course of the scan. A signal acquisition system may be used for obtaining at least one of an image, a diffraction pattern, or a spectrum from the scans, the image, diffraction pattern, or spectrum representing only information from at least one of a select subplurality or linear combination of all pixel locations comprising the image. A dataset may be produced from the information. A subsystem may be used for mathematically analyzing the dataset to predict actual information that would have been produced by each pixel location of the image.

  18. Ultra-low voltage scanning electron microscopy

    SciTech Connect

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

    1996-12-31

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

  19. Standardless atom counting in scanning transmission electron microscopy.

    PubMed

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

    2010-11-10

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

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

    NASA Astrophysics Data System (ADS)

    Yankovich, Andrew B.

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

  1. Focused ion beam scanning electron microscopy in biology.

    PubMed

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

    2014-06-01

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

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

    PubMed

    Zumelzu, E; Cabezas, C; Vera, A

    2003-01-01

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

  3. Microstress contrast in scanning electron acoustic microscopy of ceramics

    NASA Technical Reports Server (NTRS)

    Cantrell, John H.; Qian, Menglu

    1991-01-01

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

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

    PubMed

    Stupina, T A

    2016-08-01

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

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

    PubMed

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

    2017-01-01

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

  6. Microstress contrast in scanning electron acoustic microscopy of ceramics

    NASA Technical Reports Server (NTRS)

    Cantrell, John H.; Qian, Menglu

    1991-01-01

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

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

    PubMed

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

    2009-06-01

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

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

    PubMed

    Hempel, Casper

    2017-07-01

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

  9. The combination of scanning electron and scanning probe microscopy

    SciTech Connect

    Sapozhnikov, I. D.; Gorbenko, O. M. Felshtyn, M. L.; Golubok, A. O.

    2016-06-17

    We suggest the SPM module to combine SEM and SPM methods for studying surfaces. The module is based on the original mechanical moving and scanning system. The examples of studies of the steel surface microstructure in both SEM and SPM modes are presented.

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

    PubMed

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

    2010-01-01

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

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

    PubMed

    Ophus, Colin

    2017-01-01

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

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

    PubMed

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

    2012-08-01

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

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

    PubMed

    Rosso, Francesco; Papale, Ferdinando; Barbarisi, Alfonso

    2013-01-01

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

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

    PubMed Central

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

    2013-01-01

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

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

    PubMed

    Barton, Deborah A; Overall, Robyn L

    2015-01-01

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

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

    PubMed

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

    2010-11-01

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

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

    PubMed

    Konopka, K

    2006-09-01

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

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

    PubMed

    Jones, Chris G

    2012-01-01

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

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

    PubMed

    King, B F

    1991-01-01

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

  20. A dynamic scanning method based on signal-statistics for scanning electron microscopy.

    PubMed

    Timischl, F

    2014-01-01

    A novel dynamic scanning method for noise reduction in scanning electron microscopy and related applications is presented. The scanning method dynamically adjusts the scanning speed of the electron beam depending on the statistical behavior of the detector signal and gives SEM images with uniform and predefined standard deviation, independent of the signal value itself. In the case of partially saturated images, the proposed method decreases image acquisition time without sacrificing image quality. The effectiveness of the proposed method is shown and compared to the conventional scanning method and median filtering using numerical simulations.

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

    PubMed

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

    2017-10-01

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

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

    PubMed

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

    2000-05-01

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

  3. Measurement of dihedral angles by scanning electron microscopy.

    NASA Technical Reports Server (NTRS)

    Achutaramayya, G.; Scott, W. D.

    1973-01-01

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

  4. Measurement of dihedral angles by scanning electron microscopy.

    NASA Technical Reports Server (NTRS)

    Achutaramayya, G.; Scott, W. D.

    1973-01-01

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

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

    NASA Astrophysics Data System (ADS)

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

    2008-12-01

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

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

    PubMed

    Sempere, F; Santamarina, M P

    2011-03-01

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

  7. Microbial Nanowire Electronic Structure Probed by Scanning Tunneling Microscopy

    NASA Astrophysics Data System (ADS)

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

    2010-03-01

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

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

    PubMed

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

    2016-02-01

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

  9. 3D correlative light and electron microscopy of cultured cells using serial blockface scanning electron microscopy

    PubMed Central

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

    2017-01-01

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

  10. 3D correlative light and electron microscopy of cultured cells using serial blockface scanning electron microscopy.

    PubMed

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

    2017-01-01

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

  11. Scanning SQUID microscopy with single electron spin sensitivity

    NASA Astrophysics Data System (ADS)

    Vasyukov, Denis

    2014-03-01

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

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

    PubMed Central

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

    2014-01-01

    The growing use of nanoparticles in biomedical applications, including cancer diagnosis and treatment, demands the capability to exactly locate them within complex biological systems. In this work a correlative optical and scanning electron microscopy technique was developed to locate and observe multi-modal gold core nanoparticle accumulation in brain tumor models. Entire brain sections from mice containing orthotopic brain tumors injected intravenously with nanoparticles were imaged using both optical microscopy to identify the brain tumor, and scanning electron microscopy to identify the individual nanoparticles. Gold-based nanoparticles were readily identified in the scanning electron microscope using backscattered electron imaging as bright spots against a darker background. This information was then correlated to determine the exact location of the nanoparticles within the brain tissue. The nanoparticles were located only in areas that contained tumor cells, and not in the surrounding healthy brain tissue. This correlative technique provides a powerful method to relate the macro- and micro-scale features visible in light microscopy with the nanoscale features resolvable in scanning electron microscopy. PMID:25464144

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

    PubMed

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

    2015-01-01

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

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

    SciTech Connect

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

    2014-01-01

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

  15. Theory and application of scanning electron acoustic microscopy

    NASA Technical Reports Server (NTRS)

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

    1992-01-01

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

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

    PubMed

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

    2008-02-01

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

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

    PubMed

    Kohashi, Teruo; Motai, Kumi

    2013-08-01

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

  18. Theory and application of scanning electron acoustic microscopy

    NASA Technical Reports Server (NTRS)

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

    1992-01-01

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

  19. Volume scanning electron microscopy for imaging biological ultrastructure.

    PubMed

    Titze, Benjamin; Genoud, Christel

    2016-11-01

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

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

    SciTech Connect

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

    2011-01-01

    Amyloid fibrils are filamentous protein aggregates implicated in several common diseases such as Alzheimer's disease and type II diabetes. Similar structures are also the molecular principle of the infectious spongiform encephalopathies such as Creutzfeldt-Jakob disease in humans, scrapie in sheep, and of the so-called yeast prions, inherited non-chromosomal elements found in yeast and fungi. Scanning transmission electron microscopy (STEM) is often used to delineate the assembly mechanism and structural properties of amyloid aggregates. In this review we consider specifically contributions and limitations of STEM for the investigation of amyloid assembly pathways, fibril polymorphisms and structural models of amyloid fibrils. This type of microscopy provides the only method to directly measure the mass-per-length (MPL) of individual filaments. Made on both in vitro assembled and ex vivo samples, STEM mass measurements have illuminated the hierarchical relationships between amyloid fibrils and revealed that polymorphic fibrils and various globular oligomers can assemble simultaneously from a single polypeptide. The MPLs also impose strong constraints on possible packing schemes, assisting in molecular model building when combined with high-resolution methods like solid-state nuclear magnetic resonance (NMR) and electron paramagnetic resonance (EPR).

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

    PubMed Central

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

    2010-01-01

    Amyloid fibrils are filamentous protein aggregates implicated in several common diseases like Alzheimer’s disease and type II diabetes. Similar structures are also the molecular principle of the infectious spongiform encephalopathies like Creutzfeldt-Jakob disease in humans, scrapie in sheep, and of the so-called yeast prions, inherited non-chromosomal elements found in yeast and fungi. Scanning transmission electron microscopy (STEM) is often used to delineate the assembly mechanism and structural properties of amyloid aggregates. In this review we consider specifically contributions and limitations of STEM for the investigation of amyloid assembly pathways, fibril polymorphisms and structural models of amyloid fibrils. This type of microscopy provides the only method to directly measure the mass-per-length (MPL) of individual filaments. Made on both in vitro assembled and ex vivo samples, STEM mass measurements have illuminated the hierarchical relationships between amyloid fibrils and revealed that polymorphic fibrils and various globular oligomers can assemble simultaneously from a single polypeptide. The MPLs also impose strong constraints on possible packing schemes, assisting in molecular model building when combined with high-resolution methods like solid-state nuclear magnetic resonance (NMR) and electron paramagnetic resonance (EPR). PMID:20868754

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

    PubMed

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

    2011-01-01

    Amyloid fibrils are filamentous protein aggregates implicated in several common diseases such as Alzheimer's disease and type II diabetes. Similar structures are also the molecular principle of the infectious spongiform encephalopathies such as Creutzfeldt-Jakob disease in humans, scrapie in sheep, and of the so-called yeast prions, inherited non-chromosomal elements found in yeast and fungi. Scanning transmission electron microscopy (STEM) is often used to delineate the assembly mechanism and structural properties of amyloid aggregates. In this review we consider specifically contributions and limitations of STEM for the investigation of amyloid assembly pathways, fibril polymorphisms and structural models of amyloid fibrils. This type of microscopy provides the only method to directly measure the mass-per-length (MPL) of individual filaments. Made on both in vitro assembled and ex vivo samples, STEM mass measurements have illuminated the hierarchical relationships between amyloid fibrils and revealed that polymorphic fibrils and various globular oligomers can assemble simultaneously from a single polypeptide. The MPLs also impose strong constraints on possible packing schemes, assisting in molecular model building when combined with high-resolution methods like solid-state nuclear magnetic resonance (NMR) and electron paramagnetic resonance (EPR). Copyright © 2010 Elsevier Inc. All rights reserved.

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

    SciTech Connect

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

    2014-07-15

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

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

    PubMed

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

    1975-02-28

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

  5. Contamination mitigation strategies for scanning transmission electron microscopy.

    PubMed

    Mitchell, D R G

    2015-06-01

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

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

    PubMed Central

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

    2010-01-01

    A three-dimensional (3D) reconstruction of the cytoskeleton and a clathrin-coated pit in mammalian cells has been achieved from a focal-series of images recorded in an aberration-corrected scanning transmission electron microscope (STEM). The specimen was a metallic replica of the biological structure comprising Pt nanoparticles 2–3 nm in diameter, with a high stability under electron beam radiation. The 3D dataset was processed by an automated deconvolution procedure. The lateral resolution was 1.1 nm, set by pixel size. Particles differing by only 10 nm in vertical position were identified as separate objects with greater than 20% dip in contrast between them. We refer to this value as the axial resolution of the deconvolution or reconstruction, the ability to recognize two objects, which were unresolved in the original dataset. The resolution of the reconstruction is comparable to that achieved by tilt-series transmission electron microscopy. However, the focal-series method does not require mechanical tilting and is therefore much faster. 3D STEM images were also recorded of the Golgi ribbon in conventional thin sections containing 3T3 cells with a comparable axial resolution in the deconvolved dataset. PMID:20082729

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

    SciTech Connect

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

    2010-01-01

    A three-dimensional (3D) reconstruction of the cytoskeleton and a clathrin-coated pit in mammalian cells has been achieved from a focal-series of images recorded in an aberration-corrected scanning transmission electron microscope (STEM). The specimen was a metallic replica of the biological structure comprising Pt nanoparticles 2 - 3 nm in diameter, with a high stability under electron beam radiation. The 3D dataset was processed by an automated deconvolution procedure. The lateral resolution was 1.1 nm, set by pixel size. Particles differing by only 10 nm in vertical position were identified as separate objects with greater than 20% dip in contrast between them. We refer to this value as the axial resolution of the deconvolution or reconstruction, the ability to recognize two objects, which were unresolved in the original data set. The precision of the height determination was 0.2 nm. The resolution of the reconstruction is comparable to that achieved by tilt-series transmission electron microscopy (TEM). However, the focal-series method does not require mechanical tilting and is therefore much faster. 3D STEM images were also recorded of the Golgi ribbon in conventional thin sections containing 3T3 cells with a comparable axial resolution in the deconvolved data set.

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

    PubMed

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

    1978-01-01

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

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

    DOE PAGES

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

    2016-10-18

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

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

    PubMed

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

    2010-02-01

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

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

    NASA Astrophysics Data System (ADS)

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

    2016-10-01

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

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

    SciTech Connect

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

    2014-10-07

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

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

    PubMed

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

    2008-02-01

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

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

    NASA Astrophysics Data System (ADS)

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

    2016-04-01

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

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

    PubMed

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

    2015-12-01

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

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

    PubMed Central

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

    2016-01-01

    Solid-state properties such as strain or chemical composition often leave characteristic fingerprints in the angular dependence of electron scattering. Scanning transmission electron microscopy (STEM) is dedicated to probe scattered intensity with atomic resolution, but it drastically lacks angular resolution. Here we report both a setup to exploit the explicit angular dependence of scattered intensity and applications of angle-resolved STEM to semiconductor nanostructures. Our method is applied to measure nitrogen content and specimen thickness in a GaNxAs1−x layer independently at atomic resolution by evaluating two dedicated angular intervals. We demonstrate contrast formation due to strain and composition in a Si- based metal-oxide semiconductor field effect transistor (MOSFET) with GexSi1−x stressors as a function of the angles used for imaging. To shed light on the validity of current theoretical approaches this data is compared with theory, namely the Rutherford approach and contemporary multislice simulations. Inconsistency is found for the Rutherford model in the whole angular range of 16–255 mrad. Contrary, the multislice simulations are applicable for angles larger than 35 mrad whereas a significant mismatch is observed at lower angles. This limitation of established simulations is discussed particularly on the basis of inelastic scattering. PMID:27849001

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

    PubMed

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

    2014-04-01

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

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

    NASA Astrophysics Data System (ADS)

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

    2016-11-01

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

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

    PubMed

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

    2016-11-16

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

  20. Characterization of two-dimensional hexagonal boron nitride using scanning electron and scanning helium ion microscopy

    SciTech Connect

    Guo, Hongxuan E-mail: msxu@zju.edu.cn; Gao, Jianhua; Ishida, Nobuyuki; Xu, Mingsheng E-mail: msxu@zju.edu.cn; Fujita, Daisuke

    2014-01-20

    Characterization of the structural and physical properties of two-dimensional (2D) materials, such as layer number and inelastic mean free path measurements, is very important to optimize their synthesis and application. In this study, we characterize the layer number and morphology of hexagonal boron nitride (h-BN) nanosheets on a metallic substrate using field emission scanning electron microscopy (FE-SEM) and scanning helium ion microscopy (HIM). Using scanning beams of various energies, we could analyze the dependence of the intensities of secondary electrons on the thickness of the h-BN nanosheets. Based on the interaction between the scanning particles (electrons and helium ions) and h-BN nanosheets, we deduced an exponential relationship between the intensities of secondary electrons and number of layers of h-BN. With the attenuation factor of the exponential formula, we calculate the inelastic mean free path of electrons and helium ions in the h-BN nanosheets. Our results show that HIM is more sensitive and consistent than FE-SEM for characterizing the number of layers and morphology of 2D materials.

  1. Characterization of two-dimensional hexagonal boron nitride using scanning electron and scanning helium ion microscopy

    NASA Astrophysics Data System (ADS)

    Guo, Hongxuan; Gao, Jianhua; Ishida, Nobuyuki; Xu, Mingsheng; Fujita, Daisuke

    2014-01-01

    Characterization of the structural and physical properties of two-dimensional (2D) materials, such as layer number and inelastic mean free path measurements, is very important to optimize their synthesis and application. In this study, we characterize the layer number and morphology of hexagonal boron nitride (h-BN) nanosheets on a metallic substrate using field emission scanning electron microscopy (FE-SEM) and scanning helium ion microscopy (HIM). Using scanning beams of various energies, we could analyze the dependence of the intensities of secondary electrons on the thickness of the h-BN nanosheets. Based on the interaction between the scanning particles (electrons and helium ions) and h-BN nanosheets, we deduced an exponential relationship between the intensities of secondary electrons and number of layers of h-BN. With the attenuation factor of the exponential formula, we calculate the inelastic mean free path of electrons and helium ions in the h-BN nanosheets. Our results show that HIM is more sensitive and consistent than FE-SEM for characterizing the number of layers and morphology of 2D materials.

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

    PubMed

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

    2012-01-01

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

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

    NASA Astrophysics Data System (ADS)

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

    2015-02-01

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

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

    SciTech Connect

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

    2015-02-02

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

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

    PubMed

    Alves, Marcelo de Carvalho; Pozza, Edson Ampélio

    2009-07-01

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

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

    PubMed Central

    Motta, P M; Andrews, P M

    1976-01-01

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

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

    PubMed

    Ksiezak-Reding, Hanna; Wall, Joseph S

    2005-07-01

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

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

    NASA Technical Reports Server (NTRS)

    Huss, D.; Dickman, J. D.

    2003-01-01

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

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

    PubMed Central

    Valian, Azam

    2014-01-01

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

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

    NASA Technical Reports Server (NTRS)

    Huss, D.; Dickman, J. D.

    2003-01-01

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

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

    PubMed

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

    2012-04-01

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

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

    PubMed

    Banerjee, R; Bellare, J R

    2001-01-01

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

  13. Scanning electron microscopy of lung following alpha irradiation

    SciTech Connect

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

    1989-09-01

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

  14. Three-dimensional optical sectioning by scanning confocal electron microscopy with a stage-scanning system.

    PubMed

    Hashimoto, Ayako; Shimojo, Masayuki; Mitsuishi, Kazutaka; Takeguchi, Masaki

    2010-06-01

    We evaluated the depth resolution of annular dark-field (ADF) scanning confocal electron microscopy (SCEM) with a stage-scanning system by observation of nanoparticles. ADF-SCEM is a three-dimensional (3D) imaging technique that we recently proposed. An ADF-SCEM instrument involves a pinhole aperture before a detector for rejecting electrons from the out-of-focal plane in a specimen and an annular aperture under the specimen for collecting only scattered electrons. The stage-scanning system enables us to directly obtain optical slice images perpendicular and parallel to an optical axis at a desired position. In particular, the parallel slices visualize the elongation of nanoparticles along the optical axis, which depends on the depth resolution. ADF-SCEM effectively reduced the elongation length of the nanoparticles sufficiently to demonstrate depth sectioning, in comparison with scanning transmission electron microscopy and bright-field SCEM. The experimentally obtained length was nearly equal to the theoretically estimated one from the probe size considering the experimental conditions. Furthermore, we applied this ADF-SCEM technique to analysis of the 3D position of catalytic nanoparticles on carbon nanostructures.

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

    NASA Astrophysics Data System (ADS)

    Sindern, Sven; Meyer, F. Michael

    2016-09-01

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

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

    PubMed

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

    2012-01-01

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

  17. Correcting nonlinear drift distortion of scanning probe and scanning transmission electron microscopies from image pairs with orthogonal scan directions.

    PubMed

    Ophus, Colin; Ciston, Jim; Nelson, Chris T

    2016-03-01

    Unwanted motion of the probe with respect to the sample is a ubiquitous problem in scanning probe and scanning transmission electron microscopies, causing both linear and nonlinear artifacts in experimental images. We have designed a procedure to correct these artifacts by using orthogonal scan pairs to align each measurement line-by-line along the slow scan direction, by fitting contrast variation along the lines. We demonstrate the accuracy of our algorithm on both synthetic and experimental data and provide an implementation of our method. Copyright © 2015 Elsevier B.V. All rights reserved.

  18. Correcting nonlinear drift distortion of scanning probe and scanning transmission electron microscopies from image pairs with orthogonal scan directions

    DOE PAGES

    Ophus, Colin; Ciston, Jim; Nelson, Chris T.

    2015-12-10

    Unwanted motion of the probe with respect to the sample is a ubiquitous problem in scanning probe and scanning transmission electron microscopies, causing both linear and nonlinear artifacts in experimental images. We have designed a procedure to correct these artifacts by using orthogonal scan pairs to align each measurement line-by-line along the slow scan direction, by fitting contrast variation along the lines. We demonstrate the accuracy of our algorithm on both synthetic and experimental data and provide an implementation of our method.

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

    PubMed

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

    2005-04-01

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

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

    PubMed

    Stoll, Joshua D; Kolmakov, Andrei

    2012-12-21

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

  1. Direct observations of atomic diffusion by scanning transmission electron microscopy

    PubMed Central

    Isaacson, M.; Kopf, D.; Utlaut, M.; Parker, N. W.; Crewe, A. V.

    1977-01-01

    The feasibility of using a high-resolution scanning transmission electron microscope to study the diffusion of heavy atoms on thin film substrates of low atomic number has been investigated. We have shown that it is possible to visualize the diffusion of individual uranium atoms adsorbed to thin carbon film substrates and that the observed motion of the atoms does not appear to be induced by the incident electron beam. Images PMID:16592396

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

    PubMed

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

    2013-01-01

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

  3. Scanning moiré fringe imaging by scanning transmission electron microscopy.

    PubMed

    Su, Dong; Zhu, Yimei

    2010-02-01

    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.

  4. Low voltage scanning electron microscopy of interplanetary dust particles

    NASA Technical Reports Server (NTRS)

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

    1987-01-01

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

  5. Low voltage scanning electron microscopy of interplanetary dust particles

    NASA Technical Reports Server (NTRS)

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

    1987-01-01

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

  6. Visualization of Microbial Biomarkers by Scanning Electron Microscopy

    NASA Technical Reports Server (NTRS)

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

    2001-01-01

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

  7. Visualization of Microbial Biomarkers by Scanning Electron Microscopy

    NASA Technical Reports Server (NTRS)

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

    2001-01-01

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

  8. Time-resolved scanning electron microscopy with polarization analysis

    SciTech Connect

    Frömter, Robert Oepen, Hans Peter; Kloodt, Fabian; Rößler, Stefan; Frauen, Axel; Staeck, Philipp; Cavicchia, Demetrio R.; Bocklage, Lars; Röbisch, Volker; Quandt, Eckhard

    2016-04-04

    We demonstrate the feasibility of investigating periodically driven magnetization dynamics in a scanning electron microscope with polarization analysis based on spin-polarized low-energy electron diffraction. With the present setup, analyzing the time structure of the scattering events, we obtain a temporal resolution of 700 ps, which is demonstrated by means of imaging the field-driven 100 MHz gyration of the vortex in a soft-magnetic FeCoSiB square. Owing to the efficient intrinsic timing scheme, high-quality movies, giving two components of the magnetization simultaneously, can be recorded on the time scale of hours.

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

    PubMed

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

    2012-12-01

    Some four decades were needed to catch up with the vision that Albert Crewe and his group had for the scanning transmission electron microscope (STEM) in the nineteen sixties and seventies: attaining 0.5Å resolution, and identifying single atoms spectroscopically. With these goals now attained, STEM developments are turning toward new directions, such as rapid atomic resolution imaging and exploring atomic bonding and electronic properties of samples at atomic resolution. The accomplishments and the future challenges are reviewed and illustrated with practical examples. Copyright © 2012 Elsevier B.V. All rights reserved.

  10. Scanning electron microscopy of clays and clay minerals

    USGS Publications Warehouse

    Bohor, B.F.; Hughes, R.E.

    1971-01-01

    The scanning electron microscope (SEM) proves to be ideally suited for studying the configuration, texture, and fabric of clay samples. Growth mechanics of crystalline units-interpenetration and interlocking of crystallites, crystal habits, twinning, helical growth, and topotaxis-also are uniquely revealed by the SEM. Authigenic kaolins make up the bulk of the examples because their larger crystallite size, better crystallinity, and open texture make them more suited to examination by the SEM than most other clay mineral types. ?? 1971.

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

    NASA Astrophysics Data System (ADS)

    Seacor, Taylor; Howell, Carina

    2013-03-01

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

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

    NASA Astrophysics Data System (ADS)

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

    2000-06-01

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

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

    PubMed

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

    2012-10-01

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

  14. Big Data Analytics for Scanning Transmission Electron Microscopy Ptychography

    SciTech Connect

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

    2016-05-23

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

  15. Big Data Analytics for Scanning Transmission Electron Microscopy Ptychography

    DOE PAGES

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

    2016-05-23

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

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

    PubMed

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

    2016-05-23

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

  17. Big Data Analytics for Scanning Transmission Electron Microscopy Ptychography

    NASA Astrophysics Data System (ADS)

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

    2016-05-01

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

  18. Big Data Analytics for Scanning Transmission Electron Microscopy Ptychography

    PubMed Central

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

    2016-01-01

    Electron microscopy is undergoing a transition; from the model of producing only a few micrographs, through the current state where many images and spectra can be digitally recorded, to a new mode where very large volumes of data (movies, ptychographic and multi-dimensional series) can be rapidly obtained. Here, we discuss the application of so-called “big-data” methods to high dimensional microscopy data, using unsupervised multivariate statistical techniques, in order to explore salient image features in a specific example of BiFeO3 domains. Remarkably, k-means clustering reveals domain differentiation despite the fact that the algorithm is purely statistical in nature and does not require any prior information regarding the material, any coexisting phases, or any differentiating structures. While this is a somewhat trivial case, this example signifies the extraction of useful physical and structural information without any prior bias regarding the sample or the instrumental modality. Further interpretation of these types of results may still require human intervention. However, the open nature of this algorithm and its wide availability, enable broad collaborations and exploratory work necessary to enable efficient data analysis in electron microscopy. PMID:27211523

  19. Scanning Electron Microscopy with Samples in an Electric Field

    PubMed Central

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

    2012-01-01

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

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

    PubMed

    Müller, E; Gerthsen, D

    2017-02-01

    The contrast of backscattered electron (BSE) images in scanning electron microscopy (SEM) depends on material parameters which can be exploited for composition quantification if some information on the material system is available. As an example, the In-concentration in thin InxGa1-xAs layers embedded in a GaAs matrix is analyzed in this work. The spatial resolution of the technique is improved by using thin electron-transparent specimens instead of bulk samples. Although the BSEs are detected in a comparably small angular range by an annular semiconductor detector, the image intensity can be evaluated to determine the composition and local thickness of the specimen. The measured intensities are calibrated within one single image to eliminate the influence of the detection and amplification system. Quantification is performed by comparison of experimental and calculated data. Instead of using time-consuming Monte-Carlo simulations, an analytical model is applied for BSE-intensity calculations which considers single electron scattering and electron diffusion.

  1. A spin rotator for spin-polarized scanning electron microscopy

    NASA Astrophysics Data System (ADS)

    Kohashi, Teruo; Konoto, Makoto; Koike, Kazuyuki

    2004-06-01

    A Wien filter, which is a common energy analyzer, was modified as a spin rotator for use in a spin-polarized scanning electron microscope. By switching the spin rotator on and off, magnetic domain images of all three magnetization vectors can be produced in one scan. The electrodes and the magnetic pole pieces were specially designed by using a three-dimensional computer simulation for electric and magnetic fields, electron trajectories, and spin rotation; the broad beam of the secondary electrons passes through to the spin detector with a 90° rotation. The structure is simple with only two electrodes that have hyperbolically curved surfaces to create a stigmatic focusing effect, while the surfaces of the magnetic pole pieces are flat to enable a uniform rotation of all electron spins. The performance was tested and confirmed to be effective by observing the magnetic domain structures of Fe(001) with in-surface-plane magnetization and a TbFeCo magneto-optical medium with surface normal magnetization.

  2. The theory and practice of high resolution scanning electron microscopy

    SciTech Connect

    Joy, D.C. Oak Ridge National Lab., TN )

    1990-01-01

    Recent advances in instrumentation have produced the first commercial examples of what can justifiably be called High Resolution Scanning Electron Microscopes. The key components of such instruments are a cold field emission gun, a small-gap immersion probe-forming lens, and a clean dry-pumped vacuum. The performance of these microscopes is characterized by several major features including a spatial resolution, in secondary electron mode on solid specimens, which can exceed 1nm on a routine basis; an incident probe current density of the order of 10{sup 6} amps/cm{sup 2}; and the ability to maintain these levels of performance over an accelerating voltage range of from 1 to 30keV. This combination of high resolution, high probe current, low contamination and flexible electron-optical conditions provides many new opportunitites for the application of the SEM to materials science, physics, and the life sciences. 27 refs., 14 figs.

  3. [Using of scanning electron microscopy for detection of gunshot residue].

    PubMed

    Havel, J; Vajtr, D; Starý, V; Vrána, J; Zelenka, K; Adámek, T

    2006-07-01

    Scanning electron microscope improves the possibility of investigation of surroundings near of gunshot wounds in forensic medicine, it is the next subsequent method for differentiating of area of entrance and exit wound, supplemental method for determination of firing distance, permit of detection (GSR) on the hand of shooter and ensured describing of samples and their stored. Detection of GSR provides many information about composition of bullet and primer. Authors are demonstrating the possibility of detection of GSR on experimental shooting to the krupon (pigs' skin) in different situation (such as in a room and in outside area) and using of different weapon (hand gun CZ No.75 and machine gun No.58).

  4. [Scanning electron microscopy of heat-damaged bone tissue].

    PubMed

    Harsanyl, L

    1977-02-01

    Parts of diaphyses of bones were exposed to high temperature of 200-1300 degrees C. Damage to the bone tissue caused by the heat was investigated. The scanning electron microscopic picture seems to be characteristic of the temperature applied. When the bones heated to the high temperature of 700 degrees C characteristic changes appear on the periostal surface, higher temperatura on the other hand causes damage to the compact bone tissue and can be observed on the fracture-surface. Author stresses the importance of this technique in the legal medicine and anthropology.

  5. Factors influencing quantitative liquid (scanning) transmission electron microscopy

    SciTech Connect

    Abellan Baeza, Patricia; Woehl, Taylor J.; Parent, Lucas R.; Browning, Nigel D.; Evans, James E.; Arslan, Ilke

    2014-04-15

    One of the experimental challenges in the study of nanomaterials in liquids in the (scanning) transmission electron microscope ((S)TEM) is gaining quantitative information. A successful experiment in the fluid stage will depend upon the ability to plan for sensitive factors such as the electron dose applied, imaging mode, acceleration voltage, beam-induced solution chemistry changes, and the specifics of solution reactivity. In this paper, we make use of a visual approach to show the extent of damage of different instrumental and experimental factors in liquid samples imaged in the (S)TEM. Previous results as well as new insights are presented to create an overview of beam-sample interactions identified for changing imaging and experimental conditions. This work establishes procedures to understand the effect of the electron beam on a solution, provides information to allow for a deliberate choice of the optimal experimental conditions to enable quantification, and identifies the experimental factors that require further analysis for achieving fully quantitative results in the liquid (S)TEM.

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

    SciTech Connect

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

    1988-11-01

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

  7. Moessbauer spectroscopy and scanning electron microscopy of the Murchison meteorite

    NASA Technical Reports Server (NTRS)

    Brown, Christopher L.; Oliver, Frederick W.; Hammond, Ernest C., Jr.

    1989-01-01

    Meteorites provide a wealth of information about the solar system's formation, since they have similar building blocks as the Earth's crust but have been virtually unaltered since their formation. Some stony meteorites contain minerals and silicate inclusions, called chondrules, in the matrix. Utilizing Moessbauer spectroscopy, we identified minerals in the Murchison meteorite, a carbonaceous chondritic meteorite, by the gamma ray resonance lines observed. Absorption patterns of the spectra were found due to the minerals olivine and phyllosilicate. We used a scanning electron microscope to describe the structure of the chondrules in the Murchison meteorite. The chondrules were found to be deformed due to weathering of the meteorite. Diameters varied in size from 0.2 to 0.5 mm. Further enhancement of the microscopic imagery using a digital image processor was used to describe the physical characteristics of the inclusions.

  8. [Cleaning implantation burs. Observations using scanning electron microscopy].

    PubMed

    Penel, G; Iost, A; Libersa, J C

    2001-01-01

    Drastic aseptic conditions are necessary in implantological treatments. A good sterilizing procedure of the specific instrumentation, like drills, is based on an efficient cleaning. Because of their design, the cleaning of drills is a real challenge. The aim of this investigation is to evaluate two different cleaning procedures usually used by implantologists. One is based on a manual cleaning, the other on an ultra-sonic cleaning. The instrument observed by scan-electron-microscope, is a I.T.I. system drill. The results show the superiority of the ultra-sonic cleaning. The manual cleaning is especially ineffective on the inside and the cutting part of the drill. Even if ultra-sonic cleaning is definitely a better procedure, it has to be improved. A extensive study should be conducted to optimize the cleaning parameters, if not, single-use drill should be definitely preferred by implantologists.

  9. Scanning electron microscopy of human cortical bone failure surfaces.

    PubMed

    Braidotti, P; Branca, F P; Stagni, L

    1997-02-01

    Undecalcified samples extracted from human femoral shafts are fractured by bending and the fracture surfaces are examined with a scanning electron microscope (SEM). The investigation is performed on both dry and wet (hydrated with a saline solution) specimens. SEM micrographs show patterns in many respects similar to those observed in fractography studies of laminated fiber-reinforced synthetic composites. In particular, dry and wet samples behave like brittle and ductile matrix laminates, respectively. An analysis carried out on the basis of the mechanisms that dominate the fracture process of laminates shows that a reasonable cortical bone model is that of a laminated composite material whose matrix is composed of extracellular noncollagenous calcified proteins, and the reinforcement is constituted by the calcified collagen fiber system.

  10. Detector non-uniformity in scanning transmission electron microscopy.

    PubMed

    Findlay, S D; LeBeau, J M

    2013-01-01

    A non-uniform response across scanning transmission electron microscope annular detectors has been found experimentally, but is seldom incorporated into simulations. Through case study simulations, we establish the nature and scale of the discrepancies which may arise from failing to account for detector non-uniformity. If standard detectors are used at long camera lengths such that the detector is within or near to the bright field region, we find errors in contrast of the order of 10%, sufficiently small for qualitative work but non-trivial as experiments become more quantitative. In cases where the detector has been characterized in advance, we discuss the detector response normalization and how it may be incorporated in simulations. Copyright © 2012 Elsevier B.V. All rights reserved.

  11. Automated rapid particle investigation using scanning electron microscopy

    NASA Astrophysics Data System (ADS)

    Wilkins, Jerod Laurence

    The chemical composition of fly ash particles has been known to vary significantly depending on a number of factors. Current bulk methods of investigation including X-Ray Fluorescence and X-Ray Diffraction are thought to be inadequate in determining the performance of fly ash in concrete. It is the goal of this research to develop a method of Automated Rapid Particle Investigation that will not look at fly ash as a bulk material but as individual particles. By examining each particle individually scientists and engineers will have the ability to study the variation in chemical composition by comparing the chemistry present in each particle. The method of investigation developed by this research provides a practical technique that will allow the automated chemical analysis of hundreds, or even thousands, of fly ash particles in a matter of minutes upon completion of sample preparation and automated scanning electron microscope (ASEM) scanning. This research does not examine the significance of the chemical compounds discovered; rather, only the investigation methodology is discussed. Further research will be done to examine the importance of the chemistry discovered with this automated rapid particle investigation technique.

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

    PubMed

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

    2011-09-01

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

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

    NASA Astrophysics Data System (ADS)

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

    2016-12-01

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

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

    SciTech Connect

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

    2016-06-07

    Electron tomography in materials science has flourished with the demand to characterize nanoscale materials in three dimensions (3D). Access to experimental data is vital for developing and validating reconstruction methods that improve resolution and reduce radiation dose requirements. This work presents five high-quality scanning transmission electron microscope (STEM) tomography datasets in order to address the critical need for open access data in this field. The datasets represent the current limits of experimental technique, are of high quality, and contain materials with structural complexity. Included are tomographic series of a hyperbranched Co 2 P nanocrystal, platinum nanoparticles on a carbon nanofibre imaged over the complete 180° tilt range, a platinum nanoparticle and a tungsten needle both imaged at atomic resolution by equal slope tomography, and a through-focal tilt series of PtCu nanoparticles. A volumetric reconstruction from every dataset is provided for comparison and development of post-processing and visualization techniques. Researchers interested in creating novel data processing and reconstruction algorithms will now have access to state of the art experimental test data.

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

    PubMed Central

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

    2016-01-01

    Electron tomography in materials science has flourished with the demand to characterize nanoscale materials in three dimensions (3D). Access to experimental data is vital for developing and validating reconstruction methods that improve resolution and reduce radiation dose requirements. This work presents five high-quality scanning transmission electron microscope (STEM) tomography datasets in order to address the critical need for open access data in this field. The datasets represent the current limits of experimental technique, are of high quality, and contain materials with structural complexity. Included are tomographic series of a hyperbranched Co2P nanocrystal, platinum nanoparticles on a carbon nanofibre imaged over the complete 180° tilt range, a platinum nanoparticle and a tungsten needle both imaged at atomic resolution by equal slope tomography, and a through-focal tilt series of PtCu nanoparticles. A volumetric reconstruction from every dataset is provided for comparison and development of post-processing and visualization techniques. Researchers interested in creating novel data processing and reconstruction algorithms will now have access to state of the art experimental test data. PMID:27272459

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

    DOE PAGES

    Levin, Barnaby D. A.; Padgett, Elliot; Chen, Chien-Chun; ...

    2016-06-07

    Electron tomography in materials science has flourished with the demand to characterize nanoscale materials in three dimensions (3D). Access to experimental data is vital for developing and validating reconstruction methods that improve resolution and reduce radiation dose requirements. This work presents five high-quality scanning transmission electron microscope (STEM) tomography datasets in order to address the critical need for open access data in this field. The datasets represent the current limits of experimental technique, are of high quality, and contain materials with structural complexity. Included are tomographic series of a hyperbranched Co 2 P nanocrystal, platinum nanoparticles on a carbon nanofibremore » imaged over the complete 180° tilt range, a platinum nanoparticle and a tungsten needle both imaged at atomic resolution by equal slope tomography, and a through-focal tilt series of PtCu nanoparticles. A volumetric reconstruction from every dataset is provided for comparison and development of post-processing and visualization techniques. Researchers interested in creating novel data processing and reconstruction algorithms will now have access to state of the art experimental test data.« less

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

    NASA Astrophysics Data System (ADS)

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

    2016-06-01

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

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

    PubMed

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

    2016-06-07

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

  19. Scanning Electron Microscopy And Data Digitization Of Craniofacial Growth

    NASA Astrophysics Data System (ADS)

    Rice, Robert W.; Oyen, Ordean J.; Walker, Alan C.

    1980-07-01

    The scanning electron microscope (SEM), combining high resolution and large depth of focus, affords detailed observation of surface microstructure in a three-dimensional perspective. It also allows large specimen dimensions and avoids the processing and sectioning limitations of light and transmission electron microscopic procedures. For these reasons the SEM is ideally suited for analyses of bone, a rigid tissue whose surface topography and internal architecture accurately reflect the developmental, metabolic and mechanical influences exerted upon it. Furthermore, SEM photomicrographs are compatible with devices for quantification, mathematical manipulation and graphic reconstruction of the image. Features of a photo may be traced with a stylus on the electromagnetically activated surface of a data digitizer, which converts the outlined path to x and y axis coordinates. Interfaced with a programmed calculator these data undergo algebraic and geometrical computation and may be stored for statistical analyses. Alternatively, stereopairs of micrograph transparencies may be utilized in micro-stereophotogrammetric procedures in which x, y and z axis coordinates are generated for selected morphologic points. Our research concerns spatiotemporal interrelationships of primate craniofacial growth as evidenced by changes in the skeletal gross morphology and microanatomy of the orbital region, jaws and teeth during their growth and development. Applications of SEM and digitization techniques to these studies and an evaluation of the derived data will be presented.

  20. U-10Mo Sample Preparation and Examination using Optical and Scanning Electron Microscopy

    SciTech Connect

    Prabhakaran, Ramprashad; Joshi, Vineet V.; Rhodes, Mark A.; Schemer-Kohrn, Alan L.; Guzman, Anthony D.; Lavender, Curt A.

    2016-03-30

    The purpose of this document is to provide guidelines to prepare specimens of uranium alloyed with 10 weight percent molybdenum (U-10Mo) for optical metallography and scanning electron microscopy. This document also provides instructions to set up an optical microscope and a scanning electron microscope to analyze U-10Mo specimens and to obtain the required information.

  1. U-10Mo Sample Preparation and Examination using Optical and Scanning Electron Microscopy

    SciTech Connect

    Prabhakaran, Ramprashad; Joshi, Vineet V.; Rhodes, Mark A.; Schemer-Kohrn, Alan L.; Guzman, Anthony D.; Lavender, Curt A.

    2016-10-01

    The purpose of this document is to provide guidelines to prepare specimens of uranium alloyed with 10 weight percent molybdenum (U-10Mo) for optical metallography and scanning electron microscopy. This document also provides instructions to set up an optical microscope and a scanning electron microscope to analyze U-10Mo specimens and to obtain the required information.

  2. EVALUATION OF COMPUTER-CONTROLLED SCANNING ELECTRON MICROSCOPY APPLIED TO AN AMBIENT URBAN AEROSOL SAMPLE

    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...

  3. EVALUATION OF COMPUTER-CONTROLLED SCANNING ELECTRON MICROSCOPY APPLIED TO AN AMBIENT URBAN AEROSOL SAMPLE

    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...

  4. Scanning Electron Microscopy of Colonies of Six Species of Candida

    PubMed Central

    Joshi, K. R.; Wheeler, E. E.; Gavin, J. B.

    1973-01-01

    Thirty strains of six species of Candida isolated from patients were cultured for 60 h on Sabouraud agar, freeze-dried, and examined with a scanning electron microscope. The colonies were circular (Candida albicans, C. guilliermondii) or oval (C. tropicalis, C. pseudotropicalis, C. krusei, C. parakrusei) in outline, and those of C. pseudotropicalis and C. krusei had an irregular outline due to a peripheral pseudomycelium. The morphology of individual microorganisms was examined at the margins and apex of those species which lacked a surface coat (C. pseudotropicalis, C. krusei, C. parakrusei, C. guilliermondii), and through cracks in the surface coating of those which showed a surface coat (C. albicans, C. tropicalis). All species showed buds, bud scars, and interconnecting intercellular processes, but were generally spherical (C. albicans, C. tropicalis) or ovoid (C. pseudotropicalis, C. krusei, C. parakrusei, C. guilliermondii) in fixed preparations. In unfixed material, individual organisms were almost invariably indented. Fixation with 3% glutaraldehyde and washing before freeze-drying caused partial removal of the surface coating of colonies of C. albicans and C. tropicalis, which persisted only as irregular sheets or as a filamentous meshwork. This filamentous meshwork was also present among the organisms of colonies of C. albicans, C. tropicalis, and C. pseudotropicalis. It is concluded that these filaments represent the precipitation or unmasking of some component of the intercellular matrix of these organisms. Images PMID:4197906

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

    PubMed

    Prozorowska, Ewelina; Jackowiak, Hanna

    2015-03-01

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

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

    NASA Astrophysics Data System (ADS)

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

    2014-03-01

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

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

    NASA Astrophysics Data System (ADS)

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

    2012-11-01

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

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

    SciTech Connect

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

    2012-11-19

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

  9. Scanning electron microscopy of hair treated in hard water.

    PubMed

    Srinivasan, Gautham; Chakravarthy Rangachari, Srinivas

    2016-06-01

    Hardness of water is determined by the amount of calcium carbonate (CaCO3 ) and magnesium sulfate (MgSO4 ) dissolved in it. Hardness of water used for washing hair may damage the hair. The objective of this study is to observe the surface changes due to hard water usage and compare the thickness of hair between hard and soft water treated samples. Ten to 15 hair strands of length 15-20 cm, which were lost during combing, were obtained from 15 healthy female volunteers. Each hair sample was cut into two equal halves to obtain two sets per volunteer. Each hair sample was wrapped around a glass rod. One set of 15 samples was washed with hard water, and the other set was washed with distilled water for 10 minutes on alternate days and air-dried. This procedure was carried out for 30 days. The surface of hair treated in hard and soft water was examined under a scanning electron microscope. The CaCO3 and MgSO4 content of hard and distilled water samples were determined as 212.5 ppm of CaCO3 and 10 ppm of CaCO3 respectively. The mean calcium deposition in hard and distilled water treated hair was determined as 0.804% and 0.26%, respectively. The mean magnesium deposition in hard and distilled water treated hair was determined as 0.34% and 0.078%, respectively. The mean thickness of hair treated in hard water and distilled water were 72.78 and 78.14 μm, respectively. The surface of hard water treated hair has a ruffled appearance with higher mineral deposition and decreased thickness when compared with the surface of distilled water treated hair. © 2015 The International Society of Dermatology.

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

    PubMed

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

    2014-04-01

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

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

    PubMed

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

    2017-04-03

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

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

    NASA Astrophysics Data System (ADS)

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

    2011-08-01

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

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

    PubMed

    Cazaux, Jacques

    2012-01-01

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

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

    PubMed

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

    2014-03-01

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

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

    PubMed Central

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

    2017-01-01

    Atomic-resolution imaging in an aberration-corrected scanning transmission electron microscope (STEM) can enable direct correlation between atomic structure and materials functionality. The fast and precise control of the STEM probe is, however, challenging because the true beam location deviates from the assigned location depending on the properties of the deflectors. To reduce these deviations, i.e. image distortions, we use spiral scanning paths, allowing precise control of a sub-Å sized electron probe within an aberration-corrected STEM. Although spiral scanning avoids the sudden changes in the beam location (fly-back distortion) present in conventional raster scans, it is not distortion-free. “Archimedean” spirals, with a constant angular frequency within each scan, are used to determine the characteristic response at different frequencies. We then show that such characteristic functions can be used to correct image distortions present in more complicated constant linear velocity spirals, where the frequency varies within each scan. Through the combined application of constant linear velocity scanning and beam path corrections, spiral scan images are shown to exhibit less scan distortion than conventional raster scan images. The methodology presented here will be useful for in situ STEM imaging at higher temporal resolution and for imaging beam sensitive materials. PMID:28272404

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

    PubMed

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

    2017-03-08

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

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

    NASA Astrophysics Data System (ADS)

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

    2017-03-01

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

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

    PubMed

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

    2013-01-01

    The objective was to confirm the finding of "Bone microstructure is similar in osteopenic and osteoporotic patients with femoral neck fracture." obtained in previous "light microscopy study", which was new and important data. Fourteen patients (5 males, 9 females) who were admitted with proximal femoral fracture following low energy trauma (patients who participated in the light microscopy study) were included. The patients were divided into two groups based on the bone mineral density (BMD) measurement, including osteopenic group (n=7, mean age 69 years; range 63 to 74 years) and osteoporotic group (n=7, mean age 74.1 years; range 67 to 78 years). Cortical and trabecular bone samples were taken from the patients who underwent endoprosthesis during partial hip arthroplasty and these samples were analyzed using transmission electron microscopy and scanning electron microscopy evaluations which are more sophisticated higher resolution techniques. The mean cortical bone thickness was 3622.14 mm in osteopenic group and 2323.14 mm in osteoporotic group (p<0.005). Transmission electron microscopy and scanning electron microscopy evaluations revealed similar findings for both groups. Although a significant difference in cortical thickness was found between the groups, transmission and scanning electron microscopy confirmed that bone microstructure shared similar characteristics in osteopenic and osteoporotic patients with low-energy femoral neck fracture, as it was in previous light microscopy study.

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

    PubMed

    Volkenandt, Tobias; Müller, Erich; Gerthsen, Dagmar

    2014-02-01

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

  20. Advantages of Environmental Scanning Electron Microscopy in Studies of Microorganisms

    DTIC Science & Technology

    1993-01-01

    disruption of sporangia. Uncoated algal cells of Euglena gracilis and Spirogyra sp. were examined using the backscatter electron detector (BSE) and the...agar. (Fig. 7). Uncoated, freeze-dried Euglena sp. cells exam- ined with either ESD or BSE detectors did not exhibit Specimen Preparation for ESEM...P.L. (1967) Observations in the fine struc- Biofilms: An ESEM evaluation of artifacts introduced during SEM ture of the pellicle pores of Euglena

  1. Stochastic Micro-Pattern for Automated Correlative Fluorescence - Scanning Electron Microscopy

    PubMed Central

    Begemann, Isabell; Viplav, Abhiyan; Rasch, Christiane; Galic, Milos

    2015-01-01

    Studies of cellular surface features gain from correlative approaches, where live cell information acquired by fluorescence light microscopy is complemented by ultrastructural information from scanning electron micrographs. Current approaches to spatially align fluorescence images with scanning electron micrographs are technically challenging and often cost or time-intensive. Relying exclusively on open-source software and equipment available in a standard lab, we have developed a method for rapid, software-assisted alignment of fluorescence images with the corresponding scanning electron micrographs via a stochastic gold micro-pattern. Here, we provide detailed instructions for micro-pattern production and image processing, troubleshooting for critical intermediate steps, and examples of membrane ultra-structures aligned with the fluorescence signal of proteins enriched at such sites. Together, the presented method for correlative fluorescence – scanning electron microscopy is versatile, robust and easily integrated into existing workflows, permitting image alignment with accuracy comparable to existing approaches with negligible investment of time or capital. PMID:26647824

  2. Scanning electron microscopy in characterizing seeds of some leguminous trees

    NASA Astrophysics Data System (ADS)

    Ghosh, Nabarun; Chatterjee, Amiyanghshu; Smith, Don W.

    2009-05-01

    SEM has greatly increased our knowledge of the microstructure of seeds. Mature seed coats are rather thick walled and stable in a vacuum: this allows quick preparation for SEM examination, without the need of complicated dehydration techniques. The low level of technical expenditure required, in combination with the high structural diversity exhibited and the intuitive ability to understand the "three dimensional", often aesthetically appealing micro-structures visualized, has turned seed-coat studies into a favorite tool of many taxonomists. We used dry mature seeds of 26 species of 4 Leguminous genera, Acacia, Albizia, Cassia and Dalbergia to standardize a procedure for identifying the seeds through SEM on the seed surface and seed sections. We cut transverse and longitudinal sections of the seeds and observed the sections from different regions of seeds: midseed, near the hilum and two distal ends. Light microscopy showed the color, texture, pleurograms, fissures and hilum at lower magnification. The anatomical study with SEM on the seed sections revealed the size, shape, and number of tiers and cellular organization of the epidermis, hypodermis, endosperm and internal structural details. We found the ornamentation pattern of the seeds including undulations, reticulations and rugae that were species specific. Species of Dalbergia (assamica, latifolia and sissoo), Albizia (odoratissima and procera), Acaia (arabica and catechu) and Cassia (glauca, siamia and spectabilis) are difficult to distinguish externally, but SEM studies provided enough characteristic features to distinguish from the other. This technique could be valuable in identifying seeds of important plant species for conservation and trading.

  3. Scanning Electron Microscopy (SEM) Procedure for HE Powders on a LEO 438VP System

    SciTech Connect

    Zaka, Fowzia

    2016-03-08

    This method describes the characterization of HE powders by Scanning Electron Microscopy (SEM). HE particles are dispersed onto an aluminum standard SEM specimen mount. Electron micrographs are collected at various magnifications (150 to 10,000 X) depending on HE particle size.

  4. Scanning Electron Microscopy (SEM) Procedure for HE Powders on a LEO 438VP System

    SciTech Connect

    Zaka, Fowzia

    2016-03-21

    This method describes the characterization of HE powders by Scanning Electron Microscopy (SEM). HE particles are dispersed onto an aluminum standard SEM specimen mount. Electron micrographs are collected at various magnifications (150 to 10,000 X) depending on HE particle size.

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

    PubMed

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

    2011-06-01

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

  6. Some strategies for quantitative scanning Auger electron microscopy

    NASA Technical Reports Server (NTRS)

    Browning, R.; Peacock, D. C.; Prutton, M.

    1985-01-01

    The general applicability of power law forms of the background in electron spectra is pointed out and exploited for background removal from under Auger peaks. This form of B(E) is found to be extremely sensitive to instrumental alignment and to fault-free construction - an observation which can be used to set up analyser configurations in an accurate way. Also, differences between N(E) and B(E) can be used to derive a spectrometer transmission function T(E). The questions of information density in an energy-analysing spatially-resolving instrument are addressed after reliable instrumental characterization has been established. Strategies involving ratio histograms, showing the population distribution of the ratio of a pair of Auger peak heights, composition scatter diagrams and windowed imaging are discussed and illustrated.

  7. Some strategies for quantitative scanning Auger electron microscopy

    NASA Technical Reports Server (NTRS)

    Browning, R.; Peacock, D. C.; Prutton, M.

    1985-01-01

    The general applicability of power law forms of the background in electron spectra is pointed out and exploited for background removal from under Auger peaks. This form of B(E) is found to be extremely sensitive to instrumental alignment and to fault-free construction - an observation which can be used to set up analyser configurations in an accurate way. Also, differences between N(E) and B(E) can be used to derive a spectrometer transmission function T(E). The questions of information density in an energy-analysing spatially-resolving instrument are addressed after reliable instrumental characterization has been established. Strategies involving ratio histograms, showing the population distribution of the ratio of a pair of Auger peak heights, composition scatter diagrams and windowed imaging are discussed and illustrated.

  8. High-Contrast Observation of Unstained Proteins and Viruses by Scanning Electron Microscopy

    PubMed Central

    Ogura, Toshihiko

    2012-01-01

    Scanning electron microscopy (SEM) is an important tool for the nanometre-scale analysis of the various samples. Imaging of biological specimens can be difficult for two reasons: (1) Samples must often be left unstained to observe detail of the biological structures; however, lack of staining significantly decreases image contrast. (2) Samples are prone to serious radiation damage from electron beam. Herein we report a novel method for sample preparation involving placement on a new metal-coated insulator film. This method enables obtaining high-contrast images from unstained proteins and viruses by scanning electron microscopy with minimal electron radiation damage. These images are similar to those obtained by transmission electron microscopy. In addition, the method can be easily used to observe specimens of proteins, viruses and other organic samples by using SEM. PMID:23056522

  9. Strain measurement in semiconductor heterostructures by scanning transmission electron microscopy.

    PubMed

    Müller, Knut; Rosenauer, Andreas; Schowalter, Marco; Zweck, Josef; Fritz, Rafael; Volz, Kerstin

    2012-10-01

    This article deals with the measurement of strain in semiconductor heterostructures from convergent beam electron diffraction patterns. In particular, three different algorithms in the field of (circular) pattern recognition are presented that are able to detect diffracted disc positions accurately, from which the strain in growth direction is calculated. Although the three approaches are very different as one is based on edge detection, one on rotational averages, and one on cross correlation with masks, it is found that identical strain profiles result for an In x Ga1-x N y As1-y /GaAs heterostructure consisting of five compressively and tensile strained layers. We achieve a precision of strain measurements of 7-9·10-4 and a spatial resolution of 0.5-0.7 nm over the whole width of the layer stack which was 350 nm. Being already very applicable to strain measurements in contemporary nanostructures, we additionally suggest future hardware and software designs optimized for fast and direct acquisition of strain distributions, motivated by the present studies.

  10. Scanning and Transmission Electron Microscopy of High Temperature Materials

    NASA Technical Reports Server (NTRS)

    1994-01-01

    Software and hardware updates to further extend the capability of the electron microscope were carried out. A range of materials such as intermetallics, metal-matrix composites, ceramic-matrix composites, ceramics and intermetallic compounds, based on refractory elements were examined under this research. Crystal structure, size, shape and volume fraction distribution of various phases which constitute the microstructures were examined. Deformed materials were studied to understand the effect of interfacial microstructure on the deformation and fracture behavior of these materials. Specimens tested for a range of mechanical property requirements, such as stress rupture, creep, low cycle fatigue, high cycle fatigue, thermomechanical fatigue, etc. were examined. Microstructural and microchemical stability of these materials exposed to simulated operating environments were investigated. The EOIM Shuttle post-flight samples were also examined to understand the influence of low gravity processing on microstructure. In addition, fractographic analyses of Nb-Zr-W, titanium aluminide, molybdenum silicide and silicon carbide samples were carried out. Extensive characterization of sapphire fibers in the fiber-reinforced composites made by powder cloth processing was made. Finally, pressure infiltration casting of metal-matrix composites was carried out.

  11. Scanning electron microscopy of experimental Trichophyton mentagrophytes infections in guinea pig skin.

    PubMed Central

    Hutton, R D; Kerbs, S; Yee, K

    1978-01-01

    Trichophyton mentagrophytes invasion of guinea pig skin was examined by scanning electron microscopy. Biopsies were obtained daily for 12 days from experimental infection sites. Dermatophyte invasion, examined in detail by scanning electron microscopy of cross-sectioned, prefixed skin was evidenced by: the appearance of hyphae within the stratum corneum; follicular invasion by hyphae, which remained initially within the follicle wall; emergence of the hyphae from the wall into the follicular canal; proliferation of the fungus down the follicle, with furrowing of the follicle wall and hair shaft cuticle; penetration of hyphae into the hair shaft by subcuticular and transcuticular routes; and massive peripilar hyphal proliferation with arthrosporogenesis. A three-dimensional perception of the invasion sequence of a dermatophyte in guinea pig skin was obtained by scanning electron microscopy. Images PMID:711318

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

    PubMed

    Tomitori, Masahiko; Sasahara, Akira

    2014-11-01

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

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

    PubMed Central

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

    2010-01-01

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

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

    PubMed

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

    2010-07-27

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

  15. Scanning electron microscopy imaging of dislocations in bulk materials, using electron channeling contrast.

    PubMed

    Crimp, Martin A

    2006-05-01

    The imaging and characterization of dislocations is commonly carried out by thin foil transmission electron microscopy (TEM) using diffraction contrast imaging. However, the thin foil approach is limited by difficult sample preparation, thin foil artifacts, relatively small viewable areas, and constraints on carrying out in situ studies. Electron channeling imaging of electron channeling contrast imaging (ECCI) offers an alternative approach for imaging crystalline defects, including dislocations. Because ECCI is carried out with field emission gun scanning electron microscope (FEG-SEM) using bulk specimens, many of the limitations of TEM thin foil analysis are overcome. This paper outlines the development of electron channeling patterns and channeling imaging to the current state of the art. The experimental parameters and set up necessary to carry out routine channeling imaging are reviewed. A number of examples that illustrate some of the advantages of ECCI over thin foil TEM are presented along with a discussion of some of the limitations on carrying out channeling contrast analysis of defect structures. Copyright (c) 2006 Wiley-Liss, Inc.

  16. Scanning electron microscopy of the human endolymphatic sac: a preliminary report.

    PubMed

    Galey, F R; House, W F

    1980-04-01

    Scanning electron microscopy has been used to examine and compare one normal endolymphatic sac with one endolymphatic sac from a patient with Meniere's disease. The surgical procedure for obtaining these specimens and their preparation for scanning electron microscopy are described. The luminal surface of the rugose portion of both specimens was lined with two populations of epithelial cells: one with a dome-shaped apical surface, the other with a flattened polygonal surface. The surface of dome-shaped cells in both specimens was covered with microvilli. Neither specimen had observable loss of epithelial integrity or fibrosis.

  17. Correlation of live-cell imaging with volume scanning electron microscopy.

    PubMed

    Lucas, Miriam S; Günthert, Maja; Bittermann, Anne Greet; de Marco, Alex; Wepf, Roger

    2017-01-01

    Live-cell imaging is one of the most widely applied methods in live science. Here we describe two setups for live-cell imaging, which can easily be combined with volume SEM for correlative studies. The first procedure applies cell culture dishes with a gridded glass support, which can be used for any light microscopy modality. The second approach is a flow-chamber setup based on Ibidi μ-slides. Both live-cell imaging strategies can be followed up with serial blockface- or focused ion beam-scanning electron microscopy. Two types of resin embedding after heavy metal staining and dehydration are presented making best use of the particular advantages of each imaging modality: classical en-bloc embedding and thin-layer plastification. The latter can be used only for focused ion beam-scanning electron microscopy, but is advantageous for studying cell-interactions with specific substrates, or when the substrate cannot be removed. En-bloc embedding has diverse applications and can be applied for both described volume scanning electron microscopy techniques. Finally, strategies for relocating the cell of interest are discussed for both embedding approaches and in respect to the applied light and scanning electron microscopy methods. Copyright © 2017 Elsevier Inc. All rights reserved.

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

    PubMed Central

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

    2014-01-01

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

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

    PubMed

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

    2014-09-01

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

  20. An inexpensive approach for bright-field and dark-field imaging by scanning transmission electron microscopy in scanning electron microscopy.

    PubMed

    Patel, Binay; Watanabe, Masashi

    2014-02-01

    Scanning transmission electron microscopy in scanning electron microscopy (STEM-in-SEM) is a convenient technique for soft materials characterization. Various specimen-holder geometries and detector arrangements have been used for bright-field (BF) STEM-in-SEM imaging. In this study, to further the characterization potential of STEM-IN-SEM, a new specimen holder has been developed to facilitate direct detection of BF signals and indirect detection of dark-field (DF) signals without the need for substantial instrument modification. DF imaging is conducted with the use of a gold (Au)-coated copper (Cu) plate attached to the specimen holder which directs highly scattered transmitted electrons to an off-axis yttrium-aluminum-garnet (YAG) detector. A hole in the copper plate allows for BF imaging with a transmission electron (TE) detector. The inclusion of an Au-coated Cu plate enhanced DF signal intensity. Experiments validating the acquisition of true DF signals revealed that atomic number (Z) contrast may be achieved for materials with large lattice spacing. However, materials with small lattice spacing still exhibit diffraction contrast effects in this approach. The calculated theoretical fine probe size is 1.8 nm. At 30 kV, in this indirect approach, DF spatial resolution is limited to 3.2 nm as confirmed experimentally.

  1. Chemical mapping and quantification at the atomic scale by scanning transmission electron microscopy.

    PubMed

    Chu, Ming-Wen; Chen, Cheng Hsuan

    2013-06-25

    With innovative modern material-growth methods, a broad spectrum of fascinating materials with reduced dimensions-ranging from single-atom catalysts, nanoplasmonic and nanophotonic materials to two-dimensional heterostructural interfaces-is continually emerging and extending the new frontiers of materials research. A persistent central challenge in this grand scientific context has been the detailed characterization of the individual objects in these materials with the highest spatial resolution, a problem prompting the need for experimental techniques that integrate both microscopic and spectroscopic capabilities. To date, several representative microscopy-spectroscopy combinations have become available, such as scanning tunneling microscopy, tip-enhanced scanning optical microscopy, atom probe tomography, scanning transmission X-ray microscopy, and scanning transmission electron microscopy (STEM). Among these tools, STEM boasts unique chemical and electronic sensitivity at unparalleled resolution. In this Perspective, we elucidate the advances in STEM and chemical mapping applications at the atomic scale by energy-dispersive X-ray spectroscopy and electron energy loss spectroscopy with a focus on the ultimate challenge of chemical quantification with atomic accuracy.

  2. The spatial coherence function in scanning transmission electron microscopy and spectroscopy.

    PubMed

    Nguyen, D T; Findlay, S D; Etheridge, J

    2014-11-01

    We investigate the implications of the form of the spatial coherence function, also referred to as the effective source distribution, for quantitative analysis in scanning transmission electron microscopy, and in particular for interpreting the spatial origin of imaging and spectroscopy signals. These questions are explored using three different source distribution models applied to a GaAs crystal case study. The shape of the effective source distribution was found to have a strong influence not only on the scanning transmission electron microscopy (STEM) image contrast, but also on the distribution of the scattered electron wavefield and hence on the spatial origin of the detected electron intensities. The implications this has for measuring structure, composition and bonding at atomic resolution via annular dark field, X-ray and electron energy loss STEM imaging are discussed. Copyright © 2014 Elsevier B.V. All rights reserved.

  3. A compilation of cold cases using scanning electron microscopy at the University of Rhode Island

    NASA Astrophysics Data System (ADS)

    Platek, Michael J.; Gregory, Otto J.

    2015-10-01

    Scanning electron microscopy combined with microchemical analysis has evolved into one of the most widely used instruments in forensic science today. In particular, the environmental scanning electron microscope (SEM) in conjunction with energy dispersive spectroscopy (EDS), has created unique opportunities in forensic science in regard to the examination of trace evidence; i.e. the examination of evidence without altering the evidence with conductive coatings, thereby enabling criminalists to solve cases that were previously considered unsolvable. Two cold cases were solved at URI using a JEOL 5900 LV SEM in conjunction with EDS. A cold case murder and a cold missing person case will be presented from the viewpoint of the microscopist and will include sample preparation, as well as image and chemical analysis of the trace evidence using electron microscopy and optical microscopy.

  4. Morphological aspects of Angiostrongylus costaricensis by light and scanning electron microscopy.

    PubMed

    Rebello, Karina M; Menna-Barreto, Rubem F S; Chagas-Moutinho, Vanessa A; Mota, Ester M; Perales, Jonas; Neves-Ferreira, Ana Gisele C; Oliveira-Menezes, Aleksandra; Lenzi, Henrique

    2013-09-01

    Angiostrongylus costaricensis is a parasitic nematode that can cause severe gastrointestinal disease, known as abdominal angiostrongiliasis, in humans. This paper presents the characterization of first- and third-stage larvae and male and female adult worms of A. costaricensis by scanning electron and light microscopy. Several novel anatomical structures were identified by scanning electron microscopy, including details of the cuticular striations of the spicules in male worms and a protective flap of the cuticle covering the vulvar aperture in female worms. Other taxonomic features revealed by light microscopy include the gubernaculum and the esophageal-intestinal valve. The use of two microscopy techniques allowed a detailed characterization of the morphology of this nematode. A number of previously identified taxonomic features, such as the striated nature of the spicules and the lateral alae were confirmed; however, the use of scanning electron microscopy resulted in a reassessment of the correct number of papillae distributed around the oral opening and behind the cloacal opening. These observations, in combination with light microscopy-based characterization of the gubernaculum and esophageal valves, have allowed a more detailed description of this nematode taxonomy.

  5. Morphology of glass fibers in electronics workers with fiberglass dermatitis--a scanning electron microscopy study.

    PubMed

    Hsieh, M Y; Guo, Y L; Shiao, J S; Sheu, H M

    2001-04-01

    Fiberglass is used as a reinforcement filler material in printed circuit boards (PCBs) which are widely used in the electronics industry. In a recent survey, we demonstrated that fiberglass dermatitis is the most common occupational dermatosis among electronics industry workers in Taiwan. Little is known, however, about the morphologic structures of the glass fibers which induce dermatitis. The purpose of this study was to assess the morphology of fiber spicules and to determine the relationship of this structure to fiberglass dermatitis. Fourteen female patients with a diagnosis of fiberglass dermatitis were selected for study. The diagnosis was confirmed in all patients by positive skin stripping for glass fibers and matching with glass fibers from dust collected in work areas and from samples collected by scraping the edge of PCBs. Samples of collected glass fibers were analyzed by scanning electron microscopy (SEM). SEM of the fiberglass samples revealed that fibers were approximately 10 microm in diameter. In samples from both the edge of PCBs and from dust collected in work areas, SEM revealed that most of the fibers were in bundles of various sizes and lengths. All fibers collected from patients' skin by tape stripping showed a singular spicule, most had a sharp free end, and the lengths were in the range 50-150 microm . Singular glass fibers with a sharp free end and a length of 50-150 microm are most likely to induce fiberglass dermatitis.

  6. Dental wax impressions of plant tissues for viewing with scanning electron microscopy (SEM).

    PubMed

    Beermann, Anke; Hülskamp, Martin

    2010-09-01

    Scanning electron microscopy (SEM) is a valuable method for examining surface structures. Taking wax impressions of plant structures, such as leaves, is a nondestructive procedure that makes it possible to view changes in surface structures over time, such as during development. This protocol describes a method for making dental wax impressions of plant tissues.

  7. Characterizing individual particles on tree leaves using computer automated scanning electron microscopy

    Treesearch

    D. L. Johnson; D. J. Nowak; V. A. Jouraeva

    1999-01-01

    Leaves from twenty-three deciduous tree species and five conifer species were collected within a limited geographic range (1 km radius) and evaluated for possible application of scanning electron microscopy and X-ray microanalysis techniques of individual particle analysis (IPA). The goal was to identify tree species with leaves suitable for the automated...

  8. Scanning electron microscopy of Ascaridia galli (Schrank, 1788), Freeborn, 1923 and A. columbae (Linstow, 1903).

    PubMed

    Ashour, A A

    1994-08-01

    The morphology of the two ascaridoid nematodes Ascaridia galli and A. columbae was studied by scanning electron microscopy. The two nematodes were compared together and their specific characteristics were established, including lips, cephalic papillae, body cuticle, spicules and caudal papillae of the male.

  9. Electronic properties of graphene: a perspective from scanning tunneling microscopy and magnetotransport.

    PubMed

    Andrei, Eva Y; Li, Guohong; Du, Xu

    2012-05-01

    This review covers recent experimental progress in probing the electronic properties of graphene and how they are influenced by various substrates, by the presence of a magnetic field and by the proximity to a superconductor. The focus is on results obtained using scanning tunneling microscopy, spectroscopy, transport and magnetotransport techniques.

  10. EVALUATION OF COMPUTER-CONTROLLED SCANNING ELECTRON MICROSCOPY APPLIED TO AN AMBIENT URBAN AEROSOL SAMPLE

    EPA Science Inventory


    Recent interest in monitoring and speciation of particulate matter has led to increased application of scanning electron microscopy (SEM) coupled with energy-dispersive x-ray analysis (EDX) to individual particle analysis. SEM/EDX provides information on the size, shape, co...

  11. Observation of microporous cesium salts of 12-tungstosilicic acid using scanning transmission electron microscopy.

    PubMed

    Hiyoshi, Norihito; Kamiya, Yuichi

    2015-06-21

    Heteropolyanions and their arrays in microporous cesium salts of 12-tungstosilicic acid, Cs2.5H1.5[SiW12O40] and Cs4.0[SiW12O40], were observed by aberration-corrected scanning transmission electron microscopy. Microstructures that form micropores in the polyoxometalates were visualized.

  12. EVALUATION OF COMPUTER-CONTROLLED SCANNING ELECTRON MICROSCOPY APPLIED TO AN AMBIENT URBAN AEROSOL SAMPLE

    EPA Science Inventory


    Recent interest in monitoring and speciation of particulate matter has led to increased application of scanning electron microscopy (SEM) coupled with energy-dispersive x-ray analysis (EDX) to individual particle analysis. SEM/EDX provides information on the size, shape, co...

  13. The material dependence of temperature measurement resolution in thermal scanning electron microscopy

    SciTech Connect

    Wu, Xiaowei; Hull, Robert

    2013-03-18

    Thermal scanning electron microscopy is a recently developed temperature mapping technique based on thermal diffuse scattering in electron backscatter diffraction in a scanning electron microscope. It provides nano-scale and non-contact temperature mapping capabilities. Due to the specific temperature sensitive mechanism inherent to this technique, the temperature resolution is highly material dependent. A thorough investigation of what material properties affect the temperature resolution is important for realizing the inherent temperature resolution limit for each material. In this paper, three material dependent parameters-the Debye-Waller B-factor temperature sensitivity, backscatter yield, and lattice constant-are shown to control the temperature resolution.

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

    PubMed

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

    2013-12-01

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

  15. SEM, TEM and SLEEM (scanning low energy electron microscopy) of CB2 steel after creep testing

    NASA Astrophysics Data System (ADS)

    Kasl, J.; Mikmeková, Š.; Jandová, D.

    2014-03-01

    The demand to produce electrical power with higher efficiency and with lower environmental pollution is leading to the use of new advanced materials in the production of power plant equipment. To understand the processes taking place in parts produced from these materials during their operation under severe conditions (such as high temperature, high stress, and environmental corrosion) requires detailed evaluation of their substructure. It is usually necessary to use transmission electron microscopy (TEM). However, this method is very exacting and time-consuming. So there is an effort to use new scanning electron microscopy techniques instead of TEM. One of them is scanning low energy electron microscopy (SLEEM). This paper deals with an assessment of the possibility to use SLEEM for describing the substructure of creep resistant steel CB2 after long-term creep testing. In the SLEEM images more information is contained about the microstructure of the material in comparison with standard scanning electron microscopy. Study of materials using slow and very slow electrons opens the way to better understanding their microstructures.

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

    PubMed

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

    2011-08-01

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

  17. In situ tensile testing of nanofibers by combining atomic force microscopy and scanning electron microscopy.

    PubMed

    Hang, Fei; Lu, Dun; Bailey, Russell J; Jimenez-Palomar, Ines; Stachewicz, Urszula; Cortes-Ballesteros, Beatriz; Davies, Martin; Zech, Martin; Bödefeld, Christoph; Barber, Asa H

    2011-09-07

    A nanomechanical testing set-up is developed by integrating an atomic force microscope (AFM) for force measurements with a scanning electron microscope (SEM) to provide imaging capabilities. Electrospun nanofibers of polyvinyl alcohol (PVA), nylon-6 and biological mineralized collagen fibrils (MCFs) from antler bone were manipulated and tensile-tested using the AFM-SEM set-up. The complete stress-strain behavior to failure of individual nanofibers was recorded and a diversity of mechanical properties observed, highlighting how this technique is able to elucidate mechanical behavior due to structural composition at nanometer length scales.

  18. Tip surface changes in endocardial stimulation electrode, visualised by scanning electron microscopy.

    PubMed

    Hladky, M; Horn, V; Kamaryt, P; Cabanova, J; Zeman, K

    1975-01-01

    The authors have been probably the first investigators who applied scanning electron microscopy to studies of the changes occurring in the surface of the metalic tip of an endocardial stimulating electrode. They found a lowered conductivity for secondary electron emission, and describe the surface changes in a platiniridium-tipped electrode which had been used for almost four years, in comparison with an unused electrode.

  19. Epidermal growth factor receptor subunit locations determined in hydrated cells with environmental scanning electron microscopy.

    PubMed

    Peckys, Diana B; Baudoin, Jean-Pierre; Eder, Magdalena; Werner, Ulf; de Jonge, Niels

    2013-01-01

    Imaging single epidermal growth factor receptors (EGFR) in intact cells is presently limited by the available microscopy methods. Environmental scanning electron microscopy (ESEM) of whole cells in hydrated state in combination with specific labeling with gold nanoparticles was used to localize activated EGFRs in the plasma membranes of COS7 and A549 cells. The use of a scanning transmission electron microscopy (STEM) detector yielded a spatial resolution of 3 nm, sufficient to identify the locations of individual EGFR dimer subunits. The sizes and distribution of dimers and higher order clusters of EGFRs were determined. The distance between labels bound to dimers amounted to 19 nm, consistent with a molecular model. A fraction of the EGFRs was found in higher order clusters with sizes ranging from 32-56 nm. ESEM can be used for quantitative whole cell screening studies of membrane receptors, and for the study of nanoparticle-cell interactions in general.

  20. Epidermal growth factor receptor subunit locations determined in hydrated cells with environmental scanning electron microscopy

    PubMed Central

    Peckys, Diana B.; Baudoin, Jean-Pierre; Eder, Magdalena; Werner, Ulf; de Jonge, Niels

    2013-01-01

    Imaging single epidermal growth factor receptors (EGFR) in intact cells is presently limited by the available microscopy methods. Environmental scanning electron microscopy (ESEM) of whole cells in hydrated state in combination with specific labeling with gold nanoparticles was used to localize activated EGFRs in the plasma membranes of COS7 and A549 cells. The use of a scanning transmission electron microscopy (STEM) detector yielded a spatial resolution of 3 nm, sufficient to identify the locations of individual EGFR dimer subunits. The sizes and distribution of dimers and higher order clusters of EGFRs were determined. The distance between labels bound to dimers amounted to 19 nm, consistent with a molecular model. A fraction of the EGFRs was found in higher order clusters with sizes ranging from 32–56 nm. ESEM can be used for quantitative whole cell screening studies of membrane receptors, and for the study of nanoparticle-cell interactions in general. PMID:24022088

  1. Scanning electron microscopy of cells and tissues under fully hydrated conditions.

    PubMed

    Thiberge, Stephan; Nechushtan, Amotz; Sprinzak, David; Gileadi, Opher; Behar, Vered; Zik, Ory; Chowers, Yehuda; Michaeli, Shulamit; Schlessinger, Joseph; Moses, Elisha

    2004-03-09

    A capability for scanning electron microscopy of wet biological specimens is presented. A membrane that is transparent to electrons protects the fully hydrated sample from the vacuum. The result is a hybrid technique combining the ease of use and ability to see into cells of optical microscopy with the higher resolution of electron microscopy. The resolution of low-contrast materials is approximately 100 nm, whereas in high-contrast materials the resolution can reach 10 nm. Standard immunogold techniques and heavy-metal stains can be applied and viewed in the fluid to improve the contrast. Images present a striking combination of whole-cell morphology with a wealth of internal details. A possibility for direct inspection of tissue slices transpires, imaging only the external layer of cells. Simultaneous imaging with photons excited by the electrons incorporates data on material distribution, indicating a potential for multilabeling and specific scintillating markers.

  2. Visualizing Macromolecular Complexes with In Situ Liquid Scanning Transmission Electron Microscopy

    SciTech Connect

    Evans, James E.; Jungjohann, K. L.; Wong, Peony C. K.; Chiu, Po-Lin; Dutrow, Gavin H.; Arslan, Ilke; Browning, Nigel D.

    2012-11-01

    A central focus of biological research is understanding the structure/function relationship of macromolecular protein complexes. Yet conventional transmission electron microscopy techniques are limited to static observations. Here we present the first direct images of purified macromolecular protein complexes using in situ liquid scanning transmission electron microscopy. Our results establish the capability of this technique for visualizing the interface between biology and nanotechnology with high fidelity while also probing the interactions of biomolecules within solution. This method represents an important advancement towards allowing future high-resolution observations of biological processes and conformational dynamics in real-time.

  3. Phase reconstruction in annular bright-field scanning transmission electron microscopy.

    PubMed

    Ishida, Takafumi; Kawasaki, Tadahiro; Tanji, Takayoshi; Kodama, Tetsuji; Matsutani, Takaomi; Ogai, Keiko; Ikuta, Takashi

    2015-04-01

    A novel technique for reconstructing the phase shifts of electron waves was applied to Cs-corrected scanning transmission electron microscopy (STEM). To realize this method, a new STEM system equipped with an annular aperture, annularly arrayed detectors and an arrayed image processor has been developed and evaluated in experiments. We show a reconstructed phase image of graphite particles and demonstrate that this new method works effectively for high-resolution phase imaging. © The Author 2014. Published by Oxford University Press on behalf of The Japanese Society of Microscopy. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

  4. Visualizing macromolecular complexes with in situ liquid scanning transmission electron microscopy.

    PubMed

    Evans, James E; Jungjohann, Katherine L; Wong, Peony C K; Chiu, Po-Lin; Dutrow, Gavin H; Arslan, Ilke; Browning, Nigel D

    2012-11-01

    A central focus of biological research is understanding the structure/function relationship of macromolecular protein complexes. Yet conventional transmission electron microscopy techniques are limited to static observations. Here we present the first direct images of purified macromolecular protein complexes using in situ liquid scanning transmission electron microscopy. Our results establish the capability of this technique for visualizing the interface between biology and nanotechnology with high fidelity while also probing the interactions of biomolecules within solution. This method represents an important advancement towards allowing future high-resolution observations of biological processes and conformational dynamics in real-time. Published by Elsevier Ltd.

  5. Liquid scanning transmission electron microscopy: imaging protein complexes in their native environment in whole eukaryotic cells.

    PubMed

    Peckys, Diana B; de Jonge, Niels

    2014-04-01

    Scanning transmission electron microscopy (STEM) of specimens in liquid, so-called Liquid STEM, is capable of imaging the individual subunits of macromolecular complexes in whole eukaryotic cells in liquid. This paper discusses this new microscopy modality within the context of state-of-the-art microscopy of cells. The principle of operation and equations for the resolution are described. The obtained images are different from those acquired with standard transmission electron microscopy showing the cellular ultrastructure. Instead, contrast is obtained on specific labels. Images can be recorded in two ways, either via STEM at 200 keV electron beam energy using a microfluidic chamber enclosing the cells, or via environmental scanning electron microscopy at 30 keV of cells in a wet environment. The first series of experiments involved the epidermal growth factor receptor labeled with gold nanoparticles. The labels were imaged in whole fixed cells with nanometer resolution. Since the cells can be kept alive in the microfluidic chamber, it is also feasible to detect the labels in unfixed, live cells. The rapid sample preparation and imaging allows studies of multiple whole cells.

  6. Use of Low Temperature Scanning Electron Microscopy to Observe Frozen Hydrated Specimens of Nematodes

    PubMed Central

    Wergin, William P.; Sayre, Richard M.; Erbe, Eric F.

    1993-01-01

    Frozen hydrated specimens of Pratylenchus agilis and dauer larvae of Steinernema carpocapsae were observed with low-temperature field emission scanning electron microscopy. This new technique provides information about the surface features of nematodes and also allows specimens to be fractured to reveal their internal structure. Furthermore, both halves of fractured specimens can be retained, examined, and photographed either as two-dimensional micrographs or as three-dimensional images for stereo observation (stereology) or quantitative measurements (stereometry). This technique avoids artifacts normally associated with procedures required to prepare nematodes for examination in the transmission and scanning electron microscopes, such as chemical fixation, dehydration, and sectioning or critical point drying. PMID:19279761

  7. A novel approach to scanning electron microscopy at ambient atmospheric pressure.

    PubMed

    Ominami, Yusuke; Kawanishi, Shinsuke; Ushiki, Tatsuo; Ito, Sukehiro

    2015-04-01

    Scanning electron microscopy (SEM) for observing samples at ambient atmospheric pressure is introduced in this study. An additional specimen chamber with a small window is inserted in the main specimen chamber, and the window is separated with a thin membrane or diaphragm allowing electron beam propagation. Close proximity of the sample to the membrane enables the detection of back-scattered electrons sufficient for imaging. In addition to the empirical imaging data, a probability analysis of the un-scattered fraction of the incident electron beam further supports the feasibility of atmospheric SEM imaging over a controlled membrane-sample distance.

  8. Electron scattering in a multiwall carbon nanotube bend junction studied by scanning tunneling microscopy

    NASA Astrophysics Data System (ADS)

    Tapasztó, L.; Nemes-Incze, P.; Osváth, Z.; Darabont, Al.; Lambin, Ph.; Biró, L. P.

    2006-12-01

    The atomic resolution scanning tunneling microscopy investigation of a multiwall carbon nanotube bend junction is reported. Atomic resolution images taken at the junction region revealed position-dependent modulation of the electronic density of states, with a period larger than but commensurate to the underlying atomic lattice, attributed to the scattering of electrons on defect sites present in the junction region. We propose an interference model, suitable to interpret the experimentally observed electron density patterns by considering electronic states near the bands crossing points involved in the scattering processes. The model predicts that complex charge density oscillations present near defects are tunable by varying the applied bias potential.

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

    SciTech Connect

    Lansåker, Pia C. Niklasson, Gunnar A.; Granqvist, Claes G.; Hallén, Anders

    2014-10-15

    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.

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

    PubMed

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

    2001-10-01

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

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

    SciTech Connect

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

    1990-01-01

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

  12. Cytogenetic Characterization of the TM4 Mouse Sertoli Cell Line. II. Chromosome Microdissection, FISH, Scanning Electron Microscopy, and Confocal Laser Scanning Microscopy.

    PubMed

    Schmid, Michael; Guttenbach, Martina; Steinlein, Claus; Wanner, Gerhard; Houben, Andreas

    2015-01-01

    The chromosomes and interphase cell nuclei of the permanent mouse Sertoli cell line TM4 were examined by chromosome microdissection, FISH, scanning electron microscopy, and confocal laser scanning microscopy. The already known marker chromosomes m1-m5 were confirmed, and 2 new large marker chromosomes m6 and m7 were characterized. The minute heterochromatic marker chromosomes m4 and m5 were microdissected and their DNA amplified by DOP-PCR. FISH of this DNA probe on TM4 metaphase chromosomes demonstrated that the m4 and m5 marker chromosomes have derived from the centromeric regions of normal telocentric mouse chromosomes. Ectopic pairing of the m4 and m5 marker chromosomes with the centromeric region of any of the other chromosomes (centromeric associations) was apparent in ∼60% of the metaphases. Scanning electron microscopy revealed DNA-protein bridges connecting the centromeric regions of normal chromosomes and the associated m4 and m5 marker chromosomes. Interphase cell nuclei of TM4 Sertoli cells did not exhibit the characteristic morphology of Sertoli cells in the testes of adult mice as shown by fluorescence microscopy and confocal laser scanning microscopy.

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

    PubMed

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

    2017-05-01

    We analyze electron-beam induced carbon contamination in a transmission electron microscope. The study is performed on thin films potentially suitable as phase plates for phase-contrast transmission electron microscopy. Electron energy-loss spectroscopy and phase-plate imaging is utilized to analyze the contamination. The deposited contamination layer is identified as a graphitic carbon layer which is not prone to electrostatic charging whereas a non-conductive underlying substrate charges. Several methods that inhibit contamination are evaluated and the impact of carbon contamination on phase-plate imaging is discussed. The findings are in general interesting for scanning transmission electron microscopy applications. Crown Copyright © 2017. Published by Elsevier Ltd. All rights reserved.

  14. X-ray diffraction and scanning electron microscopy of galvannealed coatings on steel.

    PubMed

    Schmid, P; Uran, K; Macherey, F; Ebert, M; Ullrich, H-J; Sommer, D; Friedel, F

    2009-04-01

    The formation of Fe-Zn intermetallic compounds, as relevant in the commercial product galvannealed steel sheet, was investigated by scanning electron microscopy and different methods of X-ray diffraction. A scanning electron microscope with high resolution was applied to investigate the layers of the galvannealed coating and its topography. Grazing incidence X-ray diffraction (GID) was preferred over conventional Bragg-Brentano geometry for analysing thin crystalline layers because of its lower incidence angle alpha and its lower depth of information. Furthermore, in situ experiments at an environmental scanning electron microscope (ESEM) with an internal heating plate and at an X-ray diffractometer equipped with a high-temperature chamber were carried out. Thus, it was possible to investigate the phase evolution during heat treatment by X-ray diffraction and to display the growth of the zeta crystals in the ESEM.

  15. Electron Microscopy.

    ERIC Educational Resources Information Center

    Beer, Michael

    1980-01-01

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

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

    PubMed

    Attree, S M; Sheffield, E

    1985-08-01

    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 staining with fluorescein diacetate. With scanning electron microscopy, plasmolysed protoplast surfaces appeared smooth with no evidence of wrinkling or infolding of excess membrane. The formation of irregular-shaped protoplasts, protoplasmic threads, subprotoplasts, and protoplasmic networks covering internal wall surfaces all provided evidence for strong wall adhesion of the protoplasm. The availability of membrane for uptake into folds or vesicles is therefore thought to be minimal. Transmission electron microscopy showed some protoplasmic threads to be plasmodesmata, the remainder being cell-wall contact points. Remnants of these threads were occasionally observed on isolated protoplasts in both the light and electron microscopes.

  17. Correlative In Vivo 2 Photon and Focused Ion Beam Scanning Electron Microscopy of Cortical Neurons

    PubMed Central

    Maco, Bohumil; Holtmaat, Anthony; Cantoni, Marco; Kreshuk, Anna; Straehle, Christoph N.; Hamprecht, Fred A.; Knott, Graham W.

    2013-01-01

    Correlating in vivo imaging of neurons and their synaptic connections with electron microscopy combines dynamic and ultrastructural information. Here we describe a semi-automated technique whereby volumes of brain tissue containing axons and dendrites, previously studied in vivo, are subsequently imaged in three dimensions with focused ion beam scanning electron microcopy. These neurites are then identified and reconstructed automatically from the image series using the latest segmentation algorithms. The fast and reliable imaging and reconstruction technique avoids any specific labeling to identify the features of interest in the electron microscope, and optimises their preservation and staining for 3D analysis. PMID:23468982

  18. Scanning electron microscopy of glomerular and non glomerular red blood cells.

    PubMed

    Fassett, R G; Horgan, B; Gove, D; Mathew, T H

    1983-07-01

    Phase contrast microscopic examination of the urine has been recently shown to be of value in predicting whether hematuria is due to glomerulonephritis or lesions of the lower urinary tract. Glomerular red cells show variations in size and shape and have distorted surfaces. Non glomerular red cells are uniform in size and shape and have smooth surfaces. Scanning electron microscopy was performed on urine sediment containing either glomerular or non glomerular red cells to better define their surface characteristics. Glomerular red cells exhibited a variety of forms, most cells having lumpy projections from the surface, some showing fragmentation of the membrane and others showing gross distortion. In contrast non glomerular red cells show smooth surfaces and usually maintain the normal biconcave disc shape of peripheral red blood cells. Scanning electron microscopy can better define surface structural abnormalities of urinary glomerular and non glomerular red blood cells.

  19. Chromatin Higher-Order Structure Studied by Neutron Scattering and Scanning Transmission Electron Microscopy

    NASA Astrophysics Data System (ADS)

    Gerchman, S. E.; Ramakrishnan, V.

    1987-11-01

    Neutron scattering in solution and scanning transmission electron microscopy were simultaneously done on chicken erythrocyte chromatin at various salt and magnesium concentrations. We show that chromatin is organized into a higher-order structure even at low ionic strength and that the mass per unit length increases continuously as a function of salt concentration, reaching a limiting value of between six and seven nucleosomes per 11 nm. There is no evidence of a transition from a 10-nm to a 30-nm fiber. Fiber diameter is correlated with mass per unit length, showing that both increase during condensation. We also find that there is no essential difference between the mass per unit length measured by scanning transmission electron microscopy and neutron scattering in solution, showing that the ordered regions seen in micrographs are representative of chromatin in solution.

  20. Scanning electron microscopy as an analytical tool for the study of calcified intrauterine contraceptive devices

    SciTech Connect

    Khan, S.R.; Wilkinson, E.J.

    1985-01-01

    Within the endometrial cavity intrauterine contraceptive devices (IUDs) become encrusted with cellular, acellular, and fibrillar substances. Scanning electron microscopy was used to study the crust. Cellular material consisted mainly of blood cells and various types of bacteria. The fibrillar material appeared to be fibrin which was omnipresent in the crust and formed a thin layer immediately over the IUD surface. X-ray microanalysis of the acellular component of the crust revealed the presence of calcium. No other major peaks were identified. Near the IUD surface characteristic calcium phosphate crystals were present. Their microanalysis showed peaks for calcium and phosphorus. X-ray diffraction of the crust however, showed it to contain only calcite. It is through the use of scanning electron microscopy that calcium phosphate has been detected in the IUD crust and a fibrillar layer has been visualized on the IUD surface. This study further demonstrates the effectiveness of SEM analytical techniques in the area of biomedical research.

  1. Scanning electron microscopy analysis of experimental bone hacking trauma of the mandible.

    PubMed

    Alunni-Perret, Véronique; Borg, Cybèle; Laugier, Jean-Pierre; Bertrand, Marie-France; Staccini, Pascal; Bolla, Marc; Quatrehomme, Gérald; Muller-Bolla, Michèle

    2010-12-01

    The authors report on a macroscopic and microscopic study of human mandible bone lesions achieved by a single-blade knife and a hatchet. The aim of this work was to complete the previous data (scanning electron microscopy analysis of bone lesions made by a single-blade knife and a hatchet, on human femurs) and to compare the lesions of the femur with those of the mandible. The results indicate that the mandible is a more fragile bone, but the features observed on the mandible are quite similar to those previously observed on the femur. This work spells out the main scanning electron microscopy characteristics of sharp (bone cutting) and blunt (exerting a pressure on the bone) mechanisms on human bone. Weapon characteristics serve to explain all of these features.

  2. Picometre-precision analysis of scanning transmission electron microscopy images of platinum nanocatalysts.

    PubMed

    Yankovich, Andrew B; Berkels, Benjamin; Dahmen, W; Binev, P; Sanchez, S I; Bradley, S A; Li, Ao; Szlufarska, Izabela; Voyles, Paul M

    2014-06-11

    Measuring picometre-scale shifts in the positions of individual atoms in materials provides new insight into the structure of surfaces, defects and interfaces that influence a broad variety of materials' behaviour. Here we demonstrate sub-picometre precision measurements of atom positions in aberration-corrected Z-contrast scanning transmission electron microscopy images based on the non-rigid registration and averaging of an image series. Non-rigid registration achieves five to seven times better precision than previous methods. Non-rigidly registered images of a silica-supported platinum nanocatalyst show pm-scale contraction of atoms at a (111)/(111) corner towards the particle centre and expansion of a flat (111) facet. Sub-picometre precision and standardless atom counting with <1 atom uncertainty in the same scanning transmission electron microscopy image provide new insight into the three-dimensional atomic structure of catalyst nanoparticle surfaces, which contain the active sites controlling catalytic reactions.

  3. Efficient linear phase contrast in scanning transmission electron microscopy with matched illumination and detector interferometry

    NASA Astrophysics Data System (ADS)

    Ophus, Colin; Ciston, Jim; Pierce, Jordan; Harvey, Tyler R.; Chess, Jordan; McMorran, Benjamin J.; Czarnik, Cory; Rose, Harald H.; Ercius, Peter

    2016-02-01

    The ability to image light elements in soft matter at atomic resolution enables unprecedented insight into the structure and properties of molecular heterostructures and beam-sensitive nanomaterials. In this study, we introduce a scanning transmission electron microscopy technique combining a pre-specimen phase plate designed to produce a probe with structured phase with a high-speed direct electron detector to generate nearly linear contrast images with high efficiency. We demonstrate this method by using both experiment and simulation to simultaneously image the atomic-scale structure of weakly scattering amorphous carbon and strongly scattering gold nanoparticles. Our method demonstrates strong contrast for both materials, making it a promising candidate for structural determination of heterogeneous soft/hard matter samples even at low electron doses comparable to traditional phase-contrast transmission electron microscopy. Simulated images demonstrate the extension of this technique to the challenging problem of structural determination of biological material at the surface of inorganic crystals.

  4. Efficient linear phase contrast in scanning transmission electron microscopy with matched illumination and detector interferometry

    PubMed Central

    Ophus, Colin; Ciston, Jim; Pierce, Jordan; Harvey, Tyler R.; Chess, Jordan; McMorran, Benjamin J.; Czarnik, Cory; Rose, Harald H.; Ercius, Peter

    2016-01-01

    The ability to image light elements in soft matter at atomic resolution enables unprecedented insight into the structure and properties of molecular heterostructures and beam-sensitive nanomaterials. In this study, we introduce a scanning transmission electron microscopy technique combining a pre-specimen phase plate designed to produce a probe with structured phase with a high-speed direct electron detector to generate nearly linear contrast images with high efficiency. We demonstrate this method by using both experiment and simulation to simultaneously image the atomic-scale structure of weakly scattering amorphous carbon and strongly scattering gold nanoparticles. Our method demonstrates strong contrast for both materials, making it a promising candidate for structural determination of heterogeneous soft/hard matter samples even at low electron doses comparable to traditional phase-contrast transmission electron microscopy. Simulated images demonstrate the extension of this technique to the challenging problem of structural determination of biological material at the surface of inorganic crystals. PMID:26923483

  5. Efficient linear phase contrast in scanning transmission electron microscopy with matched illumination and detector interferometry.

    PubMed

    Ophus, Colin; Ciston, Jim; Pierce, Jordan; Harvey, Tyler R; Chess, Jordan; McMorran, Benjamin J; Czarnik, Cory; Rose, Harald H; Ercius, Peter

    2016-02-29

    The ability to image light elements in soft matter at atomic resolution enables unprecedented insight into the structure and properties of molecular heterostructures and beam-sensitive nanomaterials. In this study, we introduce a scanning transmission electron microscopy technique combining a pre-specimen phase plate designed to produce a probe with structured phase with a high-speed direct electron detector to generate nearly linear contrast images with high efficiency. We demonstrate this method by using both experiment and simulation to simultaneously image the atomic-scale structure of weakly scattering amorphous carbon and strongly scattering gold nanoparticles. Our method demonstrates strong contrast for both materials, making it a promising candidate for structural determination of heterogeneous soft/hard matter samples even at low electron doses comparable to traditional phase-contrast transmission electron microscopy. Simulated images demonstrate the extension of this technique to the challenging problem of structural determination of biological material at the surface of inorganic crystals.

  6. Efficient linear phase contrast in scanning transmission electron microscopy with matched illumination and detector interferometry

    SciTech Connect

    Ophus, Colin; Ciston, Jim; Pierce, Jordan; Harvey, Tyler R.; Chess, Jordan; McMorran, Benjamin J.; Czarnik, Cory; Rose, Harald H.; Ercius, Peter

    2016-02-29

    The ability to image light elements in soft matter at atomic resolution enables unprecedented insight into the structure and properties of molecular heterostructures and beam-sensitive nanomaterials. In this study, we introduce a scanning transmission electron microscopy technique combining a pre-specimen phase plate designed to produce a probe with structured phase with a high-speed direct electron detector to generate nearly linear contrast images with high efficiency. We demonstrate this method by using both experiment and simulation to simultaneously image the atomic-scale structure of weakly scattering amorphous carbon and strongly scattering gold nanoparticles. Our method demonstrates strong contrast for both materials, making it a promising candidate for structural determination of heterogeneous soft/hard matter samples even at low electron doses comparable to traditional phase-contrast transmission electron microscopy. Ultimately, simulated images demonstrate the extension of this technique to the challenging problem of structural determination of biological material at the surface of inorganic crystals.

  7. A Novel Approach for Automated Analysis of Cell Attachment and Spreading Based on Backscattered Electron Imaging by Scanning Electron Microscopy

    PubMed Central

    Katsen-Globa, Alisa; Peter, Leonora; Zöllner, Susan; Dörge, Thomas; Daffertshofer, Martin; Preckel, Hartwig; Schmitt, Daniel; Zimmermann, Heiko

    2009-01-01

    The development of new materials for biological application requires in vitro testing of cell/surface interactions. Cell adhesion and spreading are difficult to quantify as most materials are non-transparent and transmission microscopy cannot be used. Contrast in reflection microscopy is rather poor. We propose an alternative method for the automated screening of cell attachment and spreading using backscattered electron imaging of scanning electron microscopy. The enhanced cell contrast permits study of cell/material interactions by little differences between cells and material.

  8. Visualization of Aspergillus fumigatus biofilms with Scanning Electron Microscopy and Variable Pressure-Scanning Electron Microscopy: A comparison of processing techniques.

    PubMed

    Joubert, Lydia-Marie; Ferreira, Jose Ag; Stevens, David A; Nazik, Hasan; Cegelski, Lynette

    2017-01-01

    Aspergillus fumigatus biofilms consist of a three-dimensional network of cellular hyphae and extracellular matrix. They are involved in infections of immune-compromised individuals, particularly those with cystic fibrosis. These structures are associated with persistence of infection, resistance to host immunity, and antimicrobial resistance. Thorough understanding of structure and function is imperative in the design of therapeutic drugs. Optimization of processing parameters, including aldehyde fixation, heavy metal contrasting, drying techniques and Ionic Liquid treatment, was undertaken for an ultrastructural approach to understand cellular and extracellular biofilm components. Conventional and Variable Pressure Scanning Electron Microscopy were applied to analyze the structure of biofilms attached to plastic and formed at an air-liquid interface.

  9. Ultra structural studies of the surface of Hymenolepis nana by scanning and transmission electron microscopy.

    PubMed

    Abouzakham, A A; Romia, S A; Hegazi, M M

    1990-06-01

    Scanning electron microscopy of the surface of Hymenolepis nana indicated that dense populations of microtriches occur on scolex proper, suckers and strobila, with an average density of 20/micron2. The excellent preservation of microtriches proves the efficacy of the critical point drying method for preparing cestodes for study of SEM. The cytological structure of the tegument of H. nana corresponds in general to that of other tapeworms.

  10. Endolithic algae and micrite envelope formation in Bahamian oolites as revealed by scanning electron microscopy.

    NASA Technical Reports Server (NTRS)

    Margolis, S.; Rex, R. W.

    1971-01-01

    Examination of Holocene Bahamian ooelites by scanning electron and light microscopy has revealed the morphology and orientation of aragonite crystals in the lamellar ooelitic envelope, and their modification by the boring activities of endolithic algae. The voids produced by these algae are found in progressive stages of being lined and filled with precipitated microcrystalline aragonite, which is similar to the process of micrite envelope formation in molluscan and other skeletal carbonate grains.

  11. Scanning electron microscopy of lunar regolith from the Sea of Fertility

    NASA Technical Reports Server (NTRS)

    Antoshin, M. K.; Ilin, N. P.; Spivak, G. V.

    1974-01-01

    Scanning electron microscopy was used in studying the morphology and cathodoluminescence of lunar regolith particles. Surface and structure of two groups of particles are differentiated: (1) Crystalline with well defined facets and spalling surfaces, which are grains of minerals and rock fragments: and (2) amorphous, fused, and partially or entirely glazed particles. Local melting of particles and the round openings on their surfaces are attributed to secondary influence on the regolith of factors of lunar weathering and above all micrometeoric impacts.

  12. Automatic 3D reconstruction of quasi-planar stereo Scanning Electron Microscopy (SEM) images.

    PubMed

    Roy, S; Meunier, J; Marian, A M; Vidal, F; Brunette, I; Costantino, S

    2012-01-01

    Scanning Electron Microscopy (SEM) is widely used in science to characterize the surface roughness of materials. Three-dimensional information can be obtained with SEM based on stereovision techniques. A stereo pair is typically obtained by tilting the sample by a few degrees. In this paper we present a fully automated method for 3D reconstruction from a SEM stereo pair without any particular constraint. Results are presented for corneal stromal surfaces.

  13. Spectrometric analysis and scanning electronic microscopy of two pleural plaques from mediaeval Portuguese period.

    PubMed

    Fernandes, T; Granja, R; Thillaud, P L

    2014-01-01

    During an archaeological excavation at a mediaeval monastery (Flor da Rosa, Crato, Portugal), a skeleton of a adult woman was found with two calcifications in the thoracic cage. The location and the macroscopic analysis of the calcifications allowed them to be assigned as pleural plaques. Spectrometric analysis and scanning electronic microscopy enabled to establish that it originated with an infectious process. These results associated with the lesions found in the ribs and vertebrae strongly suggest tuberculosis as the cause of these pleural plaques.

  14. Novel Automatic Electrochemical-mechanical Polishing (ECMP) of Metals for Scanning Electron Microscopy (Postprint)

    DTIC Science & Technology

    2010-03-23

    Micron 41 (2010) 615–621 619 Fig. 4 . XPS binding energy (eV) versus sputtering time (s) results for the Ti 2p peaks for the titanium samples: (a...improved the IQ values. 4 . Conclusions The electrochemical–mechanical polishing system (ECMP) removed material from titanium and nickel alloys at a...March 2014 4 . TITLE AND SUBTITLE NOVEL AUTOMATIC ELECTROCHEMICAL-MECHANICAL POLISHING (ECMP) OF METALS FOR SCANNING ELECTRON MICROSCOPY

  15. Endolithic algae and micrite envelope formation in Bahamian oolites as revealed by scanning electron microscopy.

    NASA Technical Reports Server (NTRS)

    Margolis, S.; Rex, R. W.

    1971-01-01

    Examination of Holocene Bahamian ooelites by scanning electron and light microscopy has revealed the morphology and orientation of aragonite crystals in the lamellar ooelitic envelope, and their modification by the boring activities of endolithic algae. The voids produced by these algae are found in progressive stages of being lined and filled with precipitated microcrystalline aragonite, which is similar to the process of micrite envelope formation in molluscan and other skeletal carbonate grains.

  16. Scanning electron microscopy of lunar regolith from the Sea of Fertility

    NASA Technical Reports Server (NTRS)

    Antoshin, M. K.; Ilin, N. P.; Spivak, G. V.

    1974-01-01

    Scanning electron microscopy was used in studying the morphology and cathodoluminescence of lunar regolith particles. Surface and structure of two groups of particles are differentiated: (1) Crystalline with well defined facets and spalling surfaces, which are grains of minerals and rock fragments: and (2) amorphous, fused, and partially or entirely glazed particles. Local melting of particles and the round openings on their surfaces are attributed to secondary influence on the regolith of factors of lunar weathering and above all micrometeoric impacts.

  17. [Study of the root nodules in some species of the Papilionaceae subfamily by scanning electron microscopy].

    PubMed

    Novikova, T I; Gordienko, N Ia

    2001-01-01

    Nitrogen-fixing nodules from 16 species in 6 tribes of the sub-family Papilionaceae have been examined by scanning electron microscopy. The structure of infection threads was similar in all the studied papilionoid species except Lupinus polyphillus. In this species the infection threads were found in young nodules only. The morphology of bacterioids and the character of their "package" are determined by the host plant genotype. The obtained results are discussed in relation to the evolution of the legumes.

  18. Direct visualization of lithium via annular bright field scanning transmission electron microscopy: a review.

    PubMed

    Findlay, Scott David; Huang, Rong; Ishikawa, Ryo; Shibata, Naoya; Ikuhara, Yuichi

    2017-02-08

    Annular bright field (ABF) scanning transmission electron microscopy has proven able to directly image lithium columns within crystalline environments, offering much insight into the structure and properties of lithium-ion battery materials. We summarize the image formation mechanisms underpinning ABF imaging, review the experimental application of this technique to imaging lithium in materials and overview the conditions that help maximize the visibility of lithium columns. © The Author 2016. Published by Oxford University Press on behalf of The Japanese Society of Microscopy. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

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

    PubMed

    de Siqueira, Alexandre Fioravante; Cabrera, Flávio Camargo; Pagamisse, Aylton; Job, Aldo Eloizo

    2014-01-01

    Electronic microscopy has been used for morphology evaluation of different materials structures. However, microscopy results may be affected by several factors. Image processing methods can be used to correct and improve the quality of these results. In this article, we propose an algorithm based on starlets to perform the segmentation of scanning electron microscopy images. An application is presented in order to locate gold nanoparticles in natural rubber membranes. In this application, our method showed accuracy greater than 85% for all test images. Results given by this method will be used in future studies, to computationally estimate the density distribution of gold nanoparticles in natural rubber samples and to predict reduction kinetics of gold nanoparticles at different time periods.

  20. Challenges of microtome-based serial block-face scanning electron microscopy in neuroscience.

    PubMed

    Wanner, A A; Kirschmann, M A; Genoud, C

    2015-08-01

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

  1. Electron probe X-ray microanalysis of cultured myogenic C2C12 cells with scanning and scanning transmission electron microscopy.

    PubMed

    Tylko, G; Karasiński, J; Wróblewski, R; Roomans, G M; Kilarski, W M

    2000-01-01

    Heterogeneity of the elemental content of myogenic C2C12 cultured cells was studied by electron probe X-ray microanalysis (EPXMA) with scanning (SEM EPXMA) and scanning transmission electron microscopy (STEM EPXMA). The best plastic substrate for growing cells was Thermanox. For STEM EPXMA, a Formvar film coated with carbon was found to be suitable substrate. The cells examined by scanning transmission electron microscopy showed great heterogeneity in their elemental content in comparison with the cells examined in the scanning electron microscope despite of an almost identical preparation procedure for EPXMA. Nevertheless the K/Na ratios obtained from both methods of EPXMA were very close (4.1 and 4.3). We conclude that the observed discrepancy in the elemental content obtained by the two methods may be due to differences in instrumentation and this must be taken into account when planning a comparative study.

  2. Gold nanoparticle uptake in whole cells in liquid examined by environmental scanning electron microscopy.

    PubMed

    Peckys, Diana B; de Jonge, Niels

    2014-02-01

    The size of gold nanoparticles (AuNPs) can influence various aspects of their cellular uptake. Light microscopy is not capable of resolving most AuNPs, while electron microscopy (EM) is not practically capable of acquiring the necessary statistical data from many cells and the results may suffer from various artifacts. Here, we demonstrate the use of a fast EM method for obtaining high-resolution data from a much larger population of cells than is usually feasible with conventional EM. A549 (human lung carcinoma) cells were subjected to uptake protocols with 10, 15, or 30 nm diameter AuNPs with adsorbed serum proteins. After 20 min, 24 h, or 45 h, the cells were fixed and imaged in whole in a thin layer of liquid water with environmental scanning electron microscopy equipped with a scanning transmission electron microscopy detector. The fast preparation and imaging of 145 whole cells in liquid allowed collection of nanoscale data within an exceptionally small amount of time of ~80 h. Analysis of 1,041 AuNP-filled vesicles showed that the long-term AuNP storing lysosomes increased their average size by 80 nm when AuNPs with 30 nm diameter were uptaken, compared to lysosomes of cells incubated with AuNPs of 10 and 15 nm diameter.

  3. In-Situ Imaging of Langmuir Films using Environmental Scanning Electron Microscopy

    NASA Astrophysics Data System (ADS)

    Miller, Aline

    2002-03-01

    Environmental scanning electron microscopy (ESEM) has been applied to image the organization of spread films at the air-water interface directly. This recent extension of conventional scanning electron microscopy permits the resolution of electron microscopy to be applied to insulated and hydrated samples maintained in their natural state. We demonstrate the great potential of this technique by in-situ imaging of both synthetic and natural polymer spread films at the air-water interface. A brief synopsis of the background to ESEM will be given followed by a discussion of the pertinent experimental conditions for the study of Langmuir layers. Experimental results for two systems will be presented. Firstly results for a nylon 6 6 polymer film will be given and discussed in relation to external reflection FTIR studies and those using the more conventional analytical techniques of surface pressure isotherms and optical microscopy. Secondly the kinetics of the interaction of a cationic intercalator amphiphile with double stranded DNA at the air-water interface will be presented and the structure of the densely packed two-dimensional arrays formed will be discussed.

  4. The Integration of Scanning Electron Microscopy, Scanning Probe Microscopy, and Luminescence Spectroscopy in one Platform: New Opportunities and Applications in Photovoltaics

    NASA Astrophysics Data System (ADS)

    Romero, Manuel

    2012-02-01

    We have recently integrated scanning tunneling microscopy (STM), atomic force microscopy (AFM), and near-field scanning optical microscopy (NSOM) onto the mechanical stage of a scanning electron microscope compatible with operation under high vacuum and the use of cryogenics. This instrument is unique in the sense that is not just the assembly of different microscopes but an integrated platform in which both the electron beam and the ultrasharp tip of the AFM/STM/NSOM can be controlled simultaneously and independently as excitation or sensing elements, providing innovative modes of operation and access to optoelectronic properties in the micro and nanoscale not accessible before. Furthermore, this instrument is equipped with focused laser illumination of the tip and detection of luminescence and can be used to measure cathodoluminescence, scanning tunneling luminescence, photoluminescence, and electroluminescence, all with high resolution. In this contribution, we review the application of these techniques to the development of second- and third-generation photovoltaics (PV) beyond those commercially available today. Among these applications, we present the luminescence and electron transport across single grain boundaries in chalcopyrite and kesterite compounds, the detection of single molecule species using plasmonics, the nanoscale imaging of the exciton transport in organic semiconductors, and the insitu manipulation and measurement of nanowires.

  5. Application of low-vacuum scanning electron microscopy for renal biopsy specimens.

    PubMed

    Miyazaki, Hiroki; Uozaki, Hiroshi; Tojo, Akihiro; Hirashima, Sayuri; Inaga, Sumire; Sakuma, Kei; Morishita, Yasuyuki; Fukayama, Masashi

    2012-09-15

    Low-vacuum scanning electron microscopy (LV-SEM) has been developed which enables the observation of soft, moist, and electrically insulating materials without any pretreatment unlike conventional scanning electron microscopy, in which samples must be solid, dry and usually electrically conductive. The purpose of this study was to assess the usefulness of LV-SEM for renal biopsy specimens. We analyzed 20 renal biopsy samples obtained for diagnostic purposes. The sections were stained with periodic acid methenamine silver to enhance the contrast, and subsequently examined by LV-SEM. LV-SEM showed a precise and fine structure of the glomerulus in both formalin fixed paraffin and glutaraldehyde-osmium tetroxide-fixed epoxy resin sections up to 10,000-fold magnification. The spike formation on the basement membrane was clearly observed in the membranous nephropathy samples. Similarly to transmission electron microscopy, electron dense deposits were observed in the epoxy resin sections of the IgA nephropathy and membranous nephropathy samples. LV-SEM could accurately show various glomerular lesions at high magnification after a simple and rapid processing of the samples. We consider that this is a novel and useful diagnostic tool for renal pathologies.

  6. Specimen preparation by ion beam slope cutting for characterization of ductile damage by scanning electron microscopy.

    PubMed

    Besserer, Hans-Bernward; Gerstein, Gregory; Maier, Hans Jürgen; Nürnberger, Florian

    2016-04-01

    To investigate ductile damage in parts made by cold sheet-bulk metal forming a suited specimen preparation is required to observe the microstructure and defects such as voids by electron microscopy. By means of ion beam slope cutting both a targeted material removal can be applied and mechanical or thermal influences during preparation avoided. In combination with scanning electron microscopy this method allows to examine voids in the submicron range and thus to analyze early stages of ductile damage. In addition, a relief structure is formed by the selectivity of the ion bombardment, which depends on grain orientation and microstructural defects. The formation of these relief structures is studied using scanning electron microscopy and electron backscatter diffraction and the use of this side effect to interpret the microstructural mechanisms of voids formation by plastic deformation is discussed. A comprehensive investigation of the suitability of ion beam milling to analyze ductile damage is given at the examples of a ferritic deep drawing steel and a dual phase steel. © 2016 Wiley Periodicals, Inc.

  7. Variable Temperature Setup for Scanning Electron Microscopy in Liquids and Atmospheric Pressure Gaseous Environments

    NASA Astrophysics Data System (ADS)

    Al-Asadi, Ahmed; Zhang, Jie; Li, Jianbo; Denault, Lauraine; Potyrailo, Radislav; Kolmakov, Andrei

    2014-03-01

    A thermoelectric cooling / heating setup for commercial Quantomix QX WETSEM scanning electron microscopy environmental cells was designed and tested. This addition allows extending ambient pressure in situ studies to be conducted in a wide temperature range both in liquid and gaseous environments. Instead of cooling/heating the entire body of QX-WETCELL, ultrathin polyimide electron transparent membrane window supported by metal mesh on the top of the cell has been used as an agent for heat transfer to/ from the Pelltier element. A butterfly wing of Morph sulkowskyi has been used as a model object in the QX-WETCELL's chamber due to its unique micro/nanostructure and peculiar wettability behavior. The dynamics of the water desorption, condensation and freezing processes were observed complementary using both optical microscopy and Scanning Electron Microscopy in vivo. The observations revel that the initial droplet formation were most likely taking place on the top of the wing ridges due to the waxy component of its surface. In addition, The SEM observation showed that the high intensity electron beam can heat the butterfly wing locally delaying the water condensation and freezing processes.

  8. Novel method of simultaneous multiple immunogold localization on resin sections in high resolution scanning electron microscopy.

    PubMed

    Nebesarova, Jana; Wandrol, Petr; Vancova, Marie

    2016-01-01

    We present a new method of multiple immunolabeling that is suitable for a broad spectrum of biomedical applications. The general concept is to label both sides of the ultrathin section with the thickness of 70-80 nm with different antibodies conjugated to gold nanoparticles and to distinguish the labeled side by advanced imaging methods with high resolution scanning electron microscopy, such as by correlating images acquired at different energies of primary electrons using different signals. From the Clinical Editor: The use of transmission electron microscopy has become an indispensible tool in the detection of cellular proteins. In this short but interesting article, the authors described their new method of labeling and the identification of four different proteins simultaneously, which represents another advance in imaging technique.

  9. Charged nanoparticle dynamics in water induced by scanning transmission electron microscopy.

    PubMed

    White, E R; Mecklenburg, Matthew; Shevitski, Brian; Singer, S B; Regan, B C

    2012-02-28

    Using scanning transmission electron microscopy we image ~4 nm platinum nanoparticles deposited on an insulating membrane, where the membrane is one of two electron-transparent windows separating an aqueous environment from the microscope's high vacuum. Upon receiving a relatively moderate dose of ~10(4) e/nm(2), initially immobile nanoparticles begin to move along trajectories that are directed radially outward from the center of the field of view. With larger dose rates the particle motion becomes increasingly dramatic. These observations demonstrate that, even under mild imaging conditions, the in situ electron microscopy of aqueous environments can produce electrophoretic charging effects that dominate the dynamics of nanoparticles under observation. © 2012 American Chemical Society

  10. Scanning electron microscopy of the nail plate in onychomycosis patients with negative fungal culture.

    PubMed

    Yue, Xueping; Li, Qing; Wang, Hongwei; Sun, Yilin; Wang, Aiping; Zhang, Qi; Zhang, Cuiping

    2016-01-01

    Onychomycosis is a common dermatological problem and can be identified by direct microscopic examination and fungal culture. However, the positive rate of fungal culture is low. This study investigated the application of scanning electron microscopy in the diagnosis of onychomycosis in 20 patients with negative fungal culture. In this study, a routine glutaraldehyde fixation method was used to prepare specimens for electron microscope examination. Results showed that under the scanning electron microscope, significant structural damage was observed in the nail plate in all patients. Hyphaes were seen in 70% of cases. A mixture of scattered hyphaes, pseudohyphaes, and spores was observed in 30% of cases. A mixture of spores and bacteria was observed in 10% of cases. A mixture of hyphaes and bacteria was observed in 20% of cases. The typical hyphae pierced a thin layer or single layer of corneocytes. Hyphaes could be smooth, sleek, and straight with visible separation, or dry, bent, and folded with a smooth surface. The diameter of hyphaes was 1-2 µm. The scattered spores were the main form of spore growth, and the growth of budding spores can be seen attached to the surface of layered armor. Most of the bacteria were gathered in clumps on the ventral surface, especially in grooves. In conclusion, scanning electron microscopy can be used to preliminarily identify the pathogen involved and the degree of damage in cases where onychomycosis is clinically diagnosed, but fungal culture is negative.

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

    SciTech Connect

    Kim, Suhyun; Kim, Joong Jung; Jung, Younheum; Lee, Kyungwoo; Byun, Gwangsun; Hwang, KyoungHwan; Lee, Sunyoung; Lee, Kyupil

    2013-09-15

    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.

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

    SciTech Connect

    Cullen, David A; Koestner, Roland; Kukreja, Ratan; Minko, Sergiy; Trotsenko, Oleksandr; Tokarev, Alexander V; Guetaz, Laure; Meyer III, Harry M; Parish, Chad M; More, Karren Leslie

    2014-01-01

    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.

  13. Image formation mechanisms in scanning electron microscopy of carbon nanotubes, and retrieval of their intrinsic dimensions.

    PubMed

    Jackman, H; Krakhmalev, P; Svensson, K

    2013-01-01

    We present a detailed analysis of the image formation mechanisms that are involved in the imaging of carbon nanotubes with scanning electron microscopy (SEM). We show how SEM images can be modelled by accounting for surface enhancement effects together with the absorption coefficient for secondary electrons, and the electron-probe shape. Images can then be deconvoluted, enabling retrieval of the intrinsic nanotube dimensions. Accurate estimates of their dimensions can thereby be obtained even for structures that are comparable to the electron-probe size (on the order of 2 nm). We also present a simple and robust model for obtaining the outer diameter of nanotubes without any detailed knowledge about the electron-probe shape.

  14. Note on in situ (scanning) transmission electron microscopy study of liquid samples.

    PubMed

    Jiang, Nan

    2017-08-01

    Liquid cell (scanning) transmission electron microscopy has been developed rapidly, using amorphous SiNx membranes as electron transparent windows. The current interpretations of electron beam effects are mainly based on radiolytic processes. In this note, additional effects of the electric field due to electron-beam irradiation are discussed. The electric field can be produced by the charge accumulation due to the emission of secondary and Auger electrons. Besides various beam-induced phenomena, such as nanoparticle precipitation and gas bubble formation and motion, two other effects need to be considered; one is the change of Gibbs free energy of nucleation and the other is the violation of Brownian motion due to ion drifting driven by the electric field. Copyright © 2017 Elsevier B.V. All rights reserved.

  15. High Dynamic Range Pixel Array Detector for Scanning Transmission Electron Microscopy.

    PubMed

    Tate, Mark W; Purohit, Prafull; Chamberlain, Darol; Nguyen, Kayla X; Hovden, Robert; Chang, Celesta S; Deb, Pratiti; Turgut, Emrah; Heron, John T; Schlom, Darrell G; Ralph, Daniel C; Fuchs, Gregory D; Shanks, Katherine S; Philipp, Hugh T; Muller, David A; Gruner, Sol M

    2016-02-01

    We describe a hybrid pixel array detector (electron microscope pixel array detector, or EMPAD) adapted for use in electron microscope applications, especially as a universal detector for scanning transmission electron microscopy. The 128×128 pixel detector consists of a 500 µm thick silicon diode array bump-bonded pixel-by-pixel to an application-specific integrated circuit. The in-pixel circuitry provides a 1,000,000:1 dynamic range within a single frame, allowing the direct electron beam to be imaged while still maintaining single electron sensitivity. A 1.1 kHz framing rate enables rapid data collection and minimizes sample drift distortions while scanning. By capturing the entire unsaturated diffraction pattern in scanning mode, one can simultaneously capture bright field, dark field, and phase contrast information, as well as being able to analyze the full scattering distribution, allowing true center of mass imaging. The scattering is recorded on an absolute scale, so that information such as local sample thickness can be directly determined. This paper describes the detector architecture, data acquisition system, and preliminary results from experiments with 80-200 keV electron beams.

  16. Anisotropic Shape Changes of Silica Nanoparticles Induced in Liquid with Scanning Transmission Electron Microscopy.

    PubMed

    Zečević, Jovana; Hermannsdörfer, Justus; Schuh, Tobias; de Jong, Krijn P; de Jonge, Niels

    2017-01-01

    Liquid-phase transmission electron microscopy (TEM) is used for in-situ imaging of nanoscale processes taking place in liquid, such as the evolution of nanoparticles during synthesis or structural changes of nanomaterials in liquid environment. Here, it is shown that the focused electron beam of scanning TEM (STEM) brings about the dissolution of silica nanoparticles in water by a gradual reduction of their sizes, and that silica redeposites at the sides of the nanoparticles in the scanning direction of the electron beam, such that elongated nanoparticles are formed. Nanoparticles with an elongation in a different direction are obtained simply by changing the scan direction. Material is expelled from the center of the nanoparticles at higher electron dose, leading to the formation of doughnut-shaped objects. Nanoparticles assembled in an aggregate gradually fuse, and the electron beam exposed section of the aggregate reduces in size and is elongated. Under TEM conditions with a stationary electron beam, the nanoparticles dissolve but do not elongate. The observed phenomena are important to consider when conducting liquid-phase STEM experiments on silica-based materials and may find future application for controlled anisotropic manipulation of the size and the shape of nanoparticles in liquid. © 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

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

    PubMed

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

    2016-05-25

    Large-scale 2D electron microscopy (EM), or nanotomy, is the tissue-wide application of nanoscale resolution electron microscopy. Others and we previously applied large scale EM to human skin pancreatic islets, tissue culture and whole zebrafish larvae(1-7). Here we describe a universally applicable method for tissue-scale scanning EM for unbiased detection of sub-cellular and molecular features. Nanotomy was applied to investigate the healthy and a neurodegenerative zebrafish brain. Our method is based on standardized EM sample preparation protocols: Fixation with glutaraldehyde and osmium, followed by epoxy-resin embedding, ultrathin sectioning and mounting of ultrathin-sections on one-hole grids, followed by post staining with uranyl and lead. Large-scale 2D EM mosaic images are acquired using a scanning EM connected to an external large area scan generator using scanning transmission EM (STEM). Large scale EM images are typically ~ 5 - 50 G pixels in size, and best viewed using zoomable HTML files, which can be opened in any web browser, similar to online geographical HTML maps. This method can be applied to (human) tissue, cross sections of whole animals as well as tissue culture(1-5). Here, zebrafish brains were analyzed in a non-invasive neuronal ablation model. We visualize within a single dataset tissue, cellular and subcellular changes which can be quantified in various cell types including neurons and microglia, the brain's macrophages. In addition, nanotomy facilitates the correlation of EM with light microscopy (CLEM)(8) on the same tissue, as large surface areas previously imaged using fluorescent microscopy, can subsequently be subjected to large area EM, resulting in the nano-anatomy (nanotomy) of tissues. In all, nanotomy allows unbiased detection of features at EM level in a tissue-wide quantifiable manner.

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

    PubMed Central

    Kuipers, Jeroen; Kalicharan, Ruby D.; Wolters, Anouk H. G.

    2016-01-01

    Large-scale 2D electron microscopy (EM), or nanotomy, is the tissue-wide application of nanoscale resolution electron microscopy. Others and we previously applied large scale EM to human skin pancreatic islets, tissue culture and whole zebrafish larvae1-7. Here we describe a universally applicable method for tissue-scale scanning EM for unbiased detection of sub-cellular and molecular features. Nanotomy was applied to investigate the healthy and a neurodegenerative zebrafish brain. Our method is based on standardized EM sample preparation protocols: Fixation with glutaraldehyde and osmium, followed by epoxy-resin embedding, ultrathin sectioning and mounting of ultrathin-sections on one-hole grids, followed by post staining with uranyl and lead. Large-scale 2D EM mosaic images are acquired using a scanning EM connected to an external large area scan generator using scanning transmission EM (STEM). Large scale EM images are typically ~ 5 - 50 G pixels in size, and best viewed using zoomable HTML files, which can be opened in any web browser, similar to online geographical HTML maps. This method can be applied to (human) tissue, cross sections of whole animals as well as tissue culture1-5. Here, zebrafish brains were analyzed in a non-invasive neuronal ablation model. We visualize within a single dataset tissue, cellular and subcellular changes which can be quantified in various cell types including neurons and microglia, the brain's macrophages. In addition, nanotomy facilitates the correlation of EM with light microscopy (CLEM)8 on the same tissue, as large surface areas previously imaged using fluorescent microscopy, can subsequently be subjected to large area EM, resulting in the nano-anatomy (nanotomy) of tissues. In all, nanotomy allows unbiased detection of features at EM level in a tissue-wide quantifiable manner. PMID:27285162

  19. Thirty per cent contrast in secondary-electron imaging by scanning field-emission microscopy.

    PubMed

    Zanin, D A; De Pietro, L G; Peter, Q; Kostanyan, A; Cabrera, H; Vindigni, A; Bähler, Th; Pescia, D; Ramsperger, U

    2016-11-01

    We perform scanning tunnelling microscopy (STM) in a regime where primary electrons are field-emitted from the tip and excite secondary electrons out of the target-the scanning field-emission microscopy regime (SFM). In the SFM mode, a secondary-electron contrast as high as 30% is observed when imaging a monoatomic step between a clean W(110)- and an Fe-covered W(110)-terrace. This is a figure of contrast comparable to STM. The apparent width of the monoatomic step attains the 1 nm mark, i.e. it is only marginally worse than the corresponding width observed in STM. The origin of the unexpected strong contrast in SFM is the material dependence of the secondary-electron yield and not the dependence of the transported current on the tip-target distance, typical of STM: accordingly, we expect that a technology combining STM and SFM will highlight complementary aspects of a surface while simultaneously making electrons, selected with nanometre spatial precision, available to a macroscopic environment for further processing.

  20. Thirty per cent contrast in secondary-electron imaging by scanning field-emission microscopy

    PubMed Central

    De Pietro, L. G.; Peter, Q.; Kostanyan, A.; Cabrera, H.; Vindigni, A.; Bähler, Th.; Pescia, D.; Ramsperger, U.

    2016-01-01

    We perform scanning tunnelling microscopy (STM) in a regime where primary electrons are field-emitted from the tip and excite secondary electrons out of the target—the scanning field-emission microscopy regime (SFM). In the SFM mode, a secondary-electron contrast as high as 30% is observed when imaging a monoatomic step between a clean W(110)- and an Fe-covered W(110)-terrace. This is a figure of contrast comparable to STM. The apparent width of the monoatomic step attains the 1 nm mark, i.e. it is only marginally worse than the corresponding width observed in STM. The origin of the unexpected strong contrast in SFM is the material dependence of the secondary-electron yield and not the dependence of the transported current on the tip–target distance, typical of STM: accordingly, we expect that a technology combining STM and SFM will highlight complementary aspects of a surface while simultaneously making electrons, selected with nanometre spatial precision, available to a macroscopic environment for further processing. PMID:27956876

  1. Scanning electron microscopy study of adhesion in sea urchin blastulae. M.S. Thesis

    NASA Technical Reports Server (NTRS)

    Crowther, Susan D.

    1988-01-01

    The dissociation supernatant (DS) isolated by disaggregating Strongylocentrotus purpuratus blastulae in calcium- and magnesium-free seawater specifically promotes reaggregation of S. purpuratus blastula cells. The purpose of this study was to use scanning electron microscopy to examine the gross morphology of aggregates formed in the presence of DS to see if it resembles adhesion in partially dissociated blastulae. A new reaggregation procedure developed here, using large volumes of cell suspension and a large diameter of rotation, was utilized to obtain sufficient quantities of aggregates for scanning electron microscopy. The results indicate that aggregates formed in the presence of DS resemble partially dissociated intact embryos in terms of the direct cell-cell adhesion observed. DS did not cause aggregation to form as a result of the entrapment of cells in masses of extracellular material. These studies provide the groundwork for further studies using transmission electron microscopy to more precisely define the adhesive contacts made by cells in the presence of the putative adhesion molecules present in DS.

  2. Scanning electron microscopy study of adhesion in sea urchin blastulae. M.S. Thesis

    NASA Technical Reports Server (NTRS)

    Crowther, Susan D.

    1988-01-01

    The dissociation supernatant (DS) isolated by disaggregating Strongylocentrotus purpuratus blastulae in calcium- and magnesium-free seawater specifically promotes reaggregation of S. purpuratus blastula cells. The purpose of this study was to use scanning electron microscopy to examine the gross morphology of aggregates formed in the presence of DS to see if it resembles adhesion in partially dissociated blastulae. A new reaggregation procedure developed here, using large volumes of cell suspension and a large diameter of rotation, was utilized to obtain sufficient quantities of aggregates for scanning electron microscopy. The results indicate that aggregates formed in the presence of DS resemble partially dissociated intact embryos in terms of the direct cell-cell adhesion observed. DS did not cause aggregation to form as a result of the entrapment of cells in masses of extracellular material. These studies provide the groundwork for further studies using transmission electron microscopy to more precisely define the adhesive contacts made by cells in the presence of the putative adhesion molecules present in DS.

  3. Scanning transmission electron microscopy mass analysis of fibrillin-containing microfibrils from foetal elastic tissues.

    PubMed

    Sherratt, M J; Holmes, D F; Shuttleworth, C A; Kielty, C M

    1997-01-01

    We have applied scanning transmission electron microscopy to intact native fibrillin-containing microfibrils isolated from foetal bovine elastic tissues in order to derive new insights into microfibril organisation. This technique provides quantitative data on the mass per unit length and axial mass distribution of unstained, unshadowed macromolecules. Scanning transmission electron microscopy of microfibrils from aorta, skin and nuchal ligament revealed that the beads corresponded to peaks of mass and the interbead regions to troughs of mass. These major features of axial mass distribution were characteristic of all microfibrils examined. Tissue-specific and age-dependent variations in mass were identified in microfibrils that were structurally comparable by rotary shadowing electron microscopy. Increased microfibril mass correlated with increasing gestational age. The additional mass was associated predominantly at, or close to, the bead. Some microfibril populations exhibited pronounced assymetry in their axial mass distribution. These data indicate that intact native microfibrillar assemblies from developing elastic tissues are heterogeneous in composition. Loss of mass following chondroitinase ABC or AC lyase treatment confirmed the presence of chondroitin sulphate in nuchal ligament microfibrillar assemblies.

  4. A new method of scanning electron microscopy for imaging biological tissues.

    PubMed

    Boyde, A; Reid, S A

    1983-04-07

    The scanning electron microscope (SEM) has proved of little value in the examination of material prepared for light microscopic histology. One of the chief reasons for this is that the secondary electron signal used for image formation in routine scanning microscopy derives from the surface of the specimen. In the case of histological material this surface is one which has been severely distorted by processing and cutting procedures. Light microscopy sections can be usefully studied in te SEM if the signal used to form the image derives from a considerable portion of the thickness of the section. Thus the backscattered electron (BSE) image has been successfully used in studying the distribution of dense material or densely staining components several micrometres deep to the surface of dried sections. Such sections are, however, usually mounted on low density (poorly BSE reflecting) non-transparent substrates such as beryllium or carbon, so that matching light microscopy of the same samples is not possible. We report here a method by which histological sections mounted on glass slides can be imaged in the SEM at a resolution higher than that obtained using conventional light microscopy. The method exploits the facts that the ordinary, cheap light microscope slide is strongly cathodoluminescent, yet the standard histological (7 micrometers) section is of such a mass thickness that it absorbs a significant proportion of electrons which energies (5-20 keV) usually used in biological SEM. Thus the measure of the glass cathodoluminescence signal is the measure of the electron flux passing through the specimen.

  5. Pulsed and scanned carbon dioxide laser resurfacing 2 years after treatment: comparison by means of scanning electron microscopy.

    PubMed

    Trelles, Mario A; Garcia, Luisa; Rigau, Josepa; Allones, Inès; Velez, Marìano

    2003-05-01

    Studies have reported short-term and long-term (1-year) findings for laser skin resurfacing. Two of the most popular systems used for this procedure, the continuous-wave Sharplan 40C SilkTouch system and the pulsed Coherent 5000C UltraPulse system with a computer pattern generator, were previously compared for a range of follow-up times up to 1 year, using light microscopy and transmission electron microscopy. This study analyzed the 2-year morphological differences using scanning electron microscopy. Tissue samples were obtained from 10 patients (age range, 50 to 72 years; skin types II and III) who had undergone laser resurfacing 2 years previously. One half of the face of each patient had been treated with the continuous-wave system and the other half with the pulsed system. The samples were subjected to scanning electron microscopy. On the continuous-wave-treated side, significantly better dermal collagen organization was observed at 2 years, with plump-appearing fibers that were closely knit to form a compact structure. On the side treated with the pulsed system, the collagen fibers in the papillary dermis were more loosely arranged and appeared drier. In both the continuous-wave-treated and pulsed-treated areas, the epidermis appeared healthy and exhibited some signs of age-related deterioration, with slightly flatter plaques and somewhat more flaking keratin on the pulsed-treated side. Probably because of the greater degree of residual thermal damage associated with the continuous-wave system, at 2 years after treatment there was more prolific synthesis and better orientation of collagen fibers, which were maintained for longer times, compared with the pulsed-treated specimens.

  6. Valence electron energy-loss spectroscopy in monochromated scanning transmission electron microscopy.

    PubMed

    Erni, Rolf; Browning, Nigel D

    2005-10-01

    With the development of monochromators for (scanning) transmission electron microscopes, valence electron energy-loss spectroscopy (VEELS) is developing into a unique technique to study the band structure and optical properties of nanoscale materials. This article discusses practical aspects of spatially resolved VEELS performed in scanning transmission mode and the alignments necessary to achieve the current optimum performance of approximately 0.15 eV energy resolution with an electron probe size of approximately 1 nm. In particular, a collection of basic concepts concerning the acquisition process, the optimization of the energy resolution, the spatial resolution and the data processing are provided. A brief study of planar defects in a Y(1)Ba(2)Cu(3)O(7-)(delta) high-temperature superconductor illustrates these concepts and shows what kind of information can be accessed by VEELS.

  7. Use of fluorescence and scanning electron microscopy as tools in teaching biology

    NASA Astrophysics Data System (ADS)

    Ghosh, Nabarun; Silva, Jessica; Vazquez, Aracely; Das, A. B.; Smith, Don W.

    2011-06-01

    Recent nationwide surveys reveal significant decline in students' interest in Math and Sciences. The objective of this project was to inspire young minds in using various techniques involved in Sciences including Scanning Electron Microscopy. We used Scanning Electron Microscope in demonstrating various types of Biological samples. An SEM Tabletop model in the past decade has revolutionized the use of Scanning Electron Microscopes. Using SEM Tabletop model TM 1000 we studied biological specimens of fungal spores, pollen grains, diatoms, plant fibers, dust mites, insect parts and leaf surfaces. We also used fluorescence microscopy to view, to record and analyze various specimens with an Olympus BX40 microscope equipped with FITC and TRITC fluorescent filters, a mercury lamp source, DP-70 digital camera with Image Pro 6.0 software. Micrographs were captured using bright field microscopy, the fluoresceinisothiocyanate (FITC) filter, and the tetramethylrhodamine (TRITC) filter settings at 40X. A high pressure mercury lamp or UV source was used to excite the storage molecules or proteins which exhibited autofluorescence. We used fluorescent microscopy to confirm the localization of sugar beet viruses in plant organs by viewing the vascular bundles in the thin sections of the leaves and other tissues. We worked with the REU summer students on sample preparation and observation on various samples utilizing the SEM. Critical Point Drying (CPD) and metal coating with the sputter coater was followed before observing some cultured specimen and the samples that were soft in textures with high water content. SEM Top allowed investigating the detailed morphological features that can be used for classroom teaching. Undergraduate and graduate researchers studied biological samples of Arthropods, pollen grains and teeth collected from four species of snakes using SEM. This project inspired the research students to pursue their career in higher studies in science and 45% of the

  8. Application of low vacuum scanning electron microscopy for Papanicolaou-stained slides for cytopathology examinations.

    PubMed

    Yano, Tetsuya; Soejima, Yurie; Sawabe, Motoji

    2016-06-01

    Papanicolaou (Pap)-stained slides are usually observed using a transmitted light microscope for cytopathology. However, progress in pathological examinations has created a need for new diagnostic tools, because cytopathological preparations do not allow additional examinations without a loss of specimen, unlike histopathology. Low-vacuum scanning electron microscopy (LVSEM) can reveal the surface topography at an ultrastructual resolution without metal coating. The aim of this study was to determine the conditions required for observing Pap-stained slides of oral smears using LVSEM without any loss of specimen and to reexamine the same slides again using light microscopy, while preserving the cytopathological information. © The Author 2016. Published by Oxford University Press on behalf of The Japanese Society of Microscopy. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

  9. 4D scanning ultrafast electron microscopy: visualization of materials surface dynamics.

    PubMed

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

    2011-05-25

    The continuous electron beam of conventional scanning electron microscopes (SEM) limits the temporal resolution required for the study of ultrafast dynamics of materials surfaces. Here, we report the development of scanning ultrafast electron microscopy (S-UEM) as a time-resolved method with resolutions in both space and time. The approach is demonstrated in the investigation of the dynamics of semiconducting and metallic materials visualized using secondary-electron images and backscattering electron diffraction patterns. For probing, the electron packet was photogenerated from the sharp field-emitter tip of the microscope with a very low number of electrons in order to suppress space-charge repulsion between electrons and reach the ultrashort temporal resolution, an improvement of orders of magnitude when compared to the traditional beam-blanking method. Moreover, the spatial resolution of SEM is maintained, thus enabling spatiotemporal visualization of surface dynamics following the initiation of change by femtosecond heating or excitation. We discuss capabilities and potential applications of S-UEM in materials and biological science.

  10. Secretory glands and microvascular systems imaged in aqueous solution by atmospheric scanning electron microscopy (ASEM).

    PubMed

    Yamazawa, Toshiko; Nakamura, Naotoshi; Sato, Mari; Sato, Chikara

    2016-12-01

    Exocrine glands, e.g., salivary and pancreatic glands, play an important role in digestive enzyme secretion, while endocrine glands, e.g., pancreatic islets, secrete hormones that regulate blood glucose levels. The dysfunction of these secretory organs immediately leads to various diseases, such as diabetes or Sjögren's syndrome, by poorly understood mechanisms. Gland-related diseases have been studied by optical microscopy (OM), and at higher resolution by transmission electron microscopy (TEM) of Epon embedded samples, which necessitates hydrophobic sample pretreatment. Here, we report the direct observation of tissue in aqueous solution by atmospheric scanning electron microscopy (ASEM). Salivary glands, lacrimal glands, and pancreas were fixed, sectioned into slabs, stained with phosphotungstic acid (PTA), and inspected in radical scavenger d-glucose solution from below by an inverted scanning electron microscopy (SEM), guided by optical microscopy from above to target the tissue substructures. A 2- to 3-µm specimen thickness was visualized by the SEM. In secretory cells, cytoplasmic vesicles and other organelles were clearly imaged at high resolution, and the former could be classified according to the degree of PTA staining. In islets of Langerhans, the microvascular system used as an outlet by the secretory cells was also clearly observed. Microvascular system is also critically involved in the onset of diabetic complications and was clearly visible in subcutaneous tissue imaged by ASEM. The results suggest the use of in-solution ASEM for histology and to study vesicle secretion systems. Further, the high-throughput of ASEM makes it a potential tool for the diagnosis of exocrine and endocrine-related diseases.

  11. A differentially pumped secondary electron detector for low-vacuum scanning electron microscopy.

    PubMed

    Jacka, M; Zadrazil, M; Lopour, F

    2003-01-01

    A new design of secondary electron (SE) detector is described for use in low-vacuum scanning electron microscopes. Its distinguishing feature is a separate detector chamber, which can be maintained at a pressure independent of the pressure in the specimen chamber. The two chambers are separated by a perforated membrane or mesh across which an electric field is applied, making it relatively transparent to low-energy electrons but considerably less so to the gas molecules. The benefits of this arrangement are discussed. The final means of detecting the electrons can be a conventional scintillator and photomultiplier arrangement or any of the methods using the ambient gas as an amplifying medium. Images obtained with the detector show good SE contrast and low backscattered electron contribution.

  12. Annular dark-field scanning transmission electron microscopy (ADF-STEM) tomography of polymer systems.

    PubMed

    Lu, Kangbo; Sourty, Erwan; Loos, Joachim

    2010-08-01

    We have utilized bright-field conventional transmission electron microscopy tomography and annular dark-field scanning transmission electron microscopy (ADF-STEM) tomography to characterize a well-defined carbon black (CB)-filled polymer nanocomposite with known CB volume concentration. For both imaging methods, contrast can be generated between the CB and the surrounding polymer matrix. The involved contrast mechanisms, in particular for ADF-STEM, will be discussed in detail. The obtained volume reconstructions were analysed and the CB volume concentrations were carefully determined from the reconstructed data. For both imaging modes, the measured CB volume concentrations are substantially different and only quantification based on the ADF-STEM data revealed about the same value as the known CB loading. Moreover, when applying low-convergence angles for imaging ADF-STEM tomography, data can be obtained of micrometre-thick samples.

  13. Photon scanning tunneling microscopy

    SciTech Connect

    Goudonnet, J.P.; Salomon, L.; De Fornel, F.; Chabrier, G. . Lab. de Physique du Solide); Warmack, R.J.; Ferrell, T.L. )

    1990-01-01

    The Photon Scanning Tunneling Microscopy (PSTM) is the photon analogue of the electron Scanning Tunneling Microscope (STM). It uses the evanescent field due to the total internal reflection of a light beam in a Total Internal Reflection (TIR) prism. The sample, mounted on the base of the prism, modulates the evanescent field. A sharpened optical fiber probes this field, and the collected light is processed to generate an image of the topography and the chemical composition of the surface. We give, in this paper, a description of the microscope and discuss the influence of several parameters such as -- polarization of light, angle of incidence, shape of the end of the fiber -- on the resolution. Images of various samples -- glass samples, teflon spheres -- are presented. 8 refs., 7 figs.

  14. Accurate Nanoscale Crystallography in Real-Space Using Scanning Transmission Electron Microscopy.

    PubMed

    Dycus, J Houston; Harris, Joshua S; Sang, Xiahan; Fancher, Chris M; Findlay, Scott D; Oni, Adedapo A; Chan, Tsung-Ta E; Koch, Carl C; Jones, Jacob L; Allen, Leslie J; Irving, Douglas L; LeBeau, James M

    2015-08-01

    Here, we report reproducible and accurate measurement of crystallographic parameters using scanning transmission electron microscopy. This is made possible by removing drift and residual scan distortion. We demonstrate real-space lattice parameter measurements with <0.1% error for complex-layered chalcogenides Bi2Te3, Bi2Se3, and a Bi2Te2.7Se0.3 nanostructured alloy. Pairing the technique with atomic resolution spectroscopy, we connect local structure with chemistry and bonding. Combining these results with density functional theory, we show that the incorporation of Se into Bi2Te3 causes charge redistribution that anomalously increases the van der Waals gap between building blocks of the layered structure. The results show that atomic resolution imaging with electrons can accurately and robustly quantify crystallography at the nanoscale.

  15. Atomic-scale mapping of electronic structures across heterointerfaces by cross-sectional scanning tunneling microscopy

    NASA Astrophysics Data System (ADS)

    Chiu, Ya-Ping; Huang, Bo-Chao; Shih, Min-Chuan; Huang, Po-Cheng; Chen, Chun-Wei

    2015-09-01

    Interfacial science has received much attention recently based on the development of state-of-the-art analytical tools that can create and manipulate the charge, spin, orbital, and lattice degrees of freedom at interfaces. Motivated by the importance of nanoscale interfacial science that governs device operation, we present a technique to probe the electronic characteristics of heterointerfaces with atomic resolution. In this work, the interfacial characteristics of heteroepitaxial structures are investigated and the fundamental mechanisms that pertain in these systems are elucidated through cross-sectional scanning tunneling microscopy (XSTM). The XSTM technique is employed here to directly observe epitaxial interfacial structures and probe local electronic properties with atomic-level capability. Scanning tunneling microscopy and spectroscopy experiments with atomic precision provide insight into the origin and spatial distribution of electronic properties across heterointerfaces. The first part of this report provides a brief description of the cleavage technique and spectroscopy analysis in XSTM measurements. The second part addresses interfacial electronic structures of several model heterostructures in current condensed matter research using XSTM. Topics to be discussed include high-κ‘s/III-V’s semiconductors, polymer heterojunctions, and complex oxide heterostructures, which are all material systems whose investigation using this technique is expected to benefit the research community. Finally, practical aspects and perspectives of using XSTM in interface science are presented.

  16. Imaging three-dimensional tissue architectures by focused ion beam scanning electron microscopy.

    PubMed

    Bushby, Andrew J; P'ng, Kenneth M Y; Young, Robert D; Pinali, Christian; Knupp, Carlo; Quantock, Andrew J

    2011-06-01

    In this protocol, we describe a 3D imaging technique known as 'volume electron microscopy' or 'focused ion beam scanning electron microscopy (FIB/SEM)' applied to biological tissues. A scanning electron microscope equipped with a focused gallium ion beam, used to sequentially mill away the sample surface, and a backscattered electron (BSE) detector, used to image the milled surfaces, generates a large series of images that can be combined into a 3D rendered image of stained and embedded biological tissue. Structural information over volumes of tens of thousands of cubic micrometers is possible, revealing complex microanatomy with subcellular resolution. Methods are presented for tissue processing, for the enhancement of contrast with osmium tetroxide/potassium ferricyanide, for BSE imaging, for the preparation and platinum deposition over a selected site in the embedded tissue block, and for sequential data collection with ion beam milling; all this takes approximately 90 h. The imaging conditions, procedures for alternate milling and data acquisition and techniques for processing and partitioning the 3D data set are also described; these processes take approxiamtely 30 h. The protocol is illustrated by application to developing chick cornea, in which cells organize collagen fibril bundles into complex, multilamellar structures essential for transparency in the mature connective tissue matrix. The techniques described could have wide application in a range of fields, including pathology, developmental biology, microstructural anatomy and regenerative medicine.

  17. Atomic-scale mapping of electronic structures across heterointerfaces by cross-sectional scanning tunneling microscopy.

    PubMed

    Chiu, Ya-Ping; Huang, Bo-Chao; Shih, Min-Chuan; Huang, Po-Cheng; Chen, Chun-Wei

    2015-09-04

    Interfacial science has received much attention recently based on the development of state-of-the-art analytical tools that can create and manipulate the charge, spin, orbital, and lattice degrees of freedom at interfaces. Motivated by the importance of nanoscale interfacial science that governs device operation, we present a technique to probe the electronic characteristics of heterointerfaces with atomic resolution. In this work, the interfacial characteristics of heteroepitaxial structures are investigated and the fundamental mechanisms that pertain in these systems are elucidated through cross-sectional scanning tunneling microscopy (XSTM). The XSTM technique is employed here to directly observe epitaxial interfacial structures and probe local electronic properties with atomic-level capability. Scanning tunneling microscopy and spectroscopy experiments with atomic precision provide insight into the origin and spatial distribution of electronic properties across heterointerfaces. The first part of this report provides a brief description of the cleavage technique and spectroscopy analysis in XSTM measurements. The second part addresses interfacial electronic structures of several model heterostructures in current condensed matter research using XSTM. Topics to be discussed include high-κ's/III-V's semiconductors, polymer heterojunctions, and complex oxide heterostructures, which are all material systems whose investigation using this technique is expected to benefit the research community. Finally, practical aspects and perspectives of using XSTM in interface science are presented.

  18. Scanning electron acoustic microscopy of residual stresses in ceramics: Theory and experiment

    NASA Technical Reports Server (NTRS)

    Cantrell, John H.; Qian, Menglu

    1992-01-01

    Several reviews have highlighted a number of applications of scanning electron acoustic microscopy (SEAM) to metals and semiconductors which show that SEAM can provide new information on surface and near-surface features of such materials, but there have been few studies attempting to determine the capabilities of SEAM for characterizing ceramic materials. We have recently observed image contrast in SEAM from residual stress fields induced in brittle materials by Vickers indentations that is strongly dependent on the electron beam chopping frequency. We have also recently developed a three-dimensional mathematical model of signal generation and contrast in SEAM, appropriate to the brittle materials studied, that we use as a starting point in this paper for modeling the effect of residual stress fields on the generated electron acoustic signal. The influence of the electron beam chopping frequency is also considered under restrictive assumptions.

  19. Scanning electron acoustic microscopy of residual stresses in ceramics: Theory and experiment

    NASA Technical Reports Server (NTRS)

    Cantrell, John H.; Qian, Menglu

    1992-01-01

    Several reviews have highlighted a number of applications of scanning electron acoustic microscopy (SEAM) to metals and semiconductors which show that SEAM can provide new information on surface and near-surface features of such materials, but there have been few studies attempting to determine the capabilities of SEAM for characterizing ceramic materials. We have recently observed image contrast in SEAM from residual stress fields induced in brittle materials by Vickers indentations that is strongly dependent on the electron beam chopping frequency. We have also recently developed a three-dimensional mathematical model of signal generation and contrast in SEAM, appropriate to the brittle materials studied, that we use as a starting point in this paper for modeling the effect of residual stress fields on the generated electron acoustic signal. The influence of the electron beam chopping frequency is also considered under restrictive assumptions.

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

    SciTech Connect

    Pelliccione, M.; Bartel, J.; Goldhaber-Gordon, D.; Sciambi, A.; Pfeiffer, L. N.; West, K. W.

    2014-11-03

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

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

    NASA Astrophysics Data System (ADS)

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

    1994-11-01

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

  2. Study of fossil bones by synchrotron radiation micro-spectroscopic techniques and scanning electron microscopy.

    PubMed

    Zougrou, I M; Katsikini, M; Pinakidou, F; Paloura, E C; Papadopoulou, L; Tsoukala, E

    2014-01-01

    Earlymost Villafranchian fossil bones of an artiodactyl and a perissodactyl from the Milia excavation site in Grevena, Greece, were studied in order to evaluate diagenetic effects. Optical microscopy revealed the different bone types (fibro-lamellar and Haversian, respectively) of the two fragments and their good preservation state. The spatial distribution of bone apatite and soil-originating elements was studied using micro-X-ray fluorescence (µ-XRF) mapping and scanning electron microscopy. The approximate value of the Ca/P ratio was 2.2, as determined from scanning electron microscopy measurements. Bacterial boring was detected close to the periosteal region and Fe bearing oxides were found to fill bone cavities, e.g. Haversian canals and osteocyte lacunae. In the perissodactyl bone considerable amounts of Mn were detected close to cracks (the Mn/Fe weight ratio takes values up to 3.5). Goethite and pyrite were detected in both samples by means of metallographic microscopy. The local Ca/P ratio determined with µ-XRF varied significantly in metal-poor spots indicating spatial inhomogeneities in the ionic substitutions. XRF line scans that span the bone cross sections revealed that Fe and Mn contaminate the bones from both the periosteum and medullar cavity and aggregate around local maxima. The formation of goethite, irrespective of the local Fe concentration, was verified by the Fe K-edge X-ray absorption fine structure (XAFS) spectra. Finally, Sr K-edge extended XAFS (EXAFS) revealed that Sr substitutes for Ca in bone apatite without obvious preference to the Ca1 or Ca2 unit-cell site occupation.

  3. Scanning Transmission X-Ray, Laser Scanning, and Transmission Electron Microscopy Mapping of the Exopolymeric Matrix of Microbial Biofilms

    PubMed Central

    Lawrence, J. R.; Swerhone, G. D. W.; Leppard, G. G.; Araki, T.; Zhang, X.; West, M. M.; Hitchcock, A. P.

    2003-01-01

    Confocal laser scanning microscopy (CLSM), transmission electron microscopy (TEM), and soft X-ray scanning transmission X-ray microscopy (STXM) were used to map the distribution of macromolecular subcomponents (e.g., polysaccharides, proteins, lipids, and nucleic acids) of biofilm cells and matrix. The biofilms were developed from river water supplemented with methanol, and although they comprised a complex microbial community, the biofilms were dominated by heterotrophic bacteria. TEM provided the highest-resolution structural imaging, CLSM provided detailed compositional information when used in conjunction with molecular probes, and STXM provided compositional mapping of macromolecule distributions without the addition of probes. By examining exactly the same region of a sample with combinations of these techniques (STXM with CLSM and STXM with TEM), we demonstrate that this combination of multimicroscopy analysis can be used to create a detailed correlative map of biofilm structure and composition. We are using these correlative techniques to improve our understanding of the biochemical basis for biofilm organization and to assist studies intended to investigate and optimize biofilms for environmental remediation applications. PMID:12957944

  4. Ultrahigh-Resolution Scanning Transmission Electron Microscopy with Sub-Angstrom-Sized Electron Beams

    SciTech Connect

    Abe, E.; Pennycook, Stephen J

    2005-01-01

    The scanning transmission electron microscope (STEM) with an annular dark-field (ADF) detector provides atomic-resolution incoherent images, whose resolution is dominated, to a good approximation, by the size of convergent electron beams. Improving a spherical aberration of microscope objective lenses has been successful in converging the beam into sub-angstrom scale, promising a remarkably higher resolution for STEM. Here we describe the performance of aberration-corrected 300kV-STEM-the world-best STEM available today. The results clearly demonstrate that a sub-angstrom resolution has been indeed achieved for not only simple structures but also structurally complex systems (quasicrystals).

  5. Ultrastructural analysis of testicular tissue and sperm by transmission and scanning electron microscopy.

    PubMed

    Chemes, Hector E

    2013-01-01

    Transmission electron microscopy (TEM) studies have provided the basis for an in-depth understanding of the cell biology and normal functioning of the testis and male gametes and have opened the way to characterize the functional role played by specific organelles in spermatogenesis and sperm function. The development of the scanning electron microscope (SEM) extended these boundaries to the recognition of cell and organ surface features and the architectural array of cells and tissues. The merging of immunocytochemical and histochemical approaches with electron microscopy has completed a series of technical improvements that integrate structural and functional features to provide a broad understanding of cell biology in health and disease. With these advances the detailed study of the intricate structural and molecular organization as well as the chemical composition of cellular organelles is now possible. Immunocytochemistry is used to identify proteins or other components and localize them in specific cells or organelles with high specificity and sensitivity, and histochemistry can be used to understand their function (i.e., enzyme activity). When these techniques are used in conjunction with electron microscopy their resolving power is further increased to subcellular levels. In the present chapter we will describe in detail various ultrastructural techniques that are now available for basic or translational research in reproductive biology and reproductive medicine. These include TEM, ultrastructural immunocytochemistry, ultrastructural histochemistry, and SEM.

  6. Weak-beam scanning transmission electron microscopy for quantitative dislocation density measurement in steels.

    PubMed

    Yoshida, Kenta; Shimodaira, Masaki; Toyama, Takeshi; Shimizu, Yasuo; Inoue, Koji; Yoshiie, Toshimasa; Milan, Konstantinovic J; Gerard, Robert; Nagai, Yasuyoshi

    2017-04-01

    To evaluate dislocations induced by neutron irradiation, we developed a weak-beam scanning transmission electron microscopy (WB-STEM) system by installing a novel beam selector, an annular detector, a high-speed CCD camera and an imaging filter in the camera chamber of a spherical aberration-corrected transmission electron microscope. The capabilities of the WB-STEM with respect to wide-view imaging, real-time diffraction monitoring and multi-contrast imaging are demonstrated using typical reactor pressure vessel steel that had been used in an European nuclear reactor for 30 years as a surveillance test piece with a fluence of 1.09 × 1020 neutrons cm-2. The quantitatively measured size distribution (average loop size = 3.6 ± 2.1 nm), number density of the dislocation loops (3.6 × 1022 m-3) and dislocation density (7.8 × 1013 m m-3) were carefully compared with the values obtained via conventional weak-beam transmission electron microscopy studies. In addition, cluster analysis using atom probe tomography (APT) further demonstrated the potential of the WB-STEM for correlative electron tomography/APT experiments. © The Author 2017. Published by Oxford University Press on behalf of The Japanese Society of Microscopy. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

  7. The Effect of Electron Beam Irradiation in Environmental Scanning Transmission Electron Microscopy of Whole Cells in Liquid.

    PubMed

    Hermannsdörfer, Justus; Tinnemann, Verena; Peckys, Diana B; de Jonge, Niels

    2016-06-01

    Whole cells can be studied in their native liquid environment using electron microscopy, and unique information about the locations and stoichiometry of individual membrane proteins can be obtained from many cells thus taking cell heterogeneity into account. Of key importance for the further development of this microscopy technology is knowledge about the effect of electron beam radiation on the samples under investigation. We used environmental scanning electron microscopy (ESEM) with scanning transmission electron microscopy (STEM) detection to examine the effect of radiation for whole fixed COS7 fibroblasts in liquid. The main observation was the localization of nanoparticle labels attached to epidermal growth factor receptors (EGFRs). It was found that the relative distances between the labels remained mostly unchanged (<1.5%) for electron doses ranging from the undamaged native state at 10 e-/Å2 toward 103 e-/Å2. This dose range was sufficient to determine the EGFR locations with nanometer resolution and to distinguish between monomers and dimers. Various different forms of radiation damage became visible at higher doses, including severe dislocation, and the dissolution of labels.

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

    PubMed Central

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

    2017-01-01

    Sample preparation is critical to biological electron microscopy (EM), and there have been continuous efforts on optimizing the procedures to best preserve structures of interest in the sample. However, a quantitative characterization of the morphological changes associated with each step in EM sample preparation is currently lacking. Using correlative EM and superresolution microscopy (SRM), we have examined the effects of different drying methods as well as osmium tetroxide (OsO4) post-fixation on cell morphology during scanning electron microscopy (SEM) sample preparation. Here, SRM images of the sample acquired under hydrated conditions were used as a baseline for evaluating morphological changes as the sample went through SEM sample processing. We found that both chemical drying and critical point drying lead to a mild cellular boundary retraction of ~60 nm. Post-fixation by OsO4 causes at least 40 nm additional boundary retraction. We also found that coating coverslips with adhesion molecules such as fibronectin prior to cell plating helps reduce cell distortion from OsO4 post-fixation. These quantitative measurements offer useful information for identifying causes of cell distortions in SEM sample preparation and improving current procedures. PMID:28562683

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

    PubMed

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

    2017-01-01

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

  10. Scanning electron microscopy of scales and its taxonomic application in the fish genus Channa.

    PubMed

    Dey, Sudip; Biswas, Shyama P; Dey, Samujjwal; Bhattacharyya, Shankar P

    2014-08-01

    Scanning electron microscopy (SEM) of scales in six species of the fish genus Channa revealed certain features relevant to taxonomic significance. The location of focus, inter-radial distance and width of circuli, inter-circular space, width of radii, shape and size of lepidonts, etc. were found to be different in different species. The importance of SEM of scales in poorly understood taxonomy and phylogeny of the fish genus Channa is discussed with the help of relevant literature. Further, the role of SEM of fish scales for taxonomic applications is discussed in detail.

  11. Scanning electron microscopy of growing dental plaque: a quantitative study with different mouth rinses.

    PubMed

    Jentsch, Holger; Mozaffari, Eshan; Jonas, Ludwig

    2013-08-01

    The aim of this study was to quantify the influence of different mouth rinses on dental plaque. Wearing splints with enamel pieces 24 volunteers rinsed with essential oils, amine/stannous fluoride, or chlorhexidine digluconate (0.12%) mouth rinses. After 24, 48, 72, and 96 h the enamel pieces were analyzed by scanning electron microscopy. The counts of cocci and bacilli in different plaque layers and the plaque thickness were almost similar using essential oils and amine/stannous fluoride. These results differed significantly from those of chlorhexidine digluconate mouth rinses. The results for plaque thickness were without significant differences between the groups at any appointment.

  12. Fatal poisoning by Rumex crispus (curled dock): pathological findings and application of scanning electron microscopy.

    PubMed

    Reig, R; Sanz, P; Blanche, C; Fontarnau, R; Dominguez, A; Corbella, J

    1990-10-01

    A case of fatal poisoning due to ingestion of the plant Rumex crispus (curled dock) is described. The patient, a 53-year-old male, presented with gastrointestinal symptoms, severe hypocalcemia, metabolic acidosis and acute hepatic insufficiency. Despite therapeutic measures, the patient died 72 h after ingestion of the plant material. Noteworthy among the pathological findings were centrolobular hepatic necrosis and birefringent crystals in the liver and kidneys that were identified by histochemical techniques and scanning electron microscopy. These observations are compared with other reports in the medical literature, with an emphasis on the risk involved in the use of these plants for culinary or medicinal purposes.

  13. Liquid scanning transmission electron microscopy: Nanoscale imaging in micrometers-thick liquids

    NASA Astrophysics Data System (ADS)

    Schuh, Tobias; de Jonge, Niels

    2014-02-01

    Scanning transmission electron microscopy (STEM) of specimens in liquid is possible using a microfluidic chamber with thin silicon nitride windows. This paper includes an analytic equation of the resolution as a function of the sample thickness and the vertical position of an object in the liquid. The equipment for STEM of liquid specimen is briefly described. STEM provides nanometer resolution in micrometer-thick liquid layers with relevance for both biological research and materials science. Using this technique, we investigated tagged proteins in whole eukaryotic cells, and gold nanoparticles in liquid with time-lapse image series. Possibly future applications are discussed. xml:lang="fr"

  14. Filamentous phage pIV multimer visualized by scanning transmission electron microscopy

    SciTech Connect

    Linderoth, N.A.; Russel, M.; Simon, M.N.

    1997-11-28

    A family of homomultimeric outer-membrane proteins termed secretins mediates the secretion of large macromolecules such as enzymes and filamentous bacteriophages across bacterial outer membranes to the extracellular millieu. The secretin encoded by filamentous phage f1 was purified. Mass determination of individual molecules by scanning transmission electron microscopy revealed two forms, a unit multimer composed of about 14 subunits and a multimer dimer. The secretin is roughly cylindrical and has an internal diameter of about 80 angstroms, which is large enough to accommodate filamentous phage (diameter of 65 angstroms). 21 refs., 3 figs., 1 tab.

  15. Scanning electron microscopy of a blister roof in dystrophic epidermolysis bullosa.

    PubMed

    Almeida, Hiram Larangeira de; Monteiro, Luciane; Marques e Silva, Ricardo; Rocha, Nara Moreira; Scheffer, Hans

    2013-01-01

    In dystrophic epidermolysis bullosa the genetic defect of anchoring fibrils leads to cleavage beneath the basement membrane, with its consequent loss. We performed scanning electron microscopy of an inverted blister roof of a case of dystrophic epidermolysis bullosa, confirmed by immunomapping and gene sequencing. With a magnification of 2000 times a net attached to the blister roof could be easily identified. This net was composed of intertwined flat fibers. With higher magnifications, different fiber sizes could be observed, some thin fibers measuring around 80 nm and thicker ones measuring between 200 and 300 nm.

  16. Lanthanum Deposition in the Stomach: Usefulness of Scanning Electron Microscopy for Its Detection.

    PubMed

    Iwamuro, Masaya; Urata, Haruo; Tanaka, Takehiro; Ando, Akemi; Nada, Takahiro; Kimura, Kosuke; Yamauchi, Kenji; Kusumoto, Chiaki; Otsuka, Fumio; Okada, Hiroyuki

    2017-02-01

    After having been treated with lanthanum carbonate administration for 4 years for hyperphosphatemia, a 75-year-old Japanese woman undergoing hemodialysis was diagnosed with lanthanum phosphate deposition in the stomach. The deposition, seen as white microgranules, was observed using esophagogastroduodenoscopy with magnifying observation. To the best of our knowledge, these are the minutest endoscopy images of lanthanum phosphate deposition in the gastric mucosa. Scanning electron microscopy (SEM) observation enabled easier identification of the deposited material, which was visible as bright areas. The present case suggests the usefulness of SEM observation in the detection of lanthanum phosphate deposition in the gastrointestinal tract.

  17. Use of low-temperature scanning electron microscopy to observe icicles, ice fabric, rime and frost

    SciTech Connect

    Wergin, W.P.; Rango, A.; Erbe, E.F.

    1996-12-31

    Previous studies showed that low temperature scanning electron microscopy (SEM) can be used to record images of precipitated snow crystals, which collectively form structures that are commonly known as snowflakes. Information about the structure of snow crystals can be used to improve models that estimate the water equivalent of the winter snowpack. These models, which are based on satellite microwave data, have practical use in approximating the quantity of water that will be available for crop irrigation and hydroelectric power. Our previous success of using low temperature SEM to image snow crystals has encouraged us to utilize this technique for other types of frozen aqueous specimens.

  18. Visualizing gold nanoparticle uptake in live cells with liquid scanning transmission electron microscopy.

    PubMed

    Peckys, Diana B; de Jonge, Niels

    2011-04-13

    The intracellular uptake of 30 nm diameter gold nanoparticles (Au-NPs) was studied at the nanoscale in pristine eukaryotic cells. Live COS-7 cells were maintained in a microfluidic chamber and imaged using scanning transmission electron microscopy. A quantitative image analysis showed that Au-NPs bound to the membranes of vesicles, possibly lysosomes, and occupied 67% of the available surface area. The vesicles accumulated to form a micrometer-sized cluster after 24 h of incubation. Two clusters were analyzed and found to consist of 117 ± 9 and 164 ± 4 NP-filled vesicles.

  19. Study of environmental biodegradation of LDPE films in soil using optical and scanning electron microscopy.

    PubMed

    Mumtaz, Tabassum; Khan, M R; Hassan, Mohd Ali

    2010-07-01

    An outdoor soil burial test was carried out to evaluate the degradation of commercially available LDPE carrier bags in natural soil for up to 2 years. Biodegradability of low density polyethylene films in soil was monitored using both optical and scanning electron microscopy (SEM). After 7-9 months of soil exposure, microbial colonization was evident on the film surface. Exposed LDPE samples exhibit progressive changes towards degradation after 17-22 months. SEM images reveal signs of degradation such as exfoliation and formation of cracks on film leading to disintegration. The possible degradation mode and consequences on the use and disposal of LDPE films is discussed.

  20. Microstructure development in particulate coatings examined with high-resolution cryogenic scanning electron microscopy

    SciTech Connect

    Sheehan, J.G.; Davis, H.T.; Scriven, L.E.; Takamura, Koichi

    1993-12-01

    The authors used cryogenic scanning electron microscopy to examine the early stages of latex film formation. They visualized the influence of ionic strength and extent of carboxylation in latex-calcium carbonate formulations and in latex-only formulations. Results demonstrated that latex particles deposited on calcium carbonate surfaces creating a suspension of carboxylic acid-stabilized calcium carbonate particles. Images of consolidation fronts showed that variation of ionic strength and extent of carboxylation dramatically changes the way latex particles consolidate and form films.

  1. In situ scanning electron microscopy of graphene growth on polycrystalline Ni substrate

    NASA Astrophysics Data System (ADS)

    Takahashi, Katsuhiro; Yamada, Kazuki; Kato, Hiroki; Hibino, Hiroki; Homma, Yoshikazu

    2012-04-01

    Scanning electron microscopy (SEM) is shown to be capable of imaging a monolayer of graphene, and is employed to observe in situ the graphene growth process by segregation of bulk-dissolved carbon on a polycrystalline nickel surface. Because of a wide field of view, SEM could easily track the rapid graphene growth induced by carbon segregation. Monolayer graphene extended on (111)- and (011)-oriented nickel grains, but was excluded from the (001) grains. This is due to the difference in carbon-nickel binding energy among these crystalline faces. This work proves the usefulness of in situ SEM imaging for the investigation of large area graphene growth.

  2. Scanning electron microscopy with polarization analysis for multilayered chiral spin textures

    NASA Astrophysics Data System (ADS)

    Lucassen, Juriaan; Kloodt-Twesten, Fabian; Frömter, Robert; Oepen, Hans Peter; Duine, Rembert A.; Swagten, Henk J. M.; Koopmans, Bert; Lavrijsen, Reinoud

    2017-09-01

    We show that scanning electron microscopy with polarization analysis (SEMPA) that is sensitive to both in-plane magnetization components can be used to image the out-of-plane magnetized multi-domain state in multilayered chiral spin textures. By depositing a thin layer of Fe on top of the multilayer, we image the underlying out-of-plane domain state through the mapping of its stray fields in the Fe. We also demonstrate that SEMPA can be used to image the domain wall chirality in these systems after milling away the capping layer and imaging the topmost magnetic layer directly.

  3. Scanning electron microscopy of the orbital Harderian gland in the male Atlantic bottlenose dolphin (Tursiops truncatus).

    PubMed

    Ortiz, G G; Feria-Velasco, A; Pacheco-Moisés, F P; Rodríguez-Reinoso, S; Cruz-Ramos, J A; Rosales-Corral, S A; Reiter, R J

    2009-08-01

    The ultrastructure of the Harderian gland of Atlantic bottlenose dolphin (Tursiops truncatus) was examined by scanning electron microscopy (SEM). We found the following surface features: the typical round appearance of the ascinar glandular unit with a finely granular surface, a thin cortex and immediately below two types of cells: type I cells (characterized by small lipid vacuoles) and type II cells (characterized by large lipid vacuoles). It has been suggested that different cells forms represent a single cell type in varying activity states. Additionally, a coalescent tubular complex, a small balloon-like structures and large globular structures were observed. These structures may be reservoirs of secretion products.

  4. Sialolith characterization by scanning electron microscopy and X-ray photoelectron spectroscopy.

    PubMed

    Giray, C Bahadir; Dogan, Meral; Akalin, Ayse; Baltrusaitis, Jonas; Chan, Daniel C N; Skinner, H Catherine W; Dogan, A Umran

    2007-01-01

    The objective of this study has been to characterize sialolith, a calcium phosphate deposit that develops in the human oral cavity, by high-resolution field emission scanning electron microscopy (SEM) and X-ray photoelectron spectroscopy (XPS). The morphological and chemical data obtained helped in the determination of their formation mechanism in salivary glands. Sialoliths in the submandibular salivary glands may arise secondary to sialodenitis, but not via a luminal organic nidus. We believe this is the first study that characterizes a sialolith by XPS.

  5. Scanning electron microscopy in the investigation of the in vitro hemolytic activity of Trichomonas vaginalis.

    PubMed

    Rosset, Iveli; Tasca, Tiana; Tessele, Paola M; De Carli, Geraldo A

    2002-04-01

    The in vitro hemolytic activity of Trichomonas vaginalis has been previously demonstrated, but the mechanisms involved remain to be elucidated. In this work we used scanning electron microscopy to investigate the contact dependency of the hemolytic phenomenon caused by the parasites. The erythrocytes adhered to the parasites' surface and were phagocytosed. These observations suggest that the contact between T. vaginalis and erythrocytes may be an important mechanism in the injury caused to the erythrocytes. The hemolytic activity of T. vaginalis may be an efficient means of obtaining nutrients for the parasite and allow the investigation of the mechanism used by T. vaginalis to damage cellular membranes.

  6. Silica granuloma: scanning electron microscopy and energy dispersive X-ray microanalysis.

    PubMed

    Chun, S I; Cho, S W

    1991-02-01

    A 46-year-old woman had 1-month-old erythematous papules on the left elbow and both knees where acupuncture with gold needles had been performed twenty years earlier. She also had a 2-month-old pruritic scar lesion on the nape. Histopathologic findings showed sarcoidal granulomas. The presence of silica components in the granulomas was confirmed by scanning electron microscopy (SEM) of the lesion on the nape and energy dispersive X-ray microanalysis (EDXA) of the lesions on the nape and the left elbow. We suggest that acupuncture with gold needles may be one cause of silica granuloma.

  7. Scanning transmission electron microscopy analysis of grain structure in perpendicular magnetic recording media.

    PubMed

    Hossein-Babaei, Faraz; Sinclair, Robert; Sinclair, Robert A; Srinivasan, Kumar; Bertero, Gerardo A

    2011-09-14

    The key component of a hard disk medium is a Co-based magnetic layer (ML) grown on a Ru seed layer. The ML nanostructure, composed of less than 10 nm grains, is believed to be controlled by this seed layer. We successfully used scanning transmission electron microscopy energy dispersive spectrometry simultaneous composition-based imaging and Moiré pattern analysis for determining the mutual structural and orientation relationship between the two layers revealing a grain-to-grain agreement. The method presented here can be utilized for observing structural correlations between consecutive polycrystalline thin film layers in general.

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

    PubMed

    Schreiner, Manfred; Melcher, Michael; Uhlir, Katharina

    2007-02-01

    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.

  9. Scanning electron microscopy of a blister roof in dystrophic epidermolysis bullosa*

    PubMed Central

    de Almeida Jr., Hiram Larangeira; Monteiro, Luciane; Silva, Ricardo Marques e; Rocha, Nara Moreira; Scheffer, Hans

    2013-01-01

    In dystrophic epidermolysis bullosa the genetic defect of anchoring fibrils leads to cleavage beneath the basement membrane, with its consequent loss. We performed scanning electron microscopy of an inverted blister roof of a case of dystrophic epidermolysis bullosa, confirmed by immunomapping and gene sequencing. With a magnification of 2000 times a net attached to the blister roof could be easily identified. This net was composed of intertwined flat fibers. With higher magnifications, different fiber sizes could be observed, some thin fibers measuring around 80 nm and thicker ones measuring between 200 and 300 nm. PMID:24474107

  10. Scanning electron microscopy of the collodion membrane from a self-healing collodion baby*

    PubMed Central

    de Almeida Jr., Hiram Larangeira; Isaacsson, Henrique; Guarenti, Isabelle Maffei; Silva, Ricardo Marques e; de Castro, Luis Antônio Suita

    2015-01-01

    Abstract Self-healing collodion baby is a well-established subtype of this condition. We examined a male newborn, who was covered by a collodion membrane. The shed membrane was examined with scanning electron microscopy. The outer surface showed a very compact keratin without the normal elimination of corneocytes. The lateral view of the specimen revealed a very thick, horny layer. The inner surface showed the structure of lower corneocytes with polygonal contour. With higher magnifications villous projections were seen in the cell membrane. PMID:26375232

  11. Scanning electron microscopy and energy dispersive analysis of machined denture base surfaces.

    PubMed

    Radford, D R; Walter, J D; Challacombe, S J

    1997-01-01

    To relate the characteristics of rotary instruments to the surfaces they produce, acrylic resin, Molloplast B, and Novus were investigated with energy dispersive analysis and scanning electron microscopy (secondary and backscatter images) before and after machining. The chemical composition of cutting instruments, material surfaces, and residues was identified. Machined debris embedded in Molloplast B after machining with the Molloplast stone was found to contain a mean lead content of 45%. High concentrations of barium sulphate were discovered on the arbor band-machined surface of Novus. These differences were related to clinically appropriate instrumentation, and, therefore, biocompatibility studies that intimately relate to the in vivo situation should be considered for new materials.

  12. Atomic bonding effects in annular dark field scanning transmission electron microscopy. II. Experiments

    SciTech Connect

    Odlyzko, Michael L.; Held, Jacob T.; Mkhoyan, K. Andre

    2016-07-15

    Quantitatively calibrated annular dark field scanning transmission electron microscopy (ADF-STEM) imaging experiments were compared to frozen phonon multislice simulations adapted to include chemical bonding effects. Having carefully matched simulation parameters to experimental conditions, a depth-dependent bonding effect was observed for high-angle ADF-STEM imaging of aluminum nitride. This result is explained by computational predictions, systematically examined in the preceding portion of this study, showing the propagation of the converged STEM beam to be highly sensitive to net interatomic charge transfer. Thus, although uncertainties in experimental conditions and simulation accuracy remain, the computationally predicted experimental bonding effect withstands the experimental testing reported here.

  13. Reaction of LiD with water vapor: thermogravimetric and scanning electron microscopy studies

    SciTech Connect

    Balooch, M; Dinh, L N; LeMay, J D

    2000-09-14

    The kinetics of hydroxide film growth on LiD have been studied by the thermogravimetric method in nitrogen saturated with water vapor and by scanning electron microscopy (SEM) of samples that have been exposed to air with 50% relative humidity. The reaction probability is estimated to be 4 x 10{sup -7} for LiD exposed to ambient air with 50% relative humidity, suggesting that the diffusion through the hydroxide film is not the limiting step on the overall process at high moisture levels. The rate of growth is drastically reduced when the temperature is increased to 60 C.

  14. Three-dimensional aspects of superficial disseminated porokeratosis with scanning electron microscopy*

    PubMed Central

    de Almeida Jr., Hiram Larangeira; de Abreu, Luciana Boff; Rampon, Greice; Silva, Ricardo Marques e; Rocha, Nara Moreira

    2014-01-01

    The three-dimensional findings of the surface and from a cross section from a case of disseminated superficial porokeratois using scanning electron microscopy are reported. On the surface of the skin, irregular keratin with a serpiginous distribution was seen. A gross aspect of keratin in the hyperkeratotic wall was also observed and compared to the normal area, in which the release of corneocytes seemed normal. The cross-sectional imaging easily identified the cornoid lamella, with compact keratin surrounded by normal stratum corneum. PMID:25387509

  15. On the visibility of very thin specimens in annular bright field scanning transmission electron microscopy

    SciTech Connect

    Phillips, P. J.; Klie, R. F.

    2013-07-15

    Annular bright field (ABF) scanning transmission electron microscopy (STEM) is emerging as an important observation mode for its ability to simultaneously image both heavy and light elements. However, recent results have demonstrated that in the limit of a very thin specimen (a few atomic layers), the ABF and high angle annular dark field (HAADF) signals cease to be intuitively related: a phenomenon which is generally irrelevant when imaging 'normal' specimens. ABF/HAADF STEM observations and multislice image simulations of two catalyst samples of differing atomic weights are presented; it is shown that the nature of the ABF signal is specimen dependent.

  16. Three-dimensional structure of a parameatal urethral cyst by scanning electron microscopy.

    PubMed

    Yamada, Nanako; Yoshida, Yuichi; Ito, Ayako; Morino, Shinichi; Yamamoto, Osamu

    2013-05-01

    We report scanning electron microscopy (SEM) for a case of parameatal urethral cyst. A 6-year-old Japanese boy presented with a cyst on the right lateral side of the urethral meatus. Histological examination revealed a cyst lined with columnar epithelium. An immunohistochemical study showed positive staining for CK7, CK13, and CEA, and negative for CK20 in luminal cells. On SEM examination, the inner surface of the cyst showed ridges arranged in a gyrus-like manner at lower magnification. Higher magnification revealed luminal cells with short microvillus projections. Some cells showed apocrine, merocrine, and possibly holocrine-type secretions.

  17. High-resolution characterization of multiferroic heterojunction using aberration-corrected scanning transmission electron microscopy

    NASA Astrophysics Data System (ADS)

    Yuan, Zhoushen; Ruan, Jieji; Xie, Lin; Pan, Xiaoqing; Wu, Di; Wang, Peng

    2017-04-01

    Multiferroic tunnel junctions have been considered as potential candidates for nonvolatile memory devices. Understanding the atomic structure at the interface is crucial for optimizing the performances in such oxide electronics. Spatially resolved electron energy loss spectroscopy (EELS) combined with aberration-corrected scanning transmission electron microscopy is employed to measure the compositional profiles across the interfaces of different layers with atomic resolution. Two-dimensional elemental imaging with atomic resolution is demonstrated, and the influences of the interface sharpness, the terminal layer, and cation intermixing are investigated. An asymmetric sublattice intermixing at the Pr0.8Ca0.2MnO3/BaTiO3/La0.7Sr0.3MnO3 interface is observed, which can affect the local Mn valence and coupling. The reduction in the Mn valence at the interface is further studied using EELS near-edge fine structures.

  18. New area detector for atomic-resolution scanning transmission electron microscopy.

    PubMed

    Shibata, Naoya; Kohno, Yuji; Findlay, Scott D; Sawada, Hidetaka; Kondo, Yukihito; Ikuhara, Yuichi

    2010-01-01

    A new area detector for atomic-resolution scanning transmission electron microscopy (STEM) is developed and tested. The circular detector is divided into 16 segments which are individually optically coupled with photomultiplier tubes. Thus, 16 atomic-resolution STEM images which are sensitive to the spatial distribution of scattered electrons on the detector plane can be simultaneously obtained. This new detector can be potentially used not only for the simultaneous formation of common bright-field, low-angle annular dark-field and high-angle annular dark-field images, but also for the quantification of images by detecting the full range of scattered electrons and even for exploring novel atomic-resolution imaging modes by post-processing combination of the individual images.

  19. Field emission scanning electron microscopy (FE-SEM) as an approach for nanoparticle detection inside cells.

    PubMed

    Havrdova, M; Polakova, K; Skopalik, J; Vujtek, M; Mokdad, A; Homolkova, M; Tucek, J; Nebesarova, J; Zboril, R

    2014-12-01

    When developing new nanoparticles for bio-applications, it is important to fully characterize the nanoparticle's behavior in biological systems. The most common techniques employed for mapping nanoparticles inside cells include transmission electron microscopy (TEM) and scanning transmission electron microscopy (STEM). These techniques entail passing an electron beam through a thin specimen. STEM or TEM imaging is often used for the detection of nanoparticles inside cellular organelles. However, lengthy sample preparation is required (i.e., fixation, dehydration, drying, resin embedding, and cutting). In the present work, a new matrix (FTO glass) for biological samples was used and characterized by field emission scanning electron microscopy (FE-SEM) to generate images comparable to those obtained by TEM. Using FE-SEM, nanoparticle images were acquired inside endo/lysosomes without disruption of the cellular shape. Furthermore, the initial steps of nanoparticle incorporation into the cells were captured. In addition, the conductive FTO glass endowed the sample with high stability under the required accelerating voltage. Owing to these features of the sample, further analyses could be performed (material contrast and energy-dispersive X-ray spectroscopy (EDS)), which confirmed the presence of nanoparticles inside the cells. The results showed that FE-SEM can enable detailed characterization of nanoparticles in endosomes without the need for contrast staining or metal coating of the sample. Images showing the intracellular distribution of nanoparticles together with cellular morphology can give important information on the biocompatibility and demonstrate the potential of nanoparticle utilization in medicine. Copyright © 2014 Elsevier Ltd. All rights reserved.

  20. Investigating the use of in situ liquid cell scanning transmission electron microscopy

    SciTech Connect

    Nguy, Amanda

    2016-02-19

    Engineering nanoparticles with desired shape-dependent properties is the key to many applications in nanotechnology. Although many synthetic procedures exist to produce anisotropic gold nanoparticles, the dynamics of growth are typically unknown or hypothetical. In the case of seed-mediated growth in the presence of DNA into anisotropic nanoparticles, it is not known exactly how DNA directs growth into specific morphologies. A series of preliminary experiments were carried out to contribute to the investigation of the possible mechanism of DNA-mediated growth of gold nanoprisms into gold nanostars using liquid cell scanning transmission electron microscopy (STEM). Imaging in the liquid phase was achieved through the use of a liquid cell platform and liquid cell holder that allow the sample to be contained within a “chip sandwich” between two electron transparent windows. Ex situ growth experiments were performed using Au-T30 NPrisms (30-base thymine oligonucleotide-coated gold nanoprisms) that are expected to grow into gold nanostars. Growth to form these nanostars were imaged using TEM (transmission electron microscopy) and liquid cell STEM (scanning transmission electron microscopy). An attempt to perform in situ growth experiments with the same Au-T30 nanoprisms revealed challenges in obtaining desired morphology results due to the environmental differences within the liquid cell compared to the ex situ environment. Different parameters in the experimental method were explored including fluid line set up, simultaneous and alternating reagent addition, and the effect of different liquid cell volumes to ensure adequate flow of reagents into the liquid cell. Lastly, the binding affinities were compared for T30 and A30 DNA incubated with gold nanoparticles using zeta potential measurements, absorption spectroscopy, and isothermal titration calorimetry (ITC). It was previously reported thymine bases have a lower binding affinity to gold surfaces than adenine

  1. Morphology of the dentin structure of sloths Bradypus tridactylus: a light and scanning electron microscopy investigation.

    PubMed

    Santana, L N S; Barbosa, L V M; Teixeira, F B; Costa, A M P; Fernandes, L M P; Lima, R R

    2013-12-01

    The aim of this study was to describe the dentine morphology of sloths (Bradypus tridactylus). The sloth teeth were removed and prepared for light microscopy (LM) and scanning electron microscopy analyses (SEM). LM revealed two patterns of tubular dentins: an outer with dentinary tubules over the all tooth length and one in the inner part with larger diameter and more spaced tubules, when compared to those present in the outer dentine. These findings were confirmed by SEM, which revealed a tubular pattern in the outer dentine like in humans. The inner dentine displayed pared grouped tubules that were characterized as vascular channels. It can be concluded that this sloth species present two types of dentins: an inner dentin (ortodentin) and an outer dentin characterized as a vascular dentin. This suggests a partial evolutive/adaptive process of this dental tissue, as compared to other mammalian species.

  2. Scanning electron microscopy for flow-diverting stent research: technical tips and tricks.

    PubMed

    Iosif, Christina; Carles, Pierre; Trolliard, Gilles; Yardin, Catherine; Mounayer, Charbel

    2015-06-01

    Flow-diverting (FD) stents represent a new concept in the treatment of intracranial aneurysms with challenging anatomical dispositions. Having been introduced to clinical practice only in the last 5 years and featuring complex mechanisms of action, they are still under research. Scanning electron microscopy, as part of an animal research protocol, provides detailed surface observations of neointimal healing at the aneurysm's neck, as well as covered side branch ostia, allowing for the confirmation of scientific hypotheses and observations. Technical adaptations of preparation protocols are presented based on a pilot study on Large White pigs, stented with FD stents at carotid bifurcations. © The Author 2015. Published by Oxford University Press on behalf of The Japanese Society of Microscopy. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

  3. Direct observation of polyelectrolyte brushes under wet and dry conditions by atmospheric scanning electron microscopy.

    PubMed

    Higuchi, Takeshi; Konyuba, Yuji; Nishiyama, Hidetoshi; Suga, Mitsuo; Takahara, Atsushi; Jinnai, Hiroshi

    2016-04-01

    Polyelectrolyte brushes are polyelectrolyte polymers with one end fixed to a substrate. In this study, direct nano-scale visualization of polyelectrolyte brushes was carried out under 'aqueous conditions' by atmospheric scanning electron microscopy. The thickness of the polyelectrolyte brush layer was measured under both dry and aqueous conditions, experimentally confirming the swollen state of the brushes. These experimental findings qualitatively agreed with the results from previous neutron reflectivity experiments using similar polyelectrolyte brushes. Such direct visualization of polymer brushes in real space opens up a new route for better understanding their surface properties, such as friction, adhesion and wettability. © The Author 2015. Published by Oxford University Press on behalf of The Japanese Society of Microscopy. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

  4. Rigid gas permeable contact lenses surface roughness examined by interferential shifting phase and scanning electron microscopies.

    PubMed

    Merindano, M D; Canals, M; Saona, C; Costa, J

    1998-01-01

    The anterior surface roughness of seven factory new rigid gas permeable (RGP) contact lenses has been studied by interferential shifting phase microscopy (ISPM) and scanning electron microscopy (SEM). Five lenses were fluorsilicone acrylate and two lenses were silicone acrylate. Their material Dk ranged from 14 to 210. ISPM is shown to be a reliable and non-destructive method to observe and measure the relief of the contact lens surface. Moreover, profile and contour data are easily stored for further quantitative studies. ISPM contour patterns of the studied lenses are qualitatively compared with those obtained by SEM for the same lenses. Results point out that ISPM gives similar accuracy but it is non-destructive and cheaper than SEM. Moreover, the quantitative study of surface roughness suggests that there is a relationship between surface roughness and Dk of the lens material: surface roughness increases with Dk and allows to distinguish between lenses with low, medium and high Dk.

  5. An In vitro Study on Post Bleaching Pigmentation Susceptibility of Teeth and Scanning Electron Microscopy Analysis

    PubMed Central

    Latha, S Pushpa; Hegde, Vani; Raheel, Syed Ahmed; Tarakji, Bassel; Azzeghaiby, Saleh Nasser; Nassani, Mohammad Zakaria

    2014-01-01

    Background: To determine the susceptibility of teeth for repigmentation after bleaching. Materials and Methods: Forty premolars were assigned to three groups (n = 12). Group 1 was bleached using 30% w/v hydrogen peroxide 15 min 3 times a day every other day for 4 days. In Group 2 was bleached using 16% carbamide peroxide (Polanight), 90 min a day for 15 days. 2 days later, the shades of the bleached teeth were recorded. Remaining 4 teeth were bleached according to Group 1 and 2 and were subjected to atomic force microscopy, scanning electron microscopy analysis. Results: Specimens of athome bleaching were lighter than the specimens of inoffice bleaching. Conclusion: The susceptibility of enamel to pigmentation can be increased after bleaching, and pigmentation is greater if bleaching is performed with H2O2. The percentage change (lighter) was more for athome bleaching specimens as compared to inoffice bleaching specimens. PMID:25395800

  6. Narrow superficial injury to rabbit aortic endothelium. The healing process as observed by scanning electron microscopy.

    PubMed

    Ramsay, M M; Walker, L N; Bowyer, D E

    1982-06-01

    A study was made of the healing of aortic endothelium in rabbits following the production of a defined superficial injury. This was induced using a fine nylon filament which removed the endothelial cells without producing significant damage to underlying structures. The morphology of the injury and subsequent repair was observed using light microscopy and scanning and transmission electron microscopy. Two forms of injury were produced (a) a longitudinal injury along the full length of the aorta which was 50-80 microns wide (about 5-8 cell widths), (b) a circumferential injury approximately 80 microns wide (about 2 cell lengths). Thirty minutes after injury the exposed tissue was almost devoid of adherent cells, but after 4 h became covered by a sparse monolayer of platelets. Occasional leukocytes were also present from 7 h after injury. Injury tracks were found to repair very quickly; re-endothelialisation being complete by 48 h and there being no sign of injury by 7 days.

  7. Scanning electron microscopy of legs of two species of sucking lice (Anoplura: Phthiraptera).

    PubMed

    Soler Cruz, M D; Martin Mateo, M P

    2009-04-01

    Pretarsal, tarsal and tibial structures of the forelegs, midlegs and hindlegs of Pediculus humanus of humans and of Haematopinus apri Goureau, 1866 (Phthiraptera), a parasite of feral hogs, were studied using light microscopy and scanning electron microscopy. Details of the tibial thumb-like process (tl) with the spine of the thumb (spn), tarsal apophysis (ta) and the coupled finger-like process (cfl) can be observed in the leg photomicrograph of both species. A frontal view of the leg in open position shows the articulation of the claw: the structures of an open-closed system, a tooth row (te), rack-system (rs) and two telescopic columns (tc) which are present near the base of the claw in both species. In H. apri, we observed a pad-like structure, the flap-like tibial lobe (fl) on the ventral surface on the tarsus, the euplantulae, with several sensilla basiconica, which is present in each leg.

  8. Staining and embedding of human chromosomes for 3-d serial block-face scanning electron microscopy.

    PubMed

    Yusuf, Mohammed; Chen, Bo; Hashimoto, Teruo; Estandarte, Ana Katrina; Thompson, George; Robinson, Ian

    2014-12-01

    The high-order structure of human chromosomes is an important biological question that is still under investigation. Studies have been done on imaging human mitotic chromosomes using mostly 2-D microscopy methods. To image micron-sized human chromosomes in 3-D, we developed a procedure for preparing samples for serial block-face scanning electron microscopy (SBFSEM). Polyamine chromosomes are first separated using a simple filtration method and then stained with heavy metal. We show that the DNA-specific platinum blue provides higher contrast than osmium tetroxide. A two-step procedure for embedding chromosomes in resin is then used to concentrate the chromosome samples. After stacking the SBFSEM images, a familiar X-shaped chromosome was observed in 3-D.

  9. Probing core-electron orbitals by scanning transmission electron microscopy and measuring the delocalization of core-level excitations

    NASA Astrophysics Data System (ADS)

    Jeong, Jong Seok; Odlyzko, Michael L.; Xu, Peng; Jalan, Bharat; Mkhoyan, K. Andre

    2016-04-01

    By recording low-noise energy-dispersive x-ray spectroscopy maps from crystalline specimens using aberration-corrected scanning transmission electron microscopy, it is possible to probe core-level electron orbitals in real space. Both the 1 s and 2 p orbitals of Sr and Ti atoms in SrTi O3 are probed, and their projected excitation potentials are determined. This paper also demonstrates experimental measurement of the electronic excitation impact parameter and the delocalization of an excitation due to Coulombic beam-orbital interaction.

  10. Enhanced imaging of biomolecules with electron beam deposited tips for scanning force microscopy

    NASA Astrophysics Data System (ADS)

    Zenhausern, F.; Adrian, M.; ten Heggeler-Bordier, B.; Ardizzoni, F.; Descouts, P.

    1993-06-01

    Tip/sample interaction on the scanning force microscope (SFM) is a particularly difficult problem with biological materials. One major factor affecting image quality is the tip shape. Improved electron beam induced deposition technique with a scanning electron microscope (SEM) was used for the reproducible fabrication of carbon sharp tips on the end of commercially available silicon nitride cantilevers for scanning force microscopy. By aligning a fine focused beam of 20 nm diameter directly down the axis of the pyramidal tip at electron energy of 20 kV, carbon deposits grow with full cone angle of about 25°, cone length of 2 μm, and radii of curvature down to 10 nm, making these e-beam tips suitable for biomolecules imaging. The tip dimensions also were controlled by adjusting the beam parameters. Three different types of SFM tips were used to image tobacco mosaic virus (TMV). Conventional pyramidal tips appeared generally worse for imaging helical particles of TMV than SEM-deposited tips which were found more robust than commercially available conical tips. The use of sharper tips for SFM imaging of protein DNA revealed a 25% improvement in lateral resolution.

  11. Anatomical descriptions of silicified woods from Madagascar and Indonesia by scanning electron microscopy.

    PubMed

    Yoon, Chul Jong; Kim, Ki Woo

    2008-10-01

    Fine structure and tissue substitution by minerals were investigated in silicified woods from Madagascar and Indonesia by scanning electron microscopy and X-ray microanalysis. The silicified woods maintained the exterior morphology of once grown trees and showed typical inner structures of conifers. Radial planes of the silicified wood from Madagascar revealed tracheids as a major component of the axial system in the secondary xylem. Tracheids were mainly characterized by numerous bordered pits where a thickening in the middle (torus) was surrounded with the membrane (margo). The torus appeared to contrast with the fibrillar network of the margo. As a component of the axial system in the secondary phloem, sieve elements were found to have many sieve pores that were filled with seemingly crystalline materials. To correlate the colors of the silicified wood from Indonesia with elemental composition, energy-dispersive X-ray spectrometry was employed in this study. Silicon was present as a basic component of the silicified wood. Calcium and iron were detected from red-colored regions, whereas magnesium was found in blue-colored regions. These results suggest that tissues of silicified woods had been substituted by minerals over the past period, while retaining the inherent morphology of the tree species. Scanning electron microscopy and X-ray microanalysis could be applied to unravel structural details and composition of plant fossils in palaeobotany.

  12. Histological and scanning electron microscopy assessment of various vital pulp-therapy materials.

    PubMed

    Dominguez, Mercedes S; Witherspoon, David E; Gutmann, James L; Opperman, Lynne A

    2003-05-01

    Pulp capping and pulpotomy procedures were performed on 15 male mongrel dogs. Three materials were used: calcium hydroxide, acid-etched dentin bonding, and mineral trioxide aggregate. Six of the animals were killed at 50 days and nine were killed at 150 days. Samples from 11 dogs were used for histological evaluation, and the remaining dog samples were used for scanning electron microscopy evaluation. Each slide was graded histologically according to previously published criteria. Scanning electron microscopy analysis was performed, and the weight percentage of elements found in the dentin of a nontreated tooth versus the bridge formed in the exposed specimen was established. By evaluating pulp inflammation in vital pulp-therapy treatments, it was found that mineral trioxide aggregate was not significantly different from the untreated control group, both in pulp-capping procedures at 50 days (p = 0.357) or 150 days (p = 0.198) and pulpotomy procedures at 50 days (p = 0.357) or 150 days (p = 0.198). Moreover, histologically mineral trioxide aggregate was a considerably better material than calcium hydroxide or acid-etched dentin bonding in maintaining the integrity of the pulp.

  13. Microfluidic device for a rapid immobilization of zebrafish larvae in environmental scanning electron microscopy.

    PubMed

    Akagi, Jin; Zhu, Feng; Skommer, Joanna; Hall, Chris J; Crosier, Philip S; Cialkowski, Michal; Wlodkowic, Donald

    2015-03-01

    Small vertebrate model organisms have recently gained popularity as attractive experimental models that enhance our understanding of human tissue and organ development. Despite a large body of evidence using optical spectroscopy for the characterization of small model organism on chip-based devices, no attempts have been so far made to interface microfabricated technologies with environmental scanning electron microscopy (ESEM). Conventional scanning electron microscopy requires high vacuum environments and biological samples must be, therefore, submitted to many preparative procedures to dehydrate, fix, and subsequently stain the sample with gold-palladium deposition. This process is inherently low-throughput and can introduce many analytical artifacts. This work describes a proof-of-concept microfluidic chip-based system for immobilizing zebrafish larvae for ESEM imaging that is performed in a gaseous atmosphere, under low vacuum mode and without any need for sample staining protocols. The microfabricated technology provides a user-friendly and simple interface to perform ESEM imaging on zebrafish larvae. Presented lab-on-a-chip device was fabricated using a high-speed infrared laser micromachining in a biocompatible poly(methyl methacrylate) thermoplastic. It consisted of a reservoir with multiple semispherical microwells designed to hold the yolk of dechorionated zebrafish larvae. Immobilization of the larvae was achieved by a gentle suction generated during blotting of the medium. Trapping region allowed for multiple specimens to be conveniently positioned on the chip-based device within few minutes for ESEM imaging.

  14. New Aspidoderidae species parasite of Didelphis aurita (Mammalia: Didelphidae): a light and scanning electron microscopy approach.

    PubMed

    Chagas-Moutinho, V A; Sant'anna, V; Oliveira-Menezes, A; De Souza, W

    2014-02-01

    Nematodes of the family Aspidoderidae (Nematoda: Heterakoidea) Skrjabin and Schikobalova, 1947, are widely distributed in the Americas. The family Aspidoderidae includes the subfamilies Aspidoderinae Skrjabin and Schikobalova, 1947, and Lauroiinae Skrjabin and Schikobalova, 1951. These two subfamilies are delineated by the presence or absence of cephalic cordons at the anterior region. The nematodes in the subfamily Aspidoderinae, which includes the genus AspidoderaRailliet and Henry, 1912, are represented by nematodes with anterior cephalic cordons at the anterior end. The nematodes of the genus AspidoderaRailliet and Henry, 1912, are found in the cecum and large intestine of mammals of the orders Edentata, Marsupialia and Rodentia. Species within this genus have many morphological similarities. The use of scanning electron microscopy allows the specific characterization of the species within this genus. In the present work, we describe a new species of Aspidodera parasite of the large intestine of Didelphis aurita (Mammalia: Didelphidae) Wied-Neuwied, 1826, collected from Cachoeiras de Macacu, Rio de Janeiro. The combination of light and scanning electron microscopy allowed us a detailed analysis of this nematode.

  15. Experimental examination of the characteristics of bright-field scanning confocal electron microscopy images.

    PubMed

    Hashimoto, Ayako; Mitsuishi, Kazutaka; Shimojo, Masayuki; Zhu, Yufang; Takeguchi, Masaki

    2011-01-01

    We experimentally examined the characteristics of bright-field (BF) scanning confocal electron microscopy (SCEM) images by changing the observation conditions and comparing the images with those obtained by BF transmission electron microscopy (TEM) and BF scanning TEM (STEM) modes. The observation of 5-nm-diameter Au nanoparticles demonstrated that BF-SCEM produces object elongation of more than 2000 nm along the optical axis, as do BF-TEM and BF-STEM. We demonstrated the relationship between elongation length and geometric effects such as convergence and collection angles of a probe and the lateral size of an object; the relationship is consistent with previous theoretical prediction. Further, we observed interesting features that are seen only in the BF-SCEM images; the film contrast was strongly enhanced, compared with that of BF-STEM. In addition, a bright contrast appeared around the object position in the elongated images. Using this characteristic, we could determine the object position and structure.

  16. Scanning electron microscopy of the tongue, pharynx, and larynx of rats exposed to cigarette smoke.

    PubMed

    Martins, Regina Helena Garcia; Goncalves, Tatiana Maria; Madeira, Sergio Luis; Dias, Norimar Hernandes; de Oliveira Semenzati, Graziela

    2014-05-01

    To examine, by using scanning electron microscopy, the surface of the tongue, pharynx, and larynx of animals exposed to tobacco. Experimental study. Twenty rats were allocated to two groups: group I, control group: 10 rats not exposed to any inhaling pollutant and group II, tobacco group: 10 rats exposed to smoke from 10 cigarettes twice a day for 260 days. Animals of both groups had no restriction of food or water. After those 260 days, their aerodigestive segment was removed, and fragments of their tongue, hypopharynx, and right vocal fold were immersed in 2.5% glutaraldehyde and prepared for scanning electron microscopy. The filiform tongue papillae of the tobacco group were irregularly displayed, flattened, and adhered to each other. The hypopharynx mucosa was highly irregular, thickened, rough and had increased superficial peeling. The mucosa of the vocal folds had deep furrows surrounding the cells. These alterations were not identified for the control group. Several changes were recorded for the tongue, pharynx, and larynx of tobacco group animals, confirming the harmful effects of smoking to the respiratory and digestive epithelium. Copyright © 2014 The Voice Foundation. Published by Mosby, Inc. All rights reserved.

  17. Advancing Atomic Scale Quantification of Interface Structure and Chemistry via Scanning Transmission Electron Microscopy

    NASA Astrophysics Data System (ADS)

    Dycus, Joseph Houston

    Electron microscopy is a powerful tool to characterize materials from the micro to atomic scale. Scanning transmission electron microscopy (STEM) has become notably important with resolution within the Angstrom regime. Such resolution provides information on the relationship between atomic structure and material properties. In addition, breakthroughs in spectroscopy have provided elemental mapping of individual atom columns leading to information such as chemical segregation. Although STEM is a powerful tool, its widespread use is relatively recent, leaving many opportunities to employ STEM to discover new information in materials systems and develop techniques to further advance capabilities of the instrument. In this work, the combination of aberration-corrected STEM imaging and spectroscopy are employed to identify relationships between structure and chemistry in materials systems for interfaces and bulk materials. Further, by building upon recently developed revolving STEM (RevSTEM), a calibration scheme has been developed to reduce scan distortion in imaging, enabling highly precise and accurate determination of atom column positions. Additionally, pathways for improving the signal-to-noise in energy dispersive X-ray mapping are shown. By a combination highly accurate distance mapping and atomic scale EDS, important materials relationships can be determined such as the effects of elemental segregation on strain and changes in the local bonding environment at surface reconstructions.

  18. In vitro MR imaging of hyaline cartilage: correlation with scanning electron microscopy.

    PubMed

    Goodwin, D W; Zhu, H; Dunn, J F

    2000-02-01

    Our goal was to determine how the three-dimensional structure of hyaline cartilage affects its MR appearance and to correlate this appearance with detailed structural analysis using scanning electron microscopy and freeze-fracture sectioning techniques. In vitro 7-T spin-echo MR images of hyaline cartilage specimens from four patients undergoing above-knee amputations were obtained parallel and perpendicular to the main magnetic field. Specimens were imaged with low- and high-power scanning electron microscopy after freeze fracturing. The corresponding images from both techniques were analyzed with specific attention to the three-dimensional structure of the cartilage, collagen fibril orientation, and respective changes in the MR appearance. Freeze fracturing of cartilage reveals a curved fracture plane. Expected changes in signal intensity predicted by the magic angle effect correlated with observed changes in signal intensity across the thickness of the sample. Changes in individual collagen fibril orientation did not correspond to MR layering. The three-dimensional organization of collagen in cartilage has a strong influence on the MR appearance of cartilage. This influence is caused by the restriction of water mobility and the resulting magic angle effect caused by curvature of the collagen network, possibly because of the influence on proteoglycan orientation.

  19. Scanning Electron Microscopy Evaluation of an EX-PRESS Mini Glaucoma Shunt After Explantation.

    PubMed

    Tognetto, Daniele; Cecchini, Paolo; D'Aloisio, Rossella; Vattovani, Odilla; Turco, Gianluca

    2017-01-01

    We report a case of an explanted stainless steel miniature glaucoma drainage device (EX-PRESS) implanted under a scleral flap for pseudoexfoliation open-angle glaucoma surgical treatment. The glaucoma shunt was implanted in a 75-year-old white man with medically refractive glaucoma. Cataract surgery was performed simultaneously. After 2 years, the shunt extruded through the scleral flap and the conjunctiva and it was, therefore, explanted. Scanning electron microscopy images of the EX-PRESS mini glaucoma shunt were acquired to verify the patency of the device lumen and the presence of fibrosis or cellular adhesion on the device. Energy dispersive spectroscopy for chemical surface characterization of the EX-PRESS shunt was performed. Scanning electron microscopy-acquired images showed minimal extracellular material proliferation on the lumen device. The energy dispersive spectroscopy analysis revealed a high peak of carbon suggesting the organic nature of the residuals found on the shunt lumen. The surface showed few superficial pits, likely due to an initial corrosion process.

  20. Plasmonic Field Enhancement of Individual Nanoparticles by Correlated Scanning and Photoemission Electron Microscopy

    SciTech Connect

    Peppernick, Samuel J.; Joly, Alan G.; Beck, Kenneth M.; Hess, Wayne P.

    2011-01-21

    We present results of a combined two-photon photoemission and scanning electron microscopy investigation to determine the electromagnetic enhancement factors of silver-coated spherical nanoparticles deposited on an atomically flat mica substrate. Femtosecond laser excitation, of the nanoparticles, produces intense photoemission, attributed to near-resonant excitation of localized surface plasmons. Enhancement factors are determined by comparing the respective two-photon photoemission yield measured for equal areas between single nanoparticles to that of the surrounding flat surface. For s-polarized, 400 nm (~ 3.1 eV) femtosecond radiation a distribution of enhancement factors are found with a large percentage (77%) of the nanoparticles falling within a median range. A correlated scanning electron microscopy analysis demonstrated that the nanoparticles typifying the median of the distribution were characterized by ideal spherical shapes and defect-free morphologies. The single largest enhancement factors were in contrast produced by a very small percentage (8%) of the total, for which evidence of silver defect anomalies were found that contributed to the overall structure of the nanoparticle. Comparisons are made between the experimentally measured enhancement factors and previously reported theoretical predictions of the localized surface plasmon near-field intensities for isolated nanometer-sized silver spheres.

  1. Scanning electron microscopy analysis of hair index on Karachi's population for social and professional appearance enhancement.

    PubMed

    Ali, N; Zohra, R R; Qader, S A U; Mumtaz, M

    2015-06-01

    Hair texture, appearance and pigment play an important role in social and professional communication and maintaining an overall appearance. This study was especially designed for morphological assessment of hair damage caused to Karachi's population due to natural factors and cosmetic treatments using scanning electron microscopy (SEM) technique. Hair samples under the study of synthetic factor's effect were given several cosmetic treatments (hot straightened, bleached, synthetic dyed and henna dyed) whereas samples under natural factor's effect (variation in gender, age and pigmentation) were left untreated. Morphological assessment was performed using SEM technique. Results obtained were statistically analysed using minitab 16 and spss 18 softwares. Scanning electron microscopy images revealed less number of cuticular scales in males than females of same age although size of cuticular scales was found to be larger in males than in females. Mean hair index of white hair was greater than black hair of the same head as it is comparatively newly originated. Tukey's method revealed that among cosmetic treatments, bleaching and synthetic henna caused most of the damage to the hair. Statistical evaluation of results obtained from SEM analysis revealed that human scalp hair index show morphological variation with respect to age, gender, hair pigmentation, chemical and physical treatments. Individuals opting for cosmetic treatments could clearly visualize the extent of hair damage these may cause in long run. © 2015 Society of Cosmetic Scientists and the Société Française de Cosmétologie.

  2. Efficient linear phase contrast in scanning transmission electron microscopy with matched illumination and detector interferometry

    DOE PAGES

    Ophus, Colin; Ciston, Jim; Pierce, Jordan; ...

    2016-02-29

    The ability to image light elements in soft matter at atomic resolution enables unprecedented insight into the structure and properties of molecular heterostructures and beam-sensitive nanomaterials. In this study, we introduce a scanning transmission electron microscopy technique combining a pre-specimen phase plate designed to produce a probe with structured phase with a high-speed direct electron detector to generate nearly linear contrast images with high efficiency. We demonstrate this method by using both experiment and simulation to simultaneously image the atomic-scale structure of weakly scattering amorphous carbon and strongly scattering gold nanoparticles. Our method demonstrates strong contrast for both materials, makingmore » it a promising candidate for structural determination of heterogeneous soft/hard matter samples even at low electron doses comparable to traditional phase-contrast transmission electron microscopy. Ultimately, simulated images demonstrate the extension of this technique to the challenging problem of structural determination of biological material at the surface of inorganic crystals.« less

  3. Synthesis and Cs-Corrected Scanning Transmission Electron Microscopy Characterization of Multimetallic Nanoparticles

    NASA Astrophysics Data System (ADS)

    Khanal, Subarna; Bhattarai, Nabraj; Velázquez-Salazar, Jesus; Jose-Yacaman, Miguel; Subarna Khanal Team

    2014-03-01

    Multimetallic nanoparticles have been attracted greater attention both in materials science and nanotechnology due to its unique electronic, optical, biological, and catalytic properties lead by physiochemical interactions among different atoms and phases. The distinct features of multimetallic nanoparticles enhanced synergetic properties, large surface to volume ratio and quantum size effects ultimately lead to novel and wide range of possibilities for different applications than monometallic counterparts. For instance, PtPd, Pt/Cu, Au-Au3Cu, AgPd/Pt, AuCu/Pt and many other multimetallic nanoparticles have raised interest for their various applications in fuel cells, ethanol and methanol oxidation reactions, hydrogen storage, and so on. The nanostructures were analyzed by transmission electron microscopy (TEM) and by aberration-corrected scanning transmission electron microscopy (Cs-corrected STEM), in combination with high angle annular dark field (HAADF), bright field (BF), energy dispersive X-ray spectroscopy (EDS), and electron energy loss spectroscopy (EELS) detectors. These techniques allowed us to probe the structure at the atomic level of the nanoparticles revealing new structural information and elemental composition of the nanoparticles. The authors would like to acknowledge NSF grants DMR-1103730, ``Alloys at the Nanoscale: The Case of Nanoparticles Second Phase'' and NSF PREM Grant # DMR 0934218.

  4. Three-dimensional morphological characterization of optic nerve fibers by atomic force microscopy and by scanning electron microscopy.

    PubMed

    Melling, Mahmoud; Karimian-Teherani, Daniela; Mostler, Sascha; Hochmeister, Sonja

    2005-08-01

    A comparative study of scanning electron microscopy (SEM) and atomic force microscopy (AFM) imaging of the healthy human optic nerve was carried out to determine the similarities and the differences. In this study we compared the fine optic nerve structures as observed by SEM and AFM. The fibers of the right optic nerve of a 61-year-old man show different arrangements in transverse sections taken from the same individual 5 mm central to the optic canal and 5 mm peripheral to the optic chiasma; this difference can be recognized by light microscopy (LM), SEM, and AFM. AFM revealed such typical optic nerve fibers (taken from a point 5 mm central to the optic canal) with annular and longitudinal orientations, which were not visible by SEM in this form. By contrast, LM and SEM visualized other structures, such as pia mater and optic nerve fibers loosely arranged in bundles, none of which was visualized by AFM. The images, however, taken 5 mm peripheral from the optic chiasma show shapeless nerve fibers having a wavy course. Our results reveal that more detailed information on optic nerve morphology is obtained by exploiting the advantages of both SEM and AFM. These are the first SEM and AFM images of healthy human optic nerve fibers, containing clear representations of the three dimensions of the optic nerve.

  5. The Probe Profile and Lateral Resolution of Scanning Transmission Electron Microscopy of Thick Specimens

    PubMed Central

    Demers, Hendrix; Ramachandra, Ranjan; Drouin, Dominique; de Jonge, Niels

    2012-01-01

    Lateral profiles of the electron probe of scanning transmission electron microscopy (STEM) were simulated at different vertical positions in a micrometers-thick carbon sample. The simulations were carried out using the Monte Carlo method in the CASINO software. A model was developed to fit the probe profiles. The model consisted of the sum of a Gaussian function describing the central peak of the profile, and two exponential decay functions describing the tail of the profile. Calculations were performed to investigate the fraction of unscattered electrons as function of the vertical position of the probe in the sample. Line scans were also simulated over gold nanoparticles at the bottom of a carbon film to calculate the achievable resolution as function of the sample thickness and the number of electrons. The resolution was shown to be noise limited for film thicknesses less than 1 μm. Probe broadening limited the resolution for thicker films. The validity of the simulation method was verified by comparing simulated data with experimental data. The simulation method can be used as quantitative method to predict STEM performance or to interpret STEM images of thick specimens. PMID:22564444

  6. The probe profile and lateral resolution of scanning transmission electron microscopy of thick specimens.

    PubMed

    Demers, Hendrix; Ramachandra, Ranjan; Drouin, Dominique; de Jonge, Niels

    2012-06-01

    Lateral profiles of the electron probe of scanning transmission electron microscopy (STEM) were simulated at different vertical positions in a micrometers-thick carbon sample. The simulations were carried out using the Monte Carlo method in CASINO software. A model was developed to fit the probe profiles. The model consisted of the sum of a Gaussian function describing the central peak of the profile and two exponential decay functions describing the tail of the profile. Calculations were performed to investigate the fraction of unscattered electrons as a function of the vertical position of the probe in the sample. Line scans were also simulated over gold nanoparticles at the bottom of a carbon film to calculate the achievable resolution as a function of the sample thickness and the number of electrons. The resolution was shown to be noise limited for film thicknesses less than 1 μm. Probe broadening limited the resolution for thicker films. The validity of the simulation method was verified by comparing simulated data with experimental data. The simulation method can be used as quantitative method to predict STEM performance or to interpret STEM images of thick specimens.

  7. Facile synthesis and electron transport properties of NiO nanostructures investigated by scanning tunneling microscopy

    NASA Astrophysics Data System (ADS)

    Mallick, Govind; Labh, Jyotsna; Giri, Lily; Pandey, Avinash C.; Karna, Shashi P.

    2017-08-01

    Due to their unique chemical, thermal, electronic and photonic properties, low -dimensional transition metal oxides, especially NiO, have attracted great deal of attention for potential applications in a wide range of technologies, such as, sensors, electrochromic coatings and self-healing materials. However, their synthesis involves multi-step complex procedures that in addition to being expensive, further introduce impurities. Here we present a low cost facile approach to synthesize uniform size NiO nanoparticles (NPs) from hydrothermally grown Ni(OH)2. Detailed transmission electron microscopic analysis reveal the average size of NiO NPs to be around 29 nm. The dimension of NiO NP is also corroborated by the small area scanning tunneling microscope (STM) measurements. Further, we investigate electron transport characteristics of newly synthesized Ni(OH)2 and NiO nanoparticles on p-type Si substrate using scanning tunneling microscopy. The conductivity of Ni(OH)2 and NiO are determined to be 1.46x10-3 S/cm and 2.37x10-5 S/cm, respectively. The NiO NPs exhibit a lower voltage window (˜0.7 V) electron tunneling than the parent Ni(OH)2.

  8. Novel scanning electron microscopy methods for analyzing the 3D structure of the Golgi apparatus.

    PubMed

    Koga, Daisuke; Ushiki, Tatsuo; Watanabe, Tsuyoshi

    2017-01-01

    The structure of the Golgi apparatus has been extensively examined by light and electron microscopy, but details of its three-dimensional (3D) structure have remained unclear because of the technical limitations of conventional microscopy techniques. To overcome this problem, we have developed several novel scanning electron microscopy (SEM) methods for observing the 3D structure of subcellular organelles including the Golgi apparatus: (1) an osmium maceration method that facilitates SEM observation of membranous organelles, including the Golgi apparatus, by selectively removing soluble cytoplasmic proteins, (2) an osmium impregnation/maceration method that combines an osmium impregnation method with the osmium maceration method to determine the polarity of the Golgi apparatus by SEM, (3) a correlative light and SEM method that combines a cryosectioning technique with the osmium maceration method to enable correlation of the immunocytochemical distribution of molecules with the 3D ultrastructure of the Golgi apparatus, and (4) array tomography based on the systematic collection and integration of SEM images of serial ultrathin sections on glass slides for revealing the 3D ultrastructure of the entire Golgi apparatus. Together, the novel SEM techniques listed above can reveal the complete 3D structure of the Golgi apparatus in different cell types.

  9. A Scanning Transmission Electron Microscopy (STEM) Approach to Analyzing Large Volumes of Tissue to Detect Nanoparticles

    PubMed Central

    Kempen, Paul J.; Thakor, Avnesh S.; Zavaleta, Cristina; Gambhir, Sanjiv S.; Sinclair, Robert

    2013-01-01

    The use of nanoparticles for the diagnosis and treatment of cancer requires the complete characterization of their toxicity, including accurately locating them within biological tissues. Owing to their size, traditional light microscopy techniques are unable to resolve them. Transmission electron microscopy provides the necessary spatial resolution to image individual nanoparticles in tissue but is severely limited by the very small analysis volume, usually on the order of tens of cubic microns. In this work we developed a scanning transmission electron microscopy (STEM) approach to analyze large volumes of tissue for the presence of polyethylene glycol coated Raman-active-silica-gold-nanoparticles (PEG-R-Si-Au-NPs). This approach utilizes the simultaneous bright and dark field imaging capabilities of STEM along with careful control of the image contrast settings to readily identify PEG-R-Si-Au-NPs in mouse liver tissue without the need for additional time consuming analytical characterization. We utilized this technique to analyze 243,000 µm3 of mouse liver tissue for the presence of PEG-R-Si-Au-NPs. Nanoparticles injected into the mice intravenously via the tail-vein accumulated in the liver while those injected intrarectally did not, indicating that they remain in the colon and do not pass through the colon wall into the systemic circulation. PMID:23803218

  10. A useful method for observing intracellular structures of free and cultured cells by scanning electron microscopy.

    PubMed

    Koga, Daisuke; Nakajima, Masato; Ushiki, Tatsuo

    2012-04-01

    Scanning electron microscopy (SEM) using osmium-maceration methods has been used for analyzing the three-dimensional structure of cell organelles in tissue samples, but it has been quite difficult to observe free and cultured cells with this technique. The present study was performed to develop a method that can be applied to free and cultured cells for SEM studies of intracellular structures after osmium maceration. The method was also applied to light microscopy (LM) and to transmission electron microscopy (TEM). HeLa cells and human leukocytes were fixed with a mixture of 0.5% paraformaldehyde and 0.5% glutaraldehyde followed by an additional fixation with 1% osmium tetroxide. These cells were embedded in low-melting-point agarose. A temperature-responsive dish was also used for collection of cultured cells before embedding. For LM and TEM, the cell-embedded agarose was further embedded in epoxy resin, and semi- and ultrathin sections were examined conventionally. For SEM, the agarose was freeze-fractured in 50% dimethyl sulfoxide, processed for osmium maceration and observed in a high-resolution SEM. Low-melting-point agarose was useful as an embedding medium for SEM, because it was well preserved during prolonged osmication for SEM. Thus, the fine structure of cell organelles was clearly analyzed by SEM after osmium-maceration treatment. These SEM images could also be compared with those of LM and TEM of the agarose-embedded tissues.

  11. A scanning transmission electron microscopy approach to analyzing large volumes of tissue to detect nanoparticles.

    PubMed

    Kempen, Paul J; Thakor, Avnesh S; Zavaleta, Cristina; Gambhir, Sanjiv S; Sinclair, Robert

    2013-10-01

    The use of nanoparticles for the diagnosis and treatment of cancer requires the complete characterization of their toxicity, including accurately locating them within biological tissues. Owing to their size, traditional light microscopy techniques are unable to resolve them. Transmission electron microscopy provides the necessary spatial resolution to image individual nanoparticles in tissue, but is severely limited by the very small analysis volume, usually on the order of tens of cubic microns. In this work, we developed a scanning transmission electron microscopy (STEM) approach to analyze large volumes of tissue for the presence of polyethylene glycol-coated Raman-active-silica-gold-nanoparticles (PEG-R-Si-Au-NPs). This approach utilizes the simultaneous bright and dark field imaging capabilities of STEM along with careful control of the image contrast settings to readily identify PEG-R-Si-Au-NPs in mouse liver tissue without the need for additional time-consuming analytical characterization. We utilized this technique to analyze 243,000 mm³ of mouse liver tissue for the presence of PEG-R-Si-Au-NPs. Nanoparticles injected into the mice intravenously via the tail vein accumulated in the liver, whereas those injected intrarectally did not, indicating that they remain in the colon and do not pass through the colon wall into the systemic circulation.

  12. Immunogold scanning electron microscopy can reveal the polysaccharide architecture of xylem cell walls

    PubMed Central

    Sun, Yuliang; Juzenas, Kevin

    2017-01-01

    Abstract Immunofluorescence microscopy (IFM) and immunogold transmission electron microscopy (TEM) are the two main techniques commonly used to detect polysaccharides in plant cell walls. Both are important in localizing cell wall polysaccharides, but both have major limitations, such as low resolution in IFM and restricted sample size for immunogold TEM. In this study, we have developed a robust technique that combines immunocytochemistry with scanning electron microscopy (SEM) to study cell wall polysaccharide architecture in xylem cells at high resolution over large areas of sample. Using multiple cell wall monoclonal antibodies (mAbs), this immunogold SEM technique reliably localized groups of hemicellulosic and pectic polysaccharides in the cell walls of five different xylem structures (vessel elements, fibers, axial and ray parenchyma cells, and tyloses). This demonstrates its important advantages over the other two methods for studying cell wall polysaccharide composition and distribution in these structures. In addition, it can show the three-dimensional distribution of a polysaccharide group in the vessel lateral wall and the polysaccharide components in the cell wall of developing tyloses. This technique, therefore, should be valuable for understanding the cell wall polysaccharide composition, architecture and functions of diverse cell types. PMID:28398585

  13. Photothermal imaging scanning microscopy

    DOEpatents

    Chinn, Diane; Stolz, Christopher J.; Wu, Zhouling; Huber, Robert; Weinzapfel, Carolyn

    2006-07-11

    Photothermal Imaging Scanning Microscopy produces a rapid, thermal-based, non-destructive characterization apparatus. Also, a photothermal characterization method of surface and subsurface features includes micron and nanoscale spatial resolution of meter-sized optical materials.

  14. Studies in scanning probe microscopy

    NASA Astrophysics Data System (ADS)

    Sarid, Dror

    1995-06-01

    The following is a final report on our work in the field of Scanning Probe Microscopy (SPM), which has been funded by the AFOSR under Contract #F49620-92-J-0164. The AFOSR funding was instrumental in the establishment of a multi-lab facility at the Optical Sciences Center, which performs research in SPM using two ultrahigh vacuum (UHV) STM facilities, and several Atomic Force Microscopy (AFM) facilities. The fabrication and characterization work performed in the SPM Laboratory is supplemented by infrared (IR) spectroscopy, high resolution transmission electron microscopy (HRTEM), and scanning electron microscopy (SEM), available in other departments on campus. The report covers the following areas: (1) GaAs and CdSe Structures, (2) Optical Interactions on a nm and nsec Scales, (3) Fullerenes on Gold, (4) Fullerenes on MoS2, (5) Fullerenes on Si, (6) SiC, (7) Nanotubes, (8) Scanning Force Microscopy, and (9) Biology.

  15. Unscrambling Mixed Elements using High Angle Annular Dark Field Scanning Transmission Electron Microscopy

    NASA Astrophysics Data System (ADS)

    van den Bos, Karel H. W.; De Backer, Annick; Martinez, Gerardo T.; Winckelmans, Naomi; Bals, Sara; Nellist, Peter D.; Van Aert, Sandra

    2016-06-01

    The development of new nanocrystals with outstanding physicochemical properties requires a full three-dimensional (3D) characterization at the atomic scale. For homogeneous nanocrystals, counting the number of atoms in each atomic column from high angle annular dark field scanning transmission electron microscopy images has been shown to be a successful technique to get access to this 3D information. However, technologically important nanostructures often consist of more than one chemical element. In order to extend atom counting to heterogeneous materials, a new atomic lensing model is presented. This model takes dynamical electron diffraction into account and opens up new possibilities for unraveling the 3D composition at the atomic scale. Here, the method is applied to determine the 3D structure of Au@Ag core-shell nanorods, but it is applicable to a wide range of heterogeneous complex nanostructures.

  16. Scanning-electron-microscopy observations and mechanical characteristics of ion-beam-sputtered surgical implant alloys

    NASA Technical Reports Server (NTRS)

    Weigand, A. J.; Meyer, M. L.; Ling, J. S.

    1977-01-01

    An electron bombardment ion thruster was used as an ion source to sputter the surfaces of orthopedic prosthetic metals. Scanning electron microscopy photomicrographs were made of each ion beam textured surface. The effect of ion texturing an implant surface on its bond to bone cement was investigated. A Co-Cr-W alloy and surgical stainless steel were used as representative hard tissue implant materials to determine effects of ion texturing on bulk mechanical properties. Work was done to determine the effect of substrate temperature on the development of an ion textured surface microstructure. Results indicate that the ultimate strength of the bulk materials is unchanged by ion texturing and that the microstructure will develop more rapidly if the substrate is heated prior to ion texturing.

  17. Atomic bonding effects in annular dark field scanning transmission electron microscopy. I. Computational predictions

    SciTech Connect

    Odlyzko, Michael L.; Mkhoyan, K. Andre; Himmetoglu, Burak; Cococcioni, Matteo

    2016-07-15

    Annular dark field scanning transmission electron microscopy (ADF-STEM) image simulations were performed for zone-axis-oriented light-element single crystals, using a multislice method adapted to include charge redistribution due to chemical bonding. Examination of these image simulations alongside calculations of the propagation of the focused electron probe reveal that the evolution of the probe intensity with thickness exhibits significant sensitivity to interatomic charge transfer, accounting for observed thickness-dependent bonding sensitivity of contrast in all ADF-STEM imaging conditions. Because changes in image contrast relative to conventional neutral atom simulations scale directly with the net interatomic charge transfer, the strongest effects are seen in crystals with highly polar bonding, while no effects are seen for nonpolar bonding. Although the bonding dependence of ADF-STEM image contrast varies with detector geometry, imaging parameters, and material temperature, these simulations predict the bonding effects to be experimentally measureable.

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

    NASA Astrophysics Data System (ADS)

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

    2014-02-01

    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.

  19. Local crystal structure analysis with 10-pm accuracy using scanning transmission electron microscopy.

    PubMed

    Saito, Mitsuhiro; Kimoto, Koji; Nagai, Takuro; Fukushima, Shun; Akahoshi, Daisuke; Kuwahara, Hideki; Matsui, Yoshio; Ishizuka, Kazuo

    2009-06-01

    We demonstrate local crystal structure analysis based on annular dark-field (ADF) imaging in scanning transmission electron microscopy (STEM). Using a stabilized STEM instrument and customized software, we first realize high accuracy of elemental discrimination and atom-position determination with a 10-pm-order accuracy, which can reveal major cation displacements associated with a variety of material properties, e.g. ferroelectricity and colossal magnetoresistivity. A-site ordered/disordered perovskite manganites Tb(0.5)Ba(0.5)MnO(3) are analysed; A-site ordering and a Mn-site displacement of 12 pm are detected in each specific atomic column. This method can be applied to practical and advanced materials, e.g. strongly correlated electron materials.

  20. 4D scanning transmission ultrafast electron microscopy: Single-particle imaging and spectroscopy.

    PubMed

    Ortalan, Volkan; Zewail, Ahmed H

    2011-07-20

    We report the development of 4D scanning transmission ultrafast electron microscopy (ST-UEM). The method was demonstrated in the imaging of silver nanowires and gold nanoparticles. For the wire, the mechanical motion and shape morphological dynamics were imaged, and from the images we obtained the resonance frequency and the dephasing time of the motion. Moreover, we demonstrate here the simultaneous acquisition of dark-field images and electron energy loss spectra from a single gold nanoparticle, which is not possible with conventional methods. The local probing capabilities of ST-UEM open new avenues for probing dynamic processes, from single isolated to embedded nanostructures, without being affected by the heterogeneous processes of ensemble-averaged dynamics. Such methodology promises to have wide-ranging applications in materials science and in single-particle biological imaging.

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

    SciTech Connect

    Forslind, B.

    1984-01-01

    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.

  2. Scanning tunneling microscopy characterization of the geometric and electronic structure of hydrogen-terminated silicon surfaces

    NASA Technical Reports Server (NTRS)

    Kaiser, W. J.; Bell, L. D.; Hecht, M. H.; Grunthaner, F. J.

    1988-01-01

    Scanning tunneling microscopy (STM) methods are used to characterize hydrogen-terminated Si surfaces prepared by a novel method. The surface preparation method is used to expose the Si-SiO2 interface. STM images directly reveal the topographic structure of the Si-SiO2 interface. The dependence of interface topography on oxide preparation conditions observed by STM is compared to the results of conventional surface characterization methods. Also, the electronic structure of the hydrogen-terminated surface is studied by STM spectroscopy. The near-ideal electronic structure of this surface enables direct tunnel spectroscopy measurements of Schottky barrier phenomena. In addition, this method enables probing of semiconductor subsurface properties by STM.

  3. Identification of light elements in silicon nitride by aberration-corrected scanning transmission electron microscopy.

    PubMed

    Idrobo, Juan C; Walkosz, Weronika; Klie, Robert F; Oğüt, Serdar

    2012-12-01

    In silicon nitride structural ceramics, the overall mechanical and thermal properties are controlled by the atomic and electronic structures at the interface between the ceramic grains and the amorphous intergranular films (IGFs) formed by various sintering additives. In the last ten years the atomic arrangements of heavy elements (rare-earths) at the Si(3)N(4)/IGF interfaces have been resolved. However, the atomic position of light elements, without which it is not possible to obtain a complete description of the interfaces, has been lacking. This review article details the authors' efforts to identify the atomic arrangement of light elements such as nitrogen and oxygen at the Si(3)N(4)/SiO(2) interface and in bulk Si(3)N(4) using aberration-corrected scanning transmission electron microscopy. Published by Elsevier B.V.

  4. Unscrambling Mixed Elements using High Angle Annular Dark Field Scanning Transmission Electron Microscopy.

    PubMed

    van den Bos, Karel H W; De Backer, Annick; Martinez, Gerardo T; Winckelmans, Naomi; Bals, Sara; Nellist, Peter D; Van Aert, Sandra

    2016-06-17

    The development of new nanocrystals with outstanding physicochemical properties requires a full three-dimensional (3D) characterization at the atomic scale. For homogeneous nanocrystals, counting the number of atoms in each atomic column from high angle annular dark field scanning transmission electron microscopy images has been shown to be a successful technique to get access to this 3D information. However, technologically important nanostructures often consist of more than one chemical element. In order to extend atom counting to heterogeneous materials, a new atomic lensing model is presented. This model takes dynamical electron diffraction into account and opens up new possibilities for unraveling the 3D composition at the atomic scale. Here, the method is applied to determine the 3D structure of Au@Ag core-shell nanorods, but it is applicable to a wide range of heterogeneous complex nanostructures.

  5. Analysis of feline urinary calculi and urethral plugs by infrared spectroscopy and scanning electron microscopy.

    PubMed

    Escolar, E; Bellanato, J

    2003-05-17

    The chemical constituents of 34 feline urinary calculi and five urethral plugs were analysed by infrared spectroscopy. The analysis revealed that 18 (52.9 per cent) of the calculi contained magnesium ammonium phosphate hexahydrate (struvite) as the major component; 10 (29.4 per cent) contained complex ammonium urates (three of them also containing calcium phosphate, mainly on the surface); three were composed of calcium phosphates and three were composed mainly of calcium oxalate mono and dihydrates. The urethral plugs were composed primarily of struvite, but also contained large amounts of organic matter. The examination of 16 selected samples by scanning electron microscopy and electron dispersive x-ray analysis revealed that their crystalline structures were similar to those of canine stones.

  6. Silver methenamine staining for scanning electron microscopy of bone sections containing biomaterials.

    PubMed

    Frayssinet, P; Hanker, J S; Rouquet, N; Primout, I; Giammara, B

    1999-01-01

    Sections of tissue containing orthopedic materials are currently used to study the compatibility of those materials and to perform electron probe microanalysis at the material-tissue interface. Identification of the cells in contact with the material by Scanning electron microscopy (SEM) is of interest. We have developed a method for staining cells and tissue structures embedded in polymethyl methacrylate with silver methenamine once the sections have been obtained. Sections were prepared by grinding, and the silver methenamine was applied after oxidation with periodic acid. The procedure was carried out in a microwave oven. Backscatter SEM showed staining of the cell nucleus membrane, chromatin, the nuclear organizers, and the chromosomes of dividing cells. The cytoplasm and the cytoplasmic membrane were also stained. Collagen fibers of the extracellular matrix and the mineralized matrix of bone were labeled. Material particles in the macrophages were easily recognizable and Energy-Dispersive Spectrometer were not impaired by the presence of silver in the preparation.

  7. Three-dimensional bright-field scanning transmission electron microscopy elucidate novel nanostructure in microbial biofilms.

    PubMed

    Hickey, William J; Shetty, Ameesha R; Massey, Randall J; Toso, Daniel B; Austin, Jotham

    2017-01-01

    Bacterial biofilms play key roles in environmental and biomedical processes, and understanding their activities requires comprehension of their nanoarchitectural characteristics. Electron microscopy (EM) is an essential tool for nanostructural analysis, but conventional EM methods are limited in that they either provide topographical information alone, or are suitable for imaging only relatively thin (<300 nm) sample volumes. For biofilm investigations, these are significant restrictions. Understanding structural relations between cells requires imaging of a sample volume sufficiently large to encompass multiple cells and the capture of both external and internal details of cell structure. An emerging EM technique with such capabilities is bright-field scanning transmission electron microscopy (BF-STEM) and in the present report BF-STEM was coupled with tomography to elucidate nanostructure in biofilms formed by the polycyclic aromatic hydrocarbon-degrading soil bacterium, Delftia acidovorans Cs1-4. Dual-axis BF-STEM enabled high-resolution 3-D tomographic recontructions (6-10 nm) visualization of thick (1250 and 1500 nm) sections. The 3-D data revealed that novel extracellular structures, termed nanopods, were polymorphic and formed complex networks within cell clusters. BF-STEM tomography enabled visualization of conduits formed by nanopods that could enable intercellular movement of outer membrane vesicles, and thereby enable direct communication between cells. This report is the first to document application of dual-axis BF-STEM tomography to obtain high-resolution 3-D images of novel nanostructures in bacterial biofilms. Future work with dual-axis BF-STEM tomography combined with correlative light electron microscopy may provide deeper insights into physiological functions associated with nanopods as well as other nanostructures. © 2016 The Authors Journal of Microscopy © 2016 Royal Microscopical Society.

  8. Visualising impregnated chitosan in Pinus radiata early wood cells using light and scanning electron microscopy.

    PubMed

    Singh, Adya P; Singh, Tripti; Rickard, Catherine L

    2010-04-01

    Chitosan, a deacetylated product of an abundant naturally occurring biopolymer chitin, has been used in a range of applications, particularly in food and health areas, as an antimicrobial agent. In the work reported here Pinus radiata wood was impregnated with chitosan as an environmentally compatible organic biocide (Eikenes et al., 2005a,b) to protect wood against wood deteriorating microorganisms and to thus prolong the service life of wooden products. We developed sample preparation techniques targeted to visualise impregnated chitosan within wood tissues using light microscope and field-emission scanning electron microscope (FE-SEM). Sections were viewed with the light microscope without staining with a dye as well as after staining with the dye toluidine blue. Light microscopy was also undertaken on sections that had been stained with 1% aqueous osmium tetroxide (OsO(4)). For SEM observations, the sections were treated with OsO(4) and then examined with the FE-SEM, first in the secondary electron imaging mode (SEI) and then in the backscattered electron imaging (BEI) mode, imaging the same areas of a section in both SEI and BEI modes. The preparation techniques employed and the combined use of light and scanning electron microscopy provided valuable complementary information, revealing that chitosan had penetrated into the cavities (cell lumens, intercellular spaces) of all sizes present within wood tissues and had also impregnated early wood cell walls. The information obtained is discussed in relation to its importance in further development of chitosan formulations and refinement of impregnation technologies to optimise chitosan impregnation into and distribution within wood tissues as well as in assessing chitosan efficacy.

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

    PubMed

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

    2011-05-01

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

  10. Two-stage gas amplifier for ultrahigh resolution low vacuum scanning electron microscopy

    SciTech Connect

    Thiel, B.L.; Toth, M.; Schroemges, R.P.M.; Scholtz, J.J.; Veen, G. van; Knowles, W.R.

    2006-03-15

    We describe a magnetic field assisted, two-stage secondary electron gas amplification process for low vacuum scanning electron microscopy. The field of an ultrahigh resolution magnetic immersion objective lens and the electric field of an annular electrode configuration partition the amplification volume into two regions in which the electric and magnetic fields are parallel and crossed, respectively. The fields confine secondary electrons to axial and radial oscillations within the detector volume, until all of the kinetic energy imparted by an anode is dissipated through inelastic collisions with gas molecules. The electron confinement yields high gas amplification efficiency at short working distances and low gas pressures, facilitating high resolution imaging at low electron beam energies. Charging of insulating specimens is stabilized by positive ions produced in the gas ionization cascade. Furthermore, the signal to background level and bandwidth of this detector are superior to those of earlier generations of environmental secondary electron detectors. The combination of low vacuum, short working distance, and low beam energy is attractive to the semiconductor metrology industry, in particular, for critical dimension measurements on photolithographic masks.

  11. Two-stage gas amplifier for ultrahigh resolution low vacuum scanning electron microscopy

    NASA Astrophysics Data System (ADS)

    Thiel, B. L.; Toth, M.; Schroemges, R. P. M.; Scholtz, J. J.; van Veen, G.; Knowles, W. R.

    2006-03-01

    We describe a magnetic field assisted, two-stage secondary electron gas amplification process for low vacuum scanning electron microscopy. The field of an ultrahigh resolution magnetic immersion objective lens and the electric field of an annular electrode configuration partition the amplification volume into two regions in which the electric and magnetic fields are parallel and crossed, respectively. The fields confine secondary electrons to axial and radial oscillations within the detector volume, until all of the kinetic energy imparted by an anode is dissipated through inelastic collisions with gas molecules. The electron confinement yields high gas amplification efficiency at short working distances and low gas pressures, facilitating high resolution imaging at low electron beam energies. Charging of insulating specimens is stabilized by positive ions produced in the gas ionization cascade. Furthermore, the signal to background level and bandwidth of this detector are superior to those of earlier generations of environmental secondary electron detectors. The combination of low vacuum, short working distance, and low beam energy is attractive to the semiconductor metrology industry, in particular, for critical dimension measurements on photolithographic masks.

  12. Practical spatial resolution of electron energy loss spectroscopy in aberration corrected scanning transmission electron microscopy.

    PubMed

    Shah, A B; Ramasse, Q M; Wen, J G; Bhattacharya, A; Zuo, J M

    2011-08-01

    The resolution of electron energy loss spectroscopy (EELS) is limited by delocalization of inelastic electron scattering rather than probe size in an aberration corrected scanning transmission electron microscope (STEM). In this study, we present an experimental quantification of EELS spatial resolution using chemically modulated 2×(LaMnO(3))/2×(SrTiO(3)) and 2×(SrVO(3))/2×(SrTiO(3)) superlattices by measuring the full width at half maxima (FWHM) of integrated Ti M(2,3), Ti L(2,3), V L(2,3), Mn L(2,3), La N(4,5), La N(2,3) La M(4,5) and Sr L(3) edges over the superlattices. The EELS signals recorded using large collection angles are peaked at atomic columns. The FWHM of the EELS profile, obtained by curve-fitting, reveals a systematic trend with the energy loss for the Ti, V, and Mn edges. However, the experimental FWHM of the Sr and La edges deviates significantly from the observed experimental tendency. Copyright © 2011 Elsevier Ltd. All rights reserved.

  13. Nanoscale imaging of whole cells using a liquid enclosure and a scanning transmission electron microscopy

    SciTech Connect

    De Jonge, Niels; Peckys, Diana B; Veith, Gabriel M; Joy, David Charles

    2009-01-01

    Nanoscale imaging techniques are needed to investigate cellular function at the level of individual proteins and to study the interaction of nanomaterials with biological systems. We imaged whole fixed cells in liquid state with a scanning transmission electron microscope (STEM) using a micrometer-sized liquid enclosure with electron transparent windows providing a wet specimen environment. Wet-STEM images were obtained of fixed E. coli bacteria labeled with gold nanoparticles attached to surface membrane proteins. Mammalian cells (COS7) were incubated with gold-tagged epidermal growth factor and fixed. STEM imaging of these cells resulted in a resolution of 3 nm for the gold nanoparticles. The wet-STEM method has several advantages over conventional imaging techniques. Most important is the capability to image whole fixed cells in a wet environment with nanometer resolution, which can be used, e.g., to map individual protein distributions in/on whole cells. The sample preparation is compatible with that used for fluorescent microscopy on fixed cells for experiments involving nanoparticles. Thirdly, the system is rather simple and involves only minimal new equipment in an electron microscopy (EM) laboratory.

  14. Studying Dynamic Processes of Nano-sized Objects in Liquid using Scanning Transmission Electron Microscopy

    PubMed Central

    Hermannsdörfer, Justus; de Jonge, Niels

    2017-01-01

    Samples fully embedded in liquid can be studied at a nanoscale spatial resolution with Scanning Transmission Electron Microscopy (STEM) using a microfluidic chamber assembled in the specimen holder for Transmission Electron Microscopy (TEM) and STEM. The microfluidic system consists of two silicon microchips supporting thin Silicon Nitride (SiN) membrane windows. This article describes the basic steps of sample loading and data acquisition. Most important of all is to ensure that the liquid compartment is correctly assembled, thus providing a thin liquid layer and a vacuum seal. This protocol also includes a number of tests necessary to perform during sample loading in order to ensure correct assembly. Once the sample is loaded in the electron microscope, the liquid thickness needs to be measured. Incorrect assembly may result in a too-thick liquid, while a too-thin liquid may indicate the absence of liquid, such as when a bubble is formed. Finally, the protocol explains how images are taken and how dynamic processes can be studied. A sample containing AuNPs is imaged both in pure water and in saline. PMID:28190028

  15. Studying Dynamic Processes of Nano-sized Objects in Liquid using Scanning Transmission Electron Microscopy.

    PubMed

    Hermannsdörfer, Justus; de Jonge, Niels

    2017-02-05

    Samples fully embedded in liquid can be studied at a nanoscale spatial resolution with Scanning Transmission Electron Microscopy (STEM) using a microfluidic chamber assembled in the specimen holder for Transmission Electron Microscopy (TEM) and STEM. The microfluidic system consists of two silicon microchips supporting thin Silicon Nitride (SiN) membrane windows. This article describes the basic steps of sample loading and data acquisition. Most important of all is to ensure that the liquid compartment is correctly assembled, thus providing a thin liquid layer and a vacuum seal. This protocol also includes a number of tests necessary to perform during sample loading in order to ensure correct assembly. Once the sample is loaded in the electron microscope, the liquid thickness needs to be measured. Incorrect assembly may result in a too-thick liquid, while a too-thin liquid may indicate the absence of liquid, such as when a bubble is formed. Finally, the protocol explains how images are taken and how dynamic processes can be studied. A sample containing AuNPs is imaged both in pure water and in saline.

  16. Correlative scanning electron and confocal microscopy imaging of labeled cells coated by indium-tin-oxide.

    PubMed

    Rodighiero, Simona; Torre, Bruno; Sogne, Elisa; Ruffilli, Roberta; Cagnoli, Cinzia; Francolini, Maura; Di Fabrizio, Enzo; Falqui, Andrea

    2015-06-01

    Confocal microscopy imaging of cells allows to visualize the presence of specific antigens by using fluorescent tags or fluorescent proteins, with resolution of few hundreds of nanometers, providing their localization in a large field-of-view and the understanding of their cellular function. Conversely, in scanning electron microscopy (SEM), the surface morphology of cells is imaged down to nanometer scale using secondary electrons. Combining both imaging techniques have brought to the correlative light and electron microscopy, contributing to investigate the existing relationships between biological surface structures and functions. Furthermore, in SEM, backscattered electrons (BSE) can image local compositional differences, like those due to nanosized gold particles labeling cellular surface antigens. To perform SEM imaging of cells, they could be grown on conducting substrates, but obtaining images of limited quality. Alternatively, they could be rendered electrically conductive, coating them with a thin metal layer. However, when BSE are collected to detect gold-labeled surface antigens, heavy metals cannot be used as coating material, as they would mask the BSE signal produced by the markers. Cell surface could be then coated with a thin layer of chromium, but this results in a loss of conductivity due to the fast chromium oxidation, if the samples come in contact with air. In order to overcome these major limitations, a thin layer of indium-tin-oxide was deposited by ion-sputtering on gold-decorated HeLa cells and neurons. Indium-tin-oxide was able to provide stable electrical conductivity and preservation of the BSE signal coming from the gold-conjugated markers.

  17. Combined scanning transmission X-ray and electron microscopy for the characterization of bacterial endospores.

    PubMed

    Jamroskovic, Jan; Shao, Paul P; Suvorova, Elena; Barak, Imrich; Bernier-Latmani, Rizlan

    2014-09-01

    Endospores (also referred to as bacterial spores) are bacterial structures formed by several bacterial species of the phylum Firmicutes. Spores form as a response to environmental stress. These structures exhibit remarkable resistance to harsh environmental conditions such as exposure to heat, desiccation, and chemical oxidants. The spores include several layers of protein and peptidoglycan that surround a core harboring DNA as well as high concentrations of calcium and dipicolinic acid (DPA). A combination of scanning transmission X-ray microscopy, scanning transmission electron microscopy, and energy dispersive spectroscopy was used for the direct quantitative characterization of bacterial spores. The concentration and localization of DPA, Ca(2+) , and other elements were determined and compared for the core and cortex of spores from two distinct genera: Bacillus subtilis and Desulfotomaculum reducens. This micro-spectroscopic approach is uniquely suited for the direct study of individual bacterial spores, while classical molecular and biochemical methods access only bulk characteristics. © 2014 Federation of European Microbiological Societies. Published by John Wiley & Sons Ltd. All rights reserved.

  18. Correlative scanning-transmission electron microscopy reveals that a chimeric flavivirus is released as individual particles in secretory vesicles.

    PubMed

    Burlaud-Gaillard, Julien; Sellin, Caroline; Georgeault, Sonia; Uzbekov, Rustem; Lebos, Claude; Guillaume, Jean-Marc; Roingeard, Philippe

    2014-01-01

    The intracellular morphogenesis of flaviviruses has been well described, but flavivirus release from the host cell remains poorly documented. We took advantage of the optimized production of an attenuated chimeric yellow fever/dengue virus for vaccine purposes to study this phenomenon by microscopic approaches. Scanning electron microscopy (SEM) showed the release of numerous viral particles at the cell surface through a short-lived process. For transmission electron microscopy (TEM) studies of the intracellular ultrastructure of the small number of cells releasing viral particles at a given time, we developed a new correlative microscopy method: CSEMTEM (for correlative scanning electron microscopy - transmission electron microscopy). CSEMTEM analysis suggested that chimeric flavivirus particles were released as individual particles, in small exocytosis vesicles, via a regulated secretory pathway. Our morphological findings provide new insight into interactions between flaviviruses and cells and demonstrate that CSEMTEM is a useful new method, complementary to SEM observations of biological events by intracellular TEM investigations.

  19. Correlative Scanning-Transmission Electron Microscopy Reveals that a Chimeric Flavivirus Is Released as Individual Particles in Secretory Vesicles

    PubMed Central

    Burlaud-Gaillard, Julien; Sellin, Caroline; Georgeault, Sonia; Uzbekov, Rustem; Lebos, Claude; Guillaume, Jean-Marc; Roingeard, Philippe

    2014-01-01

    The intracellular morphogenesis of flaviviruses has been well described, but flavivirus release from the host cell remains poorly documented. We took advantage of the optimized production of an attenuated chimeric yellow fever/dengue virus for vaccine purposes to study this phenomenon by microscopic approaches. Scanning electron microscopy (SEM) showed the release of numerous viral particles at the cell surface through a short-lived process. For transmission electron microscopy (TEM) studies of the intracellular ultrastructure of the small number of cells releasing viral particles at a given time, we developed a new correlative microscopy method: CSEMTEM (for correlative scanning electron microscopy - transmission electron microscopy). CSEMTEM analysis suggested that chimeric flavivirus particles were released as individual particles, in small exocytosis vesicles, via a regulated secretory pathway. Our morphological findings provide new insight into interactions between flaviviruses and cells and demonstrate that CSEMTEM is a useful new method, complementary to SEM observations of biological events by intracellular TEM investigations. PMID:24681578

  20. Superhydrophobic and adhesive properties of surfaces: testing the quality by an elaborated scanning electron microscopy method.

    PubMed

    Ensikat, Hans J; Mayser, Matthias; Barthlott, Wilhelm

    2012-10-09

    In contrast to advancements in the fabrication of new superhydrophobic materials, the characterization of their water repellency and quality is often coarse and unsatisfactory. In view of the problems and inaccuracies, particularly in the measurement of very high contact angles, we developed alternative methods for the characterization of superhydrophobic surfaces. It was found that adhering water remnants after immersion are a useful criterion in determining the repellency quality. In this study, we introduce microscopy methods to detect traces of water-resembling test liquids on superhydrophobic surfaces by scanning electron microscopy (SEM) or fluorescence light microscopy (FLM). Diverse plant surfaces and some artificial superhydrophobic samples were examined. Instead of pure water, we used aqueous solutions containing a detectable stain and glycerol in order to prevent immediate evaporation of the microdroplets. For the SEM examinations, aqueous solutions of lead acetate were used, which could be detected in a frozen state at -90 °C with high sensitivity using a backscattered electron detector. For fluorescence microscopy, aqueous solutions of auramine were used. On different species of superhydrophobic plants, varying patterns of remaining microdroplets were found on their leaves. On some species, drop remnants occurred only on surface defects such as damaged epicuticular waxes. On others, microdroplets regularly decorated the locations of increased adhesion, particularly on hierarchically structured surfaces. Furthermore, it is demonstrated that the method is suitable for testing the limits of repellency under harsh conditions, such as drop impact or long-enduring contact. The supplementation of the visualization method by the measurement of the pull-off force between a water drop and the sample allowed us to determine the adhesive properties of superhydrophobic surfaces quantitatively. The results were in good agreement with former studies of the water

  1. Analysis of Brain Mitochondria Using Serial Block-Face Scanning Electron Microscopy.

    PubMed

    Mukherjee, Konark; Clark, Helen R; Chavan, Vrushali; Benson, Emily K; Kidd, Grahame J; Srivastava, Sarika

    2016-07-09

    Human brain is a high energy consuming organ that mainly relies on glucose as a fuel source. Glucose is catabolized by brain mitochondria via glycolysis, tri-carboxylic acid (TCA) cycle and oxidative phosphorylation (OXPHOS) pathways to produce cellular energy in the form of adenosine triphosphate (ATP). Impairment of mitochondrial ATP production causes mitochondrial disorders, which present clinically with prominent neurological and myopathic symptoms. Mitochondrial defects are also present in neurodevelopmental disorders (e.g. autism spectrum disorder) and neurodegenerative disorders (e.g. amyotrophic lateral sclerosis, Alzheimer's and Parkinson's diseases). Thus, there is an increased interest in the field for performing 3D analysis of mitochondrial morphology, structure and distribution under both healthy and disease states. The brain mitochondrial morphology is extremely diverse, with some mitochondria especially those in the synaptic region being in the range of <200 nm diameter, which is below the resolution limit of traditional light microscopy. Expressing a mitochondrially-targeted green fluorescent protein (GFP) in the brain significantly enhances the organellar detection by confocal microscopy. However, it does not overcome the constraints on the sensitivity of detection of relatively small sized mitochondria without oversaturating the images of large sized mitochondria. While serial transmission electron microscopy has been successfully used to characterize mitochondria at the neuronal synapse, this technique is extremely time-consuming especially when comparing multiple samples. The serial block-face scanning electron microscopy (SBFSEM) technique involves an automated process of sectioning, imaging blocks of tissue and data acquisition. Here, we provide a protocol to perform SBFSEM of a defined region from rodent brain to rapidly reconstruct and visualize mitochondrial morphology. This technique could also be used to provide accurate information on

  2. A scanning electron microscopy study of the macro-crystalline structure of 2-(2,4-dinitrobenzyl) pyridine

    NASA Technical Reports Server (NTRS)

    Ware, Jacqueline; Hammond, Ernest C., Jr.

    1989-01-01

    The compound, 2-(2,4-dinitrobenzyl) pyridine, was synthesized in the laboratory; an introductory level electron microscopy study of the macro-crystalline structure was conducted using the scanning electron microscope (SEM). The structure of these crystals was compared with the macrostructure of the crystal of 2-(2,4-dinitrobenzyl) pyridinium bromide, the hydrobromic salt of the compound which was also synthesized in the laboratory. A scanning electron microscopy crystal study was combined with a study of the principle of the electron microscope.

  3. Scanning Electron Microscopy Study of Hair Shaft Damage Secondary to Cosmetic Treatments of the Hair

    PubMed Central

    Kaliyadan, Feroze; Gosai, BB; Al Melhim, Walid Naief; Feroze, Kaberi; Qureshi, Habib Ahmad; Ibrahim, Sayed; Kuruvilla, Joel

    2016-01-01

    Introduction and Background: Cosmetic procedures for hair, such as bleaching, dyeing, and straightening, are commonly used around the world. It has been suggested that excessive use of such procedures can cause damage to the hair shaft. We aimed to assess hair shaft changes using scanning electron microscopy (SEM) in female volunteers who frequently use hair treatment procedures such as bleaching, dyeing, or straightening. Methods: A cross-sectional, controlled study in a sample of 25 female volunteers (19 study group and 6 controls) in the age group of 18–45 years. The study group was composed of volunteers who regularly used different cosmetic hair treatment procedures such as bleaching, dyeing, and straightening (any one of these or a combination). The control group had never used any specific hair treatment procedure. The hair shaft damage as seen on SEM was assessed using a standardized scoring system and compared among the two groups statistically. The hair shafts were also examined clinically and with light microscopy. Results: No significant differences were seen between the test and control groups with regard to normal clinical examination and light microscopy findings. A higher degree of hair shaft damage was evident under SEM in the study group as compared to the control group. This difference was statistically significant. Conclusions: Regular use of procedures such as bleaching, dyeing, or straightening can lead to subtle changes in the hair shaft which can be detected early by SEM. PMID:27601867

  4. External cervical resorption: an analysis using cone beam and microfocus computed tomography and scanning electron microscopy.

    PubMed

    Gunst, V; Mavridou, A; Huybrechts, B; Van Gorp, G; Bergmans, L; Lambrechts, P

    2013-09-01

    To provide a three-dimensional representation of external cervical resorption (ECR) with microscopy, stereo microscopy, cone beam computed tomography (CT), microfocus CT and scanning electron microscopy (SEM). External cervical resorption is an aggressive form of root resorption, leading to a loss of dental hard tissues. This is due to clastic action, activated by a damage of the covering cementum and stimulated probably by infection. Clinically, it is a challenging situation as it is characterized by a late symptomatology. This is due to the pericanalar protection from a resorption-resistant sheet, composed of pre-dentine and surrounding dentine. The clastic activity is often associated with an attempt to repair, seen by the formation of osteoid tissue. Cone beam CT is extremely useful in the diagnoses and treatment planning of ECR. SEM analyses provide a better insight into the activity of osteoclasts. The root canal is surrounded by a layer of dentine that is resistant to resorption. © 2013 International Endodontic Journal. Published by John Wiley & Sons Ltd.

  5. Scanning Electron Microscopy Study of Hair Shaft Damage Secondary to Cosmetic Treatments of the Hair.

    PubMed

    Kaliyadan, Feroze; Gosai, B B; Al Melhim, Walid Naief; Feroze, Kaberi; Qureshi, Habib Ahmad; Ibrahim, Sayed; Kuruvilla, Joel

    2016-01-01

    Cosmetic procedures for hair, such as bleaching, dyeing, and straightening, are commonly used around the world. It has been suggested that excessive use of such procedures can cause damage to the hair shaft. We aimed to assess hair shaft changes using scanning electron microscopy (SEM) in female volunteers who frequently use hair treatment procedures such as bleaching, dyeing, or straightening. A cross-sectional, controlled study in a sample of 25 female volunteers (19 study group and 6 controls) in the age group of 18-45 years. The study group was composed of volunteers who regularly used different cosmetic hair treatment procedures such as bleaching, dyeing, and straightening (any one of these or a combination). The control group had never used any specific hair treatment procedure. The hair shaft damage as seen on SEM was assessed using a standardized scoring system and compared among the two groups statistically. The hair shafts were also examined clinically and with light microscopy. No significant differences were seen between the test and control groups with regard to normal clinical examination and light microscopy findings. A higher degree of hair shaft damage was evident under SEM in the study group as compared to the control group. This difference was statistically significant. Regular use of procedures such as bleaching, dyeing, or straightening can lead to subtle changes in the hair shaft which can be detected early by SEM.

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

    NASA Astrophysics Data System (ADS)

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

    2014-03-01

    Nanomaterials including nanoparticles, nanowires and nanotubes play an important role in heterogeneous catalysis. Thanks to the rapid improvement of the electron microscopic techniques and with the advent of aberration corrected electron microscopy as well as theoretical methodologies, the potential effects induced by nanocatalysts are better understood than before by unravelling their atomic structure. A brief introduction to advanced electron microscopic techniques namely aberration corrected scanning transmission electron microscopy (Cs-STEM) is presented and subsequently two examples of nanocatalysts are considered in the present review. The first example will focus on the study of bimetallic/core-shell nanoalloys. In heterogeneous catalysis, catalysts containing two or more metals might show significantly different catalytic properties compared to the parent metals and thus are widely utilized in several catalytic reactions. Atom-by-atom insights of the nanoalloy based catalysts ex: Au-Pd will be described in the present review using a combination of advanced electron microscopic and spectroscopic techniques. A related example on the understanding of bimetallic clusters by HAADF-STEM will also be presented in addition to nanoparticles. In the second case understanding the structure of transition metal chalcogenide based nanocatalysts by HRTEM and aberration corrected STEM, for the case of MoS2 will be discussed. MoS2-based catalysts serve as model catalysts and are employed in the hydrodesulphurisations (HDS) reactions in the removal of sulphur from gasoline and related petrochemical products. They have been studied in various forms including nanowires, nanotubes and nanoplates. Their structure, atomic insights and as a consequence elucidation of their corresponding catalytic activity are thus important.

  7. Rapid imaging of mycoplasma in solution using Atmospheric Scanning Electron Microscopy (ASEM)

    SciTech Connect

    Sato, Chikara; Manaka, Sachie; Nakane, Daisuke; Nishiyama, Hidetoshi; Suga, Mitsuo; Nishizaka, Takayuki; Miyata, Makoto; Maruyama, Yuusuke

    2012-01-27

    Highlights: Black-Right-Pointing-Pointer Mycoplasma mobile was observed in buffer with the Atmospheric Scanning Electron Microscope. Black-Right-Pointing-Pointer Characteristic protein localizations were visualized using immuno-labeling. Black-Right-Pointing-Pointer M. mobile attached to sialic acid on the SiN film surface within minutes. Black-Right-Pointing-Pointer Cells were observed at low concentrations. Black-Right-Pointing-Pointer ASEM should promote study and early-stage diagnosis of mycoplasma. -- Abstract: Mycoplasma is a genus of bacterial pathogen that causes disease in vertebrates. In humans, the species Mycoplasma pneumoniae causes 15% or more of community-acquired pneumonia. Because this bacterium is tiny, corresponding in size to a large virus, diagnosis using optical microscopy is not easy. In current methods, chest X-rays are usually the first action, followed by serology, PCR amplification, and/or culture, but all of these are particularly difficult at an early stage of the disease. Using Mycoplasma mobile as a model species, we directly observed mycoplasma in buffer with the newly developed Atmospheric Scanning Electron Microscope (ASEM). This microscope features an open sample dish with a pressure-resistant thin film window in its base, through which the SEM beam scans samples in solution, from below. Because of its 2-3 {mu}m-deep scanning capability, it can observe the whole internal structure of mycoplasma cells stained with metal solutions. Characteristic protein localizations were visualized using immuno-labeling. Cells were observed at low concentrations, because suspended cells concentrate in the observable zone by attaching to sialic acid on the silicon nitride (SiN) film surface within minutes. These results suggest the applicability of the ASEM for the study of mycoplasmas as well as for early-stage mycoplasma infection diagnosis.

  8. Photon scanning tunneling microscopy

    SciTech Connect

    Reddick, R.C.; Warmack, R.J.; Chilcott, D.W.; Sharp, S.L.; Ferrell, T.L. Department of Physics and Astronomy, University of Tennessee, Knoxville, TN )

    1990-12-01

    An optical tunneling microscope is presented that operates in exactly the same way as the electron scanning tunneling microscope (ESTM). It takes advantage of evanescent fields generated by the total internal reflection (TIR) of light at the interface between materials of different optical densities. The photon scanning tunneling microscope (PSTM) employs an optically conducting probe tip to map spatial variations in the evanescent and scattered field intensity distributions adjacent to a sample surface, which forms or is placed on the TIR surface. These variations are due to the local topography, morphology, and optical activity of the surface and form the basis of imaging. Evanescent field theory is discussed and the evanescent field intensity as a function of surface-probe separation is calculated using several probe tip models. After a description of PSTM construction and operation, evanescent field intensity measurements are shown to agree with the model calculations. PSTM images of various sample surfaces demonstrate subwavelength resolution exceeding that of conventional optical microscopy, especially in the vertical dimension. Limitations and interpretation of PSTM images are discussed as well as the PSTMs applicability to other forms of surface analysis.

  9. Subsurface Examination of a Foliar Biofilm Using Scanning Electron- and Focused-Ion-Beam Microscopy

    SciTech Connect

    Wallace, Patricia K.; Arey, Bruce W.; Mahaffee, Walt F.

    2011-08-01

    The dual beam scanning electron microscope, equipped with both a focused ion- and scanning electron- beam (FIB SEM) is a novel tool for the exploration of the subsurface structure of biological tissues. The FIB can remove a predetermined amount of material from a selected site to allow for subsurface exploration and when coupled with SEM or scanning ion- beam microscopy (SIM) could be suitable to examine the subsurface structure of bacterial biofilms on the leaf surface. The suitability of chemical and cryofixation was examined for use with the FIB SEM to examine bacterial biofilms on leaf surfaces. The biological control agent, Burkholderia pyroccinia FP62, that rapidly colonizes the leaf surface and forms biofilms, was inoculated onto geranium leaves and incubated in a greenhouse for 7 or 14 days. Cryofixation was not suitable for examination of leaf biofilms because it created a frozen layer over the leaf surface that cracked when exposed to the electron beam and the protective cap required for FIB milling could not be accurately deposited. With chemically fixed samples, it was possible to precisely FIB mill a single cross section (5 µm) or sequential cross sections from a single site without any damage to the surrounding surface. Biofilms, 7 days post-inoculation (DPI), were composed of 2 to 5 bacterial cell layers while biofilms 14 DPI ranged from 5 to greater than 30 cell layers. Empty spaces between bacteria cells in the subsurface structure were observed in biofilms 7- and 14-DPI. Sequential cross sections inferred that the empty spaces were often continuous between FP62 cells and could possibly make up a network of channels throughout the biofilm. FIB SEM was a useful tool to observe the subsurface composition of a foliar biofilm.

  10. Scanning and transmission electron microscopy for evaluation of order/disorder in bone structure.

    PubMed

    Suvorova, Elena I; Petrenko, Pavel P; Buffat, Philippe A

    2007-01-01

    A comparative characterization of the structure of normal and abnormal (osteoporotic) human lumbar and thoracic vertebrae samples was carried out to reveal the type of possible disorder. Samples from the bone fragments extracted during the surgery due to vertebra fractures were examined by scanning electron microscopy (SEM), conventional and high resolution transmission electron microscopy (TEM and HRTEM), and X-ray energy dispersive spectroscopy (EDS). Contrary to what might be expected in accordance with possible processes of dissolution, formation and remineralization of hard tissues, no changes in phase composition of mineral part, crystal sizes (length, width, and thickness), and arrangement of crystals on collagen fibers were detected in abnormal bones compared to the normal ones. The following sizes were determined by HRTEM for all bone samples:

  11. Environmental scanning electron microscopy analysis of Proteus mirabilis biofilms grown on chitin and stainless steel.

    PubMed

    Fernández-Delgado, Milagro; Duque, Zoilabet; Rojas, Héctor; Suárez, Paula; Contreras, Monica; García-Amado, María A; Alciaturi, Carlos

    Proteus mirabilis is a human pathogen able to form biofilms on the surface of urinary catheters. Little is known about P. mirabilis biofilms on natural or industrial surfaces and the potential consequences for these settings. The main aim of this work was to assess and compare the adhesion and biofilm formation of P. mirabilis strains from different origins on chitin and stainless steel surfaces within 4 to 96 h. Using environmental scanning electron microscopy, the biofilms of a clinical strain grown on chitin at 4 h showed greater adhesion, aggregation, thickness, and extracellular matrix production than those grown on stainless steel, whereas biofilms of an environmental strain had less aggregation on both surfaces. Biofilms of both P. mirabilis strains developed different structures on chitin, such as pillars, mushrooms, channels, and crystalline-like precipitates between 24 and 96 h, in contrast with flat-layer biofilms produced on stainless steel. Significant differences (p < 0.05) were found in the frequency of pillars and channels. Images of transmission electron microscopy demonstrated abundant fimbriae in 100 % of cells from both strains, which could be related to surface adherence and biofilm formation. This represents the first study of P. mirabilis showing adhesion, biofilm formation, and development of different structures on surfaces found outside the human host.

  12. Scanning Electron Microscopy Reveals Two Distinct Classes of Erythroblastic Island Isolated from Adult Mammalian Bone Marrow.

    PubMed

    Yeo, Jia Hao; McAllan, Bronwyn M; Fraser, Stuart T

    2016-04-01

    Erythroblastic islands are multicellular clusters in which a central macrophage supports the development and maturation of red blood cell (erythroid) progenitors. These clusters play crucial roles in the pathogenesis observed in animal models of hematological disorders. The precise structure and function of erythroblastic islands is poorly understood. Here, we have combined scanning electron microscopy and immuno-gold labeling of surface proteins to develop a better understanding of the ultrastructure of these multicellular clusters. The erythroid-specific surface antigen Ter-119 and the transferrin receptor CD71 exhibited distinct patterns of protein sorting during erythroid cell maturation as detected by immuno-gold labeling. During electron microscopy analysis we observed two distinct classes of erythroblastic islands. The islands varied in size and morphology, and the number and type of erythroid cells interacting with the central macrophage. Assessment of femoral marrow isolated from a cavid rodent species (guinea pig, Cavis porcellus) and a marsupial carnivore species (fat-tailed dunnarts, Sminthopsis crassicaudata) showed that while the morphology of the central macrophage varied, two different types of erythroblastic islands were consistently identifiable. Our findings suggest that these two classes of erythroblastic islands are conserved in mammalian evolution and may play distinct roles in red blood cell production.

  13. Anterior lens epithelium in intumescent white cataracts - scanning and transmission electron microscopy study.

    PubMed

    Andjelic, Sofija; Drašlar, Kazimir; Hvala, Anastazija; Hawlina, Marko

    2016-02-01

    Our purpose was to study the structure of the lens epithelial cells (LECs) of intumescent white cataracts (IC) in comparison with nuclear cataracts (NC) in order to investigate possible structural reasons for development of IC. The anterior lens capsule (aLC: basement membrane and associated LECs) were obtained from cataract surgery and prepared for scanning electron microscopy (SEM) and transmission electron microscopy (TEM). We observed by SEM that in IC, LEC swelling was pronounced with the clefts surrounding the groups of LECs. Another structural feature was spherical formations, that were observed on the apical side of LEC's, towards the fibre cell layer, both by SEM and TEM. Development of these structures, bulging out from the apical cell membrane of the LEC's and disrupting it, could be followed in steps towards the sphere formation. The degeneration of the lens epithelium and the structures of the aLC in IC similar to Morgagnian globules were also observed. None of these structural changes were observed in NC. We show by SEM and TEM that, in IC, LECs have pronounced structural features not observed in NC. This supports the hypothesis that the disturbed structure of LECs plays a role in water accumulation in the IC lens. We also suggest that, in IC, LECs produce bulging spheres that represent unique structures of degenerated material, extruded from the LEC.

  14. Rare-earth-doped nanophosphors for multicolor cathodoluminescence nanobioimaging using scanning transmission electron microscopy.

    PubMed

    Furukawa, Taichi; Fukushima, Shoichiro; Niioka, Hirohiko; Yamamoto, Naoki; Miyake, Jun; Araki, Tsutomu; Hashimoto, Mamoru

    2015-05-01

    We describe rare-earth-doped nanophosphors (RE-NPs) for biological imaging using cathodoluminescence(CL) microscopy based on scanning transmission electron microscopy (STEM). We report the first demonstration of multicolor CL nanobioimaging using STEM with nanophosphors. The CL spectra of the synthesized nanophosphors (Y2O3∶Eu, Y2O3∶Tb) were sufficiently narrow to be distinguished. From CL images of RE-NPs on an elastic carbon-coated copper grid, the spatial resolution was beyond the diffraction limit of light.Y2O3∶Tb and Y2O3∶Eu RE-NPs showed a remarkable resistance against electron beam exposure even at high acceleration voltage (80 kV) and retained a CL intensity of more than 97% compared with the initial intensity for 1 min. In biological CL imaging with STEM, heavy-metal-stained cell sections containing the RE-NPs were prepared,and both the CL images of RE-NPs and cellular structures, such as mitochondria, were clearly observed from STEM images with high contrast. The cellular CL imaging using RE-NPs also had high spatial resolution even though heavy-metal-stained cells are normally regarded as highly scattering media. Moreover, since theRE-NPs exhibit photoluminescence (PL) excited by UV light, they are useful for multimodal correlative imaging using CL and PL.

  15. Antennal sensilla of two parasitoid wasps: a comparative scanning electron microscopy study.

    PubMed

    Bleeker, Maartje A K; Smid, Hans M; Van Aelst, Adriaan C; Van Loon, Joop J A; Vet, Louise E M

    2004-04-01

    Two closely related parasitoid wasp species, Cotesia glomerata (L.) and Cotesia rubecula (Marshall) (Hymenoptera:Braconidae), are different in their associative learning of plant odors. To provide a solid basis for our research on the mechanisms that underlie this difference, we described the morphology of the antennal sensilla of these two species using scanning electron microscopy complemented with transmission electron microscopy. Female and male antennae of both species have the same six types of sensilla. We classified these sensilla as sensilla trichodea without pores, sensilla trichodea with a tip pore, sensilla trichodea with wall pores, sensilla coeloconica type I, sensilla coeloconica type II, and sensilla placodea. We conclude that the morphology, numbers, and distribution of the sensory receptors are highly similar in these two closely related wasp species. Differences between species and sexes occurred only in sensilla placodea numbers. C. rubecula has more sensilla placodea than C. glomerata and males of both species have a larger number and a higher density of sensilla placodea compared to females of the same species. Copyright 2004 Wiley-Liss, Inc.

  16. Plasmon-Induced Optical Field Enhancement studied by Correlated Scanning and Photoemission Electron Microscopy

    SciTech Connect

    Peppernick, Samuel J.; Joly, Alan G.; Beck, Kenneth M.; Hess, Wayne P.

    2013-04-21

    We use multi-photon photoemission electron microscopy (PEEM) to image the enhanced electric fields of silver nanoparticles supported on a silver thin film substrate. Electromagnetic field enhancement is measured by comparing the photoelectron yield of the nanoparticles with respect to the photoelectron yield of the surrounding silver thin film. We investigate the dependence of the photoelectron yield of the nanoparticle as a function of size and shape. Multi-photon PEEM results are presented for three average nanoparticle diameters: 122 ± 6, 75 ± 6, and 34 ± 2 nm. The enhancement in photoelectron yield of single nanoparticles illuminated with femtosecond laser pulses (400 nm, ~3.1 eV) is found to be a factor of 102 to 103 times greater than that produced by the flat silver thin film. High-resolution, multi-photon PEEM images of single silver nanoparticles reveal that the greatest enhancement in photoelectron yield is localized at distinct regions on the surface of the nanoparticle whose magnitude and spatial extent is dependent on the incident electric field polarization. In conjunction with correlated scanning electron microscopy (SEM), nanoparticles that deviate from nominally spherical shapes are found to exhibit irregular spatial distributions in the multi-photon PEEM images that are correlated with the unique shape and topology of the nanoparticle.

  17. Rare-earth-doped nanophosphors for multicolor cathodoluminescence nanobioimaging using scanning transmission electron microscopy

    NASA Astrophysics Data System (ADS)

    Furukawa, Taichi; Fukushima, Shoichiro; Niioka, Hirohiko; Yamamoto, Naoki; Miyake, Jun; Araki, Tsutomu; Hashimoto, Mamoru

    2015-05-01

    We describe rare-earth-doped nanophosphors (RE-NPs) for biological imaging using cathodoluminescence (CL) microscopy based on scanning transmission electron microscopy (STEM). We report the first demonstration of multicolor CL nanobioimaging using STEM with nanophosphors. The CL spectra of the synthesized nanophosphors (Y2O3:Eu, Y2O3:Tb) were sufficiently narrow to be distinguished. From CL images of RE-NPs on an elastic carbon-coated copper grid, the spatial resolution was beyond the diffraction limit of light. Y2O3:Tb and Y2O3:Eu RE-NPs showed a remarkable resistance against electron beam exposure even at high acceleration voltage (80 kV) and retained a CL intensity of more than 97% compared with the initial intensity for 1 min. In biological CL imaging with STEM, heavy-metal-stained cell sections containing the RE-NPs were prepared, and both the CL images of RE-NPs and cellular structures, such as mitochondria, were clearly observed from STEM images with high contrast. The cellular CL imaging using RE-NPs also had high spatial resolution even though heavy-metal-stained cells are normally regarded as highly scattering media. Moreover, since the RE-NPs exhibit photoluminescence (PL) excited by UV light, they are useful for multimodal correlative imaging using CL and PL.

  18. Accessing nuclear structure for field emission, in lens, scanning electron microscopy (FEISEM).

    PubMed

    Allen, T D; Bennion, G R; Rutherford, S A; Reipert, S; Ramalho, A; Kiseleva, E; Goldberg, M W

    1996-01-01

    Scanning electron microscopy (SEM) has had a shorter time course in biology than conventional transmission electron microscopy (TEM) but has nevertheless produced a wealth of images that have significantly complemented our perception of biological structure and function from TEM information. By its nature, SEM is a surface imaging technology, and its impact at the subcellular level has been restricted by the considerably reduced resolution in conventional SEM in comparison to TEM. This restriction has been removed by the recent advent of high-brightness sources used in lensfield emission instruments (FEISEM) which have produced resolution of around 1 nanometre, which is not usually a limiting figure for biological material. This communication reviews our findings in the use of FEISEM in the imaging of nuclear surfaces, then associated structures, such as nuclear pore complexes, and the relationships of these structures with cytoplasmic and nucleoplasmic elements. High resolution SEM allows the structurally orientated cell biologist to visualise, directly and in three dimensions, subcellular structure and its modulation with a view to understanding, its functional significance. Clearly, intracellular surfaces require separation from surrounding structural elements in vivo to allow surface imaging, and we review a combination of biochemical and mechanical isolation methods for nuclear surfaces.

  19. Evaluation of the infection process by Lecanicillium fungicola in Agaricus bisporus by scanning electron microscopy.

    PubMed

    Santana Nunes, Janaira; Rocha de Brito, Manuela; Cunha Zied, Diego; Aparecida das Graças Leite, Eloisa; Souza Dias, Eustáquio; Alves, Eduardo

    Lecanicillium fungicola causes dry bubble disease in Agaricus bisporus mushrooms leading to significant economic losses in commercial production. To monitor the infection process of L. fungicola in Brazilian strains of A. bisporus. The interaction between the mycelium of L. fungicola (LF.1) and three strains of A. bisporus (ABI 7, ABI 11/14 and ABI 11/21) was studied. Electron microscopy and X-ray microanalyses of vegetative growth and basidiocarp infection were evaluated. Micrographs show that the vegetative mycelium of the Brazilian strains of A. bisporus is not infected by the parasite. The images show that the pathogen can interlace the hyphae of A. bisporus without causing damage, which contributes to the presence of L. fungicola during the substrate colonization, allowing their presence during primordial formation of A. bisporus. In the basidiocarp, germ tubes form within 16h of infection with L. fungicola and the beginning of penetration takes place within 18h, both without the formation of specialized structures. Scanning electron microscopy enabled the process of colonization and reproduction to be observed within the formation of phialides, conidiophores and verticils of L. fungicola. The formation of calcium oxalate crystals by the pathogen was also visible using the X-ray microanalysis, both at the hyphae in the Petri plate and at basidiocarp infection site. Copyright © 2016 Asociación Española de Micología. Publicado por Elsevier España, S.L.U. All rights reserved.

  20. Local crystal structure analysis with several picometer precision using scanning transmission electron microscopy.

    PubMed

    Kimoto, Koji; Asaka, Toru; Yu, Xiuzhen; Nagai, Takuro; Matsui, Yoshio; Ishizuka, Kazuo

    2010-06-01

    We report a local crystal structure analysis with a high precision of several picometers on the basis of scanning transmission electron microscopy (STEM). Advanced annular dark-field (ADF) imaging has been demonstrated using software-based experimental and data-processing techniques, such as the improvement of signal-to-noise ratio, the reduction of image distortion, the quantification of experimental parameters (e.g., thickness and defocus) and the resolution enhancement by maximum-entropy deconvolution. The accuracy in the atom position measurement depends on the validity of the incoherent imaging approximation, in which an ADF image is described as the convolution between the incident probe profile and scattering objects. Although the qualitative interpretation of ADF image contrast is possible for a wide range of specimen thicknesses, the direct observation of a crystal structure with deep-sub-angstrom accuracy requires a thin specimen (e.g., 10nm), as well as observation of the structure image by conventional high-resolution transmission electron microscopy. Copyright 2009 Elsevier B.V. All rights reserved.

  1. Macrothrombocytopenia: investigating the ultrastructure of platelets and fibrin networks using scanning and transmission electron microscopy.

    PubMed

    Pretorius, Etheresia; Oberholzer, Hester M; van der Spuy, Wendy J; Meiring, Johannes H

    2009-10-01

    Macrothrombocytopenia is a rare condition where large, circulating platelets ranging between approximately 5 and 20 microm are found (typically platelets size range from 1.5 to 2.5 microm). The condition is also characterized by the prevalence of decreased numbers of circulating platelets, bleeding, short circulating times in blood, as well as abnormal platelet destruction. The current research investigates the scanning electron microscopy (SEM) and transmission electron microscopy (TEM) of platelet aggregates and fibrin networks of a family diagnosed with macrothrombocytopenia. Although TEM analysis of macrothrombocytopenia is not novel, little is known regarding the SEM analysis of platelet aggregates and fibrin networks. Here the authors show that macrothrombocytopenia have two different variations of giant platelet aggregates: a bulbous, giant aggregate that is very similar to control aggregates, and a giant flattened aggregate, with a compressed outer rim and a centrally placed area that forms a bulbous pseudopodia-like core. TEM micrographs of controls showed that, as previously seen in the literature, an aggregate contains dense bodies and alpha granules that carry, among other compounds, fibrinogen. TEM micrographs of the individuals with macrothrombocytopenia revealed aggregates with large vacuoles and areas mostly devoid of dense bodies and alpha granules. An interesting observation was that, in the presence of added human thrombin (to initially form the clot), fibrin fiber networks, comparable to that of control fibrin networks, were formed. This might be of clinical interest in the treatment regime and should be investigated further.

  2. Scanning electron microscopy of tegument-free sensory receptor of Schistosoma mansoni.

    PubMed

    Price, Z; Voge, M

    1983-01-01

    Immersion of adult Schistosoma mansoni in buffered trypsin for a short time removed the sponge-like tegument to the level of the basal lamina, effectively uncovering the basal lamina and intact sensory receptors. Stripping the tegument from the cilium and sensory bulb exposed the crown of the bulb and its axon-like process. A cilium protrudes from the bulbs through a collar-like supporting structure that resembles the rim and spokes of a wheel. The exposed axon-like process of some bulbs penetrated the basal lamina without ramifying and disappeared into the musculature; the ramifying process of others remained on the upper surface of the lamina for some distance. Identical micromorphology of the sensory receptor by Transmission Electron Microscopy (TEM) (Silk and Spence, 1969; Hockley, 1973), and the similar appearance of the surface of the bulb and cilia and the ciliary supporting structure by Scanning Electron Microscopy (SEM) suggests that all of the receptors probably perform the same sensory function.

  3. Evaluation of enteric matrix microspheres prepared by emulsion-solvent evaporation using scanning electron microscopy.

    PubMed

    Obeidat, W M; Price, J C

    2004-02-01

    Theophylline microspheres were prepared by the emulsion-solvent evaporation method using cellulose acetate butyrate (CAB381-20) and mixtures of CAB381-20(R) and cellulose acetate phthalate. The physical state of the drug, polymers and microspheres surfaces were determined using scanning electron microscopy. For those microspheres prepared using mixtures of CAB381-20 and cellulose acetate phthalate, scanning electron micrographs were taken before dissolution and also at different stages of dissolution (in SGF, pH 1.2 and in simulated intestinal fluid, pH 7.5). Micrographs were taken of the outside surfaces of the microspheres and of the cleaved microspheres showing their interiors (core). Drug crystals were observed on or near the surface of microspheres prepared from the polymer mixtures, while no drug particles or crystals were seen on the surfaces of microspheres prepared solely from CAB381-20. An acid wash for less than 2 min was capable of extracting all drug on the surface of the microspheres prepared from a mixture of CAB381-20 and cellulose acetate phthalate. The absence of drug crystals on the surface of CAB381-20 microspheres is believed to prevent initial drug release and create a lag time in release profiles. Results suggest that in both microsphere formulations, a layer of drug-free polymer is formed outside the core matrix and is believed to be responsible for the near zero-order release profiles.

  4. Quantitative Description of Crystal Nucleation and Growth from in Situ Liquid Scanning Transmission Electron Microscopy.

    PubMed

    Ievlev, Anton V; Jesse, Stephen; Cochell, Thomas J; Unocic, Raymond R; Protopopescu, Vladimir A; Kalinin, Sergei V

    2015-12-22

    Recent advances in liquid cell (scanning) transmission electron microscopy (S)TEM has enabled in situ nanoscale investigations of controlled nanocrystal growth mechanisms. Here, we experimentally and quantitatively investigated the nucleation and growth mechanisms of Pt nanostructures from an aqueous solution of K2PtCl6. Averaged statistical, network, and local approaches have been used for the data analysis and the description of both collective particles dynamics and local growth features. In particular, interaction between neighboring particles has been revealed and attributed to reduction of the platinum concentration in the vicinity of the particle boundary. The local approach for solving the inverse problem showed that particles dynamics can be simulated by a stationary diffusional model. The obtained results are important for understanding nanocrystal formation and growth processes and for optimization of synthesis conditions.

  5. Evaluation of vermicompost maturity using scanning electron microscopy and paper chromatography analysis.

    PubMed

    Senthil Kumar, D; Satheesh Kumar, P; Rajendran, N M; Uthaya Kumar, V; Anbuganapathi, G

    2014-04-02

    Vermicompost was produced from flower waste inoculated with biofertilizers using the earthworm Eisenia fetida. Principal component analysis (PCA) and cluster analysis (CA) were carried out on the basis of physicochemical parameters of vermicomposted samples. From the results of the PCA and CA, it was possible to classify two different groups of vermicompost samples in the following categories: E2 and E5; and E1, E3, E4, and control. Scanning electron microscopy and biodynamic circular paper chromatography analysis were used to investigate the changes in surface morphology and functional groups in the control and vermicompost products. SEM analysis of E1-E5 shows more fragment and pores than the control. Chromatographic analysis of vermicompost indicated the mature condition of the compost materials.

  6. Characterization of defect growth structure in ion plated films by scanning electron microscopy

    NASA Technical Reports Server (NTRS)

    Spalvins, T.

    1979-01-01

    Copper and gold films (0.2 to 2 microns) were ion plated onto polished 304-stainless-steel surfaces. These coatings were examined by scanning electron microscopy for coating growth defects. Three types of defects were distinguished: nodular growth, abnormal or runaway growth, and spits. The cause and origin for each type of defect was traced. Nodular growth is primarily due to inherent substrate microdefects, abnormal or runaway growth is due to external surface inclusions, and spits are due to nonuniform evaporation. All these defects have adverse effects on the coatings. They induce stresses and produce porosity in the coatings and thus weaken their mechanical properties. Friction and wear characteristics are affected by coating defects, since the large nodules are pulled out and additional wear debris is generated.

  7. Imaging of surface spin textures on bulk crystals by scanning electron microscopy

    PubMed Central

    Akamine, Hiroshi; Okumura, So; Farjami, Sahar; Murakami, Yasukazu; Nishida, Minoru

    2016-01-01

    Direct observation of magnetic microstructures is vital for advancing spintronics and other technologies. Here we report a method for imaging surface domain structures on bulk samples by scanning electron microscopy (SEM). Complex magnetic domains, referred to as the maze state in CoPt/FePt alloys, were observed at a spatial resolution of less than 100 nm by using an in-lens annular detector. The method allows for imaging almost all the domain walls in the mazy structure, whereas the visualisation of the domain walls with the classical SEM method was limited. Our method provides a simple way to analyse surface domain structures in the bulk state that can be used in combination with SEM functions such as orientation or composition analysis. Thus, the method extends applications of SEM-based magnetic imaging, and is promising for resolving various problems at the forefront of fields including physics, magnetics, materials science, engineering, and chemistry. PMID:27872493

  8. Imaging of surface spin textures on bulk crystals by scanning electron microscopy.

    PubMed

    Akamine, Hiroshi; Okumura, So; Farjami, Sahar; Murakami, Yasukazu; Nishida, Minoru

    2016-11-22

    Direct observation of magnetic microstructures is vital for advancing spintronics and other technologies. Here we report a method for imaging surface domain structures on bulk samples by scanning electron microscopy (SEM). Complex magnetic domains, referred to as the maze state in CoPt/FePt alloys, were observed at a spatial resolution of less than 100 nm by using an in-lens annular detector. The method allows for imaging almost all the domain walls in the mazy structure, whereas the visualisation of the domain walls with the classical SEM method was limited. Our method provides a simple way to analyse surface domain structures in the bulk state that can be used in combination with SEM functions such as orientation or composition analysis. Thus, the method extends applications of SEM-based magnetic imaging, and is promising for resolving various problems at the forefront of fields including physics, magnetics, materials science, engineering, and chemistry.

  9. Big Data and Deep data in scanning and electron microscopies: functionality from multidimensional data sets

    SciTech Connect

    Belianinov, Alex; Vasudevan, Rama K; Strelcov, Evgheni; Steed, Chad A; Yang, Sang Mo; Tselev, Alexander; Jesse, Stephen; Biegalski, Michael D; Shipman, Galen M; Symons, Christopher T; Borisevich, Albina Y; Archibald, Richard K; Kalinin, Sergei

    2015-01-01

    The development of electron, and scanning probe microscopies in the second half of the twentieth century have produced spectacular images of internal structure and composition of matter with, at nanometer, molecular, and atomic resolution. Largely, this progress was enabled by computer-assisted methods of microscope operation, data acquisition and analysis. The progress in imaging technologies in the beginning of the twenty first century has opened the proverbial floodgates of high-veracity information on structure and functionality. High resolution imaging now allows information on atomic positions with picometer precision, allowing for quantitative measurements of individual bond length and angles. Functional imaging often leads to multidimensional data sets containing partial or full information on properties of interest, acquired as a function of multiple parameters (time, temperature, or other external stimuli). Here, we review several recent applications of the big and deep data analysis methods to visualize, compress, and translate this data into physically and chemically relevant information from imaging data.

  10. Rapid phenotypic analysis of uncoated Drosophila samples with low-vacuum scanning electron microscopy.

    PubMed

    Tardi, Nicholas J; Cook, Martha E; Edwards, Kevin A

    2012-01-01

    Research projects featuring repetitive phenotypic analysis of insects, such as taxonomic studies, quantitative genetics, and mutant screens, could be greatly facilitated by a simpler approach to scanning electron microscopy (SEM). Here, we have applied low-vacuum SEM to wild type and mutant Drosophila and demonstrate that high quality ultrastructure data can be obtained quickly using minimal preparation. Adult flies, frozen live for storage, were mounted on aluminum stubs with carbon cement and directly imaged, with no chemical treatment or sputter coating. The key imaging parameters were identified and optimized, including chamber pressure, beam size, accelerating voltage, working distance and beam exposure. Different optimal conditions were found for eyes, wings, and bristles; in particular, surface features of bristles were obscured at higher accelerating voltages. The chief difficulties were charging, beam damage, and sample movement. We conclude that our optimized protocol is well suited to large-scale ultrastructural phenotypic analysis in insects.

  11. Enterococcus Faecalis Biofilm. Formation and Development in Vitro Observed by Scanning Electron Microscopy.

    PubMed

    Bulacio, María de Los Á; Galván, Lucas R; Gaudioso, Cristina; Cangemi, Rosa; Erimbaue, Marta I

    2015-12-01

    Biofilm produced by Enterococcus faecalis isolated from root canals was detected by growing it on microplates and using 10% crystal violet stain, elution with alcohol and three procedures: no fixation, heat fixation and 10% formaldehyde fixation. The biofilm was evaluated using a Versamax Microplate Reader (USA). Twenty sterile root portions were incubated in TS broth with E. faecalis (108) for 48 hours, 4, 7, 14 and 30 days, after which they were processed and observed by scanning electron microscopy (SEM). Significantly more biofilm was found on the microplates for formaldehyde fixation than for heat fixation or no fixation (ANOVA p<0.0001). SEM showed E. faecalis growth at all times and biofilm development as from 14 days' incubation. Fixation with 10% formaldehyde was the most appropriate technique for detecting E. faecalis biofilm development on microplates. SEM confirmed biofilm formation after 14 days incubation.

  12. Devolatilization Studies of Oil Palm Biomass for Torrefaction Process through Scanning Electron Microscopy

    NASA Astrophysics Data System (ADS)

    Daud, D.; Abd. Rahman, A.; Shamsuddin, A. H.

    2016-03-01

    In this work, palm oil biomass consisting of empty fruit bunch (EFB), mesocarp fibre and palm kernel shell (PKS) were chosen as raw material for torrefaction process. Torrefaction process was conducted at various temperatures of 240 °C, 270 °C and 300 °C with a residence time of 60 minutes. The morphology of the raw and torrefied biomass was then observed through Scanning Electron Microscopy (SEM) images. Also, through this experiment the correlation between the torrefaction temperatures with the volatile gases released were studied. From the observation, the morphology structure of the biomass exhibited inter-particle gaps due to the release of volatile gases and it is obviously seen more at higher temperatures. Moreover, the change of the biomass structure is influenced by the alteration of the lignocellulose biomass.

  13. Scanning Electron Microscopy Investigation of a Sample Depth Profile Through the Martian Meteorite Nakhla

    NASA Technical Reports Server (NTRS)

    Toporski, Jan; Steele, Andrew; Westall, Frances; McKay, David S.

    2000-01-01

    The ongoing scientific debate as to whether or not the Martian meteorite ALH84001 contained evidence of possible biogenic activities showed the need to establish consistent methods to ascertain the origin of such evidence. To distinguish between terrestrial organic material/microbial contaminants and possible indigenous microbiota within meteorites is therefore crucial. With this in mind a depth profile consisting of four samples from a new sample allocation of Martian meteorite Nakhla was investigated using scanning electron microscopy (SEM) and energy dispersive X-ray analysis. SEM imaging of freshly broken fractured chips revealed structures strongly recent terrestrial microorganisms, in some cases showing evidence of active growth. This conclusion was supported by EDX analysis, which showed the presence of carbon associated with these structures, we concluded that these structures represent recent terrestrial contaminants rather than structures indigenous to the meteorite. Page

  14. Determination of the coalescence temperature of latexes by environmental scanning electron microscopy.

    PubMed

    Gonzalez, Edurne; Tollan, Christopher; Chuvilin, Andrey; Barandiaran, Maria J; Paulis, Maria

    2012-08-01

    A new methodology for quantitative characterization of the coalescence process of waterborne polymer dispersion (latex) particles by environmental scanning electron microscopy (ESEM) is proposed. The experimental setup has been developed to provide reproducible latex monolayer depositions, optimized contrast of the latex particles, and a reliable readout of the sample temperature. Quantification of the coalescence process under dry conditions has been performed by image processing based on evaluation of the image autocorrelation function. As a proof of concept the coalescence of two latexes with known and differing glass transition temperatures has been measured. It has been shown that a reproducibility of better than 1.5 °C can be obtained for the measurement of the coalescence temperature.

  15. A low-cost technique to manufacture a container to process meiofauna for scanning electron microscopy.

    PubMed

    Abolafia, J

    2015-09-01

    An easy and low-cost method to elaborate a container to dehydrate nematodes and other meiofauna in order to process them for scanning electron microscopy (SEM) is presented. Illustrations of its elaboration, step by step, are included. In addition, a brief methodology to process meiofauna, especially nematodes and kinorhynchs, and illustrations are provided. With this methodology it is possible to easily introduce the specimens, to lock them in a closed chamber allowing the infiltration of fluids and gases (ethanol, acetone, carbon dioxide) but avoiding losing the specimens. After using this meiofauna basket for SEM the results are efficient. Examples of nematode and kinorhynch SEM pictures obtained using this methodology are also included. © 2015 Wiley Periodicals, Inc.

  16. Three-dimensional architecture of the myosalpinx in the mare as revealed by scanning electron microscopy.

    PubMed

    Germanà, Antonino; Cassata, Rosa; Cristarella, Santo; Scirpo, Aurelio; Muglia, Ugo

    2002-07-01

    The three-dimensional architecture of the myosalpinx in the mare was investigated by means of scanning electron microscopy (SEM) after removal of interstitial connective tissue with NaOH digestion. In the extramural portion of the tubo-uterine junction (TUJ), isthmus, and ampulla, the myosalpinx architecture is represented by a unique muscular structure which runs from the mesosalpinx to the base of the inner mucous folds. This unique muscular structure consists mainly of bundles of muscular fibers independent of one another, which show a multiple spatial arrangement and form a complex network. Such a muscular architecture is likely more suitable for stirring rather than pushing the embryos and gametes through the Fallopian tube.

  17. Fretting wear in titanium, Monel-400, and cobalt 25-percent-molybdenum using scanning electron microscopy

    NASA Technical Reports Server (NTRS)

    Bill, R. C.

    1972-01-01

    Damage scar volume measurements taken from like metal fretting pairs combined with scanning electron microscopy observations showed that three sequentially operating mechanisms result in the fretting of titanium, Monel-400, and cobalt - 25-percent molybdenum. Initially, adhesion and plastic deformation of the surface played an important role. This was followed after a few hundred cycles by a fatigue mechanism which produced spall-like pits in the damage scar. Finally, a combination of oxidation and abrasion by debris particles became most significant. Damage scar measurements made on several elemental metals after 600,000 fretting cycles suggested that the ratio of oxide hardness to metal hardness was a measure of the susceptibility of a metal to progressive damage by fretting.

  18. Scanning electron acoustic microscopy of indentation-induced cracks and residual stresses in ceramics

    NASA Technical Reports Server (NTRS)

    Cantrell, John H.; Qian, Menglu; Ravichandran, M. V.; Knowles, K. M.

    1990-01-01

    The ability of scanning electron acoustic microscopy (SEAM) to characterize ceramic materials is assessed. SEAM images of Vickers indentations in SiC whisker-reinforced alumina clearly reveal not only the radial cracks, the length of which can be used to estimate the fracture toughness of the material, but also reveal strong contrast, interpreted as arising from the combined effects of lateral cracks and the residual stress field left in the SiC whisker-reinforced alumina by the indenter. The strong contrast is removed after the material is heat treated at 1000 C to relieve the residual stresses around the indentations. A comparison of these observations with SEAM and reflected polarized light observations of Vickers indentations in soda-lime glass both before and after heat treatment confirms the interpretation of the strong contrast.

  19. Visualizing the Dynamics of Nanoparticles in Liquids by Scanning Electron Microscopy.

    PubMed

    Kim, Paul Y; Ribbe, Alexander E; Russell, Thomas P; Hoagland, David A

    2016-06-28

    Taking advantage of ionic liquid nonvolatility, the Brownian motions of nanospheres and nanorods in free-standing liquid films were visualized in situ by scanning electron microscopy. Despite the imaging environment's high vacuum, a liquid cell was not needed. For suspensions that are dilute and films that are thick compared to the particle diameter, the translational and rotational diffusion coefficients determined by single-particle tracking agree with theoretical predictions. In thinner films, a striking dynamical pairing of nanospheres was observed, manifesting a balance of capillary and hydrodynamic interactions, the latter strongly accentuated by the two-dimensional film geometry. Nanospheres at high concentration displayed subdiffusive caged motion. Concentrated nanorods in the thinner films transiently assembled into finite stacks but did not achieve high tetratic order. The illustrated imaging protocol will broadly apply to the study of soft matter structure and dynamics with great potential impact.

  20. Identifying dislocations and stacking faults in GaN films by scanning transmission electron microscopy

    NASA Astrophysics Data System (ADS)

    Su, X. J.; Niu, M. T.; Zeng, X. H.; Huang, J.; Zhang, J. C.; Zhang, J. P.; Wang, J. F.; Xu, K.

    2016-08-01

    The application of annular bright field (ABF) and medium-angle annular dark field (MAADF) scanning transmission electron microscopy (STEM) imaging to crystalline defect analysis has been extended to dislocations and stacking faults (SFs). Dislocations and SFs have been imaged under zone-axis and two-beam diffraction conditions. Comparing to conventional two-beam diffraction contrast images, the ABF and MAADF images of dislocations and SFs not only are complementary and symmetrical with their peaks at dislocation core and SFs plane, but also show similar extinction phenomenon. It is demonstrated that conventional TEM rules for diffraction contrast, i.e. g · b and g · R invisibility criteria remain applicable. The contrast mechanism and extinction of dislocation and SFs in ABF and MAADF STEM are illuminated by zero-order Laue zone Kikuchi diffraction.

  1. Preliminary report: laboratory-induced stain removal as assessed by environmental scanning electron microscopy.

    PubMed

    Habib, C M; Kugel, G; Marcus, A

    1998-01-01

    Environmental scanning electron microscopy (ESEM) was employed to observe stain removal during brushing with Arm & Hammer Dental Care and Crest Regular Toothpaste. ESEM allows serial examinations of the same sample, and does not require a destructive preparative process. Three extracted molars were cleaned, placed into a 96-hour broth culture of Streptococcus mutans, and stain was produced with undiluted chlorhexidine rinse, concentrated coffee and tea for a period of 23 days. After staining, the teeth were examined by ESEM, then brushed using a toothbrushing machine. Imaging was repeated after 5, 10, 15 and 30 seconds of brushing. As seen with ESEM, the Arm & Hammer product had different effects than those from the distilled water control, suggesting something other than that expected from abrasive and mechanical forces alone. There were also differences from the Crest dentifrice removal on this single sample, suggesting a possible difference between the two products. Further studies are needed to confirm and explain these effects.

  2. A simple method to 'point count' silt using scanning electron microscopy aided by image analysis.

    PubMed

    Ware, C I

    2003-11-01

    This study presents a simple method to 'point count' silt-sized grains using backscattered scanning electron microscopy together with image analysis. The work materialized out of the need to determine the heavy mineral abundance within silt obtained from coastal dunes to aid in the interpretation of dune weathering. This technique allows two broad mineral groups to be quantified according to their modal abundance. The groups are characterized by their dominant atomic elements present; atomic numbers >20 are classified as 'high' (metal oxides, zircon, monazite, carbonates, pyroxenes and amphiboles) and those <20 as 'low' (quartz, feldspars and organics). As a check on this technique, X-ray fluorescence was used. This showed a strong positive correlation (r2=0.85) with the developed point counting technique.

  3. Scanning electron microscopy and calcification in amelogenesis imperfecta in anterior and posterior human teeth.

    PubMed

    Sánchez-Quevedo, M C; Ceballos, G; García, J M; Rodríguez, I A; Gómez de Ferraris, M E; Campos, A

    2001-07-01

    Teeth fragments from members of a family clinically and genetically diagnosed as having amelogenesis imperfecta were studied by scanning electron microscopy and X-ray microprobe analysis to establish the morphological patterns and the quantitative concentration of calcium in the enamel of anterior (canine, incisor) and posterior (premolar and molar) teeth. The prism patterns in the enamel of teeth from both regions were parallel or irregularly decussate, with occasional filamentous prisms accompanied by small, irregularly rounded formations. Prismless enamel showed the R- and P-type patterns. Calcium levels in enamel of amelogenesis imperfecta and control teeth differed significantly between anterior and posterior teeth, indicating that the factors that influence normal mineralization in different regions of the dental arch are not altered in the process of amelogenesis imperfecta.

  4. Applications of scanning electron microscopy and X-ray microanalysis in inner ear pathology

    SciTech Connect

    Anniko, M.; Lim, D.J.; Sobin, A.; Wroblewski, R.

    1985-01-01

    Surface pathology of inner ear structures so far described in detail concern cochlear and vestibular hair cells and the stria vascularis. In man, surgical intervention into the inner ear is very uncommon and when performed is in general with the primary objective of destroying the diseased peripheral end organs. The vast majority of inner ear tissue available for use with scanning electron microscopy (SEM) is therefore obtained from animals. The present paper reviews the progression of surface pathology caused by aminoglycoside antibiotics, acoustic overstimulation and in a guinea pig strain with genetic inner ear disease. The primary site of onset of surface pathology differs, depending on the underlying cause. Advanced surface pathology shows a similar type of morphological degeneration independent of cause. The combination of SEM and energy dispersive X-ray microanalysis (XRMA) of inner ear pathology has as yet been reported in only three studies, all concerning inner ear fluids or otoconia.

  5. Growth of vacuum evaporated ultraporous silicon studied with spectroscopic ellipsometry and scanning electron microscopy

    NASA Astrophysics Data System (ADS)

    Kaminska, Kate; Amassian, Aram; Martinu, Ludvik; Robbie, Kevin

    2005-01-01

    Using a combination of variable-angle spectroscopic ellipsometry and scanning electron microscopy, we investigated the scaling behavior of uniaxially anisotropic, ultraporous silicon manufactured with glancing angle deposition. We found that both the diameter of the nanocolumns and the spacing between them increase with film thickness according to a power-law relationship consistent with self-affine fractal growth. An ellipsometric model is proposed to fit the optical properties of the anisotropic silicon films employing an effective medium approximation mixture of Tauc-Lorentz oscillator and void. This study shows that the optical response of silicon films made at glancing incidence differs significantly from that of amorphous silicon prepared by other methods due to highly oriented nanocolumn formation and power-law scaling.

  6. Use of scanning electron microscopy to monitor nanofibre/cell interaction in digestive epithelial cells.

    PubMed

    Millaku, Agron; Drobne, Damjana; Torkar, Matjaz; Novak, Sara; Remškar, Maja; Pipan-Tkalec, Živa

    2013-09-15

    We provide data obtained by scanning electron microscopy (SEM) and energy dispersive X-ray spectroscopy (EDS) on the interaction of ingested tungsten nanofibers with epithelial cells of the digestive tubes of a test organism Porcellio scaber. Conventional toxicity endpoints including feeding behaviour, weight loss and mortality were also measured in each investigated animal. No toxicity was detected in any of exposed animals after 14 days of feeding on tungsten nanofiber dosed food, but when nanofibers enter the digestive system they can react with epithelial cells of the digestive tubes, becoming physically inserted into the cells. In this way, nanofibers can injure the epithelial cells of digestive gland tubes when they are ingested with food. Our SEM data suggest that peristaltic forces may have an important role, not predicted by in vitro experiments, in the interactions of nanomaterials with digestive intestinal cells. Copyright © 2013 Elsevier B.V. All rights reserved.

  7. Light and scanning electron microscopy of the tongue of a degu (Octodon degus).

    PubMed

    Cizek, Petr; Hamouzova, Pavla; Jekl, Vladimir; Kvapil, Pavel; Tichy, Frantisek

    2017-09-01

    The tongue of an adult degu was examined by light and scanning electron microscopy. It consists of an apex, corpus, and radix and contains a lingual prominence. The aim of this study was to describe the course of muscle fascicles of the proper lingual muscle, the presence and nature of the lingual salivary glands, and particularly the appearance and distribution of the lingual papillae. Three major types of papillae have been observed: filiform, conical, and vallate. The dorsal surface of the lingual apex extends in caudally bent filiform papillae with two spines. The lingual corpus bears long filiform papillae with a single tip. The lingual radix contains crown-like papillae in the region of the prominence and conical papillae in the remaining areas. Two oval vallate papillae were discovered caudally on the lingual radix. This first description of the lingual structures in a degu could be used for comparative studies or as basic data for differentiation of lingual morphology in this species.

  8. Imaging of surface spin textures on bulk crystals by scanning electron microscopy

    NASA Astrophysics Data System (ADS)

    Akamine, Hiroshi; Okumura, So; Farjami, Sahar; Murakami, Yasukazu; Nishida, Minoru

    2016-11-01

    Direct observation of magnetic microstructures is vital for advancing spintronics and other technologies. Here we report a method for imaging surface domain structures on bulk samples by scanning electron microscopy (SEM). Complex magnetic domains, referred to as the maze state in CoPt/FePt alloys, were observed at a spatial resolution of less than 100 nm by using an in-lens annular detector. The method allows for imaging almost all the domain walls in the mazy structure, whereas the visualisation of the domain walls with the classical SEM method was limited. Our method provides a simple way to analyse surface domain structures in the bulk state that can be used in combination with SEM functions such as orientation or composition analysis. Thus, the method extends applications of SEM-based magnetic imaging, and is promising for resolving various problems at the forefront of fields including physics, magnetics, materials science, engineering, and chemistry.

  9. Scanning electron acoustic microscopy of indentation-induced cracks and residual stresses in ceramics

    NASA Technical Reports Server (NTRS)

    Cantrell, John H.; Qian, Menglu; Ravichandran, M. V.; Knowles, K. M.

    1990-01-01

    The ability of scanning electron acoustic microscopy (SEAM) to characterize ceramic materials is assessed. SEAM images of Vickers indentations in SiC whisker-reinforced alumina clearly reveal not only the radial cracks, the length of which can be used to estimate the fracture toughness of the material, but also reveal strong contrast, interpreted as arising from the combined effects of lateral cracks and the residual stress field left in the SiC whisker-reinforced alumina by the indenter. The strong contrast is removed after the material is heat treated at 1000 C to relieve the residual stresses around the indentations. A comparison of these observations with SEAM and reflected polarized light observations of Vickers indentations in soda-lime glass both before and after heat treatment confirms the interpretation of the strong contrast.

  10. Matched Backprojection Operator for Combined Scanning Transmission Electron Microscopy Tilt- and Focal Series.

    PubMed

    Dahmen, Tim; Kohr, Holger; de Jonge, Niels; Slusallek, Philipp

    2015-06-01

    Combined tilt- and focal series scanning transmission electron microscopy is a recently developed method to obtain nanoscale three-dimensional (3D) information of thin specimens. In this study, we formulate the forward projection in this acquisition scheme as a linear operator and prove that it is a generalization of the Ray transform for parallel illumination. We analytically derive the corresponding backprojection operator as the adjoint of the forward projection. We further demonstrate that the matched backprojection operator drastically improves the convergence rate of iterative 3D reconstruction compared to the case where a backprojection based on heuristic weighting is used. In addition, we show that the 3D reconstruction is of better quality.

  11. Enhanced light element imaging in atomic resolution scanning transmission electron microscopy.

    PubMed

    Findlay, S D; Kohno, Y; Cardamone, L A; Ikuhara, Y; Shibata, N

    2014-01-01

    We show that an imaging mode based on taking the difference between signals recorded from the bright field (forward scattering region) in atomic resolution scanning transmission electron microscopy provides an enhancement of the detectability of light elements over existing techniques. In some instances this is an enhancement of the visibility of the light element columns relative to heavy element columns. In all cases explored it is an enhancement in the signal-to-noise ratio of the image at the light column site. The image formation mechanisms are explained and the technique is compared with earlier approaches. Experimental data, supported by simulation, are presented for imaging the oxygen columns in LaAlO₃. Case studies looking at imaging hydrogen columns in YH₂ and lithium columns in Al₃Li are also explored through simulation, particularly with respect to the dependence on defocus, probe-forming aperture angle and detector collection aperture angles. © 2013 Elsevier B.V. All rights reserved.

  12. Morphology of the ferritin iron core by aberration corrected scanning transmission electron microscopy

    NASA Astrophysics Data System (ADS)

    Jian, Nan; Dowle, Miriam; Horniblow, Richard D.; Tselepis, Chris; Palmer, Richard E.

    2016-11-01

    As the major iron storage protein, ferritin stores and releases iron for maintaining the balance of iron in fauna, flora, and bacteria. We present an investigation of the morphology and iron loading of ferritin (from equine spleen) using aberration-corrected high angle annular dark field scanning transmission electron microscopy. Atom counting method, with size selected Au clusters as mass standards, was employed to determine the number of iron atoms in the nanoparticle core of each ferritin protein. Quantitative analysis shows that the nuclearity of iron atoms in the mineral core varies from a few hundred iron atoms to around 5000 atoms. Moreover, a relationship between the iron loading and iron core morphology is established, in which mineral core nucleates from a single nanoparticle, then grows along the protein shell before finally forming either a solid or hollow core structure.

  13. Morphology of the ferritin iron core by aberration corrected scanning transmission electron microscopy.

    PubMed

    Jian, Nan; Dowle, Miriam; Horniblow, Richard D; Tselepis, Chris; Palmer, Richard E

    2016-11-18

    As the major iron storage protein, ferritin stores and releases iron for maintaining the balance of iron in fauna, flora, and bacteria. We present an investigation of the morphology and iron loading of ferritin (from equine spleen) using aberration-corrected high angle annular dark field scanning transmission electron microscopy. Atom counting method, with size selected Au clusters as mass standards, was employed to determine the number of iron atoms in the nanoparticle core of each ferritin protein. Quantitative analysis shows that the nuclearity of iron atoms in the mineral core varies from a few hundred iron atoms to around 5000 atoms. Moreover, a relationship between the iron loading and iron core morphology is established, in which mineral core nucleates from a single nanoparticle, then grows along the protein shell before finally forming either a solid or hollow core structure.

  14. Colorimeter and scanning electron microscopy analysis of teeth submitted to internal bleaching.

    PubMed

    Martin-Biedma, Benjamin; Gonzalez-Gonzalez, Teresa; Lopes, Manuela; Lopes, Luis; Vilar, Rui; Bahillo, José; Varela-Patiño, Purificación

    2010-02-01

    This in vitro study compared the tooth color and the ultrastructure of internal dental tissues before and after internal bleaching. Sodium perborate was placed in the pulp chamber of endodontically treated molars and sealed with intermediate restorative material. The test samples were stored in a physiologic solution, and the bleaching agent was replaced every 7 days. A control group was used. After 1 month, the colors of the test and control samples were measured with a colorimeter, and the internal surfaces were observed under field emission scanning electron microscopy (FESEM). Statistically significant differences were found between the test and control sample colors. The FESEM ultrastructure analysis of the internal enamel and dentin surfaces did not show any changes after the internal bleaching. The results of the present study show that sodium perborate is effective in bleaching nonvital teeth and does not produce ultrastructural changes in the dental tissues. Copyright 2010 American Association of Endodontists. Published by Elsevier Inc. All rights reserved.

  15. Dynamic imaging of Au-nanoparticles via scanning electron microscopy in a graphene wet cell.

    PubMed

    Yang, Wayne; Zhang, Yuning; Hilke, Michael; Reisner, Walter

    2015-08-07

    High resolution nanoscale imaging in liquid environments is crucial for studying molecular interactions in biological and chemical systems. In particular, electron microscopy is the gold-standard tool for nanoscale imaging, but its high-vacuum requirements make application to in-liquid samples extremely challenging. Here we present a new graphene based wet cell device where high resolution scanning electron microscope (SEM) and energy dispersive x-rays (EDX) analysis can be performed directly inside a liquid environment. Graphene is an ideal membrane material as its high transparancy, conductivity and mechanical strength can support the high vacuum and grounding requirements of a SEM while enabling maximal resolution and signal. In particular, we obtain high resolution ([Formula: see text] nm) SEM video images of nanoparticles undergoing Brownian motion inside the graphene wet cell and EDX analysis of nanoparticle composition in the liquid enviornment. Our obtained resolution surpasses current conventional silicon nitride devices imaged in both a SEM and transmission electron microscope under much higher electron doses.

  16. Dynamic imaging of Au-nanoparticles via scanning electron microscopy in a graphene wet cell

    NASA Astrophysics Data System (ADS)

    Yang, Wayne; Zhang, Yuning; Hilke, Michael; Reisner, Walter

    2015-08-01

    High resolution nanoscale imaging in liquid environments is crucial for studying molecular interactions in biological and chemical systems. In particular, electron microscopy is the gold-standard tool for nanoscale imaging, but its high-vacuum requirements make application to in-liquid samples extremely challenging. Here we present a new graphene based wet cell device where high resolution scanning electron microscope (SEM) and energy dispersive x-rays (EDX) analysis can be performed directly inside a liquid environment. Graphene is an ideal membrane material as its high transparancy, conductivity and mechanical strength can support the high vacuum and grounding requirements of a SEM while enabling maximal resolution and signal. In particular, we obtain high resolution (\\lt 5 nm) SEM video images of nanoparticles undergoing Brownian motion inside the graphene wet cell and EDX analysis of nanoparticle composition in the liquid enviornment. Our obtained resolution surpasses current conventional silicon nitride devices imaged in both a SEM and transmission electron microscope under much higher electron doses.

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

    PubMed

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

    2009-09-01

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

  18. 3D imaging by serial block face scanning electron microscopy for materials science using ultramicrotomy.

    PubMed

    Hashimoto, Teruo; Thompson, George E; Zhou, Xiaorong; Withers, Philip J

    2016-04-01

    Mechanical serial block face scanning electron microscopy (SBFSEM) has emerged as a means of obtaining three dimensional (3D) electron images over volumes much larger than possible by focused ion beam (FIB) serial sectioning and at higher spatial resolution than achievable with conventional X-ray computed tomography (CT). Such high resolution 3D electron images can be employed for precisely determining the shape, volume fraction, distribution and connectivity of important microstructural features. While soft (fixed or frozen) biological samples are particularly well suited for nanoscale sectioning using an ultramicrotome, the technique can also produce excellent 3D images at electron microscope resolution in a time and resource-efficient manner for engineering materials. Currently, a lack of appreciation of the capabilities of ultramicrotomy and the operational challenges associated with minimising artefacts for different materials is limiting its wider application to engineering materials. Consequently, this paper outlines the current state of the art for SBFSEM examining in detail how damage is introduced during slicing and highlighting strategies for minimising such damage. A particular focus of the study is the acquisition of 3D images for a variety of metallic and coated systems. Copyright © 2016 The Authors. Published by Elsevier B.V. All rights reserved.

  19. New Insights on Subsurface Imaging of Carbon Nanotubes in Polymer Composites via Scanning Electron Microscopy

    NASA Technical Reports Server (NTRS)

    Zhao, Minhua; Ming, Bin; Kim, Jae-Woo; Gibbons, Luke J.; Gu, Xiaohong; Nguyen, Tinh; Park, Cheol; Lillehei, Peter T.; Villarrubia, J. S.; Vladar, Andras E.; hide

    2015-01-01

    Despite many studies of subsurface imaging of carbon nanotube (CNT)-polymer composites via scanning electron microscopy (SEM), significant controversy exists concerning the imaging depth and contrast mechanisms. We studied CNT-polyimide composites and, by threedimensional reconstructions of captured stereo-pair images, determined that the maximum SEM imaging depth was typically hundreds of nanometers. The contrast mechanisms were investigated over a broad range of beam accelerating voltages from 0.3 to 30 kV, and ascribed to modulation by embedded CNTs of the effective secondary electron (SE) emission yield at the polymer surface. This modulation of the SE yield is due to non-uniform surface potential distribution resulting from current flows due to leakage and electron beam induced current. The importance of an external electric field on SEM subsurface imaging was also demonstrated. The insights gained from this study can be generally applied to SEM nondestructive subsurface imaging of conducting nanostructures embedded in dielectric matrices such as graphene-polymer composites, silicon-based single electron transistors, high resolution SEM overlay metrology or e-beam lithography, and have significant implications in nanotechnology.

  20. Sample preparation for scanning electron microscopy of plant surfaces--horses for courses.

    PubMed

    Pathan, A K; Bond, J; Gaskin, R E

    2008-12-01

    Plant tissues must be dehydrated for observation in most electron microscopes. Although a number of sample processing techniques have been developed for preserving plant tissues in their original form and structure, none of them are guaranteed artefact-free. The current paper reviews common scanning electron microscopy techniques and the sample preparation methods employed for visualisation of leaves under specific types of electron microscopes. Common artefacts introduced by specific techniques on different leaf types are discussed. Comparative examples are depicted from our lab using similar techniques; the pros and cons for specific techniques are discussed. New promising techniques and microscopes, which can alleviate some of the problems encountered in conventional methods of leaf sample processing and visualisation, are also discussed. It is concluded that the choice of technique for a specific leaf sample is dictated by the surface features that need to be preserved (such as trichomes, epidermal cells or wax microstructure), the resolution to be achieved, availability of the appropriate processing equipment and the technical capabilities of the available electron microscope.

  1. Serial Block-Face Scanning Electron Microscopy to Reconstruct Three-Dimensional Tissue Nanostructure

    PubMed Central

    Horstmann, Heinz

    2004-01-01

    Three-dimensional (3D) structural information on many length scales is of central importance in biological research. Excellent methods exist to obtain structures of molecules at atomic, organelles at electron microscopic, and tissue at light-microscopic resolution. A gap exists, however, when 3D tissue structure needs to be reconstructed over hundreds of micrometers with a resolution sufficient to follow the thinnest cellular processes and to identify small organelles such as synaptic vesicles. Such 3D data are, however, essential to understand cellular networks that, particularly in the nervous system, need to be completely reconstructed throughout a substantial spatial volume. Here we demonstrate that datasets meeting these requirements can be obtained by automated block-face imaging combined with serial sectioning inside the chamber of a scanning electron microscope. Backscattering contrast is used to visualize the heavy-metal staining of tissue prepared using techniques that are routine for transmission electron microscopy. Low-vacuum (20–60 Pa H2O) conditions prevent charging of the uncoated block face. The resolution is sufficient to trace even the thinnest axons and to identify synapses. Stacks of several hundred sections, 50–70 nm thick, have been obtained at a lateral position jitter of typically under 10 nm. This opens the possibility of automatically obtaining the electron-microscope-level 3D datasets needed to completely reconstruct the connectivity of neuronal circuits. PMID:15514700

  2. New insights into subsurface imaging of carbon nanotubes in polymer composites via scanning electron microscopy

    NASA Astrophysics Data System (ADS)

    Zhao, Minhua; Ming, Bin; Kim, Jae-Woo; Gibbons, Luke J.; Gu, Xiaohong; Nguyen, Tinh; Park, Cheol; Lillehei, Peter T.; Villarrubia, J. S.; Vladár, András E.; Liddle, J. Alexander

    2015-02-01

    Despite many studies of subsurface imaging of carbon nanotube (CNT)-polymer composites via scanning electron microscopy (SEM), significant controversy exists concerning the imaging depth and contrast mechanisms. We studied CNT-polyimide composites and, by three-dimensional reconstructions of captured stereo-pair images, determined that the maximum SEM imaging depth was typically hundreds of nanometers. The contrast mechanisms were investigated over a broad range of beam accelerating voltages from 0.3 to 30 kV, and ascribed to modulation by embedded CNTs of the effective secondary electron (SE) emission yield at the polymer surface. This modulation of the SE yield is due to non-uniform surface potential distribution resulting from current flows due to leakage and electron beam induced current. The importance of an external electric field on SEM subsurface imaging was also demonstrated. The insights gained from this study can be generally applied to SEM nondestructive subsurface imaging of conducting nanostructures embedded in dielectric matrices such as graphene-polymer composites, silicon-based single electron transistors, high resolution SEM overlay metrology or e-beam lithography, and have significant implications in nanotechnology.

  3. Comparative evaluation of topographical data of dental implant surfaces applying optical interferometry and scanning electron microscopy.

    PubMed

    Kournetas, N; Spintzyk, S; Schweizer, E; Sawada, T; Said, F; Schmid, P; Geis-Gerstorfer, J; Eliades, G; Rupp, F

    2017-08-01

    Comparability of topographical data of implant surfaces in literature is low and their clinical relevance often equivocal. The aim of this study was to investigate the ability of scanning electron microscopy and optical interferometry to assess statistically similar 3-dimensional roughness parameter results and to evaluate these data based on predefined criteria regarded relevant for a favorable biological response. Four different commercial dental screw-type implants (NanoTite Certain Prevail, TiUnite Brånemark Mk III, XiVE S Plus and SLA Standard Plus) were analyzed by stereo scanning electron microscopy and white light interferometry. Surface height, spatial and hybrid roughness parameters (Sa, Sz, Ssk, Sku, Sal, Str, Sdr) were assessed from raw and filtered data (Gaussian 50μm and 5μm cut-off-filters), respectively. Data were statistically compared by one-way ANOVA and Tukey-Kramer post-hoc test. For a clinically relevant interpretation, a categorizing evaluation approach was used based on predefined threshold criteria for each roughness parameter. The two methods exhibited predominantly statistical differences. Dependent on roughness parameters and filter settings, both methods showed variations in rankings of the implant surfaces and differed in their ability to discriminate the different topographies. Overall, the analyses revealed scale-dependent roughness data. Compared to the pure statistical approach, the categorizing evaluation resulted in much more similarities between the two methods. This study suggests to reconsider current approaches for the topographical evaluation of implant surfaces and to further seek after proper experimental settings. Furthermore, the specific role of different roughness parameters for the bioresponse has to be studied in detail in order to better define clinically relevant, scale-dependent and parameter-specific thresholds and ranges. Copyright © 2017 The Academy of Dental Materials. Published by Elsevier Ltd. All rights

  4. Evaluation of three different rotary systems during endodontic retreatment - Analysis by scanning electron microscopy

    PubMed Central

    Vidal, Flávia-Teixeira; Nunes, Eduardo; Horta, Martinho-Campolina-Rebello; Freitas, Maria-Rita-Lopes-da Silva

    2016-01-01

    Background Endodontic therapy is considered a series of important and interdependent steps, and failure of any of these steps may compromise the treatment outcome. This study aimed to evaluate the effectiveness of three different rotary systems in removing obturation materials during endodontic retreatment using scanning electron microscopy (SEM) analysis. Material and Methods Thirty-six endodontically treated teeth were selected and divided into 3 groups of 10 and 1 control group with 6 dental elements. The groups were divided according to the rotary system used for removing gutta-percha, as follows: G1: ProTaper system; G2: K3 system; G3: Mtwo system; and G4: Control group. Thereafter, the roots were split and the sections were observed under SEM, for analysis and counting of clear dentinal tubules, creating the variable “degree of dentinal tubule patency” (0: intensely clear; 1: moderately clear; 2: slightly clear; 3: completely blocked). The data were subjected to the Friedman and Kruskal-Wallis statistical tests. Results No differences were observed in the “degree of dentinal tubule patency” neither between the root thirds (to each evaluated group) nor between the groups (to each evaluated third). Nevertheless, when the three root thirds were grouped (providing evaluation of all root extension), the “degree of dentinal tubule patency” was lower in G1 than in G3 (p<0.05), but showed no differences neither between G1 and G2 nor G2 and G3. Conclusions No technique was able to completely remove the canal obturation material, despite G1 having shown better results, although without significant difference to G2 Key words:Scanning electron microscopy, NiTi, retreatment. PMID:27034750

  5. Evaluation of three different rotary systems during endodontic retreatment - Analysis by scanning electron microscopy.

    PubMed

    Vidal, Flávia-Teixeira; Nunes, Eduardo; Horta, Martinho-Campolina-Rebello; Freitas, Maria-Rita-Lopes-da Silva; Silveira, Frank-Ferreira

    2016-04-01

    Endodontic therapy is considered a series of important and interdependent steps, and failure of any of these steps may compromise the treatment outcome. This study aimed to evaluate the effectiveness of three different rotary systems in removing obturation materials during endodontic retreatment using scanning electron microscopy (SEM) analysis. Thirty-six endodontically treated teeth were selected and divided into 3 groups of 10 and 1 control group with 6 dental elements. The groups were divided according to the rotary system used for removing gutta-percha, as follows: G1: ProTaper system; G2: K3 system; G3: Mtwo system; and G4: Control group. Thereafter, the roots were split and the sections were observed under SEM, for analysis and counting of clear dentinal tubules, creating the variable "degree of dentinal tubule patency" (0: intensely clear; 1: moderately clear; 2: slightly clear; 3: completely blocked). The data were subjected to the Friedman and Kruskal-Wallis statistical tests. No differences were observed in the "degree of dentinal tubule patency" neither between the root thirds (to each evaluated group) nor between the groups (to each evaluated third). Nevertheless, when the three root thirds were grouped (providing evaluation of all root extension), the "degree of dentinal tubule patency" was lower in G1 than in G3 (p<0.05), but showed no differences neither between G1 and G2 nor G2 and G3. No technique was able to completely remove the canal obturation material, despite G1 having shown better results, although without significant difference to G2 KEY WORDS:Scanning electron microscopy, NiTi, retreatment.

  6. Thickness determination of few-layer hexagonal boron nitride films by scanning electron microscopy and Auger electron spectroscopy

    SciTech Connect

    Sutter, P. Sutter, E.

    2014-09-01

    We assess scanning electron microscopy (SEM) and Auger electron spectroscopy (AES) for thickness measurements on few-layer hexagonal boron nitride (h-BN), the layered dielectric of choice for integration with graphene and other two-dimensional materials. Observations on h-BN islands with large, atomically flat terraces show that the secondary electron intensity in SEM reflects monolayer height changes in films up to least 10 atomic layers thickness. From a quantitative analysis of AES data, the energy-dependent electron escape depth in h-BN films is deduced. The results show that AES is suitable for absolute thickness measurements of few-layer h-BN of 1 to 6 layers.

  7. The detection and influence of food soils on microorganisms on stainless steel using scanning electron microscopy and epifluorescence microscopy.

    PubMed

    Whitehead, Kathryn A; Smith, Lindsay A; Verran, Joanna

    2010-07-31

    A range of food soils and components (complex [meat extract, fish extract, and cottage cheese extract]; oils [cholesterol, fish oil, and mixed fatty acids]; proteins [bovine serum albumin (BSA), fish peptones, and casein]; and carbohydrates [glycogen, starch, and lactose]) were deposited onto 304 2B finish stainless steel surfaces at different concentrations (10-0.001%). Scanning electron microscopy (SEM) and epifluorescence microscopy were used to visualise the cell and food soil distribution across the surface. Epifluorescence microscopy was also used to quantify the percentage of a field covered by cells or soil. At 10% concentration, most soils, with the exception of BSA and fish peptone were easily visualised using SEM, presenting differences in gross soil morphology and distribution. When soil was stained with acridine orange and visualised by epifluorescence microscopy, the limit of detection of the method varied between soils, but some (meat, cottage cheese and glycogen) were detected at the lowest concentrations used (0.001%). The decrease in soil concentration did not always relate to the surface coverage measurement. When 10% food soil was applied to a surface with Escherichia coli and compared, cell attachment differed depending on the nature of the soil. The highest percentage coverage of cells was observed on surfaces with fish extract and related products (fish peptone and fish oil), followed by carbohydrates, meat extract/meat protein, cottage cheese/casein and the least to the oils (cholesterol and mixed fatty acids). Cells could not be clearly observed in the presence of some food soils using SEM. Findings demonstrate that food soils heterogeneously covered stainless steel surfaces in differing patterns. The pattern and amount of cell attachment was related to food soil type rather than to the amount of food soil detected. This work demonstrates that in the study of conditioning film and cell retention on the hygienic properties of surfaces, SEM

  8. Serial block face scanning electron microscopy for the study of cardiac muscle ultrastructure at nanoscale resolutions.

    PubMed

    Pinali, Christian; Kitmitto, Ashraf

    2014-11-01

    Electron microscopy techniques have made a significant contribution towards understanding muscle physiology since the 1950s. Subsequent advances in hardware and software have led to major breakthroughs in terms of image resolution as well as the ability to generate three-dimensional (3D) data essential for linking structure to function and dysfunction. In this methodological review we consider the application of a relatively new technique, serial block face scanning electron microscopy (SBF-SEM), for the study of cardiac muscle morphology. Employing SBF-SEM we have generated 3D data for cardiac myocytes within the myocardium with a voxel size of ~15 nm in the X-Y plane and 50 nm in the Z-direction. We describe how SBF-SEM can be used in conjunction with selective staining techniques to reveal the 3D cellular organisation and the relationship between the t-tubule (t-t) and sarcoplasmic reticulum (SR) networks. These methods describe how SBF-SEM can be used to provide qualitative data to investigate the organisation of the dyad, a specialised calcium microdomain formed between the t-ts and the junctional portion of the SR (jSR). We further describe how image analysis methods may be applied to interrogate the 3D volumes to provide quantitative data such as the volume of the cell occupied by the t-t and SR membranes and the volumes and surface area of jSR patches. We consider the strengths and weaknesses of the SBF-SEM technique, pitfalls in sample preparation together with tips and methods for image analysis. By providing a 'big picture' view at high resolutions, in comparison to conventional confocal microscopy, SBF-SEM represents a paradigm shift for imaging cellular networks in their native environment.

  9. Candida albicans biofilms: comparative analysis of room-temperature and cryofixation for scanning electron microscopy.

    PubMed

    Vila, T; Fonseca, B B; DA Cunha, M M L; Dos Santos, G R C; Ishida, K; Barreto-Bergter, E; DE Souza, W; Rozental, S

    2017-09-01

    Biofilms are frequently related to invasive fungal infections and are reported to be more resistant to antifungal drugs than planktonic cells. The structural complexity of the biofilm as well as the presence of a polymeric extracellular matrix (ECM) is thought to be associated with this resistant behavior. Scanning electron microscopy (SEM) after room temperature glutaraldehyde-based fixation, have been used to study fungal biofilm structure and drug susceptibility but they usually fail to preserve the ECM and, therefore, are not an optimised methodology to understand the complexity of the fungal biofilm. Thus, in this work, we propose a comparative analysis of room-temperature and cryofixation/freeze substitution of Candida albicans biofilms for SEM observation. Our experiments showed that room-temperature fixative protocols using glutaraldehyde and osmium tetroxide prior to alcohol dehydration led to a complete extraction of the polymeric ECM of biofilms. ECM from fixative and alcohol solutions were recovered after all processing steps and these structures were characterised by biochemistry assays, transmission electron microscopy and mass spectrometry. Cryofixation techniques followed by freeze-substitution lead to a great preservation of both ECM structure and C. albicans biofilm cells, allowing the visualisation of a more reliable biofilm structure. These findings reinforce that cryofixation should be the indicated method for SEM sample preparation to study fungal biofilms as it allows the visualisation of the EMC and the exploration of the biofilm structure to its fullest, as its structural/functional role in interaction with host cells, other pathogens and for drug resistance assays. © 2017 The Authors Journal of Microscopy © 2017 Royal Microscopical Society.

  10. An endolithic microbial community in dolomite rock in central Switzerland: characterization by reflection spectroscopy, pigment analyses, scanning electron microscopy, and laser scanning microscopy.

    PubMed

    Horath, T; Neu, T R; Bachofen, R

    2006-04-01

    A community of endolithic microorganisms dominated by phototrophs was found as a distinct band a few millimeters below the surface of bare exposed dolomite rocks in the Piora Valley in the Alps. Using in situ reflectance spectroscopy, we detected chlorophyll a (Chl a), phycobilins, carotenoids, and an unknown type of bacteriochlorophyll-like pigment absorbing in vivo at about 720 nm. In cross sections, the data indicated a defined distribution of different groups of organisms perpendicular to the rock surface. High-performance liquid chromatography analyses of pigments extracted with organic solvents confirmed the presence of two types of bacteriochlorophylls besides chlorophylls and various carotenoids. Spherical organisms of varying sizes and small filaments were observed in situ with scanning electron microscopy and confocal laser scanning microscopy (one- and two-photon technique). The latter allowed visualization of the distribution of phototrophic microorganisms by the autofluorescence of their pigments within the rock. Coccoid cyanobacteria of various sizes predominated over filamentous ones. Application of fluorescence-labeled lectins demonstrated that most cyanobacteria were embedded in an exopolymeric matrix. Nucleic acid stains revealed a wide distribution of small heterotrophs. Some biological structures emitting a green autofluorescence remain to be identified.

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

    SciTech Connect

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

    2012-06-04

    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.

  12. Direct imaging of defect formation in strained organic flexible electronics by Scanning Kelvin Probe Microscopy

    PubMed Central

    Cramer, Tobias; Travaglini, Lorenzo; Lai, Stefano; Patruno, Luca; de Miranda, Stefano; Bonfiglio, Annalisa; Cosseddu, Piero; Fraboni, Beatrice

    2016-01-01

    The development of new materials and devices for flexible electronics depends crucially on the understanding of how strain affects electronic material properties at the nano-scale. Scanning Kelvin-Probe Microscopy (SKPM) is a unique technique for nanoelectronic investigations as it combines non-invasive measurement of surface topography and surface electrical potential. Here we show that SKPM in non-contact mode is feasible on deformed flexible samples and allows to identify strain induced electronic defects. As an example we apply the technique to investigate the strain response of organic thin film transistors containing TIPS-pentacene patterned on polymer foils. Controlled surface strain is induced in the semiconducting layer by bending the transistor substrate. The amount of local strain is quantified by a mathematical model describing the bending mechanics. We find that the step-wise reduction of device performance at critical bending radii is caused by the formation of nano-cracks in the microcrystal morphology of the TIPS-pentacene film. The cracks are easily identified due to the abrupt variation in SKPM surface potential caused by a local increase in resistance. Importantly, the strong surface adhesion of microcrystals to the elastic dielectric allows to maintain a conductive path also after fracture thus providing the opportunity to attenuate strain effects. PMID:27910889

  13. Direct imaging of defect formation in strained organic flexible electronics by Scanning Kelvin Probe Microscopy.

    PubMed

    Cramer, Tobias; Travaglini, Lorenzo; Lai, Stefano; Patruno, Luca; de Miranda, Stefano; Bonfiglio, Annalisa; Cosseddu, Piero; Fraboni, Beatrice

    2016-12-02

    The development of new materials and devices for flexible electronics depends crucially on the understanding of how strain affects electronic material properties at the nano-scale. Scanning Kelvin-Probe Microscopy (SKPM) is a unique technique for nanoelectronic investigations as it combines non-invasive measurement of surface topography and surface electrical potential. Here we show that SKPM in non-contact mode is feasible on deformed flexible samples and allows to identify strain induced electronic defects. As an example we apply the technique to investigate the strain response of organic thin film transistors containing TIPS-pentacene patterned on polymer foils. Controlled surface strain is induced in the semiconducting layer by bending the transistor substrate. The amount of local strain is quantified by a mathematical model describing the bending mechanics. We find that the step-wise reduction of device performance at critical bending radii is caused by the formation of nano-cracks in the microcrystal morphology of the TIPS-pentacene film. The cracks are easily identified due to the abrupt variation in SKPM surface potential caused by a local increase in resistance. Importantly, the strong surface adhesion of microcrystals to the elastic dielectric allows to maintain a conductive path also after fracture thus providing the opportunity to attenuate strain effects.

  14. Scanning electron acoustic microscopy of residual stresses in ceramics - Theory and experiment

    NASA Technical Reports Server (NTRS)

    Cantrell, John H.; Qian, Menglu

    1992-01-01

    The paper presents a three-dimensional mathematical model of signal generation and contrast in brittle materials and uses the model to simulate the effect of residual stress fields on the scanning electron acoustic microscopy (SEAM)-generated electron acoustic signal. According to the model, a positive (tensile) strain produces an increase in the output signal, whereas a negative (compressive) strain produces a decrease in the ouput signal. Dark field contrast conditions occur at a chopping frequency at which V2 - V1 is greater than 0 (where V2 = V is the SEAM output in a region of residual stresses, and V1 is the output in a stress-free region of the sample). Under ideal conditions (maximum contrast) V1 approaches zero. It was found that tensile strains of the order 0.2-0.3 percent, possible in brittle materials, would produce a variation of the acoustic output signal of the order 10 nV (about 1 percent), well within the image contrast and signal processing capability of the SEAM electronics.

  15. Atomic-scale imaging and spectroscopy for in situ liquid scanning transmission electron microscopy.

    PubMed

    Jungjohann, Katherine L; Evans, James E; Aguiar, Jeffery A; Arslan, Ilke; Browning, Nigel D

    2012-06-01

    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. 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 that 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.

  16. Direct imaging of defect formation in strained organic flexible electronics by Scanning Kelvin Probe Microscopy

    NASA Astrophysics Data System (ADS)

    Cramer, Tobias; Travaglini, Lorenzo; Lai, Stefano; Patruno, Luca; de Miranda, Stefano; Bonfiglio, Annalisa; Cosseddu, Piero; Fraboni, Beatrice

    2016-12-01

    The development of new materials and devices for flexible electronics depends crucially on the understanding of how strain affects electronic material properties at the nano-scale. Scanning Kelvin-Probe Microscopy (SKPM) is a unique technique for nanoelectronic investigations as it combines non-invasive measurement of surface topography and surface electrical potential. Here we show that SKPM in non-contact mode is feasible on deformed flexible samples and allows to identify strain induced electronic defects. As an example we apply the technique to investigate the strain response of organic thin film transistors containing TIPS-pentacene patterned on polymer foils. Controlled surface strain is induced in the semiconducting layer by bending the transistor substrate. The amount of local strain is quantified by a mathematical model describing the bending mechanics. We find that the step-wise reduction of device performance at critical bending radii is caused by the formation of nano-cracks in the microcrystal morphology of the TIPS-pentacene film. The cracks are easily identified due to the abrupt variation in SKPM surface potential caused by a local increase in resistance. Importantly, the strong surface adhesion of microcrystals to the elastic dielectric allows to maintain a conductive path also after fracture thus providing the opportunity to attenuate strain effects.

  17. Scanning electron acoustic microscopy of residual stresses in ceramics - Theory and experiment

    NASA Technical Reports Server (NTRS)

    Cantrell, John H.; Qian, Menglu

    1992-01-01

    The paper presents a three-dimensional mathematical model of signal generation and contrast in brittle materials and uses the model to simulate the effect of residual stress fields on the scanning electron acoustic microscopy (SEAM)-generated electron acoustic signal. According to the model, a positive (tensile) strain produces an increase in the output signal, whereas a negative (compressive) strain produces a decrease in the ouput signal. Dark field contrast conditions occur at a chopping frequency at which V2 - V1 is greater than 0 (where V2 = V is the SEAM output in a region of residual stresses, and V1 is the output in a stress-free region of the sample). Under ideal conditions (maximum contrast) V1 approaches zero. It was found that tensile strains of the order 0.2-0.3 percent, possible in brittle materials, would produce a variation of the acoustic output signal of the order 10 nV (about 1 percent), well within the image contrast and signal processing capability of the SEAM electronics.

  18. Analysis of Multilayer Devices for Superconducting Electronics by High-Resolution Scanning Transmission Electron Microscopy and Energy Dispersive Spectroscopy

    DOE PAGES

    Missert, Nancy; Kotula, Paul G.; Rye, Michael; ...

    2017-02-15

    We used a focused ion beam to obtain cross-sectional specimens from both magnetic multilayer and Nb/Al-AlOx/Nb Josephson junction devices for characterization by scanning transmission electron microscopy (STEM) and energy dispersive X-ray spectroscopy (EDX). An automated multivariate statistical analysis of the EDX spectral images produced chemically unique component images of individual layers within the multilayer structures. STEM imaging elucidated distinct variations in film morphology, interface quality, and/or etch artifacts that could be correlated to magnetic and/or electrical properties measured on the same devices.

  19. Snow crystal imaging using scanning electron microscopy: III. Glacier ice, snow and biota

    USGS Publications Warehouse

    Rango, A.; Wergin, W.P.; Erbe, E.F.; Josberger, E.G.

    2000-01-01

    Low-temperature scanning electron microscopy (SEM) was used to observe metamorphosed snow, glacial firn, and glacial ice obtained from South Cascade Glacier in Washington State, USA. Biotic samples consisting of algae (Chlamydomonas nivalis) and ice worms (a species of oligochaetes) were also collected and imaged. In the field, the snow and biological samples were mounted on copper plates, cooled in liquid nitrogen, and stored in dry shipping containers which maintain a temperature of -196??C. The firn and glacier ice samples were obtained by extracting horizontal ice cores, 8 mm in diameter, at different levels from larger standard glaciological (vertical) ice cores 7.5 cm in diameter. These samples were cooled in liquid nitrogen and placed in cryotubes, were stored in the same dry shipping container, and sent to the SEM facility. In the laboratory, the samples were sputter coated with platinum and imaged by a low-temperature SEM. To image the firn and glacier ice samples, the cores were fractured in liquid nitrogen, attached to a specimen holder, and then imaged. While light microscope images of snow and ice are difficult to interpret because of internal reflection and refraction, the SEM images provide a clear and unique view of the surface of the samples because they are generated from electrons emitted or reflected only from the surface of the sample. In addition, the SEM has a great depth of field with a wide range of magnifying capabilities. The resulting images clearly show the individual grains of the seasonal snowpack and the bonding between the snow grains. Images of firn show individual ice crystals, the bonding between the crystals, and connected air spaces. Images of glacier ice show a crystal structure on a scale of 1-2 mm which is considerably smaller than the expected crystal size. Microscopic air bubbles, less than 15 ??m in diameter, clearly marked the boundaries between these crystal-like features. The life forms associated with the glacier were

  20. Regenerating titanium ventricular assist device surfaces after gold/palladium coating for scanning electron microscopy.

    PubMed

    Achneck, Hardean E; Serpe, Michael J; Jamiolkowski, Ryan M; Eibest, Leslie M; Craig, Stephen L; Lawson, Jeffrey H

    2010-01-01

    Titanium is one of the most commonly used materials for implantable devices in humans. Scanning electron microscopy (SEM) serves as an important tool for imaging titanium surfaces and analyzing cells and other organic matter adhering to titanium implants. However, high-vacuum SEM imaging of a nonconductive sample requires a conductive coating on the surface. A gold/palladium coating is commonly used and to date no method has been described to "clean" such gold/palladium covered surfaces for repeated experiments without etching the titanium itself. This constitutes a major problem with titanium-based implantable devices which are very expensive and thus in short supply. Our objective was to devise a protocol to regenerate titaniumsurfaces after SEM analysis. In a series of experiments, titanium samples from implantable cardiac assist devices were coated with fibronectin, seeded with cells and then coated with gold/palladium for SEM analysis. X-ray photoelectron spectroscopy spectra were obtained before and after five different cleaning protocols. Treatment with aqua regia (a 1:3 solution of concentrated nitric and hydrochloric acid), with or without ozonolysis, followed by sonication in soap solution and sonication in deionized water, allowed regenerating titanium surfaces to their original state. Atomic force microscopy confirmed that the established protocol did not alter the titanium microstructure. The protocol described herein is applicable to almost all titanium surfaces used in biomedical sciences and because of its short exposure time to aqua regia, will likely work for many titanium alloys as well. (c) 2009 Wiley-Liss, Inc.

  1. Multi-resolution correlative focused ion beam scanning electron microscopy: applications to cell biology.

    PubMed

    Narayan, Kedar; Danielson, Cindy M; Lagarec, Ken; Lowekamp, Bradley C; Coffman, Phil; Laquerre, Alexandre; Phaneuf, Michael W; Hope, Thomas J; Subramaniam, Sriram

    2014-03-01

    Efficient correlative imaging of small targets within large fields is a central problem in cell biology. Here, we demonstrate a series of technical advances in focused ion beam scanning electron microscopy (FIB-SEM) to address this issue. We report increases in the speed, robustness and automation of the process, and achieve consistent z slice thickness of ∼3 nm. We introduce "keyframe imaging" as a new approach to simultaneously image large fields of view and obtain high-resolution 3D images of targeted sub-volumes. We demonstrate application of these advances to image post-fusion cytoplasmic intermediates of the HIV core. Using fluorescently labeled cell membranes, proteins and HIV cores, we first produce a "target map" of an HIV infected cell by fluorescence microscopy. We then generate a correlated 3D EM volume of the entire cell as well as high-resolution 3D images of individual HIV cores, achieving correlative imaging across a volume scale of 10(9) in a single automated experimental run.

  2. Raman spectroscopy and scanning electron microscopy characterizations of fission track method datable zircon grains.

    PubMed

    Resende, Rosana Silveira; Sáenz, Carlos Alberto Tello; Curvo, Eduardo Augusto Campos; Constantino, Carlos José Leopoldo; Aroca, Ricardo F; Nakasuga, Wagner Massayuki

    2014-01-01

    Spectroscopic and morphological studies, designed to improve our understanding of the physicochemical phenomena that occur during zircon crystallization, are presented. The zircon fission track method (ZFTM) is used routinely in various laboratories around the world; however, there are some methodological difficulties needing attention. Depending on the surface fission track density observed under an optical microscope, the zircon grain surfaces are classified as homogeneous, heterogeneous, hybrid, or anomalous. In this study, zircon grain surfaces are characterized using complementary techniques such as optical microscopy (OM), micro-Raman spectroscopy, and scanning electron microscopy (SEM), both before and after chemical etching. Our results suggest that anomalous grains have subfamilies and that etching anisotropy related to heterogeneous grains is due to different crystallographic faces within the same polished surface that cannot be observed under an optical microscope. The improved methodology was used to determine the zircon fission track ages of samples collected from the Bauru Group located in the north of Paraná Basin, Brazil. A total of 514 zircon grains were analyzed, consisting of 10% homogeneous, about 10% heterogeneous, about 20% hybrid, and 60% anomalous grains. These results show that the age distributions obtained for homogeneous, heterogeneous, and hybrid grains are both statistically and geologically compatible.

  3. Microwave irradiation for shortening the processing time of samples of flagellated bacteria for scanning electron microscopy.

    PubMed

    Hernández-Chavarría, Francisco

    2004-01-01

    Microwave irradiation (MWI) has been applied to the development of rapid methods to process biological samples for scanning electron microscopy (SEM). In this paper we propose two simple and quick techniques for processing bacteria (Proteus mirabilis and Vibrio mimicus) for SEM using MWI. In the simplest methodology, the bacteria were placed on a cover-glass, air-dried, and submitted to conductivity stain. The reagent used for the conductivity stain was the mordant of a light microscopy staining method (10 ml of 5% carbolic acid solution, 2 g of tannic acid, and 10 ml of saturated aluminum sulfate 12-H2O). In the second method the samples were double fixed (glutaraldehyde and then osmium), submitted to conductivity stain, dehydrated through a series of ethanol solutions of increasing concentration, treated with hexamethyldisilazine (HMDS), and dried at 35 degrees C for 5 minutes. In both methods the steps from fixation to treatment with HMDS were done under MWI for 2 minutes in an ice-water bath, in order to dissipate the heat generated by the MWI. Although both techniques preserve bacterial morphology adequately, the latter, technique showed the best preservation, including the appearance of flagella, and that process was completed in less than 2 hours at temperatures of MWI between 4 to 5 degrees C.

  4. Vasculature of the ophthalmic rete in night herons (Nycticorax nycticorax): scanning electron microscopy of corrosion casts.

    PubMed

    Ninomiya, Hiroyoshi

    2002-09-01

    Vasculature of the ophthalmic rete (rete ophthalmicum) in the night heron (Nycticorax nycticorax) was studied using scanning electron microscopy of vascular corrosion casts and light microscopy on tissue sections. Most blood to the eyeball and a lesser volume of blood to the brain passed through the ophthalmic rete via the external ophthalmic artery. The collateral retial arterioles originated from the external ophthalmic artery forming a flat and fusiform-shaped arterial network at the ventrotemporal region of the eyeball. The arterial network was intermixed with a similar complex of the veins from the eye. The ophthalmotemporal artery, which supplied the eyeball posteriorly, and supraorbital and infraorbital arteries, which supplied the eyeball anteriorly, originated from the rete. Blood from the eye, which is a site of potential heat loss, drained into the ophthalmic rete via the ophthalmotemporal vein. On the casts of retial arterioles, slit-like cleavages at branching sites representing flap valves, which might play a role as sluice valves, were seen. In addition, marks of circularly running grooves, which might represent tufts of smooth muscle cells and might contribute to a sphincter activity, were observed. These anatomical specializations of the avian ophthalmic rete, involving parallel arrangement of arteries and veins, may function to facilitate counter-current heat exchange and to regulate blood pressure and volume to the eye and the brain.

  5. Scanning electron microscopy determination of string mozzarella cheese in gastric contents.

    PubMed

    Platek, S F; Crowe, J B; Ranieri, N; Wolnik, K A

    2001-01-01

    As part of a suspected homicide investigation, a sampling of the gastric contents from the victim was forwarded to the U.S. Food and Drug Administration's Forensic Chemistry Center (FCC) for analysis of specific, selected components. The victim was known to have consumed string mozzarella cheese, as a snack, less than 24 h before his disappearance and the subsequent discovery of the body. The investigation sought to confirm or dismiss speculation the victim may have been fed a meal or eaten additional food prior to his death. Analysis of the stomach contents involved examination by stereoscopic light microscopy (SLM) and isolation, processing, and analysis of suspect materials by scanning electron microscopy (SEM). Several wax-like, off-white to cream-colored objects were noted by SLM examination and removed from the gastric contents. Through a series of fixation, sectioning, drying, and coating steps, these objects were prepared for analysis by SEM. Comparison of the suspect material with laboratory control string mozzarella cheese showed excellent correlation between the analyzed samples, confirming the suspect material from the stomach contents as string mozzarella cheese.

  6. Chromosomes and karyotype analysis of a liver fluke, Opisthorchis viverrini, by scanning electron microscopy.

    PubMed

    Kaewkong, Worasak; Choochote, Wej; Kanla, Pipatpong; Maleewong, Wanchai; Intapan, Pewpan M; Wongkham, Sopit; Wongkham, Chaisiri

    2012-09-01

    Opisthorchis viverrini, a human liver fluke, has been categorized as the carcinogenic organism according to the strong association with carcinogenesis of cholangiocarcinoma (CCA). The infection of this food-borne parasite is a major impact on the health of humans, especially CCA patients in the northeast of Thailand. Taxonomy, morphology, epidemiology and molecular study of O. viverrini have been publicized increasingly but the precise karyotypic study is still incomplete. In this study, the chromosomes of O. viverrini were prepared from the testes of adult worms retrieved from metacercariae infected-hamsters. The chromosomes of O. viverrini were identified in haploid (n=6) meiotic metaphase and in diploid (2n=12) mitotic metaphase by light microscopy. The chromosome number, length and nomenclature of each chromosome were determined by scanning electron microscopy. The six chromosomes consist of one large-sized metacentric, one medium-sized metacentric, two small-sized metacentric, one small-sized submetacentric and one small-sized acrocentric chromosomes with the lengths of 2.84±0.03, 2.12±0.10, 1.71±0.13, 1.44±0.04, 1.23±0.03 and 0.84±0.13 μm, respectively. This is the first karyotype analysis of O. viverrini with defined complete nomenclature.

  7. Multi-resolution Correlative Focused Ion Beam Scanning Electron Microscopy: Applications to Cell Biology

    PubMed Central

    Narayan, Kedar; Danielson, Cindy M; Lagarec, Ken; Lowekamp, Bradley C; Coffman, Phil; Laquerre, Alexandre; Phaneuf, Michael W; Hope, Thomas J; Subramaniam, Sriram

    2014-01-01

    Efficient correlative imaging of small targets within large fields is a central problem in cell biology. Here, we demonstrate a series of technical advances in focused ion beam scanning electron microscopy (FIB-SEM) to address this issue. We increase the speed, robustness and automation of the process, and achieve consistent z slice thickness of ~3 nm. We introduce “keyframe imaging” to simultaneously image large fields of view and obtain high-resolution 3D images of targeted sub-volumes. We apply these advances to image post-fusion cytoplasmic intermediates of the HIV core. Using fluorescently labeled cell membranes, proteins and HIV cores, we first produce a “target map” of an HIV infected cell by fluorescence microscopy. We then generate a correlated 3D EM volume of the entire cell as well as high-resolution 3D images of individual HIV cores, achieving correlative imaging across a volume scale of 109 in a single automated experimental run. PMID:24300554

  8. Scanning electron microscopy of antennal sensory organs of the cattle grub, Hypoderma lineatum (Diptera: Oestridae).

    PubMed

    Li, X Y; Liu, X H; Ge, Y Q; Zhang, D

    2015-10-01

    Hypoderma lineatum (Villers, 1789) (Diptera: Oestridae) is a hypodermosis fly that has resulted in great economic losses worldwide. The antennae of cattle grub males and females were examined through stereoscopic microscopy and scanning electron microscopy to reveal the general morphology, combined with distribution, type, size, and ultrastructure of the antennal sensilla. All of the three antennal segments (antennal scape, pedicel, and funiculus) possess microtrichiae on their surface. Mechanoreceptors only exist on the antennal scape and pedicel. The antennal funiculus presents four types of antennal sensilla: trichoid, basiconic, coeloconic, and clavate sensilla. Three distinctive characters of H. lineatum are obvious: (1) the relatively slender, flexible, and equal-height mechanoreceptors; (2) the enlarged antennal pedicel, and numerous antennal sensory pits and pit sensilla on the antennal funiculus; and (3) all types of antennal sensilla clustered in sensory pits, respectively. Additionally, the enlarged antennal pedicel and abundant sensory pits and pit sensilla might facilitate odor detection, enhance olfactory sensitivity and accuracy, and also protect the fragile antennal sensilla from mechanical irritation or damage.

  9. Extended Depth of Field for High-Resolution Scanning Transmission Electron Microscopy

    SciTech Connect

    Hovden, Robert; Xin, Huolin L.; Muller, David A.

    2010-12-02

    Aberration-corrected scanning transmission electron microscopes (STEMs) provide sub-Angstrom lateral resolution; however, the large convergence angle greatly reduces the depth of field. For microscopes with a small depth of field, information outside of the focal plane quickly becomes blurred and less defined. It may not be possible to image some samples entirely in focus. Extended depth-of-field techniques, however, allow a single image, with all areas in focus, to be extracted from a series of images focused at a range of depths. In recent years, a variety of algorithmic approaches have been employed for bright-field optical microscopy. Here, we demonstrate that some established optical microscopy methods can also be applied to extend the ~6 nm depth of focus of a 100 kV 5th-order aberration-corrected STEM (α{sub max} = 33 mrad) to image Pt-Co nanoparticles on a thick vulcanized carbon support. These techniques allow us to automatically obtain a single image with all the particles in focus as well as a complimentary topography map.

  10. Extended depth of field for high-resolution scanning transmission electron microscopy.

    PubMed

    Hovden, Robert; Xin, Huolin L; Muller, David A

    2011-02-01

    Aberration-corrected scanning transmission electron microscopes (STEMs) provide sub-Angstrom lateral resolution; however, the large convergence angle greatly reduces the depth of field. For microscopes with a small depth of field, information outside of the focal plane quickly becomes blurred and less defined. It may not be possible to image some samples entirely in focus. Extended depth-of-field techniques, however, allow a single image, with all areas in focus, to be extracted from a series of images focused at a range of depths. In recent years, a variety of algorithmic approaches have been employed for bright-field optical microscopy. Here, we demonstrate that some established optical microscopy methods can also be applied to extend the ∼ 6 nm depth of focus of a 100 kV 5th-order aberration-corrected STEM (α max = 33 mrad) to image Pt-Co nanoparticles on a thick vulcanized carbon support. These techniques allow us to automatically obtain a single image with all the particles in focus as well as a complimentary topography map.

  11. Local sample thickness determination via scanning transmission electron microscopy defocus series.

    PubMed

    Beyer, A; Straubinger, R; Belz, J; Volz, K

    2016-05-01

    The usable aperture sizes in (scanning) transmission electron microscopy ((S)TEM) have significantly increased in the past decade due to the introduction of aberration correction. In parallel with the consequent increase of convergence angle the depth of focus has decreased severely and optical sectioning in the STEM became feasible. Here we apply STEM defocus series to derive the local sample thickness of a TEM sample. To this end experimental as well as simulated defocus series of thin Si foils were acquired. The systematic blurring of high resolution high angle annular dark field images is quantified by evaluating the standard deviation of the image intensity for each image of a defocus series. The derived dependencies exhibit a pronounced maximum at the optimum defocus and drop to a background value for higher or lower values. The full width half maximum (FWHM) of the curve is equal to the sample thickness above a minimum thickness given by the size of the used aperture and the chromatic aberration of the microscope. The thicknesses obtained from experimental defocus series applying the proposed method are in good agreement with the values derived from other established methods. The key advantages of this method compared to others are its high spatial resolution and that it does not involve any time consuming simulations. © 2015 The Authors Journal of Microscopy © 2015 Royal Microscopical Society.

  12. Subsurface examination of a foliar biofilm using scanning electron- and focused-ion-beam microscopy

    USDA-ARS?s Scientific Manuscript database

    The dual beam scanning electron microscope, equipped with both a focused ion- and scanning electron- beam (FIB SEM) is a novel tool for the exploration of the subsurface structure of biological tissues. The FIB is capable of removing small cross sections to view the subsurface features and may be s...

  13. Revolving scanning transmission electron microscopy: correcting sample drift distortion without prior knowledge.

    PubMed

    Sang, Xiahan; LeBeau, James M

    2014-03-01

    We report the development of revolving scanning transmission electron microscopy--RevSTEM--a technique that enables characterization and removal of sample drift distortion from atomic resolution images without the need for a priori crystal structure information. To measure and correct the distortion, we acquire an image series while rotating the scan coordinate system between successive frames. Through theory and experiment, we show that the revolving image series captures the information necessary to analyze sample drift rate and direction. At atomic resolution, we quantify the image distortion using the projective standard deviation, a rapid, real-space method to directly measure lattice vector angles. By fitting these angles to a physical model, we show that the refined drift parameters provide the input needed to correct distortion across the series. We demonstrate that RevSTEM simultaneously removes the need for a priori structure information to correct distortion, leads to a dramatically improved signal-to-noise ratio, and enables picometer precision and accuracy regardless of drift rate. Copyright © 2013 Elsevier B.V. All rights reserved.

  14. Direct Visualization of Local Electromagnetic Field Structures by Scanning Transmission Electron Microscopy.

    PubMed

    Shibata, Naoya; Findlay, Scott D; Matsumoto, Takao; Kohno, Yuji; Seki, Takehito; Sánchez-Santolino, Gabriel; Ikuhara, Yuichi

    2017-07-18

    The functional properties of materials and devices are critically determined by the electromagnetic field structures formed inside them, especially at nanointerface and surface regions, because such structures are strongly associated with the dynamics of electrons, holes and ions. To understand the fundamental origin of many exotic properties in modern materials and devices, it is essential to directly characterize local electromagnetic field structures at such defect regions, even down to atomic dimensions. In recent years, rapid progress in the development of high-speed area detectors for aberration-corrected scanning transmission electron microscopy (STEM) with sub-angstrom spatial resolution has opened new possibilities to directly image such electromagnetic field structures at very high-resolution. In this Account, we give an overview of our recent development of differential phase contrast (DPC) microscopy for aberration-corrected STEM and its application to many materials problems. In recent years, we have developed segmented-type STEM detectors which divide the detector plane into 16 segments and enable simultaneous imaging of 16 STEM images which are sensitive to the positions and angles of transmitted/scattered electrons on the detector plane. These detectors also have atomic-resolution imaging capability. Using these segmented-type STEM detectors, we show DPC STEM imaging to be a very powerful tool for directly imaging local electromagnetic field structures in materials and devices in real space. For example, DPC STEM can clearly visualize the local electric field variation due to the abrupt potential change across a p-n junction in a GaAs semiconductor, which cannot be observed by normal in-focus bright-field or annular type dark-field STEM imaging modes. DPC STEM is also very effective for imaging magnetic field structures in magnetic materials, such as magnetic domains and skyrmions. Moreover, real-time imaging of electromagnetic field structures can

  15. Persistence of spermatozoa on decomposing human skin: a scanning electron microscopy study.

    PubMed

    Gibelli, D; Mazzarelli, D; Rizzi, A; Kustermann, A; Cattaneo, C

    2013-09-01

    Finding spermatozoa is of the utmost importance in judicial cases involving both the living and the dead; however, most of literature actually deals with inner genitalia and does not take into consideration the chance of external deposition of semen on skin, which is not rare. In addition, the most advanced microscopic technologies such as scanning electron microscopy (SEM) have not been thoroughly investigated within this specific field of research. This study aims at applying SEM analysis to samples of decomposed skin in order to test its potential in detecting spermatozoa particularly in decomposed cadavers. A sample of skin was obtained at autopsy and divided into two thin strips; one of the samples was used as a negative control. Semen was then taken from a "donor" (with a normal spermiogram) and was spread onto the other skin sample. Every 3 days for the first 15 days (for a total of six samples), a standard slide was prepared from swabs on the treated and control skin and analyzed by standard light microscopy. In addition, every 7 days up to 91 days (3 months circa), a skin sample was taken from the positive and negative control and examined by SEM for a total of 14 samples. Results show that after 12 days, light microscopy failed in detecting spermatozoa, whereas they were still visible up to 84 days by SEM analysis. This study therefore suggests the persistence of sperm structures in time and in decomposing material as well as the possible application of SEM technology to decomposed skin in order to detect semen.

  16. Semiquantitative analysis by scanning electron microscopy of cochlear hair cell damage by ototoxic drugs.

    PubMed

    Saito, T; Manabe, Y; Honda, N; Yamada, T; Yamamoto, T; Saito, H

    1995-03-01

    The ototoxicity of cisplatin and carboplatin in the organ of Corti of the guinea pig was evaluated semiquantitatively. Damage of the stereocilia of outer hair cells (OHCs) observed by scanning electron microscopy (SEM) was classified into normal, grade 1 (10-50% loss of stereocilia), grade 2 (less than 50% remaining stereocilia), or grade 3 (missing stereocilia). The OHCs observed by light microscopy (LM) were classified as remaining or missing cells. Fifty OHCs of each row in the middle part of each turn of the cochlea were counted (a total of 150 cells per turn). Guinea pigs were administered 5 mg/kg of cisplatin or 50 mg/kg of carboplatin intraperitoneally for three consecutive days. In groups 1 and 2, in which both cochlea were fixed in 2.5% glutaraldehyde and 1% osmium tetroxide (OsO4) and observed by SEM, the percentages of damage of the OHC stereocilia were similar in each cochlear turn bilaterally. In group 3, the right cochleae were fixed in OsO4 and observed by phase contrast microscopy as surface preparations. Left cochleae were submitted for SEM observation. Missing and grade 3 cells were observed at similar percentages in each row of each turn. In group 4, succinate dehydrogenase staining was performed in the right cochleae and observed by LM. The degree of damage in the right cochleae was compared with that of the left cochleae which was observed by SEM. On average, the mean numbers of missing cells and cells showing grade 3 damage were similar in each row of each turn. From these similarities of evaluation of ototoxicity at LM and SEM levels, it was concluded that semiquantitative analysis by SEM only is appropriate for the assessment of ototoxicity.

  17. Comparison of macroscopic and microscopic (stereomicroscopy and scanning electron microscopy) features of bone lesions due to hatchet hacking trauma.

    PubMed

    Nogueira, Luísa; Quatrehomme, Gérald; Bertrand, Marie-France; Rallon, Christophe; Ceinos, Romain; du Jardin, Philippe; Adalian, Pascal; Alunni, Véronique

    2017-03-01

    This experimental study examined the lesions produced by a hatchet on human bones (tibiae). A total of 30 lesions were produced and examined macroscopically (naked eye) and by stereomicroscopy. 13 of them were also analyzed using scanning electron microscopy. The general shape of the lesion, both edges, both walls, the kerf floor and the extremities were described. The length and maximum width of the lesions were also recorded. The microscopic analysis of the lesions led to the description of a sharp-blunt mechanism. Specific criteria were identified (lateral pushing back, fragmentation of the upraising, fossa dug laterally to the edge and vertical striae) enabling the forensic expert to conclude that a hacking instrument was used. These criteria are easily identifiable using scanning electron microscopy, but can also be observed with stereomicroscopy. Overall, lateral pushing back and vertical striae visible using stereomicroscopy and scanning electron microscopy signal the use of a hacking tool.

  18. Composition analysis of coaxially grown InGaN multi quantum wells using scanning transmission electron microscopy

    SciTech Connect

    Aschenbrenner, T. Schowalter, M.; Mehrtens, T.; Müller-Caspary, K.; Rosenauer, A.; Fikry, M.; Heinz, D.; Scholz, F.; Tischer, I.; Madel, M.; Thonke, K.

    2016-05-07

    GaN nanotubes with coaxial InGaN quantum wells were analyzed by scanning transmission electron microscopy in order to determine their structural properties as well as the indium distribution across the InGaN quantum wells. For the latter, two process steps are necessary. First, a technique to prepare cross-sectional slices out of the nanotubes has been developed. Second, an existing scanning transmission electron microscopy analysis technique has been extended with respect to the special crystallographic orientation of this type of specimen. In particular, the shape of the nanotubes, their defect structure, and the incorporation of indium on different facets were investigated. The quantum wells preferentially grow on m-planes of the dodecagonally shaped nanotubes and on semipolar top facets while no significant indium signal was found on a-planes. An averaged indium concentration of 6% to 7% was found by scanning transmission electron microscopy analysis and could be confirmed by cathodoluminescence measurements.

  19. Towards the imaging of Weibel-Palade body biogenesis by serial block face-scanning electron microscopy.

    PubMed

    Mourik, M J; Faas, F G A; Zimmermann, H; Eikenboom, J; Koster, A J

    2015-08-01

    Electron microscopy is used in biological research to study the ultrastructure at high resolution to obtain information on specific cellular processes. Serial block face-scanning electron microscopy is a relatively novel electron microscopy imaging technique that allows three-dimensional characterization of the ultrastructure in both tissues and cells by measuring volumes of thousands of cubic micrometres yet at nanometre-scale resolution. In the scanning electron microscope, repeatedly an image is acquired followed by the removal of a thin layer resin embedded biological material by either a microtome or a focused ion beam. In this way, each recorded image contains novel structural information which can be used for three-dimensional analysis. Here, we explore focused ion beam facilitated serial block face-scanning electron microscopy to study the endothelial cell-specific storage organelles, the Weibel-Palade bodies, during their biogenesis at the Golgi apparatus. Weibel-Palade bodies predominantly contain the coagulation protein Von Willebrand factor which is secreted by the cell upon vascular damage. Using focused ion beam facilitated serial block face-scanning electron microscopy we show that the technique has the sensitivity to clearly reveal subcellular details like mitochondrial cristae and small vesicles with a diameter of about 50 nm. Also, we reveal numerous associations between Weibel-Palade bodies and Golgi stacks which became conceivable in large-scale three-dimensional data. We demonstrate that serial block face-scanning electron microscopy is a promising tool that offers an alternative for electron tomography to study subcellular organelle interactions in the context of a complete cell.

  20. Towards the imaging of Weibel–Palade body biogenesis by serial block face-scanning electron microscopy

    PubMed Central

    Mourik, MJ; Faas, FGA; Zimmermann, H; Eikenboom, J; Koster, AJ

    2015-01-01

    Electron microscopy is used in biological research to study the ultrastructure at high resolution to obtain information on specific cellular processes. Serial block face-scanning electron microscopy is a relatively novel electron microscopy imaging technique that allows three-dimensional characterization of the ultrastructure in both tissues and cells by measuring volumes of thousands of cubic micrometres yet at nanometre-scale resolution. In the scanning electron microscope, repeatedly an image is acquired followed by the removal of a thin layer resin embedded biological material by either a microtome or a focused ion beam. In this way, each recorded image contains novel structural information which can be used for three-dimensional analysis. Here, we explore focused ion beam facilitated serial block face-scanning electron microscopy to study the endothelial cell–specific storage organelles, the Weibel–Palade bodies, during their biogenesis at the Golgi apparatus. Weibel–Palade bodies predominantly contain the coagulation protein Von Willebrand factor which is secreted by the cell upon vascular damage. Using focused ion beam facilitated serial block face-scanning electron microscopy we show that the technique has the sensitivity to clearly reveal subcellular details like mitochondrial cristae and small vesicles with a diameter of about 50 nm. Also, we reveal numerous associations between Weibel–Palade bodies and Golgi stacks which became conceivable in large-scale three-dimensional data. We demonstrate that serial block face-scanning electron microscopy is a promising tool that offers an alternative for electron tomography to study subcellular organelle interactions in the context of a complete cell. PMID:25644989