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Sample records for block-face scanning electron

  1. 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.

  2. Three-dimensional architecture of podocytes revealed by block-face scanning electron microscopy.

    PubMed

    Ichimura, Koichiro; Miyazaki, Naoyuki; Sadayama, Shoji; Murata, Kazuyoshi; Koike, Masato; Nakamura, Kei-Ichiro; Ohta, Keisuke; Sakai, Tatsuo

    2015-03-11

    Block-face imaging is a scanning electron microscopic technique which enables easier acquisition of serial ultrastructural images directly from the surface of resin-embedded biological samples with a similar quality to transmission electron micrographs. In the present study, we analyzed the three-dimensional architecture of podocytes using serial block-face imaging. It was previously believed that podocytes are divided into three kinds of subcellular compartment: cell body, primary process, and foot process, which are simply aligned in this order. When the reconstructed podocytes were viewed from their basal side, the foot processes were branched from a ridge-like prominence, which was formed on the basal surface of the primary process and was similar to the usual foot processes in structure. Moreover, from the cell body, the foot processes were also emerged via the ridge-like prominence, as found in the primary process. The ridge-like prominence anchored the cell body and primary process to the glomerular basement membrane, and connected the foot processes to the cell body and primary process. In conclusion, serial block-face imaging is a powerful tool for clear understanding the three-dimensional architecture of podocytes through its ability to reveal novel structures which were difficult to determine by conventional transmission and scanning electron microscopes alone.

  3. Three-dimensional architecture of podocytes revealed by block-face scanning electron microscopy

    PubMed Central

    Ichimura, Koichiro; Miyazaki, Naoyuki; Sadayama, Shoji; Murata, Kazuyoshi; Koike, Masato; Nakamura, Kei-ichiro; Ohta, Keisuke; Sakai, Tatsuo

    2015-01-01

    Block-face imaging is a scanning electron microscopic technique which enables easier acquisition of serial ultrastructural images directly from the surface of resin-embedded biological samples with a similar quality to transmission electron micrographs. In the present study, we analyzed the three-dimensional architecture of podocytes using serial block-face imaging. It was previously believed that podocytes are divided into three kinds of subcellular compartment: cell body, primary process, and foot process, which are simply aligned in this order. When the reconstructed podocytes were viewed from their basal side, the foot processes were branched from a ridge-like prominence, which was formed on the basal surface of the primary process and was similar to the usual foot processes in structure. Moreover, from the cell body, the foot processes were also emerged via the ridge-like prominence, as found in the primary process. The ridge-like prominence anchored the cell body and primary process to the glomerular basement membrane, and connected the foot processes to the cell body and primary process. In conclusion, serial block-face imaging is a powerful tool for clear understanding the three-dimensional architecture of podocytes through its ability to reveal novel structures which were difficult to determine by conventional transmission and scanning electron microscopes alone. PMID:25759085

  4. 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.

  5. 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

  6. Morphological process of podocyte development revealed by block-face scanning electron microscopy.

    PubMed

    Ichimura, Koichiro; Kakuta, Soichiro; Kawasaki, Yuto; Miyaki, Takayuki; Nonami, Takahiro; Miyazaki, Naoyuki; Nakao, Tomoyo; Enomoto, Sakiko; Arai, Shigeo; Koike, Masato; Murata, Kazuyoshi; Sakai, Tatsuo

    2017-01-01

    Podocytes present a unique 3D architecture specialized for glomerular filtration. However, several 3D morphological aspects on podocyte development remain partially understood because they are difficult to reveal using conventional scanning electron microscopy (SEM). Here, we adopted serial block-face SEM imaging, a powerful tool for analyzing the 3D cellular ultrastructure, to precisely reveal the morphological process of podocyte development, such as the formation of foot processes. Development of foot processes gives rise to three morphological states: the primitive, immature and mature foot processes. Immature podocytes were columnar in shape and connected to each other by the junctional complex, which migrated toward the basal side of the cell. When the junctional complex was close to the basement membrane, immature podocytes started to interdigitate with primitive foot processes under the level of junctional complex. As primitive foot processes lengthened, the junctional complex moved between primitive foot processes to form immature foot processes. Finally, the junctional complex was gradually replaced by the slit diaphragm, resulting in the maturation of immature foot processes into mature foot processes. In conclusion, the developmental process of podocytes is now clearly visualized by block-face SEM imaging. © 2017. Published by The Company of Biologists Ltd.

  7. 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.

  8. 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.

  9. Optimizing the 3D-reconstruction technique for serial block-face scanning electron microscopy.

    PubMed

    Wernitznig, Stefan; Sele, Mariella; Urschler, Martin; Zankel, Armin; Pölt, Peter; Rind, F Claire; Leitinger, Gerd

    2016-05-01

    Elucidating the anatomy of neuronal circuits and localizing the synaptic connections between neurons, can give us important insights in how the neuronal circuits work. We are using serial block-face scanning electron microscopy (SBEM) to investigate the anatomy of a collision detection circuit including the Lobula Giant Movement Detector (LGMD) neuron in the locust, Locusta migratoria. For this, thousands of serial electron micrographs are produced that allow us to trace the neuronal branching pattern. The reconstruction of neurons was previously done manually by drawing cell outlines of each cell in each image separately. This approach was very time consuming and troublesome. To make the process more efficient a new interactive software was developed. It uses the contrast between the neuron under investigation and its surrounding for semi-automatic segmentation. For segmentation the user sets starting regions manually and the algorithm automatically selects a volume within the neuron until the edges corresponding to the neuronal outline are reached. Internally the algorithm optimizes a 3D active contour segmentation model formulated as a cost function taking the SEM image edges into account. This reduced the reconstruction time, while staying close to the manual reference segmentation result. Our algorithm is easy to use for a fast segmentation process, unlike previous methods it does not require image training nor an extended computing capacity. Our semi-automatic segmentation algorithm led to a dramatic reduction in processing time for the 3D-reconstruction of identified neurons. Copyright © 2016 Elsevier B.V. All rights reserved.

  10. 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.

  11. 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

  12. 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.

  13. 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

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

    PubMed

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

    2017-01-01

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

  15. Rapid specimen preparation to improve the throughput of electron microscopic volume imaging for three-dimensional analyses of subcellular ultrastructures with serial block-face scanning electron microscopy.

    PubMed

    Thai, Truc Quynh; Nguyen, Huy Bang; Saitoh, Sei; Wu, Bao; Saitoh, Yurika; Shimo, Satoshi; Elewa, Yaser Hosny Ali; Ichii, Osamu; Kon, Yasuhiro; Takaki, Takashi; Joh, Kensuke; Ohno, Nobuhiko

    2016-09-01

    Serial block-face imaging using scanning electron microscopy enables rapid observations of three-dimensional ultrastructures in a large volume of biological specimens. However, such imaging usually requires days for sample preparation to reduce charging and increase image contrast. In this study, we report a rapid procedure to acquire serial electron microscopic images within 1 day for three-dimensional analyses of subcellular ultrastructures. This procedure is based on serial block-face with two major modifications, including a new sample treatment device and direct polymerization on the rivets, to reduce the time and workload needed. The modified procedure without uranyl acetate can produce tens of embedded samples observable under serial block-face scanning electron microscopy within 1 day. The serial images obtained are similar to the block-face images acquired by common procedures, and are applicable to three-dimensional reconstructions at a subcellular resolution. Using this approach, regional immune deposits and the double contour or heterogeneous thinning of basement membranes were observed in the glomerular capillary loops of an autoimmune nephropathy model. These modifications provide options to improve the throughput of three-dimensional electron microscopic examinations, and will ultimately be beneficial for the wider application of volume imaging in life science and clinical medicine.

  16. Three-dimensional reconstruction of black tiger prawn (Penaeus monodon) spermatozoa using serial block-face scanning electron microscopy.

    PubMed

    Feng, Tianyi; Paterson, Brian D; Webb, Robyn; Johnston, Stephen D

    2016-05-01

    Serial Block-Face Scanning Electron Microscopy (SBF-SEM) was used in this study to examine the ultrastructural morphology of Penaeus monodon spermatozoa. SBF-SEM provided a large dataset of sequential electron-microscopic-level images that facilitated comprehensive ultrastructural observations and three-dimensional reconstructions of the sperm cell. Reconstruction divulged a nuclear region of the spermatophoral spermatozoon filled with decondensed chromatin but with two apparent levels of packaging density. In addition, the nuclear region contained, not only numerous filamentous chromatin elements with dense microregions, but also large centrally gathered granular masses. Analysis of the sperm cytoplasm revealed the presence of degenerated mitochondria and membrane-less dense granules. A large electron-lucent vesicle and "arch-like" structures were apparent in the subacrosomal area, and an acrosomal core was found in the acrosomal vesicle. The spermatozoal spike arose from the inner membrane of the acrosomal vesicle, which was slightly bulbous in the middle region of the acrosomal vesicle, but then extended distally into a broad dense plate and to a sharp point proximally. This study has demonstrated that SBF-SEM is a powerful technique for the 3D ultrastructural reconstruction of prawn spermatozoa, that will no doubt be informative for further studies of sperm assessment, reproductive pathology and the spermiocladistics of penaeid prawns, and other decapod crustaceans.

  17. Serial block face-scanning electron microscopy: a tool for studying embryonic development at the cell-matrix interface.

    PubMed

    Starborg, Tobias; Kadler, Karl E

    2015-03-01

    Studies of gene regulation, signaling pathways, and stem cell biology are contributing greatly to our understanding of early embryonic vertebrate development. However, much less is known about the events during the latter half of embryonic development, when tissues comprising mostly extracellular matrix (ECM) are formed. The matrix extends far beyond the boundaries of individual cells and is refractory to study by conventional biochemical and molecular techniques; thus major gaps exist in our knowledge of the formation and three-dimensional (3D) organization of the dense tissues that form the bulk of adult vertebrates. Serial block face-scanning electron microscopy (SBF-SEM) has the ability to image volumes of tissue containing numerous cells at a resolution sufficient to study the organization of the ECM. Furthermore, whereas light microscopy was once relatively straightforward and electron microscopy was performed in specialist laboratories, the tables are turned; SBF-SEM is relatively straightforward and is becoming routine in high-end resolution studies of embryonic structures in vivo. In this review, we discuss the emergence of SBF-SEM as a tool for studying embryonic vertebrate development.

  18. Cardiac Myocyte Diversity and a Fibroblast Network in the Junctional Region of the Zebrafish Heart Revealed by Transmission and Serial Block-Face Scanning Electron Microscopy

    PubMed Central

    Lafontant, Pascal J.; Behzad, Ali R.; Brown, Evelyn; Landry, Paul; Hu, Norman; Burns, Alan R.

    2013-01-01

    The zebrafish has emerged as an important model of heart development and regeneration. While the structural characteristics of the developing and adult zebrafish ventricle have been previously studied, little attention has been paid to the nature of the interface between the compact and spongy myocardium. Here we describe how these two distinct layers are structurally and functionally integrated. We demonstrate by transmission electron microscopy that this interface is complex and composed primarily of a junctional region occupied by collagen, as well as a population of fibroblasts that form a highly complex network. We also describe a continuum of uniquely flattened transitional cardiac myocytes that form a circumferential plate upon which the radially-oriented luminal trabeculae are anchored. In addition, we have uncovered within the transitional ring a subpopulation of markedly electron dense cardiac myocytes. At discrete intervals the transitional cardiac myocytes form contact bridges across the junctional space that are stabilized through localized desmosomes and fascia adherentes junctions with adjacent compact cardiac myocytes. Finally using serial block-face scanning electron microscopy, segmentation and volume reconstruction, we confirm the three-dimensional nature of the junctional region as well as the presence of the sheet-like fibroblast network. These ultrastructural studies demonstrate the previously unrecognized complexity with which the compact and spongy layers are structurally integrated, and provide a new basis for understanding development and regeneration in the zebrafish heart. PMID:24058412

  19. Cardiac myocyte diversity and a fibroblast network in the junctional region of the zebrafish heart revealed by transmission and serial block-face scanning electron microscopy.

    PubMed

    Lafontant, Pascal J; Behzad, Ali R; Brown, Evelyn; Landry, Paul; Hu, Norman; Burns, Alan R

    2013-01-01

    The zebrafish has emerged as an important model of heart development and regeneration. While the structural characteristics of the developing and adult zebrafish ventricle have been previously studied, little attention has been paid to the nature of the interface between the compact and spongy myocardium. Here we describe how these two distinct layers are structurally and functionally integrated. We demonstrate by transmission electron microscopy that this interface is complex and composed primarily of a junctional region occupied by collagen, as well as a population of fibroblasts that form a highly complex network. We also describe a continuum of uniquely flattened transitional cardiac myocytes that form a circumferential plate upon which the radially-oriented luminal trabeculae are anchored. In addition, we have uncovered within the transitional ring a subpopulation of markedly electron dense cardiac myocytes. At discrete intervals the transitional cardiac myocytes form contact bridges across the junctional space that are stabilized through localized desmosomes and fascia adherentes junctions with adjacent compact cardiac myocytes. Finally using serial block-face scanning electron microscopy, segmentation and volume reconstruction, we confirm the three-dimensional nature of the junctional region as well as the presence of the sheet-like fibroblast network. These ultrastructural studies demonstrate the previously unrecognized complexity with which the compact and spongy layers are structurally integrated, and provide a new basis for understanding development and regeneration in the zebrafish heart.

  20. Correlative two-photon and serial block face scanning electron microscopy in neuronal tissue using 3D near-infrared branding maps.

    PubMed

    Lees, Robert M; Peddie, Christopher J; Collinson, Lucy M; Ashby, Michael C; Verkade, Paul

    2017-01-01

    Linking cellular structure and function has always been a key goal of microscopy, but obtaining high resolution spatial and temporal information from the same specimen is a fundamental challenge. Two-photon (2P) microscopy allows imaging deep inside intact tissue, bringing great insight into the structural and functional dynamics of cells in their physiological environment. At the nanoscale, the complex ultrastructure of a cell's environment in tissue can be reconstructed in three dimensions (3D) using serial block face scanning electron microscopy (SBF-SEM). This provides a snapshot of high resolution structural information pertaining to the shape, organization, and localization of multiple subcellular structures at the same time. The pairing of these two imaging modalities in the same specimen provides key information to relate cellular dynamics to the ultrastructural environment. Until recently, approaches to relocate a region of interest (ROI) in tissue from 2P microscopy for SBF-SEM have been inefficient or unreliable. However, near-infrared branding (NIRB) overcomes this by using the laser from a multiphoton microscope to create fiducial markers for accurate correlation of 2P and electron microscopy (EM) imaging volumes. The process is quick and can be user defined for each sample. Here, to increase the efficiency of ROI relocation, multiple NIRB marks are used in 3D to target ultramicrotomy. A workflow is described and discussed to obtain a data set for 3D correlated light and electron microscopy, using three different preparations of brain tissue as examples. Copyright © 2017 Elsevier Inc. All rights reserved.

  1. Serial block-face scanning electron microscopy applied to study the trafficking of 8D3-coated gold nanoparticles at the blood-brain barrier.

    PubMed

    Cabezón, Itsaso; Augé, Elisabet; Bosch, Manel; Beckett, Alison J; Prior, Ian A; Pelegrí, Carme; Vilaplana, Jordi

    2017-07-01

    Due to the physical and physiological properties of the blood-brain barrier (BBB), the transport of neurotherapeutics from blood to brain is still a pharmaceutical challenge. We previously conducted a series of experiments to explore the potential of the anti-transferrin receptor 8D3 monoclonal antibody (mAb) to transport neurotherapeutics across the BBB. In that study, gold nanoparticles (AuNPs) were coated with the 8D3 antibody and administered intravenously to mice. Transmission electron microscopy was used and a two-dimensional (2D) image analysis was performed to detect the AuNPs in the brain capillary endothelial cells (BCECs) and brain parenchyma. In the present work, we determined that serial block-face scanning electron microscopy (SBF-SEM) is a useful tool to study the transcytosis of these AuNPs across the BBB in three dimensions and we, therefore, applied it to gain more knowledge of their transcellular trafficking. The resulting 3D reconstructions provided additional information on the endocytic vesicles containing AuNPs and the endosomal processing that occurs inside BCECs. The passage from 2D to 3D analysis reinforced the trafficking model proposed in the 2D study, and revealed that the vesicles containing AuNPs are significantly larger and more complex than described in our 2D study. We also discuss tradeoffs of using this technique for our application, and conclude that together with other volume electron microscopy imaging techniques, SBF-SEM is a powerful approach that is worth of considering for studies of drug transport across the BBB.

  2. Three-dimensional analysis of morphological changes in the malaria parasite infected red blood cell by serial block-face scanning electron microscopy.

    PubMed

    Sakaguchi, Miako; Miyazaki, Naoyuki; Fujioka, Hisashi; Kaneko, Osamu; Murata, Kazuyoshi

    2016-03-01

    The human malaria parasite, Plasmodium falciparum, exhibits morphological changes during the blood stage cycle in vertebrate hosts. Here, we used serial block-face scanning electron microscopy (SBF-SEM) to visualize the entire structures of P. falciparum-infected red blood cells (iRBCs) and to examine their morphological and volumetric changes at different stages. During developmental stages, the parasite forms Maurer's clefts and vesicles in the iRBC cytoplasm and knobs on the iRBC surface, and extensively remodels the iRBC structure for proliferation of the parasite. In our observations, the Maurer's clefts and vesicles in the P. falciparum-iRBCs, resembling the so-called tubovesicular network (TVN), were not connected to each other, and continuous membrane networks were not observed between the parasitophorous vacuole membrane (PVM) and the iRBC cytoplasmic membrane. In the volumetric analysis, the iRBC volume initially increased and then decreased to the end of the blood stage cycle. This suggests that it is necessary to absorb a substantial amount of nutrients from outside the iRBC during the initial stage, but to release waste materials from inside the iRBC at the multinucleate stage. Transportation of the materials may be through the iRBC membrane, rather than a special structure formed by the parasite, because there is no direct connection between the iRBC membrane and the parasite. These results provide new insights as to how the malaria parasite grows in the iRBC and remodels iRBC structure during developmental stages; these observation can serve as a baseline for further experiments on the effects of therapeutic agents on malaria.

  3. Microtubule organization within mitotic spindles revealed by serial block face scanning EM and image analysis.

    PubMed

    Nixon, Faye M; Honnor, Thomas R; Clarke, Nicholas I; Starling, Georgina P; Beckett, Alison J; Johansen, Adam M; Brettschneider, Julia A; Prior, Ian A; Royle, Stephen J

    2017-04-07

    Serial block face scanning electron microscopy (SBF-SEM) is a powerful method to analyze cells in 3D. Here, working at the resolution limit of the method, we describe a correlative light-SBF-SEM workflow to resolve microtubules of the mitotic spindle in human cells. We present four examples of uses for this workflow which are not practical by light microscopy and/or transmission electron microscopy. First, distinguishing closely associated microtubules within K-fibers; second, resolving bridging fibers in the mitotic spindle; third, visualizing membranes in mitotic cells, relative to the spindle apparatus; fourth, volumetric analysis of kinetochores. Our workflow also includes new computational tools for exploring the spatial arrangement of MTs within the mitotic spindle. We use these tools to show that microtubule order in mitotic spindles is sensitive to the level of TACC3 on the spindle.

  4. Mesoscale morphology at nanoscale resolution: serial block-face scanning electron microscopy reveals fine 3D detail of a novel silk spinneret system in a tube-building tanaid crustacean.

    PubMed

    Kaji, Tomonari; Kakui, Keiichi; Miyazaki, Naoyuki; Murata, Kazuyoshi; Palmer, A Richard

    2016-01-01

    The study of morphology is experiencing a renaissance due to rapid improvements in technologies for 3D visualization of complex internal and external structures. But 3D visualization of the internal structure of mesoscale objects - those in the 10-1000 μm range - remains problematic. They are too small for microCT, many lack suitable specific fluorescent markers for confocal microscopy, or they require labor-intensive stacking and smoothing of individual TEM images. Here we illustrate the first comprehensive morphological description of a complete mesoscale biological system at nanoscopic resolution using ultra-modern technology for 3D visualization - serial block-face scanning electron microscopy (SBF-SEM). The SBF-SEM machine combines an in-chamber ultramicrotome, which creates a serial array of exposed surfaces, with an SEM that images each surface as it is exposed. The serial images are then stacked automatically by 3D reconstruction software. We used SBF-SEM to study the spinneret (thread-producing) system of a small, tube-dwelling crustacean that weaves tubes of silk. Thread-producing ability is critical for the survival of many small-bodied animals but the basic morphology of these systems remains mysterious due to the limits of traditional microscopy. SBF-SEM allowed us to describe - in full 3D - well-resolved components (glands, ducts, pores, and associated nerves and muscles) of the spinneret system in the thoracic legs and body segments of Sinelobus sp. (Crustacea, Peracarida, Tanaidacea), a tube-building tanaid only 2 mm in body length. The 3D reconstruction by SBF-SEM revealed at nanoscale resolution a unique structure to the gland and duct systems: In each of three thread-producing thoracic segments, two separate ducts, derived from two separate glands located in the body, run through the entire leg and merge at the leg tip just before the spinneret pore opening. We also resolved nerves connecting to individual setae, spines and pores on the

  5. Automated in-chamber specimen coating for serial block-face electron microscopy.

    PubMed

    Titze, B; Denk, W

    2013-05-01

    When imaging insulating specimens in a scanning electron microscope, negative charge accumulates locally ('sample charging'). The resulting electric fields distort signal amplitude, focus and image geometry, which can be avoided by coating the specimen with a conductive film prior to introducing it into the microscope chamber. This, however, is incompatible with serial block-face electron microscopy (SBEM), where imaging and surface removal cycles (by diamond knife or focused ion beam) alternate, with the sample remaining in place. Here we show that coating the sample after each cutting cycle with a 1-2 nm metallic film, using an electron beam evaporator that is integrated into the microscope chamber, eliminates charging effects for both backscattered (BSE) and secondary electron (SE) imaging. The reduction in signal-to-noise ratio (SNR) caused by the film is smaller than that caused by the widely used low-vacuum method. Sample surfaces as large as 12 mm across were coated and imaged without charging effects at beam currents as high as 25 nA. The coatings also enabled the use of beam deceleration for non-conducting samples, leading to substantial SNR gains for BSE contrast. We modified and automated the evaporator to enable the acquisition of SBEM stacks, and demonstrated the acquisition of stacks of over 1000 successive cut/coat/image cycles and of stacks using beam deceleration or SE contrast. © 2013 The Authors Journal of Microscopy © 2013 Royal Microscopical Society.

  6. A novel design of a scanning probe microscope integrated with an ultramicrotome for serial block-face nanotomography

    NASA Astrophysics Data System (ADS)

    Efimov, Anton E.; Agapov, Igor I.; Agapova, Olga I.; Oleinikov, Vladimir A.; Mezin, Alexey V.; Molinari, Michael; Nabiev, Igor; Mochalov, Konstantin E.

    2017-02-01

    We present a new concept of a combined scanning probe microscope (SPM)/ultramicrotome apparatus. It enables "slice-and-view" scanning probe nanotomography measurements and 3D reconstruction of the bulk sample nanostructure from series of SPM images after consecutive ultrathin sections. The sample is fixed on a flat XYZ scanning piezostage mounted on the ultramicrotome arm. The SPM measuring head with a cantilever tip and a laser-photodiode tip detection system approaches the sample for SPM measurements of the block-face surface immediately after the ultramicrotome sectioning is performed. The SPM head is moved along guides that are also fixed on the ultramicrotome arm. Thereby, relative dysfunctional displacements of the tip, the sample, and the ultramicrotome knife are minimized. The design of the SPM head enables open frontal optical access to the sample block-face adapted for high-resolution optical lenses for correlative SPM/optical microscopy applications. The new system can be used in a wide range of applications for the study of 3D nanostructures of biological objects, biomaterials, polymer nanocomposites, and nanohybrid materials in various SPM and optical microscopy measuring modes.

  7. Pros and cons: cryo-electron microscopic evaluation of block faces versus cryo-sections from frozen-hydrated skin specimens prepared by different techniques.

    PubMed

    Richter, T; Biel, S S; Sattler, M; Wenck, H; Wittern, K-P; Wiesendanger, R; Wepf, R

    2007-02-01

    Over the last two decades, several different preparative techniques have been developed to investigate frozen-hydrated biological samples by electron microscopy. In this article, we describe an alternative approach that allows either ultrastructural investigations of frozen human skin at a resolution better than 15 nm or sample throughput that is sufficiently high enough for quantitative morphological analysis. The specimen preparation method we describe is fast, reproducible, does not require much user experience or elaborate equipment. We compare high-pressure freezing with plunge freezing, and block faces with frozen-hydrated slices (sections), to study variations in cell thickness upon hydration changes. Plunge freezing is optimal for morphological and stereological investigations of structures with low water content. By contrast, high-pressure freezing proved optimal for high-resolution studies and provided the best ultrastructural preservation. A combination of these fast-freezing techniques with cryo-ultramicrotomy yielded well-preserved block faces of the original biological material. Here we show that these block faces did not exhibit any of the artefacts normally associated with cryo-sections, and--after evaporating a heavy metal and carbon onto the surface--are stable enough in the electron beam to provide high-resolution images of large surface areas for statistical analysis in a cryo-SEM (scanning electron microscope). Because the individual preparation steps use only standard equipment and do not require much experience from the experimenter, they are generally more usable, making this approach an interesting alternative to other methods for the ultrastructural investigation of frozen-hydrated material.

  8. X-ray microscopy as an approach to increasing accuracy and efficiency of serial block-face imaging for correlated light and electron microscopy of biological specimens.

    PubMed

    Bushong, Eric A; Johnson, Donald D; Kim, Keun-Young; Terada, Masako; Hatori, Megumi; Peltier, Steven T; Panda, Satchidananda; Merkle, Arno; Ellisman, Mark H

    2015-02-01

    The recently developed three-dimensional electron microscopic (EM) method of serial block-face scanning electron microscopy (SBEM) has rapidly established itself as a powerful imaging approach. Volume EM imaging with this scanning electron microscopy (SEM) method requires intense staining of biological specimens with heavy metals to allow sufficient back-scatter electron signal and also to render specimens sufficiently conductive to control charging artifacts. These more extreme heavy metal staining protocols render specimens light opaque and make it much more difficult to track and identify regions of interest (ROIs) for the SBEM imaging process than for a typical thin section transmission electron microscopy correlative light and electron microscopy study. We present a strategy employing X-ray microscopy (XRM) both for tracking ROIs and for increasing the efficiency of the workflow used for typical projects undertaken with SBEM. XRM was found to reveal an impressive level of detail in tissue heavily stained for SBEM imaging, allowing for the identification of tissue landmarks that can be subsequently used to guide data collection in the SEM. Furthermore, specific labeling of individual cells using diaminobenzidine is detectable in XRM volumes. We demonstrate that tungsten carbide particles or upconverting nanophosphor particles can be used as fiducial markers to further increase the precision and efficiency of SBEM imaging.

  9. 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.

  10. 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

  11. Forensic Scanning Electron Microscope

    NASA Astrophysics Data System (ADS)

    Keeley, R. H.

    1983-03-01

    The scanning electron microscope equipped with an x-ray spectrometer is a versatile instrument which has many uses in the investigation of crime and preparation of scientific evidence for the courts. Major applications include microscopy and analysis of very small fragments of paint, glass and other materials which may link an individual with a scene of crime, identification of firearms residues and examination of questioned documents. Although simultaneous observation and chemical analysis of the sample is the most important feature of the instrument, other modes of operation such as cathodoluminescence spectrometry, backscattered electron imaging and direct x-ray excitation are also exploited. Marks on two bullets or cartridge cases can be compared directly by sequential scanning with a single beam or electronic linkage of two instruments. Particles of primer residue deposited on the skin and clothing when a gun is fired can be collected on adhesive tape and identified by their morphology and elemental composition. It is also possible to differentiate between the primer residues of different types of ammunition. Bullets may be identified from the small fragments left behind as they pass through the body tissues. In the examination of questioned documents the scanning electron microscope is used to establish the order in which two intersecting ink lines were written and to detect traces of chemical markers added to the security inks on official documents.

  12. 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.

  13. UAVSAR Active Electronically Scanned Array

    NASA Technical Reports Server (NTRS)

    Sadowy, Gregory, A.; Chamberlain, Neil F.; Zawadzki, Mark S.; Brown, Kyle M.; Fisher, Charles D.; Figueroa, Harry S.; Hamilton, Gary A.; Jones, Cathleen E.; Vorperian, Vatche; Grando, Maurio B.

    2011-01-01

    The Uninhabited Airborne Vehicle Synthetic Aperture Radar (UAVSAR) is a pod-based, L-band (1.26 GHz), repeatpass, interferometric, synthetic aperture radar (InSAR) used for Earth science applications. Repeat-pass interferometric radar measurements from an airborne platform require an antenna that can be steered to maintain the same angle with respect to the flight track over a wide range of aircraft yaw angles. In order to be able to collect repeat-pass InSAR data over a wide range of wind conditions, UAVSAR employs an active electronically scanned array (AESA). During data collection, the UAVSAR flight software continuously reads the aircraft attitude state measured by the Embedded GPS/INS system (EGI) and electronically steers the beam so that it remains perpendicular to the flight track throughout the data collection

  14. 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.

  15. Quantitative analysis of mouse pancreatic islet architecture by serial block-face SEM.

    PubMed

    Pfeifer, C R; Shomorony, A; Aronova, M A; Zhang, G; Cai, T; Xu, H; Notkins, A L; Leapman, R D

    2015-01-01

    We have applied serial block-face scanning electron microscopy (SBF-SEM) to measure parameters that describe the architecture of pancreatic islets of Langerhans, microscopic endocrine organs that secrete insulin and glucagon for control of blood glucose. By analyzing entire mouse islets, we show that it is possible to determine (1) the distributions of alpha and beta cells, (2) the organization of blood vessels and pericapillary spaces, and (3) the ultrastructure of the individual secretory cells. Our results show that the average volume of a beta cell is nearly twice that of an alpha cell, and the total mitochondrial volume is about four times larger. In contrast, nuclear volumes in the two cell types are found to be approximately equal. Although the cores of alpha and beta secretory granules have similar diameters, the beta granules have prominent halos resulting in overall diameters that are twice those of alpha granules. Visualization of the blood vessels revealed that every secretory cell in the islet is in contact with the pericapillary space, with an average contact area of 9±5% of the cell surface area. Our data show that consistent results can be obtained by analyzing small numbers of islets. Due to the complicated architecture of pancreatic islets, such precision cannot easily be achieved by using TEM of thin sections. Published by Elsevier Inc.

  16. Phase multiplying electronic scanning array

    NASA Technical Reports Server (NTRS)

    Seaton, A. F.

    1969-01-01

    Scanning array was designed with properties of low RF loss and phase control. The array consists of a series of special waveguides, hybrids made up of two variable reactance branch arms for input signals, an edge slot for the difference port, and a sum arm for the unradiated signal.

  17. Scanning and three-dimensional electron microscopy methods for the study of Trypanosoma brucei and Leishmania mexicana flagella.

    PubMed

    Gluenz, Eva; Wheeler, Richard John; Hughes, Louise; Vaughan, Sue

    2015-01-01

    Three-dimensional electron microscopy tools have revolutionized our understanding of cell structure and molecular complexes in biology. Here, we describe methods for studying flagellar ultrastructure and biogenesis in two unicellular parasites-Trypanosoma brucei and Leishmania mexicana. We describe methods for the preparation of these parasites for scanning electron microscopy cellular electron tomography, and serial block face scanning electron microscopy (SBFSEM). These parasites have a highly ordered cell shape and form, with a defined positioning of internal cytoskeletal structures and organelles. We show how knowledge of these can be used to dissect cell cycles in both parasites and identify the old flagellum from the new in T. brucei. Finally, we demonstrate the use of SBFSEM three-dimensional models for analysis of individual whole cells, demonstrating the excellent potential this technique has for future studies of mutant cell lines.

  18. The Scanning Electron Microscope and the Archaeologist

    ERIC Educational Resources Information Center

    Ponting, Matthew

    2004-01-01

    Images from scanning electron microscopy are now quite common and they can be of great value in archaeology. Techniques such as secondary electron imaging, backscattered electron imaging and energy-dispersive x-ray analysis can reveal information such as the presence of weevils in grain in Roman Britain, the composition of Roman coins and the…

  19. The Scanning Electron Microscope and the Archaeologist

    ERIC Educational Resources Information Center

    Ponting, Matthew

    2004-01-01

    Images from scanning electron microscopy are now quite common and they can be of great value in archaeology. Techniques such as secondary electron imaging, backscattered electron imaging and energy-dispersive x-ray analysis can reveal information such as the presence of weevils in grain in Roman Britain, the composition of Roman coins and the…

  20. Electronic imaging and scanning system

    NASA Technical Reports Server (NTRS)

    Jain, A.

    1977-01-01

    Frequency-variable illumination has been used previously to provide high resolution imaging in one dimension. The paper extends the results on this imaging by frequency scanning to derive the expression for a two-dimensional image. This is the Fourier transformation, with respect to the angle and frequency of illumination, of the electric field detected in the far-field region of the object. The case is considered of a rough object and it is shown that for roughness finer than the resolution of the imaging system, the image has a granular appearance corresponding to the classical speckle effect. Large scale phase perturbations lead to the elevation displacement effect.

  1. Gigahertz-band electronically scanned antennas

    NASA Astrophysics Data System (ADS)

    Bei, Nikolai A.

    2000-12-01

    Foundation and principles of radio lenses construction of centimeter and millimeter wave ranges with controlled refracting index, combining the quality of phased array antennas with optical devices are stated. Possibilities of the electronically scanning with wide-angle sector and high gain are maintained. Construction principles of scanning antennas with controlled lenses, combining the quality of phased array antennas with optical devices, are stated. Possibilities of electronically scanning with broad angle sector and high gain are maintained. Some examples of construction of antennas millimeter range of waves are listed here.

  2. 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

  3. Radant - New method of electronic scanning

    NASA Astrophysics Data System (ADS)

    Chekroun, C.; Herrick, D.; Michel, Y. M.; Pauchard, R.; Vidal, P.

    1981-02-01

    The paper describes a novel electronic scanning process that differs from the conventional phased array process. Called Radant (from radome antennas), the process uses an electromagnetic lens such that the direction of the optical axis can be changed electronically. The principle of this process and a working model are described.

  4. Electronic SCAN (Selected Current Aerospace Notices)

    NASA Technical Reports Server (NTRS)

    Dunbar, Rick

    1993-01-01

    The on-line version of the NASA Selected Current Aerospace Notices (SCAN) is described and the three methods for electronic access on the Internet are listed. These are (1) File Transfer Protocol (FTP), (2) Gopher, and (3) LISTSERV. An electronic address and a brief description is given for each of them.

  5. 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.

  6. Electron Beam Scanning in Industrial Applications

    NASA Astrophysics Data System (ADS)

    Jongen, Yves; Herer, Arnold

    1996-05-01

    Scanned electron beams are used within many industries for applications such as sterilization of medical disposables, crosslinking of wire and cables insulating jackets, polymerization and degradation of resins and biomaterials, modification of semiconductors, coloration of gemstones and glasses, removal of oxides from coal plant flue gasses, and the curing of advanced composites and other molded forms. X-rays generated from scanned electron beams make yet other applications, such as food irradiation, viable. Typical accelerators for these applications range in beam energy from 0.5MeV to 10 MeV, with beam powers between 5 to 500kW and scanning widths between 20 and 300 cm. Since precise control of dose delivery is required in many of these applications, the integration of beam characteristics, product conveyance, and beam scanning mechanisms must be well understood and optimized. Fundamental issues and some case examples are presented.

  7. 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.

  8. Development of Scanning Ultrafast Electron Microscope Capability.

    SciTech Connect

    Collins, Kimberlee Chiyoko; Talin, Albert Alec; Chandler, David W.; Michael, Joseph R.

    2016-11-01

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

  9. 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.

  10. 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.

  11. Development of electron moire method using a scanning electron microscope

    NASA Astrophysics Data System (ADS)

    Kishimoto, Satoshi; Egashira, Mitsuru; Shinya, Norio

    1991-12-01

    A new moire method using a scanning electron microscope (SEM) for the measurement of micro-deformation has been developed. This new method makes it possible to observe the moire fringe pattern and SEM image at the same time. In this method, a fine microgrid prepared by electron lithography is used as a model grid, and scanning exposure of the electron beam in a SEM is used as a master grid. The exposure of electron beam on the specimen with the model grid produces moire fringes of bright and dark lines formed by the different amount of the secondary electrons. This fine moire fringe pattern is fine and clear enough to measure the strain distribution in a small area. By this method, concentrated strains around a small hole in polyimide resin specimens and also the inhomogeneous micro- deformations such as grain boundary sliding in copper specimens were measured, with high accuracy.

  12. 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

  13. 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.

  14. Cryogenic Multichannel Pressure Sensor With Electronic Scanning

    NASA Technical Reports Server (NTRS)

    Hopson, Purnell, Jr.; Chapman, John J.; Kruse, Nancy M. H.

    1994-01-01

    Array of pressure sensors operates reliably and repeatably over wide temperature range, extending from normal boiling point of water down to boiling point of nitrogen. Sensors accurate and repeat to within 0.1 percent. Operate for 12 months without need for recalibration. Array scanned electronically, sensor readings multiplexed and sent to desktop computer for processing and storage. Used to measure distributions of pressure in research on boundary layers at high Reynolds numbers, achieved by low temperatures.

  15. Cryogenic Multichannel Pressure Sensor With Electronic Scanning

    NASA Technical Reports Server (NTRS)

    Hopson, Purnell, Jr.; Chapman, John J.; Kruse, Nancy M. H.

    1994-01-01

    Array of pressure sensors operates reliably and repeatably over wide temperature range, extending from normal boiling point of water down to boiling point of nitrogen. Sensors accurate and repeat to within 0.1 percent. Operate for 12 months without need for recalibration. Array scanned electronically, sensor readings multiplexed and sent to desktop computer for processing and storage. Used to measure distributions of pressure in research on boundary layers at high Reynolds numbers, achieved by low temperatures.

  16. Scanning electron microscope view of iron crystal

    NASA Technical Reports Server (NTRS)

    1972-01-01

    A scanning electron microscope photograph of iron crystals which grow in a small vug or cavity in a recrystallized breccia (fragmented rock) from the Apollo 15 Hadley-Apennino lunar landing site. The largest crystal is three microns across. Perfectly developed crystals such as these indicate slow formation from a hot vapor as the rock was cooling. The crystals are resting on an interlocking lattice of pyroxene (calsium-magnesium-iron silicate).

  17. 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

  18. Scanning electron microscope view of iron crystal

    NASA Image and Video Library

    1972-11-10

    A scanning electron microscope photograph of iron crystals which grow in a small vug or cavity in a recrystallized breccia (fragmented rock) from the Apollo 15 Hadley-Apennino lunar landing site. The largest crystal is three microns across. Perfectly developed crystals such as these indicate slow formation from a hot vapor as the rock was cooling. The crystals are resting on an interlocking lattice of pyroxene (calsium-magnesium-iron silicate).

  19. 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.

  20. 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.

  1. Cathodoluminescence in the scanning transmission electron microscope.

    PubMed

    Kociak, M; Zagonel, L F

    2016-12-19

    Cathodoluminescence (CL) is a powerful tool for the investigation of optical properties of materials. In recent years, its combination with scanning transmission electron microscopy (STEM) has demonstrated great success in unveiling new physics in the field of plasmonics and quantum emitters. Most of these results were not imaginable even twenty years ago, due to conceptual and technical limitations. The purpose of this review is to present the recent advances that broke these limitations, and the new possibilities offered by the modern STEM-CL technique. We first introduce the different STEM-CL operating modes and the technical specificities in STEM-CL instrumentation. Two main classes of optical excitations, namely the coherent one (typically plasmons) and the incoherent one (typically light emission from quantum emitters) are investigated with STEM-CL. For these two main classes, we describe both the physics of light production under electron beam irradiation and the physical basis for interpreting STEM-CL experiments. We then compare STEM-CL with its better known sister techniques: scanning electron microscope CL, photoluminescence, and electron energy-loss spectroscopy. We finish by comprehensively reviewing recent STEM-CL applications. Copyright © 2016 Elsevier B.V. All rights reserved.

  2. Cathodoluminescence in the scanning transmission electron microscope.

    PubMed

    Kociak, M; Zagonel, L F

    2017-05-01

    Cathodoluminescence (CL) is a powerful tool for the investigation of optical properties of materials. In recent years, its combination with scanning transmission electron microscopy (STEM) has demonstrated great success in unveiling new physics in the field of plasmonics and quantum emitters. Most of these results were not imaginable even twenty years ago, due to conceptual and technical limitations. The purpose of this review is to present the recent advances that broke these limitations, and the new possibilities offered by the modern STEM-CL technique. We first introduce the different STEM-CL operating modes and the technical specificities in STEM-CL instrumentation. Two main classes of optical excitations, namely the coherent one (typically plasmons) and the incoherent one (typically light emission from quantum emitters) are investigated with STEM-CL. For these two main classes, we describe both the physics of light production under electron beam irradiation and the physical basis for interpreting STEM-CL experiments. We then compare STEM-CL with its better known sister techniques: scanning electron microscope CL, photoluminescence, and electron energy-loss spectroscopy. We finish by comprehensively reviewing recent STEM-CL applications. Copyright © 2016 Elsevier B.V. All rights reserved.

  3. 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.

  4. Serial block face SEM visualization of unusual plant nuclear tubular extensions in a carnivorous plant (Utricularia, Lentibulariaceae).

    PubMed

    Plachno, Bartosz J; Swiatek, Piotr; Jobson, Richard W; Malota, Karol; Brutkowski, Wojciech

    2017-05-24

    In Utricularia nelumbifolia , the nuclei of placental nutritive tissue possess unusually shaped projections not known to occur in any other flowering plant. The main aim of the study was to document the morphology and ultrastructure of these unusual nuclei. In addition, the literature was searched to find examples of nuclear tubular projections in other plant groups, and the nuclei of closely related species of Utricularia (i.e. sects Iperua , Orchidioides , Foliosa and Utricularia ) were examined. To visualize the complexity of the nuclear structures, transmission electron microscopy (TEM) was used, and 3-D ultrastructural reconstructions were made using the serial block face scanning electron microscopy (SBEM) technique. The nuclei of 11 Utricularia species, i.e. U. nelumbifolia , U. reniformis , U. cornigera , U. nephrophylla (sect. Iperua ), U. asplundii , U. alpina , U. quelchii (sect. Orchidioides ), U. longifolia (sect. Foliosa ), U. intermedia , U. minor and U. gibba (sect. Utricularia ) were examined. Of the 11 Utricularia species examined, the spindle-like tubular projections (approx. 5 μm long) emanating from resident nuclei located in placental nutritive tissues were observed only in U. nelumbifolia . These tubular nuclear extensions contained chromatin distributed along hexagonally shaped tubules. The apices of the projections extended into the cell plasma membrane, and in many cases also made contact at the two opposing cellular poles, and with plasmodesmata via a short cisterna of the cortical endoplasmic reticulum. Images from the SBEM provide some evidence that the nuclear projections are making contact with those of neighbouring cells. The term chromatubules (chromatin-filled tubules) for the nuclear projections of U. nelumbifolia placental tissue was proposed here. Due to the apparent association with the plasma membrane and plasmodesmata, it was also speculated that chromatubules are involved in nucleus-cell-cell communication. However

  5. 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.

  6. 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.

  7. 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.

  8. 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.

  9. 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.

  10. 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.

  11. 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.

  12. 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.

  13. 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.

  14. 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

  15. 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

  16. 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.

  17. Collection of secondary electrons in scanning electron microscopes.

    PubMed

    Müllerová, I; Konvalina, I

    2009-12-01

    Collection of the secondary electrons in the scanning electron microscope was simulated and the results have been experimentally verified for two types of the objective lens and three detection systems. The aberration coefficients of both objective lenses as well as maximum axial magnetic fields in the specimen region are presented. Compared are a standard side-attached secondary electron detector, in which only weak electrostatic and nearly no magnetic field influence the signal trajectories in the specimen vicinity, and the side-attached (lower) and upper detectors in an immersion system with weak electrostatic but strong magnetic field penetrating towards the specimen. The collection efficiency was calculated for all three detection systems and several working distances. The ability of detectors to attract secondary electron trajectories for various initial azimuthal and polar angles was calculated, too. According to expectations, the lower detector of an immersion system collects no secondary electrons I and II emitted from the specimen and only backscattered electrons and secondary electrons III form the final image. The upper detector of the immersion system exhibits nearly 100% collection efficiency decreasing, however, with the working distance, but the topographical contrast is regrettably suppressed in its image. The collection efficiency of the standard detector is low for short working distances but increases with the same, preserving strong topographical contrast.

  18. 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.

  19. 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.

  20. 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.

  1. [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.

  2. Dental Wear: A Scanning Electron Microscope Study

    PubMed Central

    Levrini, Luca; Di Benedetto, Giulia

    2014-01-01

    Dental wear can be differentiated into different types on the basis of morphological and etiological factors. The present research was carried out on twelve extracted human teeth with dental wear (three teeth showing each type of wear: erosion, attrition, abrasion, and abfraction) studied by scanning electron microscopy (SEM). The study aimed, through analysis of the macro- and micromorphological features of the lesions (considering the enamel, dentin, enamel prisms, dentinal tubules, and pulp), to clarify the different clinical and diagnostic presentations of dental wear and their possible significance. Our results, which confirm current knowledge, provide a complete overview of the distinctive morphology of each lesion type. It is important to identify the type of dental wear lesion in order to recognize the contributing etiological factors and, consequently, identify other more complex, nondental disorders (such as gastroesophageal reflux, eating disorders). It is clear that each type of lesion has a specific morphology and mechanism, and further clinical studies are needed to clarify the etiological processes, particularly those underlying the onset of abfraction. PMID:25548769

  3. Dental wear: a scanning electron microscope study.

    PubMed

    Levrini, Luca; Di Benedetto, Giulia; Raspanti, Mario

    2014-01-01

    Dental wear can be differentiated into different types on the basis of morphological and etiological factors. The present research was carried out on twelve extracted human teeth with dental wear (three teeth showing each type of wear: erosion, attrition, abrasion, and abfraction) studied by scanning electron microscopy (SEM). The study aimed, through analysis of the macro- and micromorphological features of the lesions (considering the enamel, dentin, enamel prisms, dentinal tubules, and pulp), to clarify the different clinical and diagnostic presentations of dental wear and their possible significance. Our results, which confirm current knowledge, provide a complete overview of the distinctive morphology of each lesion type. It is important to identify the type of dental wear lesion in order to recognize the contributing etiological factors and, consequently, identify other more complex, nondental disorders (such as gastroesophageal reflux, eating disorders). It is clear that each type of lesion has a specific morphology and mechanism, and further clinical studies are needed to clarify the etiological processes, particularly those underlying the onset of abfraction.

  4. Electronic scan at millimetre wave frequencies

    NASA Astrophysics Data System (ADS)

    Snieder, J.

    1990-06-01

    The possibilities of realizing phased array antennas in the mm wave region are reviewed. The use of discrete components and bulk material is discussed. The MMIC (Monolithic Microwave Integrated Circuits) techniques for the realization of phased array components are excluded from this report. A survey is given of the different electronic scan possibilities. PIN diode phase shifters and different types of ferrite phase shifters are discussed, including the principle functioning of these components as well as the advantages involved in their use. The availability of various ferrite materials for phase shifters is discussed. The requirements for phased array antennas and its phase shifters are given. A discussion of array element spacing, side lobe level and related power levels, effect of pulse repetition frequency and mutual coupling are presented. An attempt is made to correlate all the available information at cm wavelength (3 to 18 GHz) and to extrapolate it to mm wavelength. All the detailed properties of the phase shifters are included. The experimental results of mm wave phase shifters are given and compared with the extrapolated results given.

  5. 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

  6. [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.

  7. 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.

  8. 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.

  9. 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.

  10. Scanning Electron Microscope Studies on Aggregation Characteristics of Alumina Nanofluids

    DTIC Science & Technology

    2013-08-01

    UNCLASSIFIED SCANNING ELECTRON MICROSCOPE STUDIES ON AGGREGATION CHARACTERISTICS OF ALUMINA NANOFLUIDS INTERIM REPORT TFLRF No. 443...UNCLASSIFIED UNCLASSIFIED UNCLASSIFIED SCANNING ELECTRON MICROSCOPE STUDIES ON AGGREGATION CHARACTERISTICS OF ALUMINA NANOFLUIDS INTERIM REPORT TFLRF...Aggregation Characteristics of Alumina Nanofluids 5a. CONTRACT NUMBER W56HZV-09-C-0100 5b. GRANT NUMBER 5c. PROGRAM ELEMENT NUMBER 6. AUTHOR(S

  11. 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.

  12. 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

  13. 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.

  14. 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.

  15. Reversed scan direction reduces electron beam damage in EBSD maps.

    PubMed

    Kidder, S; Prior, D

    2014-08-01

    The deleterious effects of electron beam damage on high-resolution electron backscatter diffraction (EBSD) maps of undeformed quartz are significantly reduced by scanning in the direction opposite to that dictated by widely used EBSD acquisition software. Higher quality electron backscatter patterns are produced when the electron beam moves progressively down the sample (the apparent 'up' direction in the resulting maps) for all step sizes where beam damage affects EBSD map quality (≤ ∼0.4 μm in this study). The relative improvement associated with downward scanning increases as step size is reduced. A comparison of high-resolution maps made in experimentally deformed quartz demonstrates that downward scanning reduces by a factor of ∼2 the lower limit in step size relative to maps scanned in the conventional direction. The electron beam damages quartz at its point of entry, forming ∼0.1-μm diameter bumps visible in Scanning electron microscope (SEM) images. Downward scanning produces better results because it minimizes the flux of electrons through these loci of damaged crystal. © 2014 The Authors Journal of Microscopy © 2014 Royal Microscopical Society.

  16. Microcircuit testing and fabrication, using scanning electron microscopes

    NASA Technical Reports Server (NTRS)

    Nicolas, D. P.

    1975-01-01

    Scanning electron microscopes are used to determine both user-induced damages and manufacturing defects subtle enough to be missed by conventional light microscopy. Method offers greater depth of field and increased working distances.

  17. 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

  18. 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.

  19. 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.

  20. 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.

  1. 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.

  2. Electronically scanned analog liquid crystal displays.

    PubMed

    Soref, R A

    1970-06-01

    A new analog display technique for liquid crystal display panels is demonstrated. The size, shape, and location of display patterns can be changed continuously using low power electronic control. The display consists of a thin liquid crystal layer sandwiched between high resistance transparent area electrodes. Transverse voltage gradients on the electrodes actuate the device. The display operates with either dynamic scattering liquids or quiescent scattering liquids. Experimental results are given for three prototype analog displays: a voltmeter, a flying spot scanner, and a null indicator.

  3. UAVSAR Active Electronically-Scanned Array

    NASA Technical Reports Server (NTRS)

    Sadowy, Gregory; Brown, Kyle; Chamberlain, Neil; Figueroa, Harry; Fisher, Charlie; Grando, Maurio; Hamilton, Gary; Vorperian, Vatche; Zawadzki, Mark

    2010-01-01

    The Uninhabited Airborne Vehicle Synthetic Aperture Radar (UAVSAR) L-band (1.2-1.3 GHz) repeat pass, interferometric synthetic aperture radar (InSAR) used for Earth science applications. Using complex radar images collected during separate passes on time scales of hours to years, changes in surface topography can be measured. The repeat-pass InSAR technique requires that the radar look angle be approximately the same on successive passes. Due to variations in aircraft attitude between passes, antenna beam steering is required to replicate the radar look angle. This paper describes an active, electronically steered array (AESA) that provides beam steering capability in the antenna azimuth plane. The array contains 24 transmit/receive modules generating 2800 W of radiated power and is capable of pulse-to-pulse beam steering and polarization agility. Designed for high reliability as well as serviceability, all array electronics are contained in single 178cm x 62cm x 12 cm air-cooled panel suitable for operation up 60,000 ft altitude.

  4. 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.

  5. UAVSAR Active Electronically-Scanned Array

    NASA Technical Reports Server (NTRS)

    Sadowy, Gregory; Brown, Kyle; Chamberlain, Neil; Figueroa, Harry; Fisher, Charlie; Grando, Maurio; Hamilton, Gary; Vorperian, Vatche; Zawadzki, Mark

    2010-01-01

    The Uninhabited Airborne Vehicle Synthetic Aperture Radar (UAVSAR) L-band (1.2-1.3 GHz) repeat pass, interferometric synthetic aperture radar (InSAR) used for Earth science applications. Using complex radar images collected during separate passes on time scales of hours to years, changes in surface topography can be measured. The repeat-pass InSAR technique requires that the radar look angle be approximately the same on successive passes. Due to variations in aircraft attitude between passes, antenna beam steering is required to replicate the radar look angle. This paper describes an active, electronically steered array (AESA) that provides beam steering capability in the antenna azimuth plane. The array contains 24 transmit/receive modules generating 2800 W of radiated power and is capable of pulse-to-pulse beam steering and polarization agility. Designed for high reliability as well as serviceability, all array electronics are contained in single 178cm x 62cm x 12 cm air-cooled panel suitable for operation up 60,000 ft altitude.

  6. Theoretical Evaluation of Compositional Contrast of Scanning Electron Microscope Images

    NASA Astrophysics Data System (ADS)

    Kotera, Masatoshi; Yamaguchi, Satoru; Fujiwara, Takafumi; Suga, Hiroshi

    1992-12-01

    The compositional contrast in the scanning electron microscope image is calculated for Al-Cu, Si-Cu and Al-Si contacts. An electron scattering phenomenon in the specimen is simulated in a direct manner. Electron refraction at the boundary, because of the agreement of each Fermi energy at the boundary, is precisely taken into account. The backscattered electron image shows better resolution than the secondary electron image in terms of the boundary contrast when the primary electron energy is 1 keV. The signal intensity varies depending on materials adjacent to the location observed. The ultimate resolution of the compositional contrast of the scanning electron microscope can be below 1 nm.

  7. 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.

  8. Comparative study of depth and lateral distributions of electron excitation between scanning ion and scanning electron microscopes.

    PubMed

    Ohya, Kaoru; Ishitani, Tohru

    2003-01-01

    In order to study the contrast difference between scanning ion microscopes (SIM) and scanning electron microscopes (SEM), the depth and lateral distributions of secondary electrons escaped from surfaces of 17 metals with atomic numbers, Z2, of 4-79 were calculated for bombardment with 30 keV Ga ions and for 10 keV electrons. For both projectiles, the excitation depth generally decreased with increasing Z2, while showing the same periodic change as the secondary-electron yield. However, an opposite trend in Z2 dependence between the Ga ion and electron bombardments was calculated with the lateral distribution of secondary electrons escaped from the surface. Except for low Z2 metals, the lateral distribution, which is much narrower for 30 keV Ga ions than for 10 keV electrons, indicates that the spatial resolution of the secondary-electron images is better for SIM than for SEM, if zero-sized probe beams are assumed. Furthermore, the present calculation reveals important effects of electron excitation by recoiled material atoms and reflected electrons on the lateral distribution, as well as the secondary-electron yield, for the Ga ion and electron bombardments, respectively.

  9. 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.

  10. 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.

  11. Ion charge neutralization effects in scanning electron microscopes.

    PubMed

    Crawford, C K

    1980-01-01

    The use of low energy ion charge neutralization to stabilize surface potentials in scanning microscopes leads to the observation of new effects. Among the most important of these, are effects which result from the primary beam being scanned in a raster. A new theory which describes raster charge-up for highly insulating specimens is presented. It is shown that the required neutralizing ion current is a surprisingly strong function of the primary electron current, the raster parameters, specimen parameters, and magnification. Contrary to intuition, the required ion current is not linearly related to the primary electron current. Methods of adjusting parameters to achieve better ion charge neutralization are discussed.

  12. Light and scanning electron microscopic report of four fractured implants.

    PubMed

    Piattelli, A; Piattelli, M; Scarano, A; Montesani, L

    1998-01-01

    Although they are fortunately rare, implant fractures can cause significant problems for both clinicians and patients. The authors present a light and scanning electron microscopic study of four fractured implants in two patients. Both patients had parafunctional habits (bruxism), hypertrophic masticatory muscles, and wear of occlusal surfaces. The scanning electron microscopic study of the fractured surfaces of all four implants showed the presence of fatigue striations. Bending overload was probably created by a combination of parafunctional forces, bone resorption, posterior location of the implants, and implant diameter.

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

    PubMed Central

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

    1969-01-01

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

  14. 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.

  15. 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

  16. 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.

  17. [Scanning electron microscope study of chemically disinfected endodontic files].

    PubMed

    Navarro, G; Mateos, M; Navarro, J L; Canalda, C

    1991-01-01

    Forty stainless steel endodontic files were observed at scanning electron microscopy after being subjected to ten disinfection cycles of 10 minutes each one, immersed in different chemical disinfectants. Corrosion was not observed on the surface of the files in circumstances that this study was made.

  18. 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.

  19. Microstress contrast in scanning electron acoustic microscopy of ceramics

    NASA Technical Reports Server (NTRS)

    Cantrell, John H.; Qian, Menglu

    1991-01-01

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

  20. 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.

  1. Scanning electron microscope facility for examination of radioactive materials

    SciTech Connect

    Gibson, J.R.; Braski, D.N.

    1985-02-01

    An AMRAY model 1200B scanning electron microscope was modified to permit remote examination of radioactive specimens. Features of the modification include pneumatic vibration isolation of the column, motorized stage controls, improvements for monitoring vacuum, and a system for changing filaments without entering the hot cell.

  2. 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.

  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. Multi-channel electronically scanned cryogenic pressure sensor

    NASA Technical Reports Server (NTRS)

    Chapman, John J. (Inventor); Hopson, Purnell, Jr. (Inventor); Kruse, Nancy M. H. (Inventor)

    1995-01-01

    A miniature, multi-channel, electronically scanned pressure measuring device uses electrostatically bonded silicon dies in a multielement array. These dies are bonded at specific sites on a glass, prepatterned substrate. Thermal data is multiplexed and recorded on each individual pressure measuring diaphragm. The device functions in a cryogenic environment without the need of heaters to keep the sensor at constant temperatures.

  5. 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.

  6. 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.

  7. Development of scanning electron and x-ray microscope

    SciTech Connect

    Matsumura, Tomokazu Hirano, Tomohiko Suyama, Motohiro

    2016-01-28

    We have developed a new type of microscope possessing a unique feature of observing both scanning electron and X-ray images under one unit. Unlike former X-ray microscopes using SEM [1, 2], this scanning electron and X-ray (SELX) microscope has a sample in vacuum, thus it enables one to observe a surface structure of a sample by SEM mode, to search the region of interest, and to observe an X-ray image which transmits the region. For the X-ray observation, we have been focusing on the soft X-ray region from 280 eV to 3 keV to observe some bio samples and soft materials. The resolutions of SEM and X-ray modes are 50 nm and 100 nm, respectively, at the electron energy of 7 keV.

  8. 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.

  9. Millimeter-wave electronically scanned reflectarray optimization and analysis

    NASA Astrophysics Data System (ADS)

    Hedden, Abigail S.; Dietlein, Charles R.; Wikner, David A.

    2012-06-01

    The development of millimeter-wave scanning reflectarrays and phased arrays provides an important path to enabling electronic scanning capabilities at high frequencies. This technology could be used to eliminate the mechanical scanners that are currently used with radar imaging systems. In this work, we analyze properties of wafer-scale two-dimensional rectangular lattice arrays that can be used with a confocal imager for 220 GHz electronic scanning of meter-sized fields of regard at 50 m. Applications include covert imaging of hidden anomalies. We examine tradeoffs between overall system size and array complexity and analyze properties of reflectarrays compatible with a system design that was chosen based on these considerations. The effects of phase quantization are considered in detail for arrays with 1- and 2- bit phase shifters and the results are compared in terms of impacts to image quality. Beam pointing accuracy, main beam energy fraction, and the number and intensity of quantization lobes that appear over the scan ranges of interest are compared. Our results indicate that arrays with 1- and 2-bit phase quantization achieve similar main beam energy efficiencies over the desired scan range. Without restricting the scan range, 1-bit phase quantization is insufficient, resulting in maximum errors that are comparable to the required minimum scan angle. Two-bit phase quantization is preferable, resulting in pointing angle errors of at most 15 % of the diffraction-limited beam-size. Both 1- and 2-bit phase quantization cases result in lobes appearing above our threshold, indicating that spurious returns are a problem that will require further attention.

  10. Miniaturized Environmental Scanning Electron Microscope for In Situ Planetary Studies

    NASA Technical Reports Server (NTRS)

    Gaskin, Jessica; Abbott, Terry; Medley, Stephanie; Gregory, Don; Thaisen, Kevin; Taylor , Lawrence; Ramsey, Brian; Jerman, Gregory; Sampson, Allen; Harvey, Ralph

    2010-01-01

    The exploration of remote planetary surfaces calls for the advancement of low power, highly-miniaturized instrumentation. Instruments of this nature that are capable of multiple types of analyses will prove to be particularly useful as we prepare for human return to the moon, and as we continue to explore increasingly remote locations in our Solar System. To this end, our group has been developing a miniaturized Environmental-Scanning Electron Microscope (mESEM) capable of remote investigations of mineralogical samples through in-situ topographical and chemical analysis on a fine scale. The functioning of an SEM is well known: an electron beam is focused to nanometer-scale onto a given sample where resulting emissions such as backscattered and secondary electrons, X-rays, and visible light are registered. Raster scanning the primary electron beam across the sample then gives a fine-scale image of the surface topography (texture), crystalline structure and orientation, with accompanying elemental composition. The flexibility in the types of measurements the mESEM is capable of, makes it ideally suited for a variety of applications. The mESEM is appropriate for use on multiple planetary surfaces, and for a variety of mission goals (from science to non-destructive analysis to ISRU). We will identify potential applications and range of potential uses related to planetary exploration. Over the past few of years we have initiated fabrication and testing of a proof-of-concept assembly, consisting of a cold-field-emission electron gun and custom high-voltage power supply, electrostatic electron-beam focusing column, and scanning-imaging electronics plus backscatter detector. Current project status will be discussed. This effort is funded through the NASA Research Opportunities in Space and Earth Sciences - Planetary Instrument Definition and Development Program.

  11. 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.

  12. 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.

  13. Influence of mechanical noise inside a scanning electron microscope

    SciTech Connect

    Gaudenzi de Faria, Marcelo; Haddab, Yassine Le Gorrec, Yann; Lutz, Philippe

    2015-04-15

    The scanning electron microscope is becoming a popular tool to perform tasks that require positioning, manipulation, characterization, and assembly of micro-components. However, some of these applications require a higher level of performance with respect to dynamics and precision of positioning. One limiting factor is the presence of unidentified noises and disturbances. This work aims to study the influence of mechanical disturbances generated by the environment and by the microscope, identifying how these can affect elements in the vacuum chamber. To achieve this objective, a dedicated setup, including a high-resolution vibrometer, was built inside the microscope. This work led to the identification and quantification of main disturbances and noise sources acting on a scanning electron microscope. Furthermore, the effects of external acoustic excitations were analysed. Potential applications of these results include noise compensation and real-time control for high accuracy tasks.

  14. Influence of mechanical noise inside a scanning electron microscope.

    PubMed

    de Faria, Marcelo Gaudenzi; Haddab, Yassine; Le Gorrec, Yann; Lutz, Philippe

    2015-04-01

    The scanning electron microscope is becoming a popular tool to perform tasks that require positioning, manipulation, characterization, and assembly of micro-components. However, some of these applications require a higher level of performance with respect to dynamics and precision of positioning. One limiting factor is the presence of unidentified noises and disturbances. This work aims to study the influence of mechanical disturbances generated by the environment and by the microscope, identifying how these can affect elements in the vacuum chamber. To achieve this objective, a dedicated setup, including a high-resolution vibrometer, was built inside the microscope. This work led to the identification and quantification of main disturbances and noise sources acting on a scanning electron microscope. Furthermore, the effects of external acoustic excitations were analysed. Potential applications of these results include noise compensation and real-time control for high accuracy tasks.

  15. 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.

  16. Cryo-scanning transmission electron tomography of vitrified cells.

    PubMed

    Wolf, Sharon Grayer; Houben, Lothar; Elbaum, Michael

    2014-04-01

    Cryo-electron tomography (CET) of fully hydrated, vitrified biological specimens has emerged as a vital tool for biological research. For cellular studies, the conventional imaging modality of transmission electron microscopy places stringent constraints on sample thickness because of its dependence on phase coherence for contrast generation. Here we demonstrate the feasibility of using scanning transmission electron microscopy for cryo-tomography of unstained vitrified specimens (CSTET). We compare CSTET and CET for the imaging of whole bacteria and human tissue culture cells, finding favorable contrast and detail in the CSTET reconstructions. Particularly at high sample tilts, the CSTET signals contain more informative data than energy-filtered CET phase contrast images, resulting in improved depth resolution. Careful control over dose delivery permits relatively high cumulative exposures before the onset of observable beam damage. The increase in acceptable specimen thickness broadens the applicability of electron cryo-tomography.

  17. Scanning electron microscope studies of human metaphase chromosomes.

    PubMed

    Shemilt, L A; Estandarte, A K C; Yusuf, M; Robinson, I K

    2014-03-06

    Scanning electron microscopy (SEM) is used to evaluate potential chromosome preparations and staining methods for application in high-resolution three-dimensional X-ray imaging. Our starting point is optical fluorescence microscopy, the standard method for chromosomes, which only gives structural detail at the 200 nm scale. In principle, with suitable sample preparation protocols, including contrast enhancing staining, the surface structure of the chromosomes can be viewed at the 1 nm level by SEM. Here, we evaluate a heavy metal nucleic-acid-specific stain, which gives strong contrast in the backscattered electron signal. This study uses SEM to examine chromosomes prepared in different ways to establish a sample preparation protocol for X-rays. Secondary electron and backscattered electron signals are compared to evaluate the effectiveness of platinum-based stains used to enhance the contrast.

  18. 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.

  19. 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.

  20. 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.

  1. A fast iterative technique for restoring scanning electron microscope images

    NASA Astrophysics Data System (ADS)

    Nakahira, Kenji; Miyamoto, Atsushi; Honda, Toshifumi

    2014-12-01

    This paper proposes a fast new technique for restoring scanning electron microscope images to improve their sharpness. The images with our approach are sharpened by deconvolution with the point spread function modeled as the intensity distribution of the electron beam at the specimen's surface. We propose an iterative technique that employs a modified cost function based on the Richardson-Lucy method to achieve faster processing. The empirical results indicate significant improvements in image quality. The proposed approach speeds up deconvolution by about 10-50 times faster than that with the conventional Richardson-Lucy method.

  2. 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.

  3. Scanning electron microscopic study on Toxascaris transfuga (Rudolphi, 1819) (Nematoda).

    PubMed

    Tenora, F; Mituch, J; Hovorka, I

    1989-01-01

    The authors present original observations on the species Toxascaris transfuga obtained by means of scanning electron microscopy. Attention was paid to the structure of head end, morphology of papillae of the head and abdominal end, specific morphological traits of cloacae edges and morphology of the egg surface. Presented are morphological criteria which apparently differentiate the species T. transfuga from T. leonina (Linstow, 1902). T. transfuga and T. multipapillata Kreis, 1938 seem to be conspecific.

  4. 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.

  5. Rapidly and continuously frequency-scanning opto-electronic oscillator.

    PubMed

    Cen, Qizhuang; Dai, Yitang; Yin, Feifei; Zhou, Yue; Li, Jianqiang; Dai, Jian; Yu, Lan; Xu, Kun

    2017-01-23

    An opto-electronic oscillator (OEO) scheme which operates at "chirp oscillation" mode and generates low-phase-noise, frequency-swept microwave is proposed and experimentally demonstrated. This frequency-swept OEO is achieved by embedding a rapidly frequency-scanning microwave filter in an opto-electronic cavity. The filter has fixed passband while its center frequency scans rapidly and periodically at cavity round-trip time, covering a large frequency range (~GHz). Experimentally, the generated frequency-swept microwave is linear frequency-modulated continuous wave (FMCW) which centers at 7 GHz with 1-GHz bandwidth. Its instantaneous frequency varies linearly from 6.5 GHz to 7.5 GHz, back and forth, in a period of 12.8 μs, resulting in a frequency scanning rate of ~156 MHz/μs. The single-side-band (SSB) noise of the generated FMCW is -104 dBc/Hz at 10 kHz offset frequency, which is much lower than that from a commercial electronic arbitrary waveform generator (E-AWG). Improvement as large as 23 dB is experimentally reported.

  6. Response function and optimum configuration of semiconductor backscattered-electron detectors for scanning electron microscopes

    SciTech Connect

    Rau, E. I.; Orlikovskiy, N. A.; Ivanova, E. S.

    2012-06-15

    A new highly efficient design for semiconductor detectors of intermediate-energy electrons (1-50 keV) for application in scanning electron microscopes is proposed. Calculations of the response function of advanced detectors and control experiments show that the efficiency of the developed devices increases on average twofold, which is a significant positive factor in the operation of modern electron microscopes in the mode of low currents and at low primary electron energies.

  7. 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.

  8. Simulation and Characterization of a Miniaturized Scanning Electron Microscope

    NASA Technical Reports Server (NTRS)

    Gaskin, Jessica A.; Jerman, Gregory A.; Medley, Stephanie; Gregory, Don; Abbott, Terry O.; Sampson, Allen R.

    2011-01-01

    A miniaturized Scanning Electron Microscope (mSEM) for in-situ lunar investigations is being developed at NASA Marshall Space Flight Center with colleagues from the University of Alabama in Huntsville (UAH), Advanced Research Systems (ARS), the University of Tennessee in Knoxville (UTK) and Case Western Reserve University (CWRU). This effort focuses on the characterization of individual components of the mSEM and simulation of the complete system. SEMs can provide information on the size, shape, morphology and chemical composition of lunar regolith. Understanding these basic properties will allow us to better estimate the challenges associated with In-Situ Resource Utilization and to improve our basic science knowledge of the lunar surface (either precluding the need for sample return or allowing differentiation of unique samples to be returned to Earth.) The main components of the mSEM prototype includes: a cold field emission electron gun (CFEG), focusing lens, deflection/scanning system and backscatter electron detector. Of these, the electron gun development is of particular importance as it dictates much of the design of the remaining components. A CFEG was chosen for use with the lunar mSEM as its emission does not depend on heating of the tungsten emitter (lower power), it offers a long operation lifetime, is orders of magnitude brighter than tungsten hairpin guns, has a small source size and exhibits low beam energy spread.

  9. 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.

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

    PubMed

    Kubota, Yoshiyuki

    2015-02-01

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

  11. In situ laser processing in a scanning electron microscope

    SciTech Connect

    Roberts, Nicholas A.; Magel, Gregory A.; Hartfield, Cheryl D.; Moore, Thomas M.; Fowlkes, Jason D.; Rack, Philip D.

    2012-07-15

    Laser delivery probes using multimode fiber optic delivery and bulk focusing optics have been constructed and used for performing materials processing experiments within scanning electron microscope/focused ion beam instruments. Controlling the current driving a 915-nm semiconductor diode laser module enables continuous or pulsed operation down to sub-microsecond durations, and with spot sizes on the order of 50 {mu}m diameter, achieving irradiances at a sample surface exceeding 1 MW/cm{sup 2}. Localized laser heating has been used to demonstrate laser chemical vapor deposition of Pt, surface melting of silicon, enhanced purity, and resistivity via laser annealing of Au deposits formed by electron beam induced deposition, and in situ secondary electron imaging of laser induced dewetting of Au metal films on SiO{sub x}.

  12. In situ laser processing in a scanning electron microscope

    SciTech Connect

    Roberts, Nicholas; Fowlkes, Jason Davidson; Rack, Prof. Philip; Moore, Tom; Magel, Greg; Hartfield, Cheryl

    2012-01-01

    Laser delivery probes using multimode fiber optic delivery and bulk focusing optics have been constructed and used for performing materials processing experiments within scanning electron microscope/focused ion beam instruments. Controlling the current driving a 915-nm semiconductor diode laser module enables continuous or pulsed operation down to sub-microsecond durations, and with spot sizes on the order of 50 {micro}m diameter, achieving irradiances at a sample surface exceeding 1 MW/cm{sup 2}. Localized laser heating has been used to demonstrate laser chemical vapor deposition of Pt, surface melting of silicon, enhanced purity, and resistivity via laser annealing of Au deposits formed by electron beam induced deposition, and in situ secondary electron imaging of laser induced dewetting of Au metal films on SiO{sub x}.

  13. 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.

  14. 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.

  15. 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.

  16. 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.

  17. 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.

  18. 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.

  19. Examination of Surveyor 3 parts with the scanning electron microscope and electron microprobe

    NASA Technical Reports Server (NTRS)

    Chodos, A. A.; Devaney, J. R.; Evens, K. C.

    1972-01-01

    Two screws and two washers, several small chips of tubing, and a fiber removed from a third screw were examined with the scanning electron microscope and the electron microprobe. The purpose of the examination was to determine the nature of the material on the surface of these samples and to search for the presence of meteoritic material.

  20. Electronic scanning pressure measuring system and transducer package

    NASA Technical Reports Server (NTRS)

    Coe, C. F. (Inventor); Parra, G. T.

    1984-01-01

    An electronic scanning pressure system that includes a plurality of pressure transducers is examined. A means obtains an electrical signal indicative of a pressure measurement from each of the plurality of pressure transducers. A multiplexing means is connected for selectivity supplying inputs from the plurality of pressure transducers to the signal obtaining means. A data bus connects the plurality of pressure transducers to the multiplexing means. A latch circuit is connected to supply control inputs to the multiplexing means. An address bus is connected to supply an address signal of a selected one of the plurality of pressure transducers to the latch circuit. In operation, each of the pressure transducers is successively scanned by the multiplexing means in response to address signals supplied on the address bus to the latch circuit.

  1. 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.

  2. 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.

  3. Phase Identification in a Scanning Electron Microscope Using Backscattered Electron Kikuchi Patterns

    PubMed Central

    Goehner, R. P.; Michael, J. R.

    1996-01-01

    Backscattered electron Kikuchi patterns (BEKP) suitable for crystallographic phase analysis can be collected in the scanning electron microscope (SEM) with a newly developed charge coupled device (CCD) based detector. Crystallographic phase identification using BEKP in the SEM is unique in that it permits high magnification images and BEKPs to be collected from a bulk specimen. The combination of scanning electron microscope (SEM) imaging, BEKP, and energy dispersive x-ray spectrometry holds the promise of a powerful new tool for materials science. PMID:27805167

  4. A new apparatus for electron tomography in the scanning electron microscope

    SciTech Connect

    Morandi, V. Maccagnani, P.; Masini, L.; Migliori, A.; Ortolani, L.; Pezza, A.; Del Marro, M.; Pallocca, G.; Vinciguerra, P.; Rossi, M.; Ferroni, M.; Sberveglieri, G.; Vittori-Antisari, M.

    2015-06-23

    The three-dimensional reconstruction of a microscopic specimen has been obtained by applying the tomographic algorithm to a set of images acquired in a Scanning Electron Microscope. This result was achieved starting from a series of projections obtained by stepwise rotating the sample under the beam raster. The Scanning Electron Microscope was operated in the scanning-transmission imaging mode, where the intensity of the transmitted electron beam is a monotonic function of the local mass-density and thickness of the specimen. The detection strategy has been implemented and tailored in order to maintain the projection requirement over the large tilt range, as required by the tomographic workflow. A Si-based electron detector and an eucentric-rotation specimen holder have been specifically developed for the purpose.

  5. [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).

  6. 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.

  7. Microcircuit failure analysis using the SEM. [Scanning Electron Microscopes

    NASA Technical Reports Server (NTRS)

    Nicolas, D. P.

    1974-01-01

    The scanning electron microscope adds a new dimension to the knowledge that can be obtained from a failed microcircuit. When used with conventional techniques, SEM assists and clarifies the analysis, but it does not replace light microscopy. The most advantageous features for microcircuit analysis are long working distances and great depth of field. Manufacturer related failure modes of microcircuits are metallization defects, poor bonding, surface and particle contamination, and design and fabrication faults. User related failure modes are caused by abuse, such as overstress. The Physics of Failure Procedure followed by the Astrionics Laboratory in failure analysis is described, which is designed to obtain maximum information available from each step.

  8. Characteristics of different frequency ranges in scanning electron microscope images

    SciTech Connect

    Sim, K. S. Nia, M. E.; Tan, T. L.; Tso, C. P.; Ee, C. S.

    2015-07-22

    We demonstrate a new approach to characterize the frequency range in general scanning electron microscope (SEM) images. First, pure frequency images are generated from low frequency to high frequency, and then, the magnification of each type of frequency image is implemented. By comparing the edge percentage of the SEM image to the self-generated frequency images, we can define the frequency ranges of the SEM images. Characterization of frequency ranges of SEM images benefits further processing and analysis of those SEM images, such as in noise filtering and contrast enhancement.

  9. Microcircuit failure analysis using the SEM. [Scanning Electron Microscopes

    NASA Technical Reports Server (NTRS)

    Nicolas, D. P.

    1974-01-01

    The scanning electron microscope adds a new dimension to the knowledge that can be obtained from a failed microcircuit. When used with conventional techniques, SEM assists and clarifies the analysis, but it does not replace light microscopy. The most advantageous features for microcircuit analysis are long working distances and great depth of field. Manufacturer related failure modes of microcircuits are metallization defects, poor bonding, surface and particle contamination, and design and fabrication faults. User related failure modes are caused by abuse, such as overstress. The Physics of Failure Procedure followed by the Astrionics Laboratory in failure analysis is described, which is designed to obtain maximum information available from each step.

  10. 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.

  11. Controller for the Electronically Scanned Thinned Array Radiometer (ESTAR) instrument

    NASA Technical Reports Server (NTRS)

    Zomberg, Brian G.; Chren, William A., Jr.

    1994-01-01

    A prototype controller for the ESTAR (electronically scanned thinned array radiometer) instrument has been designed and tested. It manages the operation of the digital data subsystem (DDS) and its communication with the Small Explorer data system (SEDS). Among the data processing tasks that it coordinates are FEM data acquisition, noise removal, phase alignment and correlation. Its control functions include instrument calibration and testing of two critical subsystems, the output data formatter and Walsh function generator. It is implemented in a Xilinx XC3064PC84-100 field programmable gate array (FPGA) and has a maximum clocking frequency of 10 MHz.

  12. Adaptive noise Wiener filter for scanning electron microscope imaging system.

    PubMed

    Sim, K S; Teh, V; Nia, M E

    2016-01-01

    Noise on scanning electron microscope (SEM) images is studied. Gaussian noise is the most common type of noise in SEM image. We developed a new noise reduction filter based on the Wiener filter. We compared the performance of this new filter namely adaptive noise Wiener (ANW) filter, with four common existing filters as well as average filter, median filter, Gaussian smoothing filter and the Wiener filter. Based on the experiments results the proposed new filter has better performance on different noise variance comparing to the other existing noise removal filters in the experiments.

  13. [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.

  14. Single cylinder in situ scanning electron microscope fatigue system

    SciTech Connect

    Smiltneek, Larry; Shinde, Sachin R.; Hoeppner, David W.

    2006-01-15

    This article introduces a single cylinder fatigue machine adaptable to a scanning electron microscope chamber. The machine includes a node control mechanism to create a still observation node at any location on the specimen as fatigue cycling occurs, thereby allowing a point of interest to remain within view. The exceptional stability of this machine enables improved in situ study of the fatigue cracking phenomenon. For example, an in situ machine enhances the researcher's ability to record material structural changes that precede crack nucleation and allows observation of the influences of microstructure (grain structure) on the early stages of crack propagation.

  15. Dynamic-scanning-electron-microscope study of friction and wear

    NASA Technical Reports Server (NTRS)

    Brainard, W. A.; Buckley, D. H.

    1974-01-01

    A friction and wear apparatus was built into a real time scanning electron microscope (SEM). The apparatus and SEM comprise a system which provides the capability of performing dynamic friction and wear experiments in situ. When the system is used in conjunction with dispersive X-ray analysis, a wide range of information on the wearing process can be obtained. The type of wear and variation with speed, load, and time can be investigated. The source, size, and distribution of wear particles can be determined and metallic transferal observed. Some typical results obtained with aluminum, copper, and iron specimens are given.

  16. 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.

  17. 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.

  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. 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.

  20. 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.

  1. 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.

  2. Imaging of vortices in superconductors by electron beam scanning

    SciTech Connect

    Martin, J.; Huebener, R.P.; le Grand, J.B.; Mears, C.A.; Labov, S.E.; Barfknecht, A.T.

    1998-07-01

    Abrikosov vortices trapped in a superconducting tunnel junction and oriented perpendicular to the barrier plane were imaged by electron beam scanning at 1.6 K. We have used NbAlO{sub x}Nb junctions. As an important feature, the top Nb electrode was covered with a SiO{sub 2} film of 300 nm thickness, absorbing most of the 5 keV beam energy. The signal generating the image is explained by a model, assuming that the beam-induced electronic excitations in the SiO{sub 2} overlay film are trapped in the local magnetic field protruding from a vortex, resulting in an increased recombination rate. In addition to providing a novel approach to the imaging of the vortices in superconductors, our results are important for understanding quasiparticle losses in tunnel junction detectors. {copyright} {ital 1998 American Institute of Physics.}

  3. 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.

  4. 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.

  5. 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.

  6. 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.

  7. 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

  8. Simulations and measurements in scanning electron microscopes at low electron energy.

    PubMed

    Walker, Christopher G H; Frank, Luděk; Müllerová, Ilona

    2016-11-01

    The advent of new imaging technologies in Scanning Electron Microscopy (SEM) using low energy (0-2 keV) electrons has brought about new ways to study materials at the nanoscale. It also brings new challenges in terms of understanding electron transport at these energies. In addition, reduction in energy has brought new contrast mechanisms producing images that are sometimes difficult to interpret. This is increasing the push for simulation tools, in particular for low impact energies of electrons. The use of Monte Carlo calculations to simulate the transport of electrons in materials has been undertaken by many authors for several decades. However, inaccuracies associated with the Monte Carlo technique start to grow as the energy is reduced. This is not simply associated with inaccuracies in the knowledge of the scattering cross-sections, but is fundamental to the Monte Carlo technique itself. This is because effects due to the wave nature of the electron and the energy band structure of the target above the vacuum energy level become important and these are properties which are difficult to handle using the Monte Carlo method. In this review we briefly describe the new techniques of scanning low energy electron microscopy and then outline the problems and challenges of trying to understand and quantify the signals that are obtained. The effects of charging and spin polarised measurement are also briefly explored. SCANNING 38:802-818, 2016. © 2016 Wiley Periodicals, Inc.

  9. 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.

  10. Creating and Probing Graphene Electron Optics with Local Scanning Probes

    NASA Astrophysics Data System (ADS)

    Stroscio, Joseph

    Ballistic propagation and the light-like dispersion of graphene charge carriers make graphene an attractive platform for optics-inspired graphene electronics where gate tunable potentials can control electron refraction and transmission. In analogy to optical wave propagation in lenses, mirrors and metamaterials, gate potentials can be used to create a negative index of refraction for Veselago lensing and Fabry-Pérot interferometers. In circular geometries, gate potentials can induce whispering gallery modes (WGM), similar to optical and acoustic whispering galleries albeit on a much smaller length scale. Klein scattering of Dirac carriers plays a central role in determining the coherent propagation of electron waves in these resonators. In this talk, I examine the probing of electron resonators in graphene confined by linear and circular gate potentials with the scanning tunneling microscope (STM). The tip in the STM tunnel junction serves both as a tunable local gate potential, and as a probe of the graphene states through tunneling spectroscopy. A combination of a back gate potential, Vg, and tip potential, Vb, creates and controls a circular pn junction that confines the WGM graphene states. The resonances are observed in two separate channels in the tunneling spectroscopy experiment: first, by directly tunneling into the state at the bias energy eVb, and, second, by tunneling from the resonance at the Fermi level as the state is gated by the tip potential. The second channel produces a fan-like set of WGM peaks, reminiscent of the fringes seen in planar geometries by transport measurements. The WGM resonances split in a small applied magnetic field, with a large energy splitting approaching the WGM spacing at 0.5 T. These results agree well with recent theory on Klein scattering in graphene electron resonators. This work is done in collaboration with Y. Zhao, J. Wyrick, F.D. Natterer, J. F. Rodriquez-Nieva, C. Lewandoswski, K. Watanabe, T. Taniguchi, N. B

  11. 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.

  12. 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

  13. Environmental Scanning Electron Microscope Imaging of Vesicle Systems.

    PubMed

    Perrie, Yvonne; Ali, Habib; Kirby, Daniel J; Mohammed, Afzal U R; McNeil, Sarah E; Vangala, Anil

    2017-01-01

    The structural characteristics of liposomes have been widely investigated and there is certainly a strong understanding of their morphological characteristics. Imaging of these systems, using techniques such as freeze-fracturing methods, transmission electron microscopy, and cryo-electron imaging, has allowed us to appreciate their bilayer structures and factors which can influence this. However, there are few methods which all us to study these systems in their natural hydrated state; commonly the liposomes are visualized after drying, staining, and/or fixation of the vesicles. Environmental Scanning Electron Microscopy (ESEM) offers the ability to image a liposome in its hydrated state without the need for prior sample preparation. Within our studies we were the first to use ESEM to study liposomes and niosomes and we have been able to dynamically follow the hydration of lipid films and changes in liposome suspensions as water condenses on to, or evaporates from, the sample in real time. This provides insight into the resistance of liposomes to coalescence during dehydration, thereby providing an alternative assay of liposome formulation and stability.

  14. 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.

  15. 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.

  16. 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

  17. 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.

  18. Electron beam detection of a Nanotube Scanning Force Microscope.

    PubMed

    Siria, Alessandro; Niguès, Antoine

    2017-09-14

    Atomic Force Microscopy (AFM) allows to probe matter at atomic scale by measuring the perturbation of a nanomechanical oscillator induced by near-field interaction forces. The quest to improve sensitivity and resolution of AFM forced the introduction of a new class of resonators with dimensions at the nanometer scale. In this context, nanotubes are the ultimate mechanical oscillators because of their one dimensional nature, small mass and almost perfect crystallinity. Coupled to the possibility of functionalisation, these properties make them the perfect candidates as ultra sensitive, on-demand force sensors. However their dimensions make the measurement of the mechanical properties a challenging task in particular when working in cavity free geometry at ambient temperature. By using a focused electron beam, we show that the mechanical response of nanotubes can be quantitatively measured while approaching to a surface sample. By coupling electron beam detection of individual nanotubes with a custom AFM we image the surface topography of a sample by continuously measuring the mechanical properties of the nanoresonators. The combination of very small size and mass together with the high resolution of the electron beam detection method offers unprecedented opportunities for the development of a new class of nanotube-based scanning force microscopy.

  19. 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.

  20. Smart flexible microrobots for scanning electron microscope (SEM) applications

    NASA Astrophysics Data System (ADS)

    Schmoeckel, Ferdinand; Fatikow, Sergej

    2000-06-01

    In the scanning electron microscope (SEM), specially designed microrobots can act as a flexible assembly facility for hybrid microsystems, as probing devices for in-situ tests on IC structures or just as a helpful teleoperated tool for the SEM operator when examining samples. Several flexible microrobots of this kind have been developed and tested. Driven by piezoactuators, these few cubic centimeters small mobile robots perform manipulations with a precision of up to 10 nm and transport the gripped objects at speeds of up to 3 cm/s. In accuracy, flexibility and price they are superior to conventional precision robots. A new SEM-suited microrobot prototype is described in this paper. The SEM's vacuum chamber has been equipped with various elements like flanges and CCD cameras to enable the robot to operate. In order to use the SEM image for the automatic real-time control of the robots, the SEM's electron beam is actively controlled by a PC. The latter submits the images to the robots' control computer system. For obtaining three-dimensional information in real time, especially for the closed-loop control of a robot endeffector, e.g. microgripper, a triangulation method with the luminescent spot of the SEM's electron beam is being investigated.

  1. 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.

  2. 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.

  3. Ultrahigh vacuum scanning electron microscope system combined with wide-movable scanning tunneling microscope

    SciTech Connect

    Kaneko, A.; Homma, Y.; Hibino, H.; Ogino, T.

    2005-08-15

    A surface analysis system has been newly developed with combination of ultrahigh vacuum scanning electron microscope (SEM) and wide-movable scanning tunneling microscope (STM). The basic performance is experimentally demonstrated. These SEM and STM images are clear enough to obtain details of surface structures. The STM unit moves horizontally over several millimeters by sliding motion of PZT actuators. The motion resolution is proved to be submicrometers. The STM tip mounted on another PZT scanner can be guided to a specific object on the sample surface during SEM observation. In the observation of a Si(111) surface rapidly cooled from high temperature, the STM tip was accurately guided to an isolated atomic step and slightly moved along it during SEM observation. The STM observation shows an asymmetry of the (7x7)-transformed region along the step between the upper and lower terraces. (7x7) bands continuously formed along the edge of terraces, while (7x7) domains distributed on the terraces slightly far from the step. These experiments show the wide-movable STM unit resolves a gap of observation area between SEM and STM and the system enables a specific object found in the SEM image to be observed easily by STM.

  4. Scanning electron microscopic observations of Anopheles albimanus (Diptera: Culicidae) eggs.

    PubMed

    Rodriguez, M H; Chavez, B; Orozco, A; Loyola, E G; Martinez-Palomo, A

    1992-05-01

    To investigate the existence of subspecies of Anopheles albimanus Wiedeman in southern Mexico, the egg morphology of specimens obtained from several field populations and from insectary-adapted colonies of uniform pupal phenotype was examined. Scanning electron microscopic observations have shown that the eggs of An. albimanus are polymorphic in respect to the size and shape of their floats, but not in their ornamentation. Four types of eggs were found. Differences in the proportion of the various morphological types were statistically significant, although proportions of egg types were variable among individuals within the same population. These observations are suggestive of distinctive populations and warrant further studies using more sensitive methods to investigate sibling species in An. albimanus sensu lato.

  5. Cylindrical Lens-Array Antenna for Wideband Electronic Scanning.

    DTIC Science & Technology

    1985-03-01

    Radome 3-21 Report No. 6546j vi WOW oft ma~ - I. Figure 3-14. Line-Feed And Housing Structure Wi thout Radome 3-20 -Zr -W -N .- .’ o - K -w .. - c - K ...I -7: 7 T Fr I~ ~ 7 .4 C 1I-7. 1,1 IF 7I1- -20 - o -----H Ii K 1 2 71 - ~ .~K14.L K ~j~jtK T ’,-. < 1 ’ IL .,, .,. I4 2 4 0 ANGL (DEGREES Fiur A-8...RD- f156 085 CYLINDRICAL LENS-RRAY ANEENNA FOR WIDEBAND ELECTRONIC 112SCAN ING(U) HAZELTINE CORP GREENL WN NY NEWAN ET AL. MAR 85 6546 F19628-80-C

  6. 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.

  7. [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.

  8. 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.

  9. An improved visual tracking method in scanning electron microscope.

    PubMed

    Ru, Changhai; Zhang, Yong; Huang, Haibo; Chen, Tao

    2012-06-01

    Since their invention, nanomanipulation systems in scanning electron microscopes (SEMs) have provided researchers with an increasing ability to interact with objects at the nanoscale. However, most nanomanipulators that are capable of generating nanometer displacement operate in an open-loop without suitable feedback mechanisms. In this article, a robust and effective tracking framework for visual servoing applications is presented inside an SEM to achieve more precise tracking manipulation and measurement. A subpixel template matching tracking algorithm based on contour models in the SEM has been developed to improve the tracking accuracy. A feed-forward controller is integrated into the control system to improve the response time. Experimental results demonstrate that a subpixel tracking accuracy is realized. Furthermore, the robustness against clutter can be achieved even in a challenging tracking environment.

  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. 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.

  12. 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.

  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. 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.

  15. 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.

  16. 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.

  17. 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

  18. 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

  19. 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.

  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. 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

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

    PubMed Central

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

    2010-01-01

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

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

    NASA Astrophysics Data System (ADS)

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

    2016-11-01

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

  4. 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.

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

    PubMed

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

    2016-11-16

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

  6. 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.

  7. 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.

  8. 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.

  9. 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.

  10. 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.

  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. 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.

  13. Scanning electron microscope automatic defect classification of process induced defects

    NASA Astrophysics Data System (ADS)

    Wolfe, Scott; McGarvey, Steve

    2017-03-01

    With the integration of high speed Scanning Electron Microscope (SEM) based Automated Defect Redetection (ADR) in both high volume semiconductor manufacturing and Research and Development (R and D), the need for reliable SEM Automated Defect Classification (ADC) has grown tremendously in the past few years. In many high volume manufacturing facilities and R and D operations, defect inspection is performed on EBeam (EB), Bright Field (BF) or Dark Field (DF) defect inspection equipment. A comma separated value (CSV) file is created by both the patterned and non-patterned defect inspection tools. The defect inspection result file contains a list of the inspection anomalies detected during the inspection tools' examination of each structure, or the examination of an entire wafers surface for non-patterned applications. This file is imported into the Defect Review Scanning Electron Microscope (DRSEM). Following the defect inspection result file import, the DRSEM automatically moves the wafer to each defect coordinate and performs ADR. During ADR the DRSEM operates in a reference mode, capturing a SEM image at the exact position of the anomalies coordinates and capturing a SEM image of a reference location in the center of the wafer. A Defect reference image is created based on the Reference image minus the Defect image. The exact coordinates of the defect is calculated based on the calculated defect position and the anomalies stage coordinate calculated when the high magnification SEM defect image is captured. The captured SEM image is processed through either DRSEM ADC binning, exporting to a Yield Analysis System (YAS), or a combination of both. Process Engineers, Yield Analysis Engineers or Failure Analysis Engineers will manually review the captured images to insure that either the YAS defect binning is accurately classifying the defects or that the DRSEM defect binning is accurately classifying the defects. This paper is an exploration of the feasibility of the

  14. 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

  15. Visualizing bone porosities using a tabletop scanning electron microscope

    NASA Astrophysics Data System (ADS)

    Krishnamoorthy, D.; DaPonte, J.; Broadbridge, C. C.; Daniel, D.; Alter, L.

    2010-04-01

    Pores are naturally occurring entities in bone. Changes in pore size and number are often associated with diseases such as Osteoporosis and even microgravity during spaceflight. Studying bone perforations may yield great insight into bone's material properties, including bone density and may contribute to identifying therapies to halt or potentially reverse bone loss. Current technologies used in this field include nuclear magnetic resonance, micro-computed tomography and the field emission scanning electron microscope (FE-SEM) 2, 5. However, limitations in each method limit further advancement. The objective of this study was to assess the effectiveness of using a new generation of analytical instruments, the TM-1000 tabletop, SEM with back-scatter electron (BSE) detector, to analyze cortical bone porosities. Hind limb unloaded and age-based controlled mouse femurs were extracted and tested in vitro for changes in pores on the periosteal surface. An important advantage of using the tabletop is the simplified sample preparation that excludes extra coatings, dehydration and fixation steps that are otherwise required for conventional SEM. For quantitative data, pores were treated as particles in order to use an analyze particles feature in the NIH ImageJ software. Several image-processing techniques for background smoothing, thresholding and filtering were employed to produce a binary image suitable for particle analysis. It was hypothesized that the unloaded bones would show an increase in pore area, as the lack of mechanical loading would affect bone-remodeling processes taking place in and around pores. Preliminary results suggest only a slight different in frequency but not in size of pores between unloaded and control femurs.

  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. Environmental scanning electron microscope imaging examples related to particle analysis.

    PubMed

    Wight, S A; Zeissler, C J

    1993-08-01

    This work provides examples of some of the imaging capabilities of environmental scanning electron microscopy applied to easily charged samples relevant to particle analysis. Environmental SEM (also referred to as high pressure or low vacuum SEM) can address uncoated samples that are known to be difficult to image. Most of these specimens are difficult to image by conventional SEM even when coated with a conductive layer. Another area where environmental SEM is particularly applicable is for specimens not compatible with high vacuum, such as volatile specimens. Samples from which images were obtained that otherwise may not have been possible by conventional methods included fly ash particles on an oiled plastic membrane impactor substrate, a one micrometer diameter fiber mounted on the end of a wire, uranium oxide particles embedded in oil-bearing cellulose nitrate, teflon and polycarbonate filter materials with collected air particulate matter, polystyrene latex spheres on cellulosic filter paper, polystyrene latex spheres "loosely" sitting on a glass slide, and subsurface tracks in an etched nuclear track-etch detector. Surface charging problems experienced in high vacuum SEMs are virtually eliminated in the low vacuum SEM, extending imaging capabilities to samples previously difficult to use or incompatible with conventional methods.

  18. 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.

  19. 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

  20. 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.

  1. 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.

  2. 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

  3. In situ fatigue loading stage inside scanning electron microscope

    NASA Technical Reports Server (NTRS)

    Telesman, Jack; Kantzos, Peter; Brewer, David

    1988-01-01

    A fatigue loading stage inside a scanning electron microscopy (SEM) was developed. The stage allows dynamic and static high-magnification and high-resolution viewing of the fatigue crack initiation and crack propagation processes. The loading stage is controlled by a closed-loop servohydraulic system. Maximum load is 1000 lb (4450 N) with test frequencies ranging up to 30 Hz. The stage accommodates specimens up to 2 inches (50 mm) in length and tolerates substantial specimen translation to view the propagating crack. At room temperature, acceptable working resolution is obtainable for magnifications ranging up to 10,000X. The system is equipped with a high-temperature setup designed for temperatures up to 2000 F (1100 C). The signal can be videotaped for further analysis of the pertinent fatigue damage mechanisms. The design allows for quick and easy interchange and conversion of the SEM from a loading stage configuration to its normal operational configuration and vice versa. Tests are performed entirely in the in-situ mode. In contrast to other designs, the NASA design has greatly extended the life of the loading stage by not exposing the bellows to cyclic loading. The loading stage was used to investigate the fatigue crack growth mechanisms in the (100)-oriented PWA 1480 single-crystal, nickel-based supperalloy. The high-magnification observations revealed the details of the crack growth processes.

  4. Scanning electron microscopic and profilometric study of different sharpening stones.

    PubMed

    Andrade Acevedo, Roberto Antonio; Cardozo, Ana Karina Veloso; Sampaio, José Eduardo César

    2006-01-01

    Scaling and root planing contribute to the recovery of periodontal health. All periodontal instruments loose their fine cutting angle after use. To maintain this angle, correct sharpening is required using specifically designed stones. The characteristics of sharpening stones can be compared to the blade of the instruments and also transported to root surface during instrumentation. Root smoothness is related to the quality of the blade. Therefore, the purpose of this study was to evaluate the characteristics of 9 sharpening stones by scanning electron microscopic and profilometric analyses. Ceramic and Neumar stones were very fine and both may be recommended to maintain the sharpness of the instruments. Arkansas, Thompson and CE stones presented greater roughness with very regular and round particles, and are suitable for maintenance of the cutting angle. In addition, these stones may be indicated for the routine sharpening of the instruments that are partly dull. Oxide Aluminum, Carborundum and JON stones were the coarsest with large irregular particles and may be indicated for initial sharpening of totally dull instruments with completion of sharpening with finer stones.

  5. 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.

  6. 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.

  7. 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

  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. 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.

  10. 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.

  11. Semiautomatic classification of cementitious materials using scanning electron microscope images

    NASA Astrophysics Data System (ADS)

    Drumetz, Lucas; Mura, Mauro Dalla; Meulenyzer, Samuel; Lombard, Sébastien; Chanussot, Jocelyn

    2015-11-01

    Segmentation and classification are prolific research topics in the image processing community. These topics have been increasingly used in the context of analysis of cementitious materials on images acquired with a scanning electron microscope. Indeed, there is a need to be able to detect and to quantify the materials present in a cement paste in order to follow the chemical reactions occurring in the material even days after the solidification. We propose a new approach for segmentation and classification of cementitious materials based on the denoising of the data with a block-matching three-dimensional (3-D) algorithm, binary partition tree (BPT) segmentation, support vector machines (SVM) classification, and interactivity with the user. The BPT provides a hierarchical representation of the spatial regions of the data, allowing a segmentation to be selected among the admissible partitions of the image. SVMs are used to obtain a classification map of the image. This approach combines state-of-the-art image processing tools with user interactivity to allow a better segmentation to be performed, or to help the classifier discriminate the classes better. We show that the proposed approach outperforms a previous method when applied to synthetic data and several real datasets coming from cement samples, both qualitatively with visual examination and quantitatively with the comparison of experimental results with theoretical ones.

  12. 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.

  13. Ultra low-K shrinkage behavior when under electron beam in a scanning electron microscope

    SciTech Connect

    Lorut, F.; Imbert, G.; Roggero, A.

    2013-08-28

    In this paper, we investigate the tendency of porous low-K dielectrics (also named Ultra Low-K, ULK) behavior to shrink when exposed to the electron beam of a scanning electron microscope. Various experimental electron beam conditions have been used for irradiating ULK thin films, and the resulting shrinkage has been measured through use of an atomic force microscope tool. We report the shrinkage to be a fast, cumulative, and dose dependent effect. Correlation of the shrinkage with incident electron beam energy loss has also been evidenced. The chemical modification of the ULK films within the interaction volume has been demonstrated, with a densification of the layer and a loss of carbon and hydrogen elements being observed.

  14. Non-destructive imaging of buried electronic interfaces using a decelerated scanning electron beam

    PubMed Central

    Hirohata, Atsufumi; Yamamoto, Yasuaki; Murphy, Benedict A.; Vick, Andrew J.

    2016-01-01

    Recent progress in nanotechnology enables the production of atomically abrupt interfaces in multilayered junctions, allowing for an increase in the number of transistors in a processor. However, uniform electron transport has not yet been achieved across the entire interfacial area in junctions due to the existence of local defects, causing local heating and reduction in transport efficiency. To date, junction uniformity has been predominantly assessed by cross-sectional transmission electron microscopy, which requires slicing and milling processes that can potentially introduce additional damage and deformation. It is therefore essential to develop an alternative non-destructive method. Here we show a non-destructive technique using scanning electron microscopy to map buried junction properties. By controlling the electron-beam energy, we demonstrate the contrast imaging of local junction resistances at a controlled depth. This technique can be applied to any buried junctions, from conventional semiconductor and metal devices to organic devices. PMID:27586090

  15. Non-destructive imaging of buried electronic interfaces using a decelerated scanning electron beam

    NASA Astrophysics Data System (ADS)

    Hirohata, Atsufumi; Yamamoto, Yasuaki; Murphy, Benedict A.; Vick, Andrew J.

    2016-09-01

    Recent progress in nanotechnology enables the production of atomically abrupt interfaces in multilayered junctions, allowing for an increase in the number of transistors in a processor. However, uniform electron transport has not yet been achieved across the entire interfacial area in junctions due to the existence of local defects, causing local heating and reduction in transport efficiency. To date, junction uniformity has been predominantly assessed by cross-sectional transmission electron microscopy, which requires slicing and milling processes that can potentially introduce additional damage and deformation. It is therefore essential to develop an alternative non-destructive method. Here we show a non-destructive technique using scanning electron microscopy to map buried junction properties. By controlling the electron-beam energy, we demonstrate the contrast imaging of local junction resistances at a controlled depth. This technique can be applied to any buried junctions, from conventional semiconductor and metal devices to organic devices.

  16. Non-destructive imaging of buried electronic interfaces using a decelerated scanning electron beam.

    PubMed

    Hirohata, Atsufumi; Yamamoto, Yasuaki; Murphy, Benedict A; Vick, Andrew J

    2016-09-02

    Recent progress in nanotechnology enables the production of atomically abrupt interfaces in multilayered junctions, allowing for an increase in the number of transistors in a processor. However, uniform electron transport has not yet been achieved across the entire interfacial area in junctions due to the existence of local defects, causing local heating and reduction in transport efficiency. To date, junction uniformity has been predominantly assessed by cross-sectional transmission electron microscopy, which requires slicing and milling processes that can potentially introduce additional damage and deformation. It is therefore essential to develop an alternative non-destructive method. Here we show a non-destructive technique using scanning electron microscopy to map buried junction properties. By controlling the electron-beam energy, we demonstrate the contrast imaging of local junction resistances at a controlled depth. This technique can be applied to any buried junctions, from conventional semiconductor and metal devices to organic devices.

  17. Electron channeling contrast imaging studies of nonpolar nitrides using a scanning electron microscope

    SciTech Connect

    Naresh-Kumar, G.; Kraeusel, S.; Bruckbauer, J.; Edwards, P. R.; Hourahine, B.; Trager-Cowan, C.; Mauder, C.; Heuken, M.; Wang, K. R.; Trampert, A.; Kalisch, H.; Vescan, A.; Giesen, C.; Day, A. P.

    2013-04-08

    Threading dislocations, stacking faults, and associated partial dislocations significantly degrade the optical and electrical properties of materials such as non-polar III-nitride semiconductor thin films. Stacking faults are generally difficult to detect and quantify with existing characterization techniques. We demonstrate the use of electron channeling contrast imaging in the scanning electron microscope to non-destructively reveal basal plane stacking faults terminated by partial dislocations in m-plane GaN and InGaN/GaN multiple quantum well structures grown on {gamma}-LiAlO{sub 2} by metal organic vapor phase epitaxy.

  18. 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).

  19. Treatment of in-vivo bladder tissue with electronically scanned high-intensity focused ultrasound

    NASA Astrophysics Data System (ADS)

    Feuillu, Benoit; Lacoste, Francois; Schlosser, Jacques; Vallancien, Guy

    1998-04-01

    Introduction: The efficacy of extracorporeal High Intensity Focused Ultrasound (HIFU) on bladder wall has been demonstrated. However, the treatment is still slow, needing about 15 min to treat 1 cm2. Objectives: Demonstrate the feasibility of HIFU with electronic scanning and reduce the during of HIFU treatments. Conclusions: Necrotic lesions on bladder posterior wall can be obtained with HIFU treatments using electronic scanning of the focal point. Average treatment duration with electronic scanning is reduced to 3 min 30 sec/cm2.

  20. 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.

  1. SCAN+

    SciTech Connect

    Kenneth Krebs, John Svoboda

    2009-11-01

    SCAN+ is a software application specifically designed to control the positioning of a gamma spectrometer by a two dimensional translation system above spent fuel bundles located in a sealed spent fuel cask. The gamma spectrometer collects gamma spectrum information for the purpose of spent fuel cask fuel loading verification. SCAN+ performs manual and automatic gamma spectrometer positioning functions as-well-as exercising control of the gamma spectrometer data acquisitioning functions. Cask configuration files are used to determine the positions of spent fuel bundles. Cask scanning files are used to determine the desired scan paths for scanning a spent fuel cask allowing for automatic unattended cask scanning that may take several hours.

  2. 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.

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

    PubMed

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

    2015-08-01

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

  4. Improved specimen coating technique for scanning electron microscope observation of decomposer microorganisms.

    PubMed

    Draggan, S

    1976-02-01

    Sputter coating of leaf litter microbe samples provides scanning electron microscope images with greater information content than either vacuum evaporation of thin metal coatings or tissue conductance.

  5. Visualization of Microbial Biomarkers by Scanning Electron Microscopy

    NASA Technical Reports Server (NTRS)

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

    2001-01-01

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

  6. Visualization of Microbial Biomarkers by Scanning Electron Microscopy

    NASA Technical Reports Server (NTRS)

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

    2001-01-01

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

  7. 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

  8. Autofocus on moving object in scanning electron microscope.

    PubMed

    Kudryavtsev, Andrey V; Dembélé, Sounkalo; Piat, Nadine

    2017-07-12

    The sharpness of the images coming from a Scanning Electron Microscope (SEM) is a very important property for many computer vision applications at micro- and nanoscale. It represents how much object details are distinctive in the images: the object may be perceived sharp or blurred. Image sharpness highly depends on the value of focal distance, or working distance in the case of the SEM. Autofocus is the technique allowing to automatically adjust the working distance to maximize the sharpness. Most of the existing algorithms allows working only with a static object which is enough for the tasks of visualization, manual microanalysis or microcharacterization. These applications work with a low frame rate, less than 1 Hz, that guarantees a low level of noise. However, static autofocus can not be used for samples performing continuous 3D motion, which is the case of robotic applications where it is required to carry out a continuous 3D position measurement, e.g., nano-assembly or nanomanipulation. Moreover, in addition to constantly keeping object in focus while it is moving, it is required to perform the operation at high frame rate. The approach offering both these possibilities is presented in this paper and is referred as dynamic autofocus. The presented solution is based on stochastic optimization techniques. It allows tracking the maximum of the sharpness of the images without sweep and without training. It works under uncertainty conditions: presence of noise in images, unknown maximal sharpness and unknown 3D motion of the specimen. The experiments, that were performed with noisy images at high frame rate (5 Hz), were conducted on a Carl Zeiss Auriga 60 FE-SEM. They prove the robustness of the algorithm with respect to the variation of optimization parameters, object speed and magnification. Moreover, it is invariant to the object structure and its variation in time. Copyright © 2017 Elsevier B.V. All rights reserved.

  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. 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.

  11. Contrast curves for low energy electron exposures of an EUV resist in a scanning electron microscope

    NASA Astrophysics Data System (ADS)

    Bhattarai, Suchit; Neureuther, Andrew R.; Naulleau, Patrick P.

    2017-03-01

    We present an experimental technique for determining the energy delivery efficiency of secondary electrons in an EUV resist, by directly exposing a positive tone chemically amplified resist with 29- 91 eV electrons created by utilizing the deceleration technology in a scanning electron microscope. Charging is an important problem associated with thin film exposure experiments. We assess the feasibility of using the SEM frame rate as a knob for controlling charging related artifacts. Preliminary measurements of secondary electron emission signal from an unexposed region in the resist provide clues about the time domain surface potentials that may form while the sample charges during exposures. These signals are found to change as a function of the SEM frame rate and landing energies. We provide contrast curve data for resist exposures with 29 eV, 49 eV and 91 eV electrons at three frame rates of 33 ms/frame, 8 s/frame and 30 s/frame. The energy delivery efficiency of electrons estimated for all three frame rates are also provided.

  12. Sparse sampling and reconstruction for electron and scanning probe microscope imaging

    DOEpatents

    Anderson, Hyrum; Helms, Jovana; Wheeler, Jason W.; Larson, Kurt W.; Rohrer, Brandon R.

    2015-07-28

    Systems and methods for conducting electron or scanning probe microscopy are provided herein. In a general embodiment, the systems and methods for conducting electron or scanning probe microscopy with an undersampled data set include: driving an electron beam or probe to scan across a sample and visit a subset of pixel locations of the sample that are randomly or pseudo-randomly designated; determining actual pixel locations on the sample that are visited by the electron beam or probe; and processing data collected by detectors from the visits of the electron beam or probe at the actual pixel locations and recovering a reconstructed image of the sample.

  13. Electronic Single Molecule Measurements with the Scanning Tunneling Microscope

    NASA Astrophysics Data System (ADS)

    Im, Jong One

    Richard Feynman said "There's plenty of room at the bottom". This inspired the techniques to improve the single molecule measurements. Since the first single molecule study was in 1961, it has been developed in various field and evolved into powerful tools to understand chemical and biological property of molecules. This thesis demonstrates electronic single molecule measurement with Scanning Tunneling Microscopy (STM) and two of applications of STM; Break Junction (BJ) and Recognition Tunneling (RT). First, the two series of carotenoid molecules with four different substituents were investigated to show how substituents relate to the conductance and molecular structure. The measured conductance by STM-BJ shows that Nitrogen induces molecular twist of phenyl distal substituents and conductivity increasing rather than Carbon. Also, the conductivity is adjustable by replacing the sort of residues at phenyl substituents. Next, amino acids and peptides were identified through STM-RT. The distribution of the intuitive features (such as amplitude or width) are mostly overlapped and gives only a little bit higher separation probability than random separation. By generating some features in frequency and cepstrum domain, the classification accuracy was dramatically increased. Because of large data size and many features, supporting vector machine (machine learning algorithm for big data) was used to identify the analyte from a data pool of all analytes RT data. The STM-RT opens a possibility of molecular sequencing in single molecule level. Similarly, carbohydrates were studied by STM-RT. Carbohydrates are difficult to read the sequence, due to their huge number of possible isomeric configurations. This study shows that STM-RT can identify not only isomers of mono-saccharides and disaccharides, but also various mono-saccharides from a data pool of eleven analytes. In addition, the binding affinity between recognition molecule and analyte was investigated by comparing with

  14. 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.

  15. Suppressing Electron Exposure Artifacts: An Electron Scanning Paradigm with Bayesian Machine Learning.

    PubMed

    Hujsak, Karl; Myers, Benjamin D; Roth, Eric; Li, Yue; Dravid, Vinayak P

    2016-08-01

    Electron microscopy of biological, polymeric, and other beam-sensitive structures is often hampered by deleterious electron beam interactions. In fact, imaging of such beam-sensitive materials is limited by the allowable radiation dosage rather that capabilities of the microscope itself, which has been compounded by the availability of high brightness electron sources. Reducing dwell times to overcome dose-related artifacts, such as radiolysis and electrostatic charging, is challenging due to the inherently low contrast in imaging of many such materials. These challenges are particularly exacerbated during dynamic time-resolved, fluidic cell imaging, or three-dimensional tomographic reconstruction-all of which undergo additional dosage. Thus, there is a pressing need for the development of techniques to produce high-quality images at ever lower electron doses. In this contribution, we demonstrate direct dose reduction and suppression of beam-induced artifacts through under-sampling pixels, by as much as 80% reduction in dosage, using a commercial scanning electron microscope with an electrostatic beam blanker and a dictionary learning in-painting algorithm. This allows for multiple sparse recoverable images to be acquired at the cost of one fully sampled image. We believe this approach may open new ways to conduct imaging, which otherwise require compromising beam current and/or exposure conditions.

  16. 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...

  17. 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.

  18. 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.

  19. Three-dimensional shape of the Golgi apparatus in different cell types: serial section scanning electron microscopy of the osmium-impregnated Golgi apparatus.

    PubMed

    Koga, Daisuke; Kusumi, Satoshi; Ushiki, Tatsuo

    2016-04-01

    Although many studies of the Golgi apparatus structure have been performed by light and electron microscopy, the full shape of the Golgi apparatus remained unclear due to the technical limitations of the previously applied microscopy techniques. In this study, we used serial section scanning electron microscopy (SEM) for the morphological study of the Golgi apparatus. This method is useful for three-dimensional (3D) reconstruction of cellular structures without requiring specialized instruments, unlike focused ion beam SEM (FIB-SEM) and serial block face SEM (SBF-SEM). Using the serial section SEM method developed by our laboratory, we investigate the 3D shape of the osmium-impregnated Golgi apparatus in rat epididymal cells, pancreatic acinar cells and gonadotropes. The combination of serial section SEM and a 3D reconstruction technique enabled us to elucidate the entire shape of the Golgi apparatus in these cells. The full shape of the Golgi apparatus in epididymal cells formed a basket-like structure with oval-shaped cisterns, while the Golgi apparatus in an acinar cell from the pancreas was composed of elongated ribbon-like structures that were connected to each other, making a coarse network. The overall image of the Golgi apparatus cisterns from a gonadotrope looked like a spherical cage. This study has clearly shown that entire 3D shape of the Golgi apparatus varies depending on the cell type and that the Golgi cisterns network appears as a single mass located in the large region of the cytoplasm. © 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.

  20. 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.

  1. 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.

  2. 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.

  3. 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.

  4. Unveiling nanometric plasmons optical properties with advanced electron spectroscopy in the Scanning Transmission Electron Microscope

    NASA Astrophysics Data System (ADS)

    Kociak, Mathieu

    Since the pioneering work of Yamamoto, the use of electron spectroscopy such as Cathodoluminescence (CL) and Electron Energy Loss Spectroscopy (EELS) in a Scanning (Transmission) Electron Microscope (STEM) has considerably helped improving our understanding of the optical properties of metallic nanoparticles. The resemblance of spectroscopic signals from electron and pure optical techniques leads to the intuition that both types of techniques are very close, an idea theoretically discussed by F.J. Garcia de Abajo and coworkers. However, it is also quite intuitive that CL and EELS should be different. For example, EELS helps detecting any sort of modes while CL can only detect radiative ones. On the other hand, even between optical spectroscopy techniques, clear differences such as energy shifts or spectral shapes changes are expected in the case of plasmons. The lack of adapted instrumentation capable of performing combined EELS and CL, as well as theoretical developments allowing to account for the generic difference between EELS and CL and their optical counterparts impeached a comprehensive understanding of plasmons physics with the otherwise amazing electron spectroscopies. In this talk, I will present recent experimental results showing combined EELS and CL spectral mapping of plasmonic properties for nanoparticles with several shapes (triangles, cubes, stars...) and composition (gold, silver, aluminum...). Helped with different theoretical tools, I will try to show how these results can be related to their optical counterparts (extinction, scattering), and what type of physical insights can be gained from these combined measurements. Finally, if time allows, pointing the weaknesses of state-of-the-art CL and EELS (in terms of spectral range and/or spectral resolution), I will present EELS results obtained on highly monochromated electron beams that could cope with these limitations

  5. Single-electron tunneling. [Microwave scanning tunneling microscope

    SciTech Connect

    Ruggiero, S.T.

    1993-01-01

    Pictures using the low-temperature microwave scanning tunneling microscope, have been made of particles and tunneling IV characteristics determined. Strong, sometimes periodic negative differential resistance was observed in small-particle systems. Au and Ag droplets and particles were studied. 4 figs.

  6. A method of dynamic chromatic aberration correction in low-voltage scanning electron microscopes.

    PubMed

    Khursheed, Anjam

    2005-07-01

    A time-of-flight concept that dynamically corrects for chromatic aberration effects in scanning electron microscopes (SEMs) is presented. The method is predicted to reduce the microscope's chromatic aberration by an order of magnitude. The scheme should significantly improve the spatial resolution of low-voltage scanning electron microscopes (LVSEMs). The dynamic means of correcting for chromatic aberration also allows for the possibility of obtaining high image resolution from electron guns that have relatively large energy spreads.

  7. 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.

  8. 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.

  9. Cryo-FIB-SEM serial milling and block face imaging: Large volume structural analysis of biological tissues preserved close to their native state.

    PubMed

    Vidavsky, Netta; Akiva, Anat; Kaplan-Ashiri, Ifat; Rechav, Katya; Addadi, Lia; Weiner, Steve; Schertel, Andreas

    2016-12-01

    Many important biological questions can be addressed by studying in 3D large volumes of intact, cryo fixed hydrated tissues (⩾10,000μm(3)) at high resolution (5-20nm). This can be achieved using serial FIB milling and block face surface imaging under cryo conditions. Here we demonstrate the unique potential of the cryo-FIB-SEM approach using two extensively studied model systems; sea urchin embryos and the tail fin of zebrafish larvae. We focus in particular on the environment of mineral deposition sites. The cellular organelles, including mitochondria, Golgi, ER, nuclei and nuclear pores are made visible by the image contrast created by differences in surface potential of different biochemical components. Auto segmentation and/or volume rendering of the image stacks and 3D reconstruction of the skeleton and the cellular environment, provides a detailed view of the relative distribution in space of the tissue/cellular components, and thus of their interactions. Simultaneous acquisition of secondary and back-scattered electron images adds additional information. For example, a serial view of the zebrafish tail reveals the presence of electron dense mineral particles inside mitochondrial networks extending more than 20μm in depth in the block. Large volume imaging using cryo FIB SEM, as demonstrated here, can contribute significantly to the understanding of the structures and functions of diverse biological tissues. Copyright © 2016 Elsevier Inc. All rights reserved.

  10. Reproducible strain measurement in electronic devices by applying integer multiple to scanning grating in scanning moiré fringe imaging

    SciTech Connect

    Kim, Suhyun Jung, Younheum; Kim, Joong Jung; Lee, Sunyoung; Lee, Haebum; Kondo, Yukihito

    2014-10-15

    Scanning moiré fringe (SMF) imaging by high-angle annular dark field scanning transmission electron microscopy was used to measure the strain field in the channel of a transistor with a CoSi{sub 2} source and drain. Nanometer-scale SMFs were formed with a scanning grating size of d{sub s} at integer multiples of the Si crystal lattice spacing d{sub l} (d{sub s} ∼ nd{sub l}, n = 2, 3, 4, 5). The moiré fringe formula was modified to establish a method for quantifying strain measurement. We showed that strain fields in a transistor measured by SMF images were reproducible with an accuracy of 0.02%.

  11. Design of a fast electron beam scanning system for compact synchrotron light sources

    NASA Astrophysics Data System (ADS)

    Moser, H. O.; Lehr, H.

    1989-07-01

    The design of an electron beam scanning system for compact storage ring synchrotron light sources is described. The main features are a scan frequency of 100 Hz and an angular amplitude of ±5 mrad. Different configurations of scan dipoles permit confining the scan to one cell using four dipoles or to repeat the scan periodically along the whole circumference by means of two scan dipoles per cell. Combinations of these basic configurations are possible. The location of the nodes of the pivoting electron beam can be optimized with respect to the maximum scan angle by slightly unbalancing the field strength in different scan dipoles. The scan dipoles are H-shaped magnets made from laminated iron. Their gap width is 68 mm. They are powered by fast transistor-bridge supplies which are controlled by freely programmable function generators capable of realizing a triangular current waveform with a deviation of less than 0.1% except for a 1% neighborhood of the apex. Estimates of the influence of the scanning on both quantum and Coulomb lifetime indicate acceptable lifetime reductions provided the minimum distance between distorted closed orbit and aperture exceeds about six standard deviations of the spatial electron distribution.

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

    DOEpatents

    de Jonge, Niels [Oak Ridge, TN

    2010-08-17

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

  13. 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

  14. 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.

  15. 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

  16. Scanning-electron-microscope used in real-time study of friction and wear

    NASA Technical Reports Server (NTRS)

    Brainard, W. A.; Buckley, D. H.

    1975-01-01

    Small friction and wear apparatus built directly into scanning-electron-microscope provides both dynamic observation and microscopic view of wear process. Friction and wear tests conducted using this system have indicated that considerable information can readily be gained.

  17. Improved Specimen Coating Technique for Scanning Electron Microscope Observation of Decomposer Microorganisms 1

    PubMed Central

    Draggan, Sidney

    1976-01-01

    Sputter coating of leaf litter microbe samples provides scanning electron microscope images with greater information content than either vacuum evaporation of thin metal coatings or tissue conductance. Images PMID:16345150

  18. 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...

  19. 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...

  20. Using the scanning electron microscope on the production line to assure quality semiconductors

    NASA Technical Reports Server (NTRS)

    Adolphsen, J. W.; Anstead, R. J.

    1972-01-01

    The use of the scanning electron microscope to detect metallization defects introduced during batch processing of semiconductor devices is discussed. A method of determining metallization integrity was developed which culminates in a procurement specification using the scanning microscope on the production line as a quality control tool. Batch process control of the metallization operation is monitored early in the manufacturing cycle.

  1. Scanning Electron Microscopy (SEM) Procedure for HE Powders on a Zeiss Sigma HD VP SEM

    SciTech Connect

    Zaka, F.

    2016-11-15

    This method describes the characterization of inert and HE materials by the Zeiss Sigma HD VP field emission Scanning Electron Microscope (SEM). The SEM uses an accelerated electron beam to generate high-magnification images of explosives and other materials. It is fitted with five detectors (SE, Inlens, STEM, VPSE, HDBSD) to enable imaging of the sample via different secondary electron signatures, angles, and energies. In addition to imaging through electron detection, the microscope is also fitted with two Oxford Instrument Energy Dispersive Spectrometer (EDS) 80 mm detectors to generate elemental constituent spectra and two-dimensional maps of the material being scanned.

  2. Nondestructive determination of the depth of planar p-n junctions by scanning electron microscopy

    NASA Technical Reports Server (NTRS)

    Chi, J.-Y.; Gatos, H. C.

    1977-01-01

    A method was developed for measuring nondestructively the depth of planar p-n junctions in simple devices as well as in integrated-circuit structures with the electron-beam induced current (EBIC) by scanning parallel to the junction in a scanning electron microscope (SEM). The results were found to be in good agreement with those obtained by the commonly used destructive method of lapping at an angle to the junction and staining to reveal the junction.

  3. Nondestructive determination of the depth of planar p-n junctions by scanning electron microscopy

    NASA Technical Reports Server (NTRS)

    Chi, J.-Y.; Gatos, H. C.

    1977-01-01

    A method was developed for measuring nondestructively the depth of planar p-n junctions in simple devices as well as in integrated-circuit structures with the electron-beam induced current (EBIC) by scanning parallel to the junction in a scanning electron microscope (SEM). The results were found to be in good agreement with those obtained by the commonly used destructive method of lapping at an angle to the junction and staining to reveal the junction.

  4. 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

  5. Large area fabrication of plasmonic nanoparticle grating structure by conventional scanning electron microscope

    SciTech Connect

    Sudheer, Tiwari, P.; Rai, V. N.; Srivastava, A. K.; Mukharjee, C.

    2015-06-24

    Plasmonic nanoparticle grating (PNG) structure of different periods has been fabricated by electron beam lithography using silver halide based transmission electron microscope film as a substrate. Conventional scanning electron microscope is used as a fabrication tool for electron beam lithography. Optical microscope and energy dispersive spectroscopy (EDS) have been used for its morphological and elemental characterization. Optical characterization is performed by UV-Vis absorption spectroscopic technique.

  6. 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.

  7. Modeling a Miniaturized Scanning Electron Microscope Focusing Column - Lessons Learned in Electron Optics Simulation

    NASA Technical Reports Server (NTRS)

    Loyd, Jody; Gregory, Don; Gaskin, Jessica

    2016-01-01

    This presentation discusses work done to assess the design of a focusing column in a miniaturized Scanning Electron Microscope (SEM) developed at the NASA Marshall Space Flight Center (MSFC) for use in-situ on the Moon-in particular for mineralogical analysis. The MSFC beam column design uses purely electrostatic fields for focusing, because of the severe constraints on mass and electrical power consumption imposed by the goals of lunar exploration and of spaceflight in general. The resolution of an SEM ultimately depends on the size of the focused spot of the scanning beam probe, for which the stated goal here is a diameter of 10 nanometers. Optical aberrations are the main challenge to this performance goal, because they blur the ideal geometrical optical image of the electron source, effectively widening the ideal spot size of the beam probe. In the present work the optical aberrations of the mini SEM focusing column were assessed using direct tracing of non-paraxial rays, as opposed to mathematical estimates of aberrations based on paraxial ray-traces. The geometrical ray-tracing employed here is completely analogous to ray-tracing as conventionally understood in the realm of photon optics, with the major difference being that in electron optics the lens is simply a smoothly varying electric field in vacuum, formed by precisely machined electrodes. Ray-tracing in this context, therefore, relies upon a model of the electrostatic field inside the focusing column to provide the mathematical description of the "lens" being traced. This work relied fundamentally on the boundary element method (BEM) for this electric field model. In carrying out this research the authors discovered that higher accuracy in the field model was essential if aberrations were to be reliably assessed using direct ray-tracing. This led to some work in testing alternative techniques for modeling the electrostatic field. Ultimately, the necessary accuracy was attained using a BEM

  8. Scanning Probe Evaluation of Electronic, Mechanical and Structural Material Properties

    NASA Astrophysics Data System (ADS)

    Virwani, Kumar

    2011-03-01

    We present atomic force microscopy (AFM) studies of a range of properties from three different classes of materials: mixed ionic electronic conductors, low-k dielectrics, and polymer-coated magnetic nanoparticles. (1) Mixed ionic electronic conductors are being investigated as novel diodes to drive phase-change memory elements. Their current-voltage characteristics are measured with direct-current and pulsed-mode conductive AFM (C-AFM). The challenges to reliability of the C-AFM method include the electrical integrity of the probe, the sample and the contacts, and the minimization of path capacitance. The role of C-AFM in the optimization of these electro-active materials will be presented. (2) Low dielectric constant (low-k) materials are used in microprocessors as interlayer insulators, a role directly affected by their mechanical performance. The mechanical properties of nanoporous silicate low-k thin films are investigated in a comparative study of nanomechanics measured by AFM and by traditional nanoindentation. Both methods are still undergoing refinement as reliable analytical tools for determining nanomechanical properties. We will focus on AFM, the faster of the two methods, and its developmental challenges of probe shape, cantilever force constant, machine compliance and calibration standards. (3) Magnetic nanoparticles are being explored for their use in patterned media for magnetic storage. Current methods for visualizing the core-shell structure of polymer-coated magnetic nanoparticles include dye-staining the polymer shell to provide contrast in transmission electron microscopy. AFM-based fast force-volume measurements provide direct visualization of the hard metal oxide core within the soft polymer shell based on structural property differences. In particular, the monitoring of adhesion and deformation between the AFM tip and the nanoparticle, particle-by-particle, provides a reliable qualitative tool to visualize core-shell contrast without the use

  9. 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.

  10. 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

  11. 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.

  12. 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.

  13. 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

  14. 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.

  15. 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.

  16. 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.

  17. Measurement of boundary plane inclination in a scanning electron microscope

    SciTech Connect

    Randle, V.; Dingley, D. )

    1989-09-01

    Recently, a method has been devised for measuring the boundary orientations using backscattered Kikuchi diffraction (BKD, otherwise known as electron backscattering, EBS). The work reported demonstrates that BKD can be efficiently used to measure both the misorientation across grain boundaries and also the orientation of boundary planes. In nickel it has been shown that the boundaries of grains which are situated along the corner of a rectangular specimen rotate so as to minimize their interfacial energy. For non-coincidence site lattices related grains, boundaries tend to align normal to the edge of the specimen, while {Sigma} = 3 and {Sigma} = 9 CSLs tend to rotate to tilt configuration, particularly asymmetric tilts such as {l brace}111{r brace}/ {l brace}115{r brace} or {l brace}110{r brace}/{l brace}114{r brace}.

  18. Space-time ambiguity functions for electronically scanned ISR applications

    NASA Astrophysics Data System (ADS)

    Swoboda, John; Semeter, Joshua; Erickson, Philip

    2015-05-01

    Electronically steerable array (ESA) technology has recently been applied to incoherent scatter radar (ISR) systems. These arrays allow for pulse-to-pulse steering of the antenna beam to collect data in a three-dimensional region. This is in direct contrast to dish-based antennas, where ISR acquisition is limited at any one time to observations in a two-dimensional slice. This new paradigm allows for more flexibility in the measurement of ionospheric plasma parameters. Multiple ESA-based ISR systems operate currently in the high-latitude region where the ionosphere is highly variable in both space and time. Because of the highly dynamic nature of the ionosphere in this region, it is important to differentiate between measurement-induced artifacts and the true behavior of the plasma. Often, three-dimensional ISR data produced by ESA systems are fitted in a spherical coordinate space and then the parameters are interpolated to a Cartesian grid, potentially introducing error and impacting the reconstructions of the plasma parameters. To take advantage of the new flexibility inherent in ESA systems, we present a new way of analyzing ISR observations through use of the space-time ambiguity function. The use of this new measurement ambiguity function allows us to pose the ISR observational problem in terms of a linear inverse problem whose goal is the estimate of the time domain lags of the intrinsic plasma autocorrelation function used for parameter fitting. The framework allows us to explore the impact of nonuniformity in plasma parameters in both time and space. We discuss examples of possible artifacts in high-latitude situations and discuss possible ways of reducing them and improving the quality of data products from electronically steerable ISRs.

  19. The current state of electronic consultation and electronic referral systems in Canada: an environmental scan.

    PubMed

    Liddy, Clare; Hogel, Matthew; Blazkho, Valerie; Keely, Erin

    2015-01-01

    Access to specialist care is a point of concern for patients, primary care providers, and specialists in Canada. Innovative e-health platforms such as electronic consultation (eConsultation) and referral (eReferral) can improve access to specialist care. These systems allow physicians to communicate asynchronously and could reduce the number of unnecessary referrals that clog wait lists, provide a record of the patient's journey through the referral system, and lead to more efficient visits. Little is known about the current state of eConsultation and eReferral in Canada. The purpose of this work was to identify current systems and gain insight into the design and implementation process of existing systems. An environmental scan approach was used, consisting of a systematic and grey literature review, and targeted semi-structured key informant interviews. Only three eConsultation/eReferral systems are currently in operation in Canada. Four themes emerged from the interviews: eReferral is an end goal for those provinces without an active eReferral system, re-organization of the referral process is a necessity prior to automation, engaging the end-user is essential, and technological incompatibilities are major impediments to progress. Despite the acknowledged need to improve the referral system and increase government spending on health information technology, eConsultation and eReferral systems remain scarce as Canada lags behind the rest of the developed world.

  20. 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.

  1. 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.

  2. Correction of image drift and distortion in a scanning electron microscopy.

    PubMed

    Jin, P; Li, X

    2015-12-01

    Continuous research on small-scale mechanical structures and systems has attracted strong demand for ultrafine deformation and strain measurements. Conventional optical microscope cannot meet such requirements owing to its lower spatial resolution. Therefore, high-resolution scanning electron microscope has become the preferred system for high spatial resolution imaging and measurements. However, scanning electron microscope usually is contaminated by distortion and drift aberrations which cause serious errors to precise imaging and measurements of tiny structures. This paper develops a new method to correct drift and distortion aberrations of scanning electron microscope images, and evaluates the effect of correction by comparing corrected images with scanning electron microscope image of a standard sample. The drift correction is based on the interpolation scheme, where a series of images are captured at one location of the sample and perform image correlation between the first image and the consequent images to interpolate the drift-time relationship of scanning electron microscope images. The distortion correction employs the axial symmetry model of charged particle imaging theory to two images sharing with the same location of one object under different imaging fields of view. The difference apart from rigid displacement between the mentioned two images will give distortion parameters. Three-order precision is considered in the model and experiment shows that one pixel maximum correction is obtained for the employed high-resolution electron microscopic system.

  3. 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.

  4. 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.

  5. 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.

  6. Monte Carlo modeling of cavity imaging in pure iron using back-scatter electron scanning microscopy

    NASA Astrophysics Data System (ADS)

    Yan, Qiang; Gigax, Jonathan; Chen, Di; Garner, F. A.; Shao, Lin

    2016-11-01

    Backscattered electrons (BSE) in a scanning electron microscope (SEM) can produce images of subsurface cavity distributions as a nondestructive characterization technique. Monte Carlo simulations were performed to understand the mechanism of void imaging and to identify key parameters in optimizing void resolution. The modeling explores an iron target of different thicknesses, electron beams of different energies, beam sizes, and scan pitch, evaluated for voids of different sizes and depths below the surface. The results show that the void image contrast is primarily caused by discontinuity of energy spectra of backscattered electrons, due to increased outward path lengths for those electrons which penetrate voids and are backscattered at deeper depths. Size resolution of voids at specific depths, and maximum detection depth of specific voids sizes are derived as a function of electron beam energy. The results are important for image optimization and data extraction.

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

    PubMed Central

    EBERLE, AL; MIKULA, S; SCHALEK, R; LICHTMAN, J; TATE, ML KNOTHE; ZEIDLER, D

    2015-01-01

    Electron–electron interactions and detector bandwidth limit the maximal imaging speed of single-beam scanning electron microscopes. We use multiple electron beams in a single column and detect secondary electrons in parallel to increase the imaging speed by close to two orders of magnitude and demonstrate imaging for a variety of samples ranging from biological brain tissue to semiconductor wafers. Lay Description The composition of our world and our bodies on the very small scale has always fascinated people, making them search for ways to make this visible to the human eye. Where light microscopes reach their resolution limit at a certain magnification, electron microscopes can go beyond. But their capability of visualizing extremely small features comes at the cost of a very small field of view. Some of the questions researchers seek to answer today deal with the ultrafine structure of brains, bones or computer chips. Capturing these objects with electron microscopes takes a lot of time – maybe even exceeding the time span of a human being – or new tools that do the job much faster. A new type of scanning electron microscope scans with 61 electron beams in parallel, acquiring 61 adjacent images of the sample at the same time a conventional scanning electron microscope captures one of these images. In principle, the multibeam scanning electron microscope’s field of view is 61 times larger and therefore coverage of the sample surface can be accomplished in less time. This enables researchers to think about large-scale projects, for example in the rather new field of connectomics. A very good introduction to imaging a brain at nanometre resolution can be found within course material from Harvard University on http://www.mcb80x.org/# as featured media entitled ‘connectomics’. PMID:25627873

  8. Mass-mapping of ECM macromolecules by scanning transmission electron microscopy.

    PubMed

    Sherratt, Michael J; Graham, Helen K; Kielty, Cay M; Holmes, David F

    2009-01-01

    In the scanning transmission electron microscope, the degree of electron scattering induced by biological specimens, such as ECM macromolecules, is dependent on the molecular mass. By calibrating the ratio of scattered to non-scattered electrons against a known mass standard, such as tobacco mosaic virus, it is possible to quantify absolute changes in both mass and mass distribution. These mass mapping approaches can provide important information on ECM assembly, organisation, and interactions which is not obtainable by other means.

  9. 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

  10. 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.

  11. Scanning electron microscope/energy dispersive x ray analysis of impact residues in LDEF tray clamps

    NASA Technical Reports Server (NTRS)

    Bernhard, Ronald P.; Durin, Christian; Zolensky, Michael E.

    1993-01-01

    Detailed optical scanning of tray clamps is being conducted in the Facility for the Optical Inspection of Large Surfaces at JSC to locate and document impacts as small as 40 microns in diameter. Residues from selected impacts are then being characterized by Scanning Electron Microscopy/Energy Dispersive X-ray Analysis at CNES. Results from this analysis will be the initial step to classifying projectile residues into specific sources.

  12. 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.

  13. Imaging of Nitroxides at 250 MHz using Rapid-Scan Electron Paramagnetic Resonance

    PubMed Central

    Biller, Joshua R.; Tseitlin, Mark; Quine, Richard W.; Rinard, George A.; Weismiller, Hilary A.; Elajaili, Hanan; Rosen, Gerald M.; Kao, Joseph P. Y.; Eaton, Sandra S.; Eaton, Gareth R.

    2014-01-01

    Projections for 2D spectral-spatial images were obtained by continuous wave and rapid-scan electron paramagnetic resonance using a bimodal cross-loop resonator at 251 MHz. The phantom consisted of three 4 mm tubes containing different 15N,2H-substituted nitroxides. Rapid-scan and continuous wave images were obtained with 5 min total acquisition times. For comparison, images also were obtained with 29 s acquisition time for rapid scan and 15 min for continuous wave. Relative to continuous wave projections obtained for the same data acquisition time, rapid-scan projections had significantly less low-frequency noise and substantially higher signal-to-noise at higher gradients. Because of the improved image quality for the same data acquisition time, linewidths could be determined more accurately from the rapid-scan images than from the continuous wave images. PMID:24650729

  14. Imaging of nitroxides at 250MHz using rapid-scan electron paramagnetic resonance.

    PubMed

    Biller, Joshua R; Tseitlin, Mark; Quine, Richard W; Rinard, George A; Weismiller, Hilary A; Elajaili, Hanan; Rosen, Gerald M; Kao, Joseph P Y; Eaton, Sandra S; Eaton, Gareth R

    2014-05-01

    Projections for 2D spectral-spatial images were obtained by continuous wave and rapid-scan electron paramagnetic resonance using a bimodal cross-loop resonator at 251MHz. The phantom consisted of three 4mm tubes containing different (15)N,(2)H-substituted nitroxides. Rapid-scan and continuous wave images were obtained with 5min total acquisition times. For comparison, images also were obtained with 29s acquisition time for rapid scan and 15min for continuous wave. Relative to continuous wave projections obtained for the same data acquisition time, rapid-scan projections had significantly less low-frequency noise and substantially higher signal-to-noise at higher gradients. Because of the improved image quality for the same data acquisition time, linewidths could be determined more accurately from the rapid-scan images than from the continuous wave images.

  15. 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

  16. Nanoelectrical probing with multiprobe SPM Systems compatible with scanning electron microscopes

    NASA Astrophysics Data System (ADS)

    Lewis, Aaron; Ignatov, Andrey; Taha, Hesham; Zhinoviev, Oleg; Komissar, Anatoly; Krol, Alexander; Lewis, David

    2011-03-01

    A scanning electron microscope compatible platform that permits multiprobe atomic force microscopy based nanoelectrical characterization will be described. To achieve such multiple parameter nanocharacterization with scanning electron microscope compatibility involves a number of innovations both in instrument and probe design. This presentation will focus on how these advances were achieved and the results obtained with such instrumentation on electrical nano-characterization and electrical nano-manipulation. The advances include: 1. Specialized scanners; 2. An ultrasensitive feedback mechanism based on tuning forks with no optical feedback interference that can induce carriers in semiconductor devices; and 3. Unique probes compatible with multiprobe geometries in which the probe tips can be brought into physical contact with one another. Experiments will be described with such systems that will include multiprobe electrical measurements with metal and glass coated coaxial nanowires of platinum. This combination of scanning electron microscopes integrated with multiprobe instrumentation allows for important applications not available today in the field of semiconductor processing technology.

  17. 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.

  18. Electrical characteristics of silicon nanowire transistors fabricated by scanning probe and electron beam lithographies.

    PubMed

    Ryu, Yu Kyoung; Chiesa, Marco; Garcia, Ricardo

    2013-08-09

    Silicon nanowire (SiNW) field-effect transistors have been fabricated by oxidation scanning probes and electron beam lithographies. The analysis and comparison of the electron mobility and subthreshold swing shows that the device performance is not affected by the top-down fabrication method. The two methods produce silicon nanowire transistors with similar electrical features, although oxidation scanning probe lithography generates nanowires with smaller channel widths. The values of the electron mobility and the subthreshold swing, 200 cm(2) V(-1) s(-1) and 500 mV dec(-1), respectively, are similar to those obtained from bottom-up methods. The compatibility of top-down methods with CMOS (complementary metal-oxide-semiconductor) procedures, the good electrical properties of the nanowire devices and the potential for making sub-10 nanowires, in particular by using oxidation scanning probe lithography, make those methods attractive for device fabrication.

  19. 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.

  20. Two-dimensional simulation and modeling in scanning electron microscope imaging and metrology research.

    PubMed

    Postek, Michael T; Vladár, András E; Lowney, Jeremiah R; Keery, William J

    2002-01-01

    Traditional Monte Carlo modeling of the electron beam-specimen interactions in a scanning electron microscope (SEM) produces information about electron beam penetration and output signal generation at either a single beam-landing location, or multiple landing positions. If the multiple landings lie on a line, the results can be graphed in a line scan-like format. Monte Carlo results formatted as line scans have proven useful in providing one-dimensional information about the sample (e.g., linewidth). When used this way, this process is called forward line scan modeling. In the present work, the concept of image simulation (or the first step in the inverse modeling of images) is introduced where the forward-modeled line scan data are carried one step further to construct theoretical two-dimensional (2-D) micrographs (i.e., theoretical SEM images) for comparison with similar experimentally obtained micrographs. This provides an ability to mimic and closely match theory and experiment using SEM images. Calculated and/or measured libraries of simulated images can be developed with this technique. The library concept will prove to be very useful in the determination of dimensional and other properties of simple structures, such as integrated circuit parts, where the shape of the features is preferably measured from a single top-down image or a line scan. This paper presents one approach to the generation of 2-D simulated images and presents some suggestions as to their application to critical dimension metrology.

  1. Tracing the Electronic Pairing Glue in Unconventional Superconductors via Inelastic Scanning Tunneling Spectroscopy

    NASA Astrophysics Data System (ADS)

    Hlobil, Patrik; Jandke, Jasmin; Wulfhekel, Wulf; Schmalian, Jörg

    2017-04-01

    Scanning tunneling microscopy has been shown to be a powerful experimental probe to detect electronic excitations and further allows us to deduce fingerprints of bosonic collective modes in superconductors. Here, we demonstrate that the inclusion of inelastic tunnel events is crucial for the interpretation of tunneling spectra of unconventional superconductors and allows us to directly probe electronic and bosonic excitations via scanning tunneling microscopy. We apply the formalism to the iron based superconductor LiFeAs. With the inclusion of inelastic contributions, we find strong evidence for a nonconventional pairing mechanism, likely via magnetic excitations.

  2. 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

  3. Long range electronic transport in microbial nanowires bridging an electrode and scanned probe

    NASA Astrophysics Data System (ADS)

    Veazey, Joshua; Lampa-Pastirk, Sanela; Walsh, Kathy; Sun, Jiebing; Zhang, Pengpeng; Reguera, Gemma; Tessmer, Stuart

    2011-03-01

    The filament-like appendages known as pili, expressed by the bacterium Geobacter sulfurreducens, are believed to act as electrically conductive nanowires. Previously, we used scanning tunneling microscopy to study the local density of states at different positions along the wire. However, the long range electron transfer believed to occur in this protein has not been directly observed. Here we discuss a system for verifying long range transport using a scanning probe technique. Transport at distances of more than a few nanometers would require a novel biological electron transfer process. The authors gratefully acknowledge support from the National Science Foundation (MCB-1021948) and the Michigan State University Foundation (Strategic Partnership Grant).

  4. A compact multipurpose nanomanipulator for use inside a scanning electron microscope

    NASA Astrophysics Data System (ADS)

    Heeres, E. C.; Katan, A. J.; van Es, M. H.; Beker, A. F.; Hesselberth, M.; van der Zalm, D. J.; Oosterkamp, T. H.

    2010-02-01

    A compact, two-stage nanomanipulator was designed and built for use inside a scanning electron microscope. It consists of a fine stage employing piezostacks that provide a 15 μm range in three dimensions and a coarse stage based on commercially available stick-slip motors. Besides the fabrication of enhanced probes for scanning probe microscopy and the enhancement of electron field emitters, other novel manipulation processes were developed, such as locating, picking up, and positioning small nanostructures with an accuracy of ˜10 nm. In combination with in situ I-V experiments, welding, and etching, this results in a multipurpose nanofactory, enabling a new range of experiments.

  5. 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.

  6. 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.

  7. Imaging correlated wave functions of few-electron quantum dots: Theory and scanning tunneling spectroscopy experimentsa)

    NASA Astrophysics Data System (ADS)

    Rontani, Massimo; Molinari, Elisa; Maruccio, Giuseppe; Janson, Martin; Schramm, Andreas; Meyer, Christian; Matsui, Tomohiro; Heyn, Christian; Hansen, Wolfgang; Wiesendanger, Roland

    2007-04-01

    We show both theoretically and experimentally that scanning tunneling spectroscopy (STS) images of semiconductor quantum dots may display clear signatures of electron-electron correlation. We apply many-body tunneling theory to a realistic model, which fully takes into account correlation effects and dot anisotropy. Comparing measured STS images of freestanding InAs quantum dots with those calculated by the full configuration interaction method, we explain the wave-function sequence in terms of images of one- and two-electron states. The STS map corresponding to double charging is significantly distorted by electron correlation with respect to the noninteracting case.

  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. 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

  10. 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.

  11. Semi-empirical model for the generation of dose distributions produced by a scanning electron beam

    SciTech Connect

    Nath, R.; Gignac, C.E.; Agostinelli, A.G.; Rothberg, S.; Schulz, R.J.

    1980-01-01

    There are linear accelerators (Sagittaire and Saturne accelerators produced by Compagnie Generale de Radiologie (CGR/MeV) Corporation) which produce broad, flat electron fields by magnetically scanning the relatively narrow electron beam as it emerges from the accelerator vacuum system. A semi-empirical model, which mimics the scanning action of this type of accelerator, was developed for the generation of dose distributions in homogeneous media. The model employs the dose distributions of the scanning electron beams. These were measured with photographic film in a polystyrene phantom by turning off the magnetic scanning system. The mean deviation calculated from measured dose distributions is about 0.2%; a few points have deviations as large as 2 to 4% inside of the 50% isodose curve, but less than 8% outside of the 50% isodose curve. The model has been used to generate the electron beam library required by a modified version of a commercially-available computerized treatment-planning system. (The RAD-8 treatment planning system was purchased from the Digital Equipment Corporation. It is currently available from Electronic Music Industries (EMI), Ltd.)

  12. Evolution of MEMS scanning mirrors for laser projection in compact consumer electronics

    NASA Astrophysics Data System (ADS)

    Tauscher, Jason; Davis, Wyatt O.; Brown, Dean; Ellis, Matt; Ma, Yunfei; Sherwood, Michael E.; Bowman, David; Helsel, Mark P.; Lee, Sung; Coy, John Wyatt

    2010-02-01

    The applicability of MOEMS scanning mirrors towards the creation of "flying spot" scanned laser displays is well established. The extension of this concept towards compact embedded pico-projectors has required an evolution of scanners and packaging to accommodate the needs of the consumer electronics space. This paper describes the progression of the biaxial MOEMS scanning mirrors developed by Microvision over recent years. Various aspects of the individual designs are compared. Early devices used a combination of magnetic quasistatic actuation and resonant electrostatic operation in an evacuated atmosphere to create a projection engine for retinal scanned displays. Subsequent designs realized the elimination of both the high voltage electrostatic drive and the vacuum package, and a simplification of the actuation scheme through proprietary technical advances. Additional advances have doubled the scan angle capability and greatly miniaturized the MOEMS component while not incurring significant increase in power consumption, making it an excellent fit for the consumer pico-projector application. The simplicity of the scanned laser-based pico-projector optical design enables high resolution and a large effective image size in a thin projection engine, all of which become critical both to the viability of the technology and adoption by consumers. Microvision's first scanned laser pico-projector is built around a MOEMS scanning mirror capable of projecting 16:9 aspect ratio, WVGA display within a 6.6 mm high package. Further evolution on this path promises continued improvement in resolution, size, and power.

  13. 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.

  14. 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

  15. 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.

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

    PubMed

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

    2016-08-01

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

  17. Scanning precession electron tomography for three-dimensional nanoscale orientation imaging and crystallographic analysis

    PubMed Central

    Eggeman, Alexander S.; Krakow, Robert; Midgley, Paul A.

    2015-01-01

    Three-dimensional (3D) reconstructions from electron tomography provide important morphological, compositional, optical and electro-magnetic information across a wide range of materials and devices. Precession electron diffraction, in combination with scanning transmission electron microscopy, can be used to elucidate the local orientation of crystalline materials. Here we show, using the example of a Ni-base superalloy, that combining these techniques and extending them to three dimensions, to produce scanning precession electron tomography, enables the 3D orientation of nanoscale sub-volumes to be determined and provides a one-to-one correspondence between 3D real space and 3D reciprocal space for almost any polycrystalline or multi-phase material. PMID:26028514

  18. Phonon focusing in quartz and sapphire imaged by electron beam scanning

    NASA Astrophysics Data System (ADS)

    Eichele, R.; Huebener, R. P.; Seifert, H.

    1982-06-01

    Anisotropic phonon propagation and phonon focusing in single-crystalline α-quartz and sapphire has been investigated using electron beam scanning for imaging the ballistic phonon propagation. The samples were circular disks of 20 mm diameter and 2.0 mm thickness. The phonons generated at the upper flat sample surface by the electron beam were detected with a small-size (43 μm×41μm) bolometer at the center of the opposite surface. The experiments were performed using a scanning electron microscope with a low-termperature stage. During electron irradiation of the upper sample surface the bottom of the sample was in direct contact with the liquid-He bath. For α-quartz, also time-resolved imaging of the phonon intensity has been performed in addition to imaging of the time-integrated bolometer signal. Our results agree remarkably well with calculations by Rösch and Weis.

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

    SciTech Connect

    Li, Meng; Xu, Chunkai E-mail: xjun@ustc.edu.cn; Zhang, Panke; Li, Zhean; Chen, Xiangjun E-mail: xjun@ustc.edu.cn

    2016-08-15

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

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

    NASA Astrophysics Data System (ADS)

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

    2016-08-01

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

  1. Study on the parameters of the scanning system for the 300 keV electron accelerator

    SciTech Connect

    Leo, K. W.; Chulan, R. M. Hashim, S. A.; Baijan, A. H.; Sabri, R. M.; Mohtar, M.; Glam, H.; Lojius, L.; Zahidee, M.; Azman, A.; Zaid, M.

    2016-01-22

    This paper describes the method to identify the magnetic coil parameters of the scanning system. This locally designed low energy electron accelerator with the present energy of 140 keV will be upgraded to 300 keV. In this accelerator, scanning system is required to deflect the energetic electron beam across a titanium foil in vertical and horizontal direction. The excitation current of the magnetic coil is determined by the energy of the electron beam. Therefore, the magnetic coil parameters must be identified to ensure the matching of the beam energy and excitation coil current. As the result, the essential parameters of the effective lengths for X-axis and Y-axis have been found as 0.1198 m and 0.1134 m and the required excitation coil currents which is dependenton the electron beam energies have be identified.

  2. Micro-column Scanning Electron Microscope and X-ray Spectrometer (MSEMS) for Planetary Exploration

    NASA Astrophysics Data System (ADS)

    Ribaya, B.; Niemann, D.; Makarewicz, J.; Clevenson, H.; McKenzie, C.; Nguyen, C.; Blake, D. F.

    2009-12-01

    Scanning Electron Microscopy combined with electron-induced X-ray Fluorescence Spectroscopy (SEM-EDX) is one of the most powerful techniques for characterizing sub-µm surface morphology and composition. In terrestrial laboratories, SEM-EDX is used to elucidate natural processes such as low-temperature diagenesis, thermal or pressure induced metamorphism, volcanism/magmatism, atmosphere/crust interaction and biological activity. Such information would be highly useful for investigating the natural history of the terrestrial planets, satellites and primitive bodies, providing morphological and elemental information that is 2 orders of magnitude higher in resolution than optical techniques. Below we describe the development of a Micro-column Scanning Electron Microscope and X-ray Spectrometer (MSEMS) for flight. The enabling technology of the MSEMS is a carbon nanotube field emission (CNTFE) electron source that is integrated with micro-electro-mechanical-systems (MEMS) - based electron gun and electron optical structures. A hallmark of CNTFE electron sources is their low chromatic aberration, which reduces the need for high accelerating voltages to obtain small spot size. The CNTFE also offers exceptional brightness and nanometer source size, eliminating the need for condenser lenses, making simple electrostatic focusing optics possible. Moreover, the CNT field emission gun (CFEG) at low operating voltage dissipates 103 less power than thermally-assisted Schottky emitters. A key feature of the MSEMS design is the lack of scanning coils. Rather, a piezoelectric sample stage capable of sub-nanometer resolution scans the sample past the fixed crossover of the MSEMS electron beam. We will describe a MEMS-based templating technique for fabricating mechanically and electrically stable miniature CFEGs. Using existing silicon (Si) technology, we fabricated highly controlled and precise MEMS structures for both the CNT cathode and focusing optics for the micro-column. The

  3. 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.

  4. Modeling for accurate dimensional scanning electron microscope metrology: then and now.

    PubMed

    Postek, Michael T; Vladár, András E

    2011-01-01

    A review of the evolution of modeling for accurate dimensional scanning electron microscopy is presented with an emphasis on developments in the Monte Carlo technique for modeling the generation of the electrons used for imaging and measurement. The progress of modeling for accurate metrology is discussed through a schematic technology timeline. In addition, a discussion of a future vision for accurate SEM dimensional metrology and the requirements to achieve it are presented.

  5. The scanning electron microscope in microbiology and diagnosis of infectious disease

    PubMed Central

    Golding, Christine G.; Lamboo, Lindsey L.; Beniac, Daniel R.; Booth, Timothy F.

    2016-01-01

    Despite being an excellent tool for investigating ultrastructure, scanning electron microscopy (SEM) is less frequently used than transmission electron microscopy for microbes such as viruses or bacteria. Here we describe rapid methods that allow SEM imaging of fully hydrated, unfixed microbes without using conventional sample preparation methods. We demonstrate improved ultrastructural preservation, with greatly reduced dehydration and shrinkage, for specimens including bacteria and viruses such as Ebola virus using infiltration with ionic liquid on conducting filter substrates for SEM. PMID:27212232

  6. Investigating the optical properties of dislocations by scanning transmission electron microscopy.

    PubMed

    Pennycook, S J

    2008-01-01

    The scanning transmission electron microscope (STEM) allows collection of a number of simultaneous signals, such as cathodoluminescence (CL), transmitted electron intensity and spectroscopic information from individual localized defects. This review traces the development of CL and atomic resolution imaging from their early inception through to the possibilities that exist today for achieving a true atomic-scale understanding of the optical properties of individual dislocations cores. This review is dedicated to Professor David Holt, a pioneer in this field.

  7. 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.

  8. 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.

  9. 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.

  10. Scanning electron microscopy of dentition: methodology and ultrastructural morphology of tooth wear.

    PubMed

    Shkurkin, G V; Almquist, A J; Pfeihofer, A A; Stoddard, E L

    1975-01-01

    Scanning electron micrographs were taken of sets of human molars-those of paleo-Indians used in mastication of, ostensibly, a highly abrasive diet, and those of contemporary Americans. Different ultrastructural patterns of enamel wear were observed between the groups.

  11. Practical application of HgI2 detectors to a space-flight scanning electron microscope

    NASA Technical Reports Server (NTRS)

    Bradley, J. G.; Conley, J. M.; Albee, A. L.; Iwanczyk, J. S.; Dabrowski, A. J.

    1989-01-01

    Mercuric iodide X-ray detectors have been undergoing tests in a prototype scanning electron microscope system being developed for unmanned space flight. The detector program addresses the issues of geometric configuration in the SEM, compact packaging that includes separate thermoelectric coolers for the detector and FET, X-ray transparent hermetic encapsulation and electrical contacts, and a clean vacuum environment.

  12. 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...

  13. The Sensura Neglecta in the Pigeon: A Scanning Electron and Light Microscope Study.

    DTIC Science & Technology

    Both scanning electron and light microscopy have been used to observe the tiny sensura neglecta which is located in the inferior utricula sinus of...the vestibular labyrinth in the pigeon. Observations indicate that the sensura neglecta in the pigeon has both crista- and macula-like features. It is

  14. Imaging of ballistic phonon propagation in quartz by electron beam scanning

    NASA Astrophysics Data System (ADS)

    Eichele, R.; Huebener, R. P.; Seifert, H.; Selig, K. P.

    1982-02-01

    Electron beam scanning represents a convenient method for imaging ballistic phonon propagation in a crystal. Using a SEM with a low-temperature stage, the method is demonstrated for single-crystalline α-quartz. Our results agree remarkably well with calculations by Rösch and Weis.

  15. 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.

  16. 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.

  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. Scanning Electron Microanalysis and Analytical Challenges of Mapping Elements in Urban Atmospheric Particles

    EPA Science Inventory

    Elemental mapping with energy-dispersive X-ray spectroscopy (EDX) associated with scanning electron microscopy is highly useful for studying internally mixed atmospheric particles. Presented is a study of individual particles from urban airsheds and the analytical challenges in q...

  19. 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...

  20. 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.

  1. A study of the small intestinal mucosa using the scanning electron microscope

    PubMed Central

    Marsh, M. N.; Swift, J. A.

    1969-01-01

    In this paper we describe the features of small intestinal structure in normal control subjects using the scanning electron microscope. ImagesFIGS. 2a and 2bFIG. 3FIG. 4FIG. 5FIG. 6FIGS. 7a and 7bFIG. 8FIG. 9FIG. 10FIG. 11FIG. 12FIG. 13FIG. 14FIG. 15 PMID:5358588

  2. Scanning Electron Microanalysis and Analytical Challenges of Mapping Elements in Urban Atmospheric Particles

    EPA Science Inventory

    Elemental mapping with energy-dispersive X-ray spectroscopy (EDX) associated with scanning electron microscopy is highly useful for studying internally mixed atmospheric particles. Presented is a study of individual particles from urban airsheds and the analytical challenges in q...

  3. Scanning electron microscopic study of an anterior chamber intraocular lens: latent endophthalmitis.

    PubMed

    Schémann, J F

    1987-01-01

    Two years after intracapsular cataract extraction and intraocular lens implantation, an anterior chamber lens was removed. The lens was studied by scanning electron microscope which demonstrated the presence of colonies of cocci, a thin acellular membrane covering part of the lens and some modifications of the lens surface.

  4. 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...

  5. Scanning electron microscope view of iron crystal growing on pyroxene crystal

    NASA Technical Reports Server (NTRS)

    1972-01-01

    A scanning electron microscope photograph of a four-micron size iron crystal growing on a pyroxene crystal (calcium-magnesium-iron silicate) from the Apollo 15 Hadley-Apennino lunar landing site. The well developed crystal faces indicate that the crystal was formed from a hot vapor as the rock was cooling.

  6. 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.

  7. Enhanced contrast separation in scanning electron microscopes via a suspended-thin sample approach.

    PubMed

    Ji, Yuan; Wang, Li; Guo, Zhenxi; Wei, Bin; Zhao, Jie; Wang, Xiaodong; Zhang, Yinqi; Sui, Manling; Han, Xiaodong

    2014-11-01

    A suspended-thin-sample (STS) approach for signal selection and contrast separation is developed in scanning electron microscopes with commonly used primary beam energies and traditional detectors. Topography contrast, electron channeling contrast and composition contrast are separated and largely enhanced from suspended thin samples of several hundred nanometers in thickness, which is less than the escape depth of backscattered electrons. This imaging technique enables to detect relatively pure secondary electron and elastic backscattered electron singles, whereas suppress multiple inelastic scattering effects. The provided contrast features are different from those of bulk samples, which are largely mixed with inelastic scattering effects. The STS imaging concept and method could be expected to have more applications in distinguishing materials of nanostructures, multilayers, compounds and composites, as well as in SEM-based electron backscatter diffraction, cathodoluminesence, and x-ray microanalysis.

  8. Analytical Formulae for Calculation of X-Ray Detector Solid Angles in the Scanning and Scanning/Transmission Analytical Electron Microscope

    SciTech Connect

    Zaluzec, Nestor J.

    2014-08-01

    Closed form analytical equations used to calculate the collection solid angle of six common geometries of solid-state X-ray detectors in scanning and scanning/transmission analytical electron microscopy are presented. Using these formulae one can make realistic comparisons of the merits of the different detector geometries in modern electron column instruments. This work updates earlier formulations and adds new detector configurations.

  9. Micro-Scanning Electron Microscope and X-ray Spectrometer for Planetary Exploration

    NASA Astrophysics Data System (ADS)

    Blake, D. F.; Nguyen, C. V.; Dholakia, G.; Ribaya, B. P.; Niemann, D.; Ngo, V.; McKenzie, C.; Rahman, M.; Alam, A.; Joy, D.; Espinosa, B.

    2007-12-01

    Scanning Electron Microscopy combined with electron-induced X-ray Fluorescence Spectroscopy (SEM-EDX) is one of the most powerful techniques for characterizing surface morphology and composition with spatial resolution of a micrometer or better. SEM-EDX can elucidate natural processes such as low-temperature diagenesis, thermal or pressure induced metamorphism, volcanism/magmatism, atmosphere/crust interaction and the like. This information is useful for the investigation of the natural history of solar system objects. We are developing a prototype micromachined scanning electron microscope with X-ray spectrometer (MSEMS) for solar system exploration. The MSEMS is comprised of a carbon nanotube field emission (CNTFE) electron source integrated with a micro-electro-mechanical-system (MEMS) based electron gun and electron optics structure. The MSEMS system will utilize a piezoelectric sample stage, having scan ranges from a few angstroms to several hundreds of microns. Compared with conventional electron sources, the CNTFE source offers advantages of low power usage, ultra-small source size and simplicity of electrostatic focusing. The MSEMS instrument, including CNTFE source, MEMS electron optic column and piezoelectric sample stage, is envisioned to be 1-2 cm in height and will operate in the range of 500 eV to 15 KeV. The imaging resolution of MEMS is predicted to be ~10 nm at 5 KeV and the spatial resolution of the X-ray spectrometer will be ~1 μm at 15 KeV. We will present field emission data from our CNTFE source as well as the MEMS electron gun and piezostage designs.

  10. 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

  11. 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.

  12. 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.

  13. 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.

  14. Tunneling rates in electron transport through double-barrier molecular junctions in a scanning tunneling microscope.

    PubMed

    Nazin, G V; Wu, S W; Ho, W

    2005-06-21

    The scanning tunneling microscope enables atomic-scale measurements of electron transport through individual molecules. Copper phthalocyanine and magnesium porphine molecules adsorbed on a thin oxide film grown on the NiAl(110) surface were probed. The single-molecule junctions contained two tunneling barriers, vacuum gap, and oxide film. Differential conductance spectroscopy shows that electron transport occurs via vibronic states of the molecules. The intensity of spectral peaks corresponding to the individual vibronic states depends on the relative electron tunneling rates through the two barriers of the junction, as found by varying the vacuum gap tunneling rate by changing the height of the scanning tunneling microscope tip above the molecule. A simple, sequential tunneling model explains the observed trends.

  15. Three-dimensional optical transfer functions in the aberration-corrected scanning transmission electron microscope.

    PubMed

    Jones, L; Nellist, P D

    2014-05-01

    In the scanning transmission electron microscope, hardware aberration correctors can now correct for the positive spherical aberration of round electron lenses. These correctors make use of nonround optics such as hexapoles or octupoles, leading to the limiting aberrations often being of a nonround type. Here we explore the effect of a number of potential limiting aberrations on the imaging performance of the scanning transmission electron microscope through their resulting optical transfer functions. In particular, the response of the optical transfer function to changes in defocus are examined, given that this is the final aberration to be tuned just before image acquisition. The resulting three-dimensional optical transfer functions also allow an assessment of the performance of a system for focal-series experiments or optical sectioning applications. © 2014 The Authors Journal of Microscopy © 2014 Royal Microscopical Society.

  16. Scanning and transmission electron microscopic study of equine infectious anemia virus.

    PubMed

    Gonda, M A; Charman, H P; Walker, J L; Coggins, L

    1978-05-01

    Scanning and transmission electron microscopy were used to study in detail the morphogenesis and replication of equine infectious anemia virus (EIAV) in cultured, persistently infected equine fetal kidney fibroblasts. The EIAV was shown by thin-section electron microscopy to resemble morphologically more closely the members of the genus Lenti-virus in the family Retroviridae than other genera. Scanning electron microscopy demonstrated budding virus on only about 5% of the equine fetal kidney fibroblasts; however, the entire surface of these cells was involved in viral replication. Except where virus budding was observed, EIAV-infected cells were smooth and free of the topographic surface alterations characteristic of cells transformed by type C retroviruses. The morphologic relationship of EIAV and pathologic manifestations of EIAV infection to those of other Retroviridae are discussed.

  17. 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.

  18. 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.

  19. 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.

  20. 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.

  1. 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.

  2. Quality improvement of environmental secondary electron detector signal using helium gas in variable pressure scanning electron microscopy.

    PubMed

    Oho, Eisaku; Suzuki, Kazuhiko; Yamazaki, Sadao

    2007-01-01

    The quality of the image signal obtained from the environmental secondary electron detector (ESED) employed in a variable pressure (VP) SEM can be dramatically improved by using helium gas. The signal-to-noise ratio (SNR) increases gradually in the range of the pressures that can be used in our modified SEM. This method is especially useful in low-voltage VP SEM as well as in a variety of SEM operating conditions, because helium gas can more or less maintain the amount of unscattered primary electrons. In order to measure the SNR precisely, a digital scan generator system for obtaining two images with identical views is employed as a precondition.

  3. 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.

  4. 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.

  5. 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.

  6. Fabrication and characterization of solid-state nanopores using a field emission scanning electron microscope

    SciTech Connect

    Chang Hung; Iqbal, Samir M.; Stach, Eric A.; King, Alexander H.; Zaluzec, Nestor J.; Bashir, Rashid

    2006-03-06

    The fabrication of solid-state nanopores using the electron beam of a transmission electron microscope (TEM) has been reported in the past. Here, we report a similar method to fabricate solid-state nanopores using the electron source of a conventional field-emission scanning electron microscope (FESEM) instead. Micromachining was used to create initial pore diameters between 50 nm and 200 nm, and controlled pore shrinking to sub 10 nm diameters was performed subsequently during in situ processing in the FESEM. Noticeably, different shrinking behavior was observed when using irradiation from the electron source of the FESEM than the TEM. Unlike previous reports of TEM mediated pore shrinkage, the mechanism of pore shrinkage when using the FESEM could be a result of surface defects generated by radiolysis and subsequent motion of silicon atoms to the pore periphery.

  7. 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.

  8. 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.

  9. An electro-conductive organic coating for scanning electron microscopy (déjà vu)

    NASA Astrophysics Data System (ADS)

    Burnett, Bryan R.

    2014-09-01

    An organic compound, originally marketed as an antistatic, can form an extremely thin electro-conductive coating upon drying. A scanning electron microscope (SEM) application for this compound was first explored in the late 1960s. A coating of this compound eliminates the need for carbon or gold coating in some applications. It is well suited for the viewing of fabric samples and associated gunshot residue (GSR) in the SEM and makes it possible to quickly analyze fabric bullet wipe and bore wipe GSR. Fabric samples can also be examined for GSR from intermediate-range shots to estimate muzzle-target distances. Scanning

  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. Image formation in the scanning transmission electron microscope using object-conjugate detectors.

    PubMed

    Dwyer, C; Lazar, S; Chang, L Y; Etheridge, J

    2012-03-01

    This work presents a theoretical analysis of image formation in a scanning transmission electron microscope equipped with electron detectors in a plane conjugate to the specimen. This optical geometry encompasses both the three-dimensional imaging technique of scanning confocal electron microscopy (SCEM) and a recently developed atomic resolution imaging technique coined real-space scanning transmission electron microscopy (R-STEM). Image formation in this geometry is considered from the viewpoints of both wave optics and geometric optics, and the validity of the latter is analysed by means of Wigner distributions. Relevant conditions for the validity of a geometric interpretation of image formation are provided. For R-STEM, where a large detector is used, it is demonstrated that a geometric optics description of image formation provides an accurate approximation to wave optics, and that this description offers distinct advantages for interpretation and numerical implementation. The resulting description of R-STEM is also demonstrated to be in good agreement with experiment. For SCEM, it is emphasized that a geometric optics description of image formation is valid provided that higher-order aberrations can be ignored and the detector size is large enough to average out diffraction from the angle-limiting aperture.

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

    NASA Astrophysics Data System (ADS)

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

    2017-08-01

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

  13. Observation of Live Ticks (Haemaphysalis flava) by Scanning Electron Microscopy under High Vacuum Pressure

    PubMed Central

    Ishigaki, Yasuhito; Nakamura, Yuka; Oikawa, Yosaburo; Yano, Yasuhiro; Kuwabata, Susumu; Nakagawa, Hideaki; Tomosugi, Naohisa; Takegami, Tsutomu

    2012-01-01

    Scanning electron microscopes (SEM), which image sample surfaces by scanning with an electron beam, are widely used for steric observations of resting samples in basic and applied biology. Various conventional methods exist for SEM sample preparation. However, conventional SEM is not a good tool to observe living organisms because of the associated exposure to high vacuum pressure and electron beam radiation. Here we attempted SEM observations of live ticks. During 1.5×10−3 Pa vacuum pressure and electron beam irradiation with accelerated voltages (2–5 kV), many ticks remained alive and moved their legs. After 30-min observation, we removed the ticks from the SEM stage; they could walk actively under atmospheric pressure. When we tested 20 ticks (8 female adults and 12 nymphs), they survived for two days after SEM observation. These results indicate the resistance of ticks against SEM observation. Our second survival test showed that the electron beam, not vacuum conditions, results in tick death. Moreover, we describe the reaction of their legs to electron beam exposure. These findings open the new possibility of SEM observation of living organisms and showed the resistance of living ticks to vacuum condition in SEM. These data also indicate, for the first time, the usefulness of tick as a model system for biology under extreme condition. PMID:22431980

  14. 2D MEMS scanning for LIDAR with sub-Nyquist sampling, electronics, and measurement procedure

    NASA Astrophysics Data System (ADS)

    Giese, Thorsten; Janes, Joachim

    2015-05-01

    Electrostatic driven 2D MEMS scanners resonantly oscillate in both axes leading to Lissajous trajectories of a digitally modulated laser beam reflected from the micro mirror. A solid angle of about 0.02 is scanned by a 658nm laser beam with a maximum repetition rate of 350MHz digital pulses. Reflected light is detected by an APD with a bandwidth of 80MHz. The phase difference between the scanned laser light and the light reflected from an obstacle is analyzed by sub-Nyquist sampling. The FPGA-based electronics and software for the evaluation of distance and velocity of objects within the scanning range are presented. Furthermore, the measures to optimize the Lidar accuracy of about 1mm and the dynamic range of up to 2m are examined. First measurements demonstrating the capability of the system and the evaluation algorithms are discussed.

  15. Gastroesophageal junction of Anatolian shepherd dog; a study by topographic anatomy, scanning electron and light microscopy.

    PubMed

    Alsafy, M A M; El-Gendy, S A A

    2012-03-01

    The aim of this study was to cast a spotlight on the topography and to point out the clinical importance of the gastroesophageal junction (GEJ) in Anatolian Shepherd dogs. Nine Anatolian Shepherd dogs were used to study the morphology of the GEJ. The esophagus was appeared has a portion within the thoracic cavity while no portion of the esophagus presented within the abdominal cavity that documented the absence of the intra-abdominal portion in all studied dogs. The topographic anatomy, scanning electron and light microscopic examinations revealed that the gastroesophageal junction was located at the level of the phrenico-esophageal ligament (PEL) inside the esophageal hiatus. Our results were distinguished the morphology of the esophageal and gastric cardiac mucosa at the level of the gastroesophageal junction by the scanning electron micrographs. The light microscopical examination was explained the PEL attached to the esophageal side in one dog and to the gastric cardiac side in three dogs.

  16. 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.

  17. 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.

  18. 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.

  19. Depth Sectioning with the Aberration-Corrected Scanning Transmission Electron Microscope

    SciTech Connect

    Borisevich, Albina Y; Lupini, Andrew R; Pennycook, Stephen J

    2006-01-01

    The ability to correct the aberrations of the probe-forming lens in the scanning transmission electron microscope provides not only a significant improvement in transverse resolution but in addition brings depth resolution at the nanometer scale. Aberration correction therefore opens up the possibility of 3D imaging by optical sectioning. Here we develop a definition for the depth resolution for scanning transmission electron microscope depth sectioning and present initial results from this method. Objects such as catalytic metal clusters and single atoms on various support materials are imaged in three dimensions with a resolution of several nanometers. Effective focal depth is determined by statistical analysis and the contributing factors are discussed. Finally, current challenges and future capabilities available through new instruments are discussed.

  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. 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

  2. The removal of the smear layer using EGTA: a scanning electron microscopic study.

    PubMed

    Viswanath, Deepak; Hegde, Amitha M; Munshi, A K

    2003-01-01

    The smear layer associated with endodontic instrumentation is currently thought to be a thin layer that occludes the orifices of the dentinal tubules and covers the intertubular dentin of the prepared canal wall. The purpose of this study was to compare the effect of ethylene glycol bis (beta-amino ethyl ether)-N, N, N', N'-tetra acetic acid (EGTA) and ethylene diamine tetra acetic acid (EDTA) on removal of the smear layer through the scanning electron microscopy. Twenty four single rooted teeth were selected, instrumented and irrigated with various solutions and the specimens were processed for scanning electron microscopy. It was found that though both EGTA and EDTA completely removed the smear layer, EDTA caused erosion and conjugation of the tubules, whereas, EGTA effectively removed the smear layer without inducing any erosion. It was thus concluded that EGTA can be effectively used as an alternative chelator for the removal of the smear layer.

  3. Scanning Electron Microscope Calibration Using a Multi-Image Non-Linear Minimization Process

    NASA Astrophysics Data System (ADS)

    Cui, Le; Marchand, Éric

    2015-04-01

    A scanning electron microscope (SEM) calibrating approach based on non-linear minimization procedure is presented in this article. A part of this article has been published in IEEE International Conference on Robotics and Automation (ICRA), 2014. . Both the intrinsic parameters and the extrinsic parameters estimations are achieved simultaneously by minimizing the registration error. The proposed approach considers multi-images of a multi-scale calibration pattern view from different positions and orientations. Since the projection geometry of the scanning electron microscope is different from that of a classical optical sensor, the perspective projection model and the parallel projection model are considered and compared with distortion models. Experiments are realized by varying the position and the orientation of a multi-scale chessboard calibration pattern from 300× to 10,000×. The experimental results show the efficiency and the accuracy of this approach.

  4. 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.

  5. 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.

  6. 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.

  7. Fracture study of quasi-brittle material using a fast-scanning electron microscope

    NASA Astrophysics Data System (ADS)

    Wang, M. L.; Ross, T. J.; Tan, L. Z.; Macy, R. J.

    1992-02-01

    A fast-scanning electron microscope (FSEM) capable of 381 Hz framing rates has been developed. An in situ tension and compression stage was designed for studying the dynamic microstructure of cementitious materials. Fast scanning of the specimen results in fewer secondary electrons produced per unit time and a reduced signal-to-noise ratio, where the image appears 'snowy'. An image correlation technique also developed to correct the deficiencies of a reduced signal-to-noise ratio in order to measure the displacement distribution in front of the tip of the propagating crack. It is concluded that the FSEM and image techniques under consideration are a viable tool for moderate dynamic fracture studies of quasi-brittle and cementitious materials.

  8. Scanning electron microscopic study of the tongue in the Oriental scops owl (Otus scops).

    PubMed

    Emura, Shoichi; Okumura, Toshihiko; Chen, Huayue

    2009-05-01

    The dorsal lingual surface of an adult owl (Otus scops) was examined by scanning electron microscopy. The tongue of the adult owl was about 1 cm long. Three parts were distinguished in the dorsal surface of the tongue: the apex, the body and the root of the tongue. The conical region between the lingual apex and lingual root was very wide area. The conical papillae of the lingual body were inclined toward the posterior of the tongue. At low magnification of scanning electron microscopy, the desquamated cells were observed in the entire dorsal surface of the lingual apex. The connective tissue cores of the epithelium of the lingual apex showed the rod-shaped protrusions. The border between the lingual apex and body was clear and the small conical papillae were observed in the lingual body. The small and large conical papillae were observed on the lingual body. The many openings of the lingual glands existed in the lingual body and lingual root.

  9. Morphological changes of the hair roots in alopecia areata: a scanning electron microscopic study.

    PubMed

    Karashima, Tadashi; Tsuruta, Daisuke; Hamada, Takahiro; Ishii, Norito; Ono, Fumitake; Ueda, Akihiro; Abe, Toshifumi; Nakama, Takekuni; Dainichi, Teruki; Hashimoto, Takashi

    2013-12-01

    Alopecia areata is a chronic inflammatory condition causing non-scarring patchy hair loss. Diagnosis of alopecia areata is made by clinical observations, hair pluck test and dermoscopic signs. However, because differentiation from other alopecia diseases is occasionally difficult, an invasive diagnostic method using a punch biopsy is performed. In this study, to develop a reliable, less invasive diagnostic method for alopecia areata, we performed scanning electron microscopy of the hair roots of alopecia areata patients. This study identified four patterns of hair morphology specific to alopecia areata: (I) long tapering structure with no accumulation of scales; (II) club-shaped hair root with fine scales; (III) proximal accumulation of scales; and (IV) sharp tapering of the proximal end of hair. On the basis of these results, we can distinguish alopecia areata by scanning electron microscopic observation of the proximal end of the hair shafts. © 2013 Japanese Dermatological Association.

  10. Image restoration for TV-scan moving images acquired through a semiconductor backscattered electron detector.

    PubMed

    Oho, Eisaku; Suzuki, Kazuhiko

    2009-01-01

    A semiconductor backscattered electron (BSE) detector has become popular in scanning electron microscopy session. However, detectors of semiconductor type have a serious disadvantage on the frequency characteristics. As a result, fast scan (e.g. TV-scan) BSE image should be blurred remarkably. It is the purpose of this study to restore this degradation by using digital image processing technology. In order to improve it practically, we have to settle several problems, such as noise, undesirable processing artifacts, and ease of use. Image processing techniques in an impromptu manner like a conventional mask processing are unhelpful for this study, because a complicated degradation of output signal affects severely the phase response as well as the amplitude response of our SEM system. Hence, based on the characteristics of an SEM signal obtained from the semiconductor BSE detector, a proper inverse filter in Fourier domain is designed successfully. Finally, the inverse filter is converted to a special convolution mask, which is skillfully designed, and applied for TV-scan moving BSE images. The improved BSE image is very effective in the work for finding important objects. (c) 2010 Wiley Periodicals, Inc.

  11. Simulation of multicomponent losses in electron beam melting and refining at varying scan frequencies

    SciTech Connect

    Powell, A.; Szekely, J.; Van Den Avyle, J.; Damkroger, B.

    1995-10-12

    A two-stage model is presented to describe alloy element evaporation rates from molten metal due to transient local heating by an electron beam. The first stage is a simulation of transient phenomena near the melt surface due to periodic heating by a scanning beam, the output of which is the relationship between operating parameters, surface temperature, and evaporation rate. At high scan rates, this can be done using a simple one-dimensional heat transfer model of the surface layer; at lower scan rates, a more complex three-dimensional model with fluid flow and periodic boundary conditions is necessary. The second stage couples this evaporation-surface temperature relationship with a larger steady state heat transfer and fluid flow model of an entire melting hearth or mold, in order to calculate local and total evaporation rates. Predictions are compared with experimental results from Sandia`s 310-kW electron beam melting furnace, in which evaporation rates and vapor compositions were studied in pure titanium and Ti-6%Al-4%V alloy. Evaporation rates were estimated from rate of condensation on a substrate held over the hearth, and were characterized as a function of beam power (150 and 225 kW), scan frequency (30, 115 and 450 Hz) and background pressure (10{sup {minus}3}, 10{sup {minus}4} and 10{sup {minus}5} torr).

  12. 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.

  13. 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.

  14. 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.

  15. Transient of scanning electron microscopic images for a buried microstructure in insulators

    SciTech Connect

    Zhang Haibo; Li Daoyu; Li Weiqin

    2007-12-15

    We clarify the transient process and its mechanism of scanning electron microscope (SEM) images of a trench microstructure buried in insulators. First, interface charges of primary electrons trapped on the trench are derived from the charging model of a capacitor considering the electron beam induced current, and the surface potential is therefore assumed. The SEM signal current is then determined from its simplified relation with the surface potential. Calculated profiles of the secondary electron (SE) signal current and their time-evolution behaviors can well fit the transient of the experimental SEM images. Results show that the variation of the surface potential due to the transient interface charges and the effect of SE redistribution result in transients of the SEM imaging signal and the image width of the buried trench.

  16. 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.

  17. 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

  18. Scanning tunneling spectroscopy of inhomogeneous electronic structure in monolayer and bilayer graphene on SiC

    NASA Astrophysics Data System (ADS)

    Brar, Victor W.; Zhang, Yuanbo; Yayon, Yossi; Ohta, Taisuke; McChesney, Jessica L.; Bostwick, Aaron; Rotenberg, Eli; Horn, Karsten; Crommie, Michael F.

    2007-09-01

    The authors present a scanning tunneling spectroscopy (STS) study of the local electronic structure of single and bilayer graphene grown epitaxially on a SiC(0001) surface. Low voltage topographic images reveal fine, atomic-scale carbon networks, whereas higher bias images are dominated by emergent spatially inhomogeneous large-scale structure similar to a carbon-rich reconstruction of SiC(0001). STS spectroscopy shows an ˜100meV gaplike feature around zero bias for both monolayer and bilayer graphene/SiC, as well as significant spatial inhomogeneity in electronic structure above the gap edge. Nanoscale structure at the SiC/graphene interface is seen to correlate with observed electronic spatial inhomogeneity. These results are relevant for potential devices involving electronic transport or tunneling in graphene/SiC.

  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. 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.

  1. 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.

  2. Comparison of Electron Imaging Modes for Dimensional Measurements in the Scanning Electron Microscope.

    PubMed

    Postek, Michael T; Vladár, András E; Villarrubia, John S; Muto, Atsushi

    2016-08-01

    Dimensional measurements from secondary electron (SE) images were compared with those from backscattered electron (BSE) and low-loss electron (LLE) images. With the commonly used 50% threshold criterion, the lines consistently appeared larger in the SE images. As the images were acquired simultaneously by an instrument with the capability to operate detectors for both signals at the same time, the differences cannot be explained by the assumption that contamination or drift between images affected the SE, BSE, or LLE images differently. Simulations with JMONSEL, an electron microscope simulator, indicate that the nanometer-scale differences observed on this sample can be explained by the different convolution effects of a beam with finite size on signals with different symmetry (the SE signal's characteristic peak versus the BSE or LLE signal's characteristic step). This effect is too small to explain the >100 nm discrepancies that were observed in earlier work on different samples. Additional modeling indicates that those discrepancies can be explained by the much larger sidewall angles of the earlier samples, coupled with the different response of SE versus BSE/LLE profiles to such wall angles.

  3. Comparison of Electron Imaging Modes for Dimensional Measurements in the Scanning Electron Microscope†

    PubMed Central

    Postek, Michael T.; Vladár, András E.; Villarrubia, John S.; Muto, Atsushi

    2016-01-01

    Dimensional measurements from secondary electron (SE) images were compared with those from backscattered electron (BSE) and low-loss electron (LLE) images. With the commonly used 50% threshold criterion, the lines consistently appeared larger in the SE images. As the images were acquired simultaneously by an instrument with the capability to operate detectors for both signals at the same time, the differences cannot be explained by the assumption that contamination or drift between images affected the SE, BSE, or LLE images differently. Simulations with JMONSEL, an electron microscope simulator, indicate that the nanometer-scale differences observed on this sample can be explained by the different convolution effects of a beam with finite size on signals with different symmetry (the SE signal’s characteristic peak versus the BSE or LLE signal’s characteristic step). This effect is too small to explain the > 100 nm discrepancies that were observed in earlier work on different samples. Additional modeling indicates that those discrepancies can be explained by the much larger sidewall angles of the earlier samples, coupled with the different response of SE versus BSE/LLE profiles to such wall angles. PMID:27452278

  4. SEM analysis of ionizing radiation effects in linear integrated circuits. [Scanning Electron Microscope

    NASA Technical Reports Server (NTRS)

    Stanley, A. G.; Gauthier, M. K.

    1977-01-01

    A successful diagnostic technique was developed using a scanning electron microscope (SEM) as a precision tool to determine ionization effects in integrated circuits. Previous SEM methods radiated the entire semiconductor chip or major areas. The large area exposure methods do not reveal the exact components which are sensitive to radiation. To locate these sensitive components a new method was developed, which consisted in successively irradiating selected components on the device chip with equal doses of electrons /10 to the 6th rad (Si)/, while the whole device was subjected to representative bias conditions. A suitable device parameter was measured in situ after each successive irradiation with the beam off.

  5. 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.

  6. New views of materials through aberration-corrected scanning transmission electron microscopy.

    PubMed

    Pennycook, S J; Varela, M

    2011-01-01

    The successful correction of third-order and, more recently, fifth-order aberrations has enormously enhanced the capabilities of the scanning transmission electron microscope (STEM), by not only achieving record resolution, but also allowing near 100% efficiency for electron energy loss spectroscopy, and higher currents for two-dimensional spectrum imaging. These advances have meant that the intrinsic advantages of the STEM, incoherent imaging and simultaneous collection of multiple complementary images can now give new insights into many areas of materials physics. Here, we review a number of examples, mostly from the field of complex oxides, and look towards new directions for the future.

  7. Ultrahigh-resolution Scanning Transmission Microscopy with Sub-?ngstrom-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 aberra- tion of microscope objective lenses has been successful in converging the beam into sub- 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- ngstrom resolution has been indeed achieved for not only simple structures but also structurally complex systems(quasicrystals).

  8. 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.

  9. The Scanning Electron Microscope As An Accelerator For The Undergraduate Advanced Physics Laboratory

    SciTech Connect

    Peterson, Randolph S.; Berggren, Karl K.; Mondol, Mark

    2011-06-01

    Few universities or colleges have an accelerator for use with advanced physics laboratories, but many of these institutions have a scanning electron microscope (SEM) on site, often in the biology department. As an accelerator for the undergraduate, advanced physics laboratory, the SEM is an excellent substitute for an ion accelerator. Although there are no nuclear physics experiments that can be performed with a typical 30 kV SEM, there is an opportunity for experimental work on accelerator physics, atomic physics, electron-solid interactions, and the basics of modern e-beam lithography.

  10. Electronic states of InAs/GaAs quantum dots by scanning tunneling spectroscopy

    SciTech Connect

    Gaan, S.; He, Guowei; Feenstra, R. M.; Walker, J.; Towe, E.

    2010-09-20

    InAs/GaAs quantum-dot (QD) heterostructures grown by molecular-beam epitaxy are studied using cross-sectional scanning tunneling microscopy and spectroscopy. Individual InAs QDs are resolved in the images. Tunneling spectra acquired 3-4 nm from the QDs show a peak located in the upper part of the GaAs band gap originating from the lowest electron confined state, together with a tail extending out from the valence band from hole confined states. A line-shape analysis is used to deduce the binding energies of the electron and hole QD states.

  11. 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.

  12. Energy dispersive X-ray analysis on an absolute scale in scanning transmission electron microscopy.

    PubMed

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

    2015-10-01

    We demonstrate absolute scale agreement between the number of X-ray counts in energy dispersive X-ray spectroscopy using an atomic-scale coherent electron probe and first-principles simulations. Scan-averaged spectra were collected across a range of thicknesses with precisely determined and controlled microscope parameters. Ionization cross-sections were calculated using the quantum excitation of phonons model, incorporating dynamical (multiple) electron scattering, which is seen to be important even for very thin specimens. Copyright © 2015 Elsevier B.V. All rights reserved.

  13. 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.

  14. Characterization of LiBC by phase-contrast scanning transmission electron microscopy.

    PubMed

    Krumeich, Frank; Wörle, Michael; Reibisch, Philipp; Nesper, Reinhard

    2014-08-01

    LiBC was used as a model compound for probing the applicability of phase-contrast (PC) imaging in an aberration-corrected scanning transmission electron microscope (STEM) to visualize lithium distributions. In the LiBC structure, boron and carbon are arranged to hetero graphite layers between which lithium is incorporated. The crystal structure is reflected in the PC-STEM images recorded perpendicular to the layers. The experimental images and their defocus dependence match with multi-slice simulations calculated utilizing the reciprocity principle. The observation that a part of the Li positions is not occupied is likely an effect of the intense electron beam triggering Li displacement.

  15. Scanning electron microscopy of egg hatching of Anopheles albimanus (Diptera: Culicidae).

    PubMed

    Rodriguez, M H; Orozco, A; Chavez, B; Martinez-Palomo, A

    1992-09-01

    Scanning electron and light microscopic observations showed that egg hatching in Anopheles albimanus Wiedemann is aided by a chisel-shaped spine. This hatching tooth is surrounded by a thin flexible membrane fixed to a groove in the head of the larvae. Increased intracranial pressure may force the spine against the egg shell until a fissure is produced. Further opening of the egg is achieved by movements of the head and the entire body of the larva.

  16. 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.

  17. 3D scanning electron microscopy applied to surface characterization of fluorosed dental enamel.

    PubMed

    Limandri, Silvina; Galván Josa, Víctor; Valentinuzzi, María Cecilia; Chena, María Emilia; Castellano, Gustavo

    2016-05-01

    The enamel surfaces of fluorotic teeth were studied by scanning electron stereomicroscopy. Different whitening treatments were applied to 25 pieces to remove stains caused by fluorosis and their surfaces were characterized by stereomicroscopy in order to obtain functional and amplitude parameters. The topographic features resulting for each treatment were determined through these parameters. The results obtained show that the 3D reconstruction achieved from the SEM stereo pairs is a valuable potential alternative for the surface characterization of this kind of samples.

  18. Bacterial Biofilm Morphology on a Failing Implant with an Oxidized Surface: A Scanning Electron Microscope Study.

    PubMed

    Simion, Massimo; Kim, David M; Pieroni, Stefano; Nevins, Myron; Cassinelli, Clara

    2016-01-01

    This case report provided a unique opportunity to investigate the extent of microbiota infiltration on the oxidized implant surface that has been compromised by peri-implantitis. Scanning electron microscopic analysis confirmed the etiologic role of the bacteria on the loss of supporting structure and the difficulty in complete removal of bacterial infiltration on the implant surface. This case report emphasizes the need to perform definitive surface decontamination on failing dental implants prior to a regeneration procedure.

  19. Multi-Channel Electronically Scanned Cryogenic Pressure Sensor And Method For Making Same

    NASA Technical Reports Server (NTRS)

    Chapman, John J. (Inventor); Hopson, Purnell, Jr. (Inventor); Holloway, Nancy M. (Inventor)

    2001-01-01

    A miniature, multi-channel, electronically scanned pressure measuring device uses electrostatically bonded silicon dies in a multi-element array. These dies are bonded at specific sites on a glass, pre-patterned substrate. Thermal data is multiplexed and recorded on each individual pressure measuring diaphragm. The device functions in a cryogenic environment without the need of heaters to keep the sensor at constant temperatures.

  20. 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.

  1. 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.

  2. 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.

  3. 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.

  4. 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.

  5. 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

  6. A new method using Scanning Electron Microscopy (SEM) for preparation of anisopterous odonates.

    PubMed

    Del Palacio, Alejandro; Sarmiento, Patricia Laura; Javier, Muzón

    2017-10-01

    Anisopterous odonate male's secondary genitalia is a complex of several structures, among them the vesica spermalis is the most informative with important specific characters. The observation of those characters, mostly of membranous nature, is difficult in the Scanning Electron Microscope due to dehydration and metallization processes. In this contribution, we discuss a new and low cost procedure for the observation of these characters in the SEM, compatible with the most common agents used for preserving specimens. © 2017 Wiley Periodicals, Inc.

  7. 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.

  8. 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.

  9. [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.

  10. Diffusion length measurements using the scanning electron microscope. [in semiconductor devices

    NASA Technical Reports Server (NTRS)

    Weizer, V. G.

    1975-01-01

    A measurement technique employing the scanning electron microscope is described in which values of the true bulk diffusion length are obtained. It is shown that surface recombination effects can be eliminated through the application of highly doped surface field layers. The influence of high injection level effects and low-high junction current generation on the resulting measurement was investigated. Close agreement is found between the diffusion lengths measured by this method and those obtained using a penetrating radiation technique.

  11. Scanning electron microscopy of a pink inclusion from the Allende meteorite

    NASA Technical Reports Server (NTRS)

    Grossman, L.; Fruland, R. M.; Mckay, D. S.

    1975-01-01

    A scanning electron microscope study of a fine-grained, pin, Ca-rich inclusion from the Allende meteorite has revealed strong evidence for direct condensation of its constituent minerals from a vapor. This observation extends to the alkali-bearing phases in addition to the Ca-, Al-silicates and suggests that the feldspathoids as well as the refractory silicates are solar nebular condensates.

  12. Adaptation of electronically scanned pressure measurement technology at Langley Research Center

    NASA Astrophysics Data System (ADS)

    Mitchell, Michael

    The use of pressure-measurement devices based on electronic scanning in wind-tunnel and other settings with harsh conditions is discussed in terms of adaptations required to protect the integrity of the devices and output data. The applications considered include thermal extremes, long cable lengths, very low pressure, high-line pressure, and variable-reference calibrations. Some of the adaptation techniques are thermal enclosures, remote power supplies, low-pressure standards, and biasing regulators.

  13. Defect tracking for nanoimprint lithography using optical surface scanner and scanning electron microscope

    NASA Astrophysics Data System (ADS)

    Yu, Zhaoning; Kurataka, Nobuo; Tran, Hieu; Gauzner, Gene

    2016-09-01

    Fast optical surface scanners are used in combination with high-resolution scanning electron microscopes to facilitate the identification and tracking of nanoimprint defects. We have confirmed that hard particles cause permanent template damages during imprint, resulting in repeating imprint defects. Since contaminants encountered during imprint are dominated by hard metal oxide particles capable of causing such damage, stringent pre-imprint substrate screening is a critical requirement in a manufacturing environment.

  14. Steel Hardness Effects in Boundary Lubricated Sliding: An In-Situ SEM (Scanning Electron Microscope) Study

    DTIC Science & Technology

    1988-01-01

    in a scanning electron microscope (SEM). Through modifications to theSEM, these experiments could be run with a thin film of hydrocarbon oil applied to...as the number of sliding passes increases, plus agglomeration of wear debris interspersed with oil around the contact. Both of these effects lead to...24). [lhcse iments could be run with a thin film of hyvdrocarbon oil applied to were all basically unlubricated experiments that insol.t.d the sliding

  15. Confocal laser scanning, scanning electron, and transmission electron microscopy investigation of Enterococcus faecalis biofilm degradation using passive and active sodium hypochlorite irrigation within a simulated root canal model.

    PubMed

    Mohmmed, Saifalarab A; Vianna, Morgana E; Penny, Matthew R; Hilton, Stephen T; Mordan, Nicola; Knowles, Jonathan C

    2017-08-01

    Root canal irrigation is an important adjunct to control microbial infection. The aim of this study was to investigate the effect of 2.5% (wt/vol) sodium hypochlorite (NaOCl) agitation on the removal, killing, and degradation of Enterococcus faecalis biofilm. A total of 45 root canal models were manufactured using 3D printing with each model comprising an 18 mm length simulated root canal of apical size 30 and taper 0.06. E. faecalis biofilms were grown on the apical 3 mm of the models for 10 days. A total of 60 s of 9 ml of 2.5% NaOCl irrigation using syringe and needle was performed, the irrigant was either left stagnant in the canal or agitated using manual (Gutta-percha), sonic, and ultrasonic methods for 30 s. Following irrigation, the residual biofilms were observed using confocal laser scanning, scanning electron, and transmission electron microscopy. The data were analyzed using one-way ANOVA with Dunnett post hoc tests at a level of significance p ≤ .05. Consequence of root canal irrigation indicate that the reduction in the amount of biofilm achieved with the active irrigation groups (manual, sonic, and ultrasonic) was significantly greater when compared with the passive and untreated groups (p < .05). Collectively, finding indicate that passive irrigation exhibited more residual biofilm on the model surface than irrigant agitated by manual or automated (sonic, ultrasonic) methods. Total biofilm degradation and nonviable cells were associated with the ultrasonic group. © 2017 The Authors. MicrobiologyOpen published by John Wiley & Sons Ltd.

  16. A high-voltage scanning electron microscopy system for in situ electromigration testing

    NASA Astrophysics Data System (ADS)

    Doan, J. C.; Lee, S.; Lee, S.-H.; Meier, N. E.; Bravman, J. C.; Flinn, P. A.; Marieb, T. N.; Madden, M. C.

    2000-07-01

    An apparatus has been constructed to conduct electromigration tests on realistic specimens while simultaneously observing them at relatively high magnification. A scanning transmission electron microscope has been converted into a high-voltage scanning electron microscope (HVSEM) with a large specimen chamber. By imaging with high-energy electrons (120 keV) and detecting backscattered electrons, voids in metal lines can be viewed through passivation layers. The HVSEM has a resolution of 50 nm through 1 μm of passivation. We have constructed instrumentation to heat and pass current through interconnect structures, while they are inside the electron microscope. Presently, the specimen temperature can be as high as 350 °C and is maintained constant to within 0.1 °C. The resistances of interconnects are measured with a precision of 0.05% during an experiment. Testing the lines at moderately accelerated conditions requires great stability of the microscope and instrumentation as well as full automation of the data collection. These requirements have been met, and metallization lines can be tested for several weeks with minimal operator intervention. Digital images of an entire 300-μm-long test structure as well as electrical data are stored automatically every few minutes during a test. The hundreds to thousands of pictures are analyzed using digital image processing techniques to extract void positions and sizes as a function of time. We use this system to characterize electromigration failure in advanced interconnect structures and to test existing theories on electromigration phenomenon.

  17. 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.

  18. 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

  19. Implementation of subcellular water mapping by electron energy loss spectroscopy in a medium-voltage scanning transmission electron microscope.

    PubMed

    Terryn, C; Michel, J; Thomas, X; Laurent-Maquin, D; Balossier, G

    2004-07-01

    The water concentration in biological cells plays a predominant role in cellular life. Using electron energy loss spectroscopy, the feasibility to measure the water content in cells has already been demonstrated. In this paper, we present an upgrade of water measurement in hydrated cryosections by spectrum imaging mode in a medium-voltage scanning transmission electron microscope. The electron energy loss spectra are recorded in spectrum imaging mode in a 2(n)x2(n) pixels array. Each spectrum is processed in order to determine the water mass content in the corresponding pixel. Then a parametric image is obtained in which grey levels are related to water concentration. In this image, it is possible to recognize the different subcellular compartments. By averaging the water concentration over the relevant pixels, we can determine the water mass content in the concerned subcellular compartment. As an example, we present water mass content measurement at subcellular level in rat hepatocytes.

  20. High-energy-resolution monochromator for aberration-corrected scanning transmission electron microscopy/electron energy-loss spectroscopy.

    PubMed

    Krivanek, Ondrej L; Ursin, Jonathan P; Bacon, Neil J; Corbin, George J; Dellby, Niklas; Hrncirik, Petr; Murfitt, Matthew F; Own, Christopher S; Szilagyi, Zoltan S

    2009-09-28

    An all-magnetic monochromator/spectrometer system for sub-30 meV energy-resolution electron energy-loss spectroscopy in the scanning transmission electron microscope is described. It will link the energy being selected by the monochromator to the energy being analysed by the spectrometer, without resorting to decelerating the electron beam. This will allow it to attain spectral energy stability comparable to systems using monochromators and spectrometers that are raised to near the high voltage of the instrument. It will also be able to correct the chromatic aberration of the probe-forming column. It should be able to provide variable energy resolution down to approximately 10 meV and spatial resolution less than 1 A.

  1. Electron--Vibron Interaction Effects on Scanning Tunneling Microscopy Current through Melamine Adsorbed on Cu(100)

    NASA Astrophysics Data System (ADS)

    Sarhan, Abdulla; Sakaue, Mamoru; Nakanishi, Hiroshi; Kasai, Hideaki

    2012-10-01

    Electron transport through the melamine molecule was studied. Melamine molecules adsorbed on a Cu(100) surface were investigated by density functional theory (DFT) calculations with the dynamical matrix method. On the basis of calculation results, a model Hamiltonian for a system composed of scanning tunneling microscope (STM), a melamine molecule, and a Cu surface was proposed, taking into account electron--vibron (electron--molecular vibrations) interactions within the melamine molecule. Then, the electronic current was formulated by the nonequilibrium Green's function (NEGF) method. Results show that current is affected by the electron--vibron interactions defined in the melamine molecule through its controllable structural changes. The rectification and fluctuation of current are triggered by low-energy electron--vibron interactions. Furthermore, the electron--vibron interaction effect is found to be enhanced as temperature increases to where higher-energy vibrons begin to be excited at lower energies. However, current becomes uniform at higher temperatures, which shows an undesired sensitivity. The weakening of the electron--vibron interaction of the out-of-molecular-plane vibrational motion can transfer the melamine molecule in its tautomerization state into a current rectifier. The reduction or induction of the repulsion of lone pairs of consecutive N atoms causes the induction or reduction of the low-energy in-plane vibrational motion, which in turn causes the switching of the I--V characteristics between less stable melamine tautomers.

  2. Design of 220 GHz electronically scanned reflectarrays for confocal imaging systems

    NASA Astrophysics Data System (ADS)

    Hedden, Abigail S.; Dietlein, Charles R.; Wikner, David A.

    2012-09-01

    The authors analyze properties of a 220 GHz imaging system that uses a scanned reflectarray to perform electronic beam scanning of a confocal imager for applications including imaging meter-sized fields of view at 50 m standoff. Designs incorporating reflectarrays with confocal imagers have not been examined previously at these frequencies. We examine tradeoffs between array size, overall system size, and number of achievable image pixels resulting in a realistic architecture capable of meeting the needs of our application. Impacts to imaging performance are assessed through encircled energy calculations, beam pointing accuracy, and examining the number and intensity of quantization lobes that appear over the scan ranges of interest. Over the desired scan range, arrays with 1 and 2-bit phase quantization showed similar array main beam energy efficiencies. Two-bit phase quantization is advantageous in terms of pointing angle error, resulting in errors of at most 15% of the diffraction-limited beam size. However, both phase quantization cases considered resulted in spurious returns over the scan range of interest and other array layouts should be examined to eliminate potential imaging artifacts.

  3. 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.

  4. 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.

  5. Atomic imaging using secondary electrons in a scanning transmission electron microscope: experimental observations and possible mechanisms.

    PubMed

    Inada, H; Su, D; Egerton, R F; Konno, M; Wu, L; Ciston, J; Wall, J; Zhu, Y

    2011-06-01

    We report detailed investigation of high-resolution imaging using secondary electrons (SE) with a sub-nanometer probe in an aberration-corrected transmission electron microscope, Hitachi HD2700C. This instrument also allows us to acquire the corresponding annular dark-field (ADF) images both simultaneously and separately. We demonstrate that atomic SE imaging is achievable for a wide range of elements, from uranium to carbon. Using the ADF images as a reference, we studied the SE image intensity and contrast as functions of applied bias, atomic number, crystal tilt, and thickness to shed light on the origin of the unexpected ultrahigh resolution in SE imaging. We have also demonstrated that the SE signal is sensitive to the terminating species at a crystal surface. A possible mechanism for atomic-scale SE imaging is proposed. The ability to image both the surface and bulk of a sample at atomic-scale is unprecedented, and can have important applications in the field of electron microscopy and materials characterization.

  6. [Localization of the apical foramen using the newest electronic instruments: stereomicroscopy and SEM (scanning electron microscopy)].

    PubMed

    Pagavino, G; Diamante, D; Marri, M; Pace, R

    1995-11-01

    Introduction of double impedence as new parameter in root canal length electronic measurement allowed first and second generation electronic apical localizers main problems overcoming: precision failure in presence of conducting fluids. Our study's purpose was an in vitro evaluation of two third generation instruments (Apit-Osada and Root ZX-Morita Corp.) ability in apical foramen localization using sodium hypoclorite as irrigating solution. 40 human monorooted teeth with immature apex were studied. 20 samples were measured by Apit and 20 by Root ZX; measurements were recorded when apical foramen was reached. Samples were fixed for stereomicroscope observation before and after apical 3 mm worn and prepared for SEM observation. Evaluations about each system's precision were made by calculating difference between foramen position determined by electronic localizer and its real anatomical position determined by a computed image analizing system linked to SEM. All measurements were included between a minimum value of -0.45 mm and a maximum value of 0.26 mm. Mann Whithney U test was performed to compare average values of the two sample groups but his was not meaningful (p = 0.18) showing that there is no valuable difference in accuracy between Apit and Root ZX. According to most researchers, who consider a +/- 0.5 mm error range clinically acceptable, and considering that in vitro measurements never exceded this limit value we conclude confirming both instruments' safety.

  7. Direct-write liquid phase transformations with a scanning transmission electron microscope

    SciTech Connect

    Unocic, Raymond R.; Lupini, Andrew R.; Borisevich, Albina Y.; Cullen, David A.; Kalinin, Sergei V.; Jesse, Stephen

    2016-08-03

    The highly energetic electron beam from a scanning transmission electron microscope (STEM) can induce local changes in the state of matter, ranging from local knock-out and atomic movement, to amorphization/crystallization, and chemical/electrochemical reactions occuring at localized liquid-solid and gas-solid interfaces. To date, fundamental studies of e-beam induced phenomena and practical applications have been limited by conventional e-beam rastering modes that allow only for uniform e-beam exposures. Here we develop an automated liquid phase nanolithography method that is capable of directly writing nanometer scaled features within silicon nitride encapsulated liquid cells. An external beam control system, connected to the scan coils of an aberration-corrected STEM, is used to precisely control the position, dwell time, and scan velocity of a sub-nanometer STEM probe. Site-specific locations in a sealed liquid cell containing an aqueous solution of H2PdCl4 are irradiated to controllably deposit palladium onto silicon nitride membranes. We determine the threshold electron dose required for the radiolytic deposition of metallic palladium, explore the influence of electron dose on the feature size and morphology of nanolithographically patterned nanostructures, and propose a feedback-controlled monitoring method for active control of the nanofabricated structures through STEM detector signal monitoring. As a result, this approach enables both fundamental studies of electron beam induced interactions with matter, as well as opens a pathway to fabricate nanostructures with tailored architectures and chemistries via shape-controlled nanolithographic patterning from liquid phase precursors.

  8. Direct-write liquid phase transformations with a scanning transmission electron microscope

    DOE PAGES

    Unocic, Raymond R.; Lupini, Andrew R.; Borisevich, Albina Y.; ...

    2016-08-03

    The highly energetic electron beam from a scanning transmission electron microscope (STEM) can induce local changes in the state of matter, ranging from local knock-out and atomic movement, to amorphization/crystallization, and chemical/electrochemical reactions occuring at localized liquid-solid and gas-solid interfaces. To date, fundamental studies of e-beam induced phenomena and practical applications have been limited by conventional e-beam rastering modes that allow only for uniform e-beam exposures. Here we develop an automated liquid phase nanolithography method that is capable of directly writing nanometer scaled features within silicon nitride encapsulated liquid cells. An external beam control system, connected to the scan coilsmore » of an aberration-corrected STEM, is used to precisely control the position, dwell time, and scan velocity of a sub-nanometer STEM probe. Site-specific locations in a sealed liquid cell containing an aqueous solution of H2PdCl4 are irradiated to controllably deposit palladium onto silicon nitride membranes. We determine the threshold electron dose required for the radiolytic deposition of metallic palladium, explore the influence of electron dose on the feature size and morphology of nanolithographically patterned nanostructures, and propose a feedback-controlled monitoring method for active control of the nanofabricated structures through STEM detector signal monitoring. As a result, this approach enables both fundamental studies of electron beam induced interactions with matter, as well as opens a pathway to fabricate nanostructures with tailored architectures and chemistries via shape-controlled nanolithographic patterning from liquid phase precursors.« less

  9. Direct-write liquid phase transformations with a scanning transmission electron microscope

    SciTech Connect

    Unocic, Raymond R.; Lupini, Andrew R.; Borisevich, Albina Y.; Cullen, David A.; Kalinin, Sergei V.; Jesse, Stephen

    2016-08-03

    The highly energetic electron beam from a scanning transmission electron microscope (STEM) can induce local changes in the state of matter, ranging from local knock-out and atomic movement, to amorphization/crystallization, and chemical/electrochemical reactions occuring at localized liquid-solid and gas-solid interfaces. To date, fundamental studies of e-beam induced phenomena and practical applications have been limited by conventional e-beam rastering modes that allow only for uniform e-beam exposures. Here we develop an automated liquid phase nanolithography method that is capable of directly writing nanometer scaled features within silicon nitride encapsulated liquid cells. An external beam control system, connected to the scan coils of an aberration-corrected STEM, is used to precisely control the position, dwell time, and scan velocity of a sub-nanometer STEM probe. Site-specific locations in a sealed liquid cell containing an aqueous solution of H2PdCl4 are irradiated to controllably deposit palladium onto silicon nitride membranes. We determine the threshold electron dose required for the radiolytic deposition of metallic palladium, explore the influence of electron dose on the feature size and morphology of nanolithographically patterned nanostructures, and propose a feedback-controlled monitoring method for active control of the nanofabricated structures through STEM detector signal monitoring. As a result, this approach enables both fundamental studies of electron beam induced interactions with matter, as well as opens a pathway to fabricate nanostructures with tailored architectures and chemistries via shape-controlled nanolithographic patterning from liquid phase precursors.

  10. Construction of a new type of low-energy scanning electron microscope with atomic resolution

    NASA Astrophysics Data System (ADS)

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

    2009-05-01

    We describe a new type of scanning electron microscope which works by directly imaging the electron field-emission sites on a nanotip. Electrons are extracted from the nanotip through a nanoscale aperture, accelerated in a high electric field and focussed to a spot using a microscale einzel lens. If the whole microscope (accelerating section and lens) and the focal length are both restricted in size to below 10 microns, then computer simulations show that the effects of aberration are extremely small and it is possible to have a system with approximately unit magnification, at electron energies as low as 300 eV. Thus a typical emission site of 1 nm diameter will produce an image of the same size and an atomic emission site with give a resolution of 0.1-0.2 nm (1-2 Å), and because the beam is not allowed to expand beyond 100nm in diameter the depth of field is large and the contribution to the beam spot size from chromatic aberrations is less than 0.02 nm (0.2 Å) for 500 eV electrons. Since it is now entirely possible to make stable atomic sized emitters (nanopyramids) it is expected that this instrument will have atomic resolution. Furthermore the brightness of the beam is determined only by the field-emission and can be up to a million times larger than in a typical (high-energy) electron microscope. The construction of this microscope, based on using a nanotip electron source which is mounted on a nanopositioner so that it can be positioned at the correct point adjacent to the microscope, entrance aperture, is described. In this geometry the scanning is achieved by moving the sample using piezos. Two methods for the construction of the microscope column are reviewed and the results of preliminary tests are described. The advantages of this low energy, bright-beam, electron microscope with atomic resolution are described. It can be used in either scanning mode or diffraction mode. The major advantage over existing microscopes is that because it works at very low

  11. Dose-rate-dependent damage of cerium dioxide in the scanning transmission electron microscope.

    PubMed

    Johnston-Peck, Aaron C; DuChene, Joseph S; Roberts, Alan D; Wei, Wei David; Herzing, Andrew A

    2016-11-01

    Beam damage caused by energetic electrons in the transmission electron microscope is a fundamental constraint limiting the collection of artifact-free information. Through understanding the influence of the electron beam, experimental routines may be adjusted to improve the data collection process. Investigations of CeO2 indicate that there is not a critical dose required for the accumulation of electron beam damage. Instead, measurements using annular dark field scanning transmission electron microscopy and electron energy loss spectroscopy demonstrate that the onset of measurable damage occurs when a critical dose rate is exceeded. The mechanism behind this phenomenon is that oxygen vacancies created by exposure to a 300keV electron beam are actively annihilated as the sample re-oxidizes in the microscope environment. As a result, only when the rate of vacancy creation exceeds the recovery rate will beam damage begin to accumulate. This observation suggests that dose-intensive experiments can be accomplished without disrupting the native structure of the sample when executed using dose rates below the appropriate threshold. Furthermore, the presence of an encapsulating carbonaceous layer inhibits processes that cause beam damage, markedly increasing the dose rate threshold for the accumulation of damage. Published by Elsevier B.V.

  12. 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

  13. 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.

  14. 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.

  15. 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.

  16. A scanning and transmission electron microscopic analysis of the cerebral aqueduct in the rabbit.

    PubMed

    Meller, S T; Dennis, B J

    1993-09-01

    An examination of the surface of the cerebral aqueduct with the scanning electron microscope revealed that the walls of the cerebral aqueduct were so heavily ciliated that most of the ependymal surface was obscured, yet certain specialized supraependymal structures could be discerned lying on (or embedded within) this matt of cilia. These structures were determined by transmission electron microscopy and Golgi analysis to be either macrophages, supraependymal neurons, dendrites from medial periaqueductal gray neurons, or axons of unknown origin. Some axons, which were found to contain vesicles, appeared to make synaptic contacts with ependymal cells. Using the transmission electron microscope, the ependymal lining was found to consist of two different cell types: normal ependymal cells and tanycytes which have a long tapering basal process that was observed to contact blood vessels or, more rarely, seemed to terminate in relation to neuronal elements. While there have been previous reports on the structure of the third and lateral ventricles in other species, there are limited reports in the rabbit. The present report is not only the first description for the rabbit, but it is the first complete scanning and transmission electron microscopic analysis of the cerebral aqueduct in any species.

  17. 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.

  18. Non-Destructive Observations of Internal Micro-Defects Using Scanning Electron-Induced Acoustic Microscope

    NASA Astrophysics Data System (ADS)

    Koyama, Atsuhiro; Shibutani, Yoji

    Scanning electron-induced acoustic microscope (SEAM) has been developed as a new tool for non-destructive observations of the internal microstructures of materials. It consists of the electric chopper to pulse the high current electron beam and the detector of the longitudinal acoustic waves, both being attached to the commercial scanning electron microscope (SEM). The cyclic chopping of electron beam with extremely high frequency of a few hundred kilohertz makes the thermal wave due to the cyclic temperature rise with the short period. The wavelength of thermal wave may determine the essential SEAM resolution, because it's much smaller than the thermal stress wave (that is, the acoustic wave), which has just the role of conveying the information of thermal wave disturbance due to unexpected change as defects. Our own-built SEAM gives the best performance for observing the internal defects like the micro-voids, because it susceptibly senses the local difference of thermal properties in the sample. The paper indicates that some non-destructive observations for the micro-voids with a few microns order existing in the sintered materials are exhibited in conjunction with their destructive observations using focused-ion beam (FIB) technique to make certain of those as the proof.

  19. 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.

  20. 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.