Investigations in space-related molecular biology. [cryo-electron microscopic and diffraction studies on terrestrial and extraterrestrial specimens

NASA Technical Reports Server (NTRS)

Fernandez-Moran, H.; Pritzker, A. N.

1974-01-01

Improved instrumentation and preparation techniques for high resolution, high voltage cryo-electron microscopic and diffraction studies on terrestrial and extraterrestrial specimens are reported. Computer correlated ultrastructural and biochemical work on hydrated and dried cell membranes and related biological systems provided information on membrane organization, ice crystal formation and ordered water, RNA virus linked to cancer, lunar rock samples, and organometallic superconducting compounds. Apollo 11, 12, 14, and 15 specimens were analyzed

Specimen preparation for high-resolution cryo-EM

PubMed Central

Passmore, Lori A.; Russo, Christopher J.

2016-01-01

Imaging a material with electrons at near-atomic resolution requires a thin specimen that is stable in the vacuum of the transmission electron microscope. For biological samples, this comprises a thin layer of frozen aqueous solution containing the biomolecular complex of interest. The process of preparing a high-quality specimen is often the limiting step in the determination of structures by single-particle electron cryomicroscopy (cryo-EM). Here we describe a systematic approach for going from a purified biomolecular complex in aqueous solution to high-resolution electron micrographs that are suitable for 3D structure determination. This includes a series of protocols for the preparation of vitrified specimens on various specimen supports, including all-gold and graphene. We also describe techniques for troubleshooting when a preparation fails to yield suitable specimens, and common mistakes to avoid during each part of the process. Finally, we include recommendations for obtaining the highest quality micrographs from prepared specimens with current microscope, detector and support technology. PMID:27572723

In Situ Microstructural Control and Mechanical Testing Inside the Transmission Electron Microscope at Elevated Temperatures

NASA Astrophysics Data System (ADS)

Wang, Baoming; Haque, M. A.

2015-08-01

With atomic-scale imaging and analytical capabilities such as electron diffraction and energy-loss spectroscopy, the transmission electron microscope has allowed access to the internal microstructure of materials like no other microscopy. It has been mostly a passive or post-mortem analysis tool, but that trend is changing with in situ straining, heating and electrical biasing. In this study, we design and demonstrate a multi-functional microchip that integrates actuators, sensors, heaters and electrodes with freestanding electron transparent specimens. In addition to mechanical testing at elevated temperatures, the chip can actively control microstructures (grain growth and phase change) of the specimen material. Using nano-crystalline aluminum, nickel and zirconium as specimen materials, we demonstrate these novel capabilities inside the microscope. Our approach of active microstructural control and quantitative testing with real-time visualization can influence mechanistic modeling by providing direct and accurate evidence of the fundamental mechanisms behind materials behavior.

Tomography experiment of an integrated circuit specimen using 3 MeV electrons in the transmission electron microscope.

PubMed

Zhang, Hai-Bo; Zhang, Xiang-Liang; Wang, Yong; Takaoka, Akio

2007-01-01

The possibility of utilizing high-energy electron tomography to characterize the micron-scale three dimensional (3D) structures of integrated circuits has been demonstrated experimentally. First, electron transmission through a tilted SiO(2) film was measured with an ultrahigh-voltage electron microscope (ultra-HVEM) and analyzed from the point of view of elastic scattering of electrons, showing that linear attenuation of the logarithmic electron transmission still holds valid for effective specimen thicknesses up to 5 microm under 2 MV accelerating voltages. Electron tomography of a micron-order thick integrated circuit specimen including the Cu/via interconnect was then tried with 3 MeV electrons in the ultra-HVEM. Serial projection images of the specimen tilted at different angles over the range of +/-90 degrees were acquired, and 3D reconstruction was performed with the images by means of the IMOD software package. Consequently, the 3D structures of the Cu lines, via and void, were revealed by cross sections and surface rendering.

Structure of Wet Specimens in Electron Microscopy

ERIC Educational Resources Information Center

Parsons, D. F.

1974-01-01

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

Nanometres-resolution Kikuchi patterns from materials science specimens with transmission electron forward scatter diffraction in the scanning electron microscope.

PubMed

Brodusch, N; Demers, H; Gauvin, R

2013-04-01

A charge-coupled device camera of an electron backscattered diffraction system in a scanning electron microscope was positioned below a thin specimen and transmission Kikuchi patterns were collected. Contrary to electron backscattered diffraction, transmission electron forward scatter diffraction provides phase identification and orientation mapping at the nanoscale. The minimum Pd particle size for which a Kikuchi diffraction pattern was detected and indexed reliably was 5.6 nm. An orientation mapping resolution of 5 nm was measured at 30 kV. The resolution obtained with transmission electron forward scatter diffraction was of the same order of magnitude than that reported in electron nanodiffraction in the transmission electron microscope. An energy dispersive spectrometer X-ray map and a transmission electron forward scatter diffraction orientation map were acquired simultaneously. The high-resolution chemical, phase and orientation maps provided at once information on the chemical form, orientation and coherency of precipitates in an aluminium-lithium 2099 alloy. © 2013 The Authors Journal of Microscopy © 2013 Royal Microscopical Society.

Simulation of transmission electron microscope images of biological specimens.

PubMed

Rullgård, H; Ofverstedt, L-G; Masich, S; Daneholt, B; Oktem, O

2011-09-01

We present a new approach to simulate electron cryo-microscope images of biological specimens. The framework for simulation consists of two parts; the first is a phantom generator that generates a model of a specimen suitable for simulation, the second is a transmission electron microscope simulator. The phantom generator calculates the scattering potential of an atomic structure in aqueous buffer and allows the user to define the distribution of molecules in the simulated image. The simulator includes a well defined electron-specimen interaction model based on the scalar Schrödinger equation, the contrast transfer function for optics, and a noise model that includes shot noise as well as detector noise including detector blurring. To enable optimal performance, the simulation framework also includes a calibration protocol for setting simulation parameters. To test the accuracy of the new framework for simulation, we compare simulated images to experimental images recorded of the Tobacco Mosaic Virus (TMV) in vitreous ice. The simulated and experimental images show good agreement with respect to contrast variations depending on dose and defocus. Furthermore, random fluctuations present in experimental and simulated images exhibit similar statistical properties. The simulator has been designed to provide a platform for development of new instrumentation and image processing procedures in single particle electron microscopy, two-dimensional crystallography and electron tomography with well documented protocols and an open source code into which new improvements and extensions are easily incorporated. © 2011 The Authors Journal of Microscopy © 2011 Royal Microscopical Society.

Foucault imaging by using non-dedicated transmission electron microscope

DOE Office of Scientific and Technical Information (OSTI.GOV)

Taniguchi, Yoshifumi; Matsumoto, Hiroaki; Harada, Ken

2012-08-27

An electron optical system for observing Foucault images was constructed using a conventional transmission electron microscope without any special equipment for Lorentz microscopy. The objective lens was switched off and an electron beam was converged by a condenser optical system to the crossover on the selected area aperture plane. The selected area aperture was used as an objective aperture to select the deflected beam for Foucault mode, and the successive image-forming lenses were controlled for observation of the specimen images. The irradiation area on the specimen was controlled by selecting the appropriate diameter of the condenser aperture.

Development of a SEM-based low-energy in-line electron holography microscope for individual particle imaging.

PubMed

Adaniya, Hidehito; Cheung, Martin; Cassidy, Cathal; Yamashita, Masao; Shintake, Tsumoru

2018-05-01

A new SEM-based in-line electron holography microscope has been under development. The microscope utilizes conventional SEM and BF-STEM functionality to allow for rapid searching of the specimen of interest, seamless interchange between SEM, BF-STEM and holographic imaging modes, and makes use of coherent low-energy in-line electron holography to obtain low-dose, high-contrast images of light element materials. We report here an overview of the instrumentation and first experimental results on gold nano-particles and carbon nano-fibers for system performance tests. Reconstructed images obtained from the holographic imaging mode of the new microscope show substantial image contrast and resolution compared to those acquired by SEM and BF-STEM modes, demonstrating the feasibility of high-contrast imaging via low-energy in-line electron holography. The prospect of utilizing the new microscope to image purified biological specimens at the individual particle level is discussed and electron optical issues and challenges to further improve resolution and contrast are considered. Copyright © 2018 Elsevier B.V. All rights reserved.

Microscopic investigation of cavitation erosion damage in metals

NASA Technical Reports Server (NTRS)

Hackworh, J. V.; Adler, W. F.

1974-01-01

The results of research to identify the cavitation erosion damage mechanisms at the microscopic level for three metals (aluminum, stainless steel, and titanium) representing a range of properties and microstructure are presented. The metals were exposed to cavitation generated in distilled water by a 20-kHz ultrasonic facility operating at a vibration amplitude of 2 mils. Representative properties of the metals and experimental details are summarized. Replicas of the eroded surfaces of the specimens obtained periodically during exposure were examined with a transmission electron microscope to follow progression of the erosion damage and identify dominant erosion mechanisms as a function of exposure time. Eroded surfaces of selected specimens were also examined with a scanning electron microscope to assist in the interpretation.

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

PubMed

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

2014-01-01

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

Phase-Contrast versus Off-Axis Illumination: Is a More Complex Microscope Always More Powerful?

ERIC Educational Resources Information Center

Hostounsky, Zdenek; Pelc, Radek

2007-01-01

In this article, a practical demonstration suitable for any biology college classroom is presented. With the examples of a complex biological specimen (slug's radula) and a simple reference specimen (electron microscopical grid imprint in gelatin), both of which can be easily prepared, the capabilities of two imaging modes commonly used in optical…

X-ray microanalysis in the scanning electron microscope.

PubMed

Roomans, Godfried M; Dragomir, Anca

2014-01-01

X-ray microanalysis conducted using the scanning electron microscope is a technique that allows the determination of chemical elements in bulk or semi-thick specimens. The lowest concentration of an element that can be detected is in the order of a few mmol/kg or a few hundred parts per million, and the smallest amount is in the order of 10(-18) g. The spatial resolution of the analysis depends on the thickness of the specimen. For biological specimen analysis, care must be taken to prevent displacement/loss of the element of interest (usually ions). Protocols are presented for the processing of frozen-hydrated and freeze-dried specimens, as well as for the analysis of small volumes of fluid, cell cultures, and other specimens. Aspects of qualitative and quantitative analysis are covered, including limitations of the technique.

X-ray microanalysis in the scanning electron microscope.

PubMed

Roomans, Godfried M; Dragomir, Anca

2007-01-01

X-ray microanalysis conducted using the scanning electron microscope is a technique that allows the determination of chemical elements in bulk or semithick specimens. The lowest concentration of an element that can be detected is in the order of a few mmol/kg or a few hundred parts per million, and the smallest amount is in the order of 10(-18) g. The spatial resolution of the analysis depends on the thickness of the specimen. For biological specimen analysis, care must be taken to prevent displacement/loss of the element of interest (usually ions). Protocols are presented for the processing of frozen-hydrated and freeze-dried specimens, as well as for the analysis of small volumes of fluid, cell cultures and other specimens. Aspects of qualitative and quantitative analysis are covered, including limitations of the technique.

Report on the Installation and Preparedness of a Protochips Fusion in-situ Heating Holder for TEM

DOE Office of Scientific and Technical Information (OSTI.GOV)

Edmondson, Philip D.

2017-03-01

This brief report documents the procurement and installation of a Protochips Fusion (formerly Aduro) high-temperature, high stability transmission electron microscopy (TEM) specimen holder that allows for the high spatial resolution characterization of material specimens at high temperature in situ of an electron microscope. This specimen holder was specifically procured for use with The FEI Talos F200X Scanning/Transmission Electron Microscope (STEM) in Oak Ridge National Laboratory’s (ORNL’s) Low Activation Materials Development and Analysis (LAMDA) Laboratory. The Protochips Fusion holder will enable high-resolution structural and chemical analysis of irradiated materials at high temperature, becoming a unique capability worldwide, and would encourage high-qualitymore » in situ experiments to be conducted on irradiated materials.« less

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.

Relative merits and limiting factors for x-ray and electron microscopy of thick, hydrated organic materials

DOE PAGES

Du, Ming; Jacobsen, Chris

2017-10-07

Electron and x-ray microscopes allow one to image the entire, unlabeled structure of hydrated materials at a resolution well beyond what visible light microscopes can achieve. However, both approaches involve ionizing radiation, so that radiation damage must be considered as one of the limits to imaging. Drawing upon earlier work, we describe here a unified approach to estimating the image contrast (and thus the required exposure and corresponding radiation dose) in both x-ray and electron microscopy. This approach accounts for factors such as plural and inelastic scattering, and (in electron microscopy) the use of energy filters to obtain so-called "zeromore » loss" images. As expected, it shows that electron microscopy offers lower dose for specimens thinner than about 1 mu m (such as for studies of macromolecules, viruses, bacteria and archaebacteria, and thin sectioned material), while x-ray microscopy offers superior characteristics for imaging thicker specimen such as whole eukaryotic cells, thick-sectioned tissues, and organs. The required radiation dose scales strongly as a function of the desired spatial resolution, allowing one to understand the limits of live and frozen hydrated specimen imaging. Lastly, we consider the factors limiting x-ray microscopy of thicker materials, suggesting that specimens as thick as a whole mouse brain can be imaged with x-ray microscopes without significant image degradation should appropriate image reconstruction methods be identified.« less

Relative merits and limiting factors for x-ray and electron microscopy of thick, hydrated organic materials

DOE Office of Scientific and Technical Information (OSTI.GOV)

Du, Ming; Jacobsen, Chris

Electron and x-ray microscopes allow one to image the entire, unlabeled structure of hydrated materials at a resolution well beyond what visible light microscopes can achieve. However, both approaches involve ionizing radiation, so that radiation damage must be considered as one of the limits to imaging. Drawing upon earlier work, we describe here a unified approach to estimating the image contrast (and thus the required exposure and corresponding radiation dose) in both x-ray and electron microscopy. This approach accounts for factors such as plural and inelastic scattering, and (in electron microscopy) the use of energy filters to obtain so-called "zeromore » loss" images. As expected, it shows that electron microscopy offers lower dose for specimens thinner than about 1 mu m (such as for studies of macromolecules, viruses, bacteria and archaebacteria, and thin sectioned material), while x-ray microscopy offers superior characteristics for imaging thicker specimen such as whole eukaryotic cells, thick-sectioned tissues, and organs. The required radiation dose scales strongly as a function of the desired spatial resolution, allowing one to understand the limits of live and frozen hydrated specimen imaging. Lastly, we consider the factors limiting x-ray microscopy of thicker materials, suggesting that specimens as thick as a whole mouse brain can be imaged with x-ray microscopes without significant image degradation should appropriate image reconstruction methods be identified.« less

The influence of structure depth on image blurring of micrometres-thick specimens in MeV transmission electron imaging.

PubMed

Wang, Fang; Sun, Ying; Cao, Meng; Nishi, Ryuji

2016-04-01

This study investigates the influence of structure depth on image blurring of micrometres-thick films by experiment and simulation with a conventional transmission electron microscope (TEM). First, ultra-high-voltage electron microscope (ultra-HVEM) images of nanometer gold particles embedded in thick epoxy-resin films were acquired in the experiment and compared with simulated images. Then, variations of image blurring of gold particles at different depths were evaluated by calculating the particle diameter. The results showed that with a decrease in depth, image blurring increased. This depth-related property was more apparent for thicker specimens. Fortunately, larger particle depth involves less image blurring, even for a 10-μm-thick epoxy-resin film. The quality dependence on depth of a 3D reconstruction of particle structures in thick specimens was revealed by electron tomography. The evolution of image blurring with structure depth is determined mainly by multiple elastic scattering effects. Thick specimens of heavier materials produced more blurring due to a larger lateral spread of electrons after scattering from the structure. Nevertheless, increasing electron energy to 2MeV can reduce blurring and produce an acceptable image quality for thick specimens in the TEM. Copyright © 2016 Elsevier Ltd. All rights reserved.

The Development of a Scanning Soft X-Ray Microscope.

NASA Astrophysics Data System (ADS)

Rarback, Harvey Miles

We have developed a scanning soft X-ray microscope, which can be used to image natural biological specimens at high resolution and with less damage than electron microscopy. The microscope focuses a monochromatic beam of synchrotron radiation to a nearly diffraction limited spot with the aid of a high resolution Fresnel zone plate, specially fabricated for us at the IBM Watson Research Center. The specimen at one atmosphere is mechanically scanned through the spot and the transmitted radiation is efficiently detected with a flow proportional counter. A computer forms a realtime transmission image of the specimen which is displayed on a color monitor. Our first generation optics have produced images of natural wet specimens at a resolution of 300 nm.

Automated 100-Position Specimen Loader and Image Acquisition System for Transmission Electron Microscopy

PubMed Central

Lefman, Jonathan; Morrison, Robert; Subramaniam, Sriram

2007-01-01

We report the development of a novel, multi-specimen imaging system for high-throughput transmission electron microscopy. Our cartridge-based loading system, called the “Gatling”, permits the sequential examination of as many as 100 specimens in the microscope for room temperature electron microscopy using mechanisms for rapid and automated specimen exchange. The software for the operation of the Gatling and automated data acquisition has been implemented in an updated version of our in-house program AutoEM. In the current implementation of the system, the time required to deliver 95 specimens into the microscope and collect overview images from each is about 13 hours. Regions of interest are identified from a low magnification atlas generation from each specimen and an unlimited number of higher magnifications images can be subsequently acquired from these regions using fully automated data acquisition procedures that can be controlled from a remote interface. We anticipate that the availability of the Gatling will greatly accelerate the speed of data acquisition for a variety of applications in biology, materials science and nanotechnology that require rapid screening and image analysis of multiple specimens. PMID:17240161

Structural Characterization and Gas Reactions of Small Metal Particles by High Resolution In-situ TEM and TED. [Transmission Electron Microscopy and Transmission Electron Diffraction

NASA Technical Reports Server (NTRS)

Heinemann, K.

1985-01-01

A commercial electron microscope with flat-plate upper pole piece configuration of the objective lens and top entry specimen introduction was modified to obtain 5 x 10 to the minus 10th power mbar pressure at the site of the specimen while maintaining the convenience of a specimen airlock system that allows operation in the 10 to the 10th power mbar range within 15 minutes after specimen change. The specimen chamber contains three wire evaporation sources, a specimen heater, and facilities for oxygen or hydrogen plasma treatment to clean as-introduced specimens. Evacuation is achieved by dural differential pumping, with fine entrance and exit apertures for the electron beam. With the microscope operating at .000001 mbar, the first differential pumping stage features a high-speed cryopump operating in a stainless steel chamber that can be mildly baked and reaches 1 x 10 to the minus 8th power mbar. The second stage, containing the evaporation sources and a custom ionization gauge within 10 cm from the specimen, is a rigorously uncompromised all-metal uhv-system that is bakable to above 200 C throughout and is pumped with an 80-liter ion pump. Design operating pressures and image quality (resolution of metal particles smaller than 1 nm in size) was achieved.

Water without windows: Evaluating the performance of open cell transmission electron microscopy under saturated water vapor conditions, and assessing its potential for microscopy of hydrated biological specimens.

PubMed

Cassidy, Cathal; Yamashita, Masao; Cheung, Martin; Kalale, Chola; Adaniya, Hidehito; Kuwahara, Ryusuke; Shintake, Tsumoru

2017-01-01

We have performed open cell transmission electron microscopy experiments through pure water vapor in the saturation pressure regime (>0.6 kPa), in a modern microscope capable of sub-Å resolution. We have systematically studied achievable pressure levels, stability and gas purity, effective thickness of the water vapor column and associated electron scattering processes, and the effect of gas pressure on electron optical resolution and image contrast. For example, for 1.3 kPa pure water vapor and 300kV electrons, we report pressure stability of ± 20 Pa over tens of minutes, effective thickness of 0.57 inelastic mean free paths, lattice resolution of 0.14 nm on a reference Au specimen, and no significant degradation in contrast or stability of a biological specimen (M13 virus, with 6 nm body diameter). We have also done some brief experiments to confirm feasibility of loading specimens into an in situ water vapor ambient without exposure to intermediate desiccating conditions. Finally, we have also checked if water experiments had any discernible impact on the microscope performance, and report pertinent vacuum and electron optical data, for reference purposes.

In situ TEM Raman spectroscopy and laser-based materials modification.

PubMed

Allen, F I; Kim, E; Andresen, N C; Grigoropoulos, C P; Minor, A M

2017-07-01

We present a modular assembly that enables both in situ Raman spectroscopy and laser-based materials processing to be performed in a transmission electron microscope. The system comprises a lensed Raman probe mounted inside the microscope column in the specimen plane and a custom specimen holder with a vacuum feedthrough for a tapered optical fiber. The Raman probe incorporates both excitation and collection optics, and localized laser processing is performed using pulsed laser light delivered to the specimen via the tapered optical fiber. Precise positioning of the fiber is achieved using a nanomanipulation stage in combination with simultaneous electron-beam imaging of the tip-to-sample distance. Materials modification is monitored in real time by transmission electron microscopy. First results obtained using the assembly are presented for in situ pulsed laser ablation of MoS 2 combined with Raman spectroscopy, complimented by electron-beam diffraction and electron energy-loss spectroscopy. Copyright © 2016 Elsevier B.V. All rights reserved.

New hardware and workflows for semi-automated correlative cryo-fluorescence and cryo-electron microscopy/tomography.

PubMed

Schorb, Martin; Gaechter, Leander; Avinoam, Ori; Sieckmann, Frank; Clarke, Mairi; Bebeacua, Cecilia; Bykov, Yury S; Sonnen, Andreas F-P; Lihl, Reinhard; Briggs, John A G

2017-02-01

Correlative light and electron microscopy allows features of interest defined by fluorescence signals to be located in an electron micrograph of the same sample. Rare dynamic events or specific objects can be identified, targeted and imaged by electron microscopy or tomography. To combine it with structural studies using cryo-electron microscopy or tomography, fluorescence microscopy must be performed while maintaining the specimen vitrified at liquid-nitrogen temperatures and in a dry environment during imaging and transfer. Here we present instrumentation, software and an experimental workflow that improves the ease of use, throughput and performance of correlated cryo-fluorescence and cryo-electron microscopy. The new cryo-stage incorporates a specially modified high-numerical aperture objective lens and provides a stable and clean imaging environment. It is combined with a transfer shuttle for contamination-free loading of the specimen. Optimized microscope control software allows automated acquisition of the entire specimen area by cryo-fluorescence microscopy. The software also facilitates direct transfer of the fluorescence image and associated coordinates to the cryo-electron microscope for subsequent fluorescence-guided automated imaging. Here we describe these technological developments and present a detailed workflow, which we applied for automated cryo-electron microscopy and tomography of various specimens. Copyright © 2016 The Authors. Published by Elsevier Inc. All rights reserved.

Combined reflection and transmission microscope for telemedicine applications in field settings.

PubMed

Biener, Gabriel; Greenbaum, Alon; Isikman, Serhan O; Lee, Kelvin; Tseng, Derek; Ozcan, Aydogan

2011-08-21

We demonstrate a field-portable upright and inverted microscope that can image specimens in both reflection and transmission modes. This compact and cost-effective dual-mode microscope weighs only ∼135 grams (<4.8 ounces) and utilizes a simple light emitting diode (LED) to illuminate the sample of interest using a beam-splitter cube that is positioned above the object plane. This LED illumination is then partially reflected from the sample to be collected by two lenses, creating a reflection image of the specimen onto an opto-electronic sensor-array that is positioned above the beam-splitter cube. In addition to this, the illumination beam is also partially transmitted through the same specimen, which then casts lensfree in-line holograms of the same objects onto a second opto-electronic sensor-array that is positioned underneath the beam-splitter cube. By rapid digital reconstruction of the acquired lensfree holograms, transmission images (both phase and amplitude) of the same specimen are also created. We tested the performance of this field-portable microscope by imaging various micro-particles, blood smears as well as a histopathology slide corresponding to skin tissue. Being compact, light-weight and cost-effective, this combined reflection and transmission microscope might especially be useful for telemedicine applications in resource limited settings. This journal is © The Royal Society of Chemistry 2011

Relative merits and limiting factors for x-ray and electron microscopy of thick, hydrated organic materials.

PubMed

Du, Ming; Jacobsen, Chris

2018-01-01

Electron and x-ray microscopes allow one to image the entire, unlabeled structure of hydrated materials at a resolution well beyond what visible light microscopes can achieve. However, both approaches involve ionizing radiation, so that radiation damage must be considered as one of the limits to imaging. Drawing upon earlier work, we describe here a unified approach to estimating the image contrast (and thus the required exposure and corresponding radiation dose) in both x-ray and electron microscopy. This approach accounts for factors such as plural and inelastic scattering, and (in electron microscopy) the use of energy filters to obtain so-called "zero loss" images. As expected, it shows that electron microscopy offers lower dose for specimens thinner than about 1 µm (such as for studies of macromolecules, viruses, bacteria and archaebacteria, and thin sectioned material), while x-ray microscopy offers superior characteristics for imaging thicker specimen such as whole eukaryotic cells, thick-sectioned tissues, and organs. The required radiation dose scales strongly as a function of the desired spatial resolution, allowing one to understand the limits of live and frozen hydrated specimen imaging. Finally, we consider the factors limiting x-ray microscopy of thicker materials, suggesting that specimens as thick as a whole mouse brain can be imaged with x-ray microscopes without significant image degradation should appropriate image reconstruction methods be identified. Copyright © 2017 Elsevier B.V. All rights reserved.

Scanning electron microscope observation of dislocations in semiconductor and metal materials.

PubMed

Kuwano, Noriyuki; Itakura, Masaru; Nagatomo, Yoshiyuki; Tachibana, Shigeaki

2010-08-01

Scanning electron microscope (SEM) image contrasts have been investigated for dislocations in semiconductor and metal materials. It is revealed that single dislocations can be observed in a high contrast in SEM images formed by backscattered electrons (BSE) under the condition of a normal configuration of SEM. The BSE images of dislocations were compared with those of the transmission electron microscope and scanning transmission electron microscope (STEM) and the dependence of BSE image contrast on the tilting of specimen was examined to discuss the origin of image contrast. From the experimental results, it is concluded that the BSE images of single dislocations are attributed to the diffraction effect and related with high-angle dark-field images of STEM.

Water without windows: Evaluating the performance of open cell transmission electron microscopy under saturated water vapor conditions, and assessing its potential for microscopy of hydrated biological specimens

PubMed Central

Yamashita, Masao; Cheung, Martin; Kalale, Chola; Adaniya, Hidehito; Kuwahara, Ryusuke; Shintake, Tsumoru

2017-01-01

We have performed open cell transmission electron microscopy experiments through pure water vapor in the saturation pressure regime (>0.6 kPa), in a modern microscope capable of sub-Å resolution. We have systematically studied achievable pressure levels, stability and gas purity, effective thickness of the water vapor column and associated electron scattering processes, and the effect of gas pressure on electron optical resolution and image contrast. For example, for 1.3 kPa pure water vapor and 300kV electrons, we report pressure stability of ± 20 Pa over tens of minutes, effective thickness of 0.57 inelastic mean free paths, lattice resolution of 0.14 nm on a reference Au specimen, and no significant degradation in contrast or stability of a biological specimen (M13 virus, with 6 nm body diameter). We have also done some brief experiments to confirm feasibility of loading specimens into an in situ water vapor ambient without exposure to intermediate desiccating conditions. Finally, we have also checked if water experiments had any discernible impact on the microscope performance, and report pertinent vacuum and electron optical data, for reference purposes. PMID:29099843

Secondary signal imaging (SSI) electron tomography (SSI-ET): A new three-dimensional metrology for mesoscale specimens in transmission electron microscope.

PubMed

Han, Chang Wan; Ortalan, Volkan

2015-09-01

We have demonstrated a new electron tomography technique utilizing the secondary signals (secondary electrons and backscattered electrons) for ultra thick (a few μm) specimens. The Monte Carlo electron scattering simulations reveal that the amount of backscattered electrons generated by 200 and 300keV incident electrons is a monotonic function of the sample thickness and this causes the thickness contrast satisfying the projection requirement for the tomographic reconstruction. Additional contribution of the secondary electrons emitted from the edges of the specimens enhances the visibility of the surface features. The acquired SSI tilt series of the specimen having mesoscopic dimensions are successfully reconstructed verifying that this new technique, so called the secondary signal imaging electron tomography (SSI-ET), can directly be utilized for 3D structural analysis of mesoscale structures. Published by Elsevier Ltd.

Transmission electron microscope studies of extraterrestrial materials

NASA Technical Reports Server (NTRS)

Keller, Lindsay P.

1995-01-01

Transmission Electron Microscopy, X-Ray spectrometry and electron-energy-loss spectroscopy are used to analyse carbon in interplanetary dust particles. Optical micrographs are shown depicting cross sections of the dust particles embedded in sulphur. Selected-area electron diffraction patterns are shown. Transmission Electron Microscope specimens of lunar soil were prepared using two methods: ion-milling and ultramicrotomy. A combination of high resolution TEM imaging and electron diffraction is used to characterize the opaque assemblages. The opaque assemblages analyzed in this study are dominated by ilmenite with lesser rutile and spinel exsolutions, and traces of Fe metal.

Transmission Electron Microscope Measures Lattice Parameters

NASA Technical Reports Server (NTRS)

Pike, William T.

1996-01-01

Convergent-beam microdiffraction (CBM) in thermionic-emission transmission electron microscope (TEM) is technique for measuring lattice parameters of nanometer-sized specimens of crystalline materials. Lattice parameters determined by use of CBM accurate to within few parts in thousand. Technique developed especially for use in quantifying lattice parameters, and thus strains, in epitaxial mismatched-crystal-lattice multilayer structures in multiple-quantum-well and other advanced semiconductor electronic devices. Ability to determine strains in indivdual layers contributes to understanding of novel electronic behaviors of devices.

Orientation and phase mapping in the transmission electron microscope using precession-assisted diffraction spot recognition: state-of-the-art results.

PubMed

Viladot, D; Véron, M; Gemmi, M; Peiró, F; Portillo, J; Estradé, S; Mendoza, J; Llorca-Isern, N; Nicolopoulos, S

2013-10-01

A recently developed technique based on the transmission electron microscope, which makes use of electron beam precession together with spot diffraction pattern recognition now offers the possibility to acquire reliable orientation/phase maps with a spatial resolution down to 2 nm on a field emission gun transmission electron microscope. The technique may be described as precession-assisted crystal orientation mapping in the transmission electron microscope, precession-assisted crystal orientation mapping technique-transmission electron microscope, also known by its product name, ASTAR, and consists in scanning the precessed electron beam in nanoprobe mode over the specimen area, thus producing a collection of precession electron diffraction spot patterns, to be thereafter indexed automatically through template matching. We present a review on several application examples relative to the characterization of microstructure/microtexture of nanocrystalline metals, ceramics, nanoparticles, minerals and organics. The strengths and limitations of the technique are also discussed using several application examples. ©2013 The Authors. Journal of Microscopy published by John Wiley & Sons Ltd on behalf of Royal Microscopical Society.

New innovations for contrast enhancement in electron microscopy

NASA Astrophysics Data System (ADS)

Mohan, A.

In this study two techniques for producing and improving contrast in Electron Microscopy are discussed. The first technique deals with the production of secondary contrast in a Variable Pressure SEM under poor vacuum conditions using the specimen current signal. A review of the prior work in this field shows that the presence of the gas ions in the microscope column results in the amplification of the specimen current signal which is enriched in secondary content. The focus of this study is to establish practical conditions for imaging samples in the microscope using specimen current with gas amplification. This is done by understanding the different variables in the microscope which affect the image formation process and then finding out optimum conditions for obtaining the best possible image, i.e., the image most enhanced in secondary contrast. A few 'real life' samples analyzed using this technique show that the gas amplified specimen current images contain secondary information and, in some cases, provide clear advantages to imaging with conventional secondary and backscattered detectors. The second technique dealing with the production of phase contrast in the TEM for extremely thin, electron transparent samples, is analyzed. A review of the literature regarding prior work in the field shows that, while the theoretical aspects of production of phase contrast in the TEM using a phase plate are well understood, there have been problems in practically implementing this in the microscope. One major assumption with most of the studies is that a fiber, partially coated with gold, results in the formation of point charges which is an essential requirement for symmetrically shifting the phase of the electron beam. The focus of this portion of the dissertation is to image the type of fields associated with such a phase plate using the technique of electron holography. It is found that there are two types of fields associated with a phase plate of this sort. One is a cylindrical field which extends along the length of the fiber while the other is a localized spherically symmetric field. A series of simulations show that the spherical field can produce phase contrast in the TEM and also improve the contrast transfer properties of the microscope.

Computer synthesis of high resolution electron micrographs

NASA Technical Reports Server (NTRS)

Nathan, R.

1976-01-01

Specimen damage, spherical aberration, low contrast and noisy sensors combine to prevent direct atomic viewing in a conventional electron microscope. The paper describes two methods for obtaining ultra-high resolution in biological specimens under the electron microscope. The first method assumes the physical limits of the electron objective lens and uses a series of dark field images of biological crystals to obtain direct information on the phases of the Fourier diffraction maxima; this information is used in an appropriate computer to synthesize a large aperture lens for a 1-A resolution. The second method assumes there is sufficient amplitude scatter from images recorded in focus which can be utilized with a sensitive densitometer and computer contrast stretching to yield fine structure image details. Cancer virus characterization is discussed as an illustrative example. Numerous photographs supplement the text.

Development of an adaptable coherent x-ray diffraction microscope with the emphasis on imaging hydrated specimens.

PubMed

Nam, Daewoong; Park, Jaehyun; Gallagher-Jones, Marcus; Shimada, Hiroki; Kim, Sangsoo; Kim, Sunam; Kohmura, Yoshiki; Ishikawa, Tetsuya; Song, Changyong

2013-11-01

This paper describes the development of a versatile coherent x-ray diffraction microscope capable of imaging biological specimens in solution. The microscope is a flexible platform accommodating various conditions, from low vacuum (10(-2) Pa) to helium gas filled ambient pressure. This flexibility greatly expands the application area, from in situ materials science to biology systems in their native state, by significantly relaxing restrictions to the sample environment. The coherent diffraction microscope has been used successfully to image a yeast cell immersed in buffer solution. We believe that the design of this coherent diffraction microscope can be directly adapted to various platforms such as table top soft x-ray laser, synchrotron x-ray sources, and x-ray free electron laser with minor relevant adjustments.

Development of an adaptable coherent x-ray diffraction microscope with the emphasis on imaging hydrated specimens

NASA Astrophysics Data System (ADS)

Nam, Daewoong; Park, Jaehyun; Gallagher-Jones, Marcus; Shimada, Hiroki; Kim, Sangsoo; Kim, Sunam; Kohmura, Yoshiki; Ishikawa, Tetsuya; Song, Changyong

2013-11-01

This paper describes the development of a versatile coherent x-ray diffraction microscope capable of imaging biological specimens in solution. The microscope is a flexible platform accommodating various conditions, from low vacuum (10-2 Pa) to helium gas filled ambient pressure. This flexibility greatly expands the application area, from in situ materials science to biology systems in their native state, by significantly relaxing restrictions to the sample environment. The coherent diffraction microscope has been used successfully to image a yeast cell immersed in buffer solution. We believe that the design of this coherent diffraction microscope can be directly adapted to various platforms such as table top soft x-ray laser, synchrotron x-ray sources, and x-ray free electron laser with minor relevant adjustments.

Acquisition parameters optimization of a transmission electron forward scatter diffraction system in a cold-field emission scanning electron microscope for nanomaterials characterization.

PubMed

Brodusch, Nicolas; Demers, Hendrix; Trudeau, Michel; Gauvin, Raynald

2013-01-01

Transmission electron forward scatter diffraction (t-EFSD) is a new technique providing crystallographic information with high resolution on thin specimens by using a conventional electron backscatter diffraction (EBSD) system in a scanning electron microscope. In this study, the impact of tilt angle, working distance, and detector distance on the Kikuchi pattern quality were investigated in a cold-field emission scanning electron microscope (CFE-SEM). We demonstrated that t-EFSD is applicable for tilt angles ranging from -20° to -40°. Working distance (WD) should be optimized for each material by choosing the WD for which the EBSD camera screen illumination is the highest, as the number of detected electrons on the screen is directly dependent on the scattering angle. To take advantage of the best performances of the CFE-SEM, the EBSD camera should be close to the sample and oriented towards the bottom to increase forward scattered electron collection efficiency. However, specimen chamber cluttering and beam/mechanical drift are important limitations in the CFE-SEM used in this work. Finally, the importance of t-EFSD in materials science characterization was illustrated through three examples of phase identification and orientation mapping. © Wiley Periodicals, Inc.

Highly Sophisticated Virtual Laboratory Instruments in Education

NASA Astrophysics Data System (ADS)

Gaskins, T.

2006-12-01

Many areas of Science have advanced or stalled according to the ability to see what can not normally be seen. Visual understanding has been key to many of the world's greatest breakthroughs, such as discovery of DNAs double helix. Scientists use sophisticated instruments to see what the human eye can not. Light microscopes, scanning electron microscopes (SEM), spectrometers and atomic force microscopes are employed to examine and learn the details of the extremely minute. It's rare that students prior to university have access to such instruments, or are granted full ability to probe and magnify as desired. Virtual Lab, by providing highly authentic software instruments and comprehensive imagery of real specimens, provides them this opportunity. Virtual Lab's instruments let explorers operate virtual devices on a personal computer to examine real specimens. Exhaustive sets of images systematically and robotically photographed at thousands of positions and multiple magnifications and focal points allow students to zoom in and focus on the most minute detail of each specimen. Controls on each Virtual Lab device interactively and smoothly move the viewer through these images to display the specimen as the instrument saw it. Users control position, magnification, focal length, filters and other parameters. Energy dispersion spectrometry is combined with SEM imagery to enable exploration of chemical composition at minute scale and arbitrary location. Annotation capabilities allow scientists, teachers and students to indicate important features or areas. Virtual Lab is a joint project of NASA and the Beckman Institute at the University of Illinois at Urbana- Champaign. Four instruments currently compose the Virtual Lab suite: A scanning electron microscope and companion energy dispersion spectrometer, a high-power light microscope, and a scanning probe microscope that captures surface properties to the level of atoms. Descriptions of instrument operating principles and uses are also part of Virtual Lab. The Virtual Lab software and its increasingly rich collection of specimens are free to anyone. This presentation describes Virtual Lab and its uses in formal and informal education.

Fermi level pinning characterisation on ammonium fluoride-treated surfaces of silicon by energy-filtered doping contrast in the scanning electron microscope

PubMed Central

Chee, Augustus K. W.

2016-01-01

Two-dimensional dopant profiling using the secondary electron (SE) signal in the scanning electron microscope (SEM) is a technique gaining impulse for its ability to enable rapid and contactless low-cost diagnostics for integrated device manufacturing. The basis is doping contrast from electrical p-n junctions, which can be influenced by wet-chemical processing methods typically adopted in ULSI technology. This paper describes the results of doping contrast studies by energy-filtering in the SEM from silicon p-n junction specimens that were etched in ammonium fluoride solution. Experimental SE micro-spectroscopy and numerical simulations indicate that Fermi level pinning occurred on the surface of the treated-specimen, and that the doping contrast can be explained in terms of the ionisation energy integral for SEs, which is a function of the dopant concentration, and surface band-bending effects that prevail in the mechanism for doping contrast as patch fields from the specimen are suppressed. PMID:27576347

Self-consistent method for quantifying indium content from X-ray spectra of thick compound semiconductor specimens in a transmission electron microscope.

PubMed

Walther, T; Wang, X

2016-05-01

Based on Monte Carlo simulations of X-ray generation by fast electrons we calculate curves of effective sensitivity factors for analytical transmission electron microscopy based energy-dispersive X-ray spectroscopy including absorption and fluorescence effects, as a function of Ga K/L ratio for different indium and gallium containing compound semiconductors. For the case of InGaN alloy thin films we show that experimental spectra can thus be quantified without the need to measure specimen thickness or density, yielding self-consistent values for quantification with Ga K and Ga L lines. The effect of uncertainties in the detector efficiency are also shown to be reduced. © 2015 The Authors Journal of Microscopy © 2015 Royal Microscopical Society.

A simple method for environmental cell depressurization for use with an electron microscope.

PubMed

Ogawa, Naoki; Mizokawa, Ryo; Saito, Minoru; Ishikawa, Akira

2017-12-01

With the aid of the environmental cell (EC) in electron microscopy, hydrated specimens have been observed at high resolutions that optical microscopy cannot attain. Due to the ultra-high vacuum conditions of the inner column of the electron microscope, the EC requires sealing films that are sufficiently thin to allow electron transmission and that are sufficiently tough to withstand the pressure difference between the inside and outside of the EC. However, most hydrated specimens can be observed at low vacuum because the saturated vapor pressure of water is known to be 0.02 atm at room temperature. These concepts have been used in the differential pumping system, but it is complicated and relatively expensive. In this work, we propose a simple method for depressurization of the EC using a 'balloon structure' and demonstrate the theoretical benefits and practical improvement for specimen observations in low-vacuum conditions. © The Author 2017. Published by Oxford University Press on behalf of The Japanese Society of Microscopy. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

Microscopic Observation of the Side Surface of Dynamically-Tensile-Fractured 6061-T6 and 2219-T87 Aluminum Alloys with Pre-Fatigue

NASA Astrophysics Data System (ADS)

Itabashi, Masaaki; Nakajima, Shigeru; Fukuda, Hiroshi

After unexpected failure of metallic structure, microscopic investigation will be performed. Generally, such an investigation is limited to search striation pattern with a SEM (scanning electron microscope). But, when the cause of the failure was not severe repeated stress, this investigation is ineffective. In this paper, new microscopic observation technique is proposed to detect low cycle fatigue-impact tensile loading history. Al alloys, 6061-T6 and 2219-T87, were fractured in dynamic tension, after severe pre-fatigue. The side surface of the fractured specimens was observed with a SEM. Neighboring fractured surface, many opened cracks on the side surface have been generated. For each specimen, the number of the cracks was counted together with information of individual sizes and geometric features. For 6061-T6 alloy specimen with the pre-fatigue, the number of the cracks is greater than that for the specimen without the pre-fatigue. For 2219-T87 alloy, the same tendency can be found after a certain screening of the crack counting. Therefore, the crack counting technique may be useful to detect the existence of the pre-fatigue from the dynamically fractured specimen surface.

Scanning Tunneling Microscope For Use In Vacuum

NASA Technical Reports Server (NTRS)

Abel, Phillip B.

1993-01-01

Scanning tunneling microscope with subangstrom resolution developed to study surface structures. Although instrument used in air, designed especially for use in vacuum. Scanning head is assembly of small, mostly rigid components made of low-outgassing materials. Includes coarse-positioning mechanical-translation stage, on which specimen mounted by use of standard mounting stub. Tunneling tip mounted on piezoelectric fine-positioning tube. Application of suitable voltages to electrodes on piezoelectric tube controls scan of tunneling tip across surface of specimen. Electronic subsystem generates scanning voltages and collects data.

Development of SEM/STEM-WDX for highly sensitive detection of light elements

NASA Astrophysics Data System (ADS)

Anan, Y.; Koguchi, M.; Kimura, T.; Sekiguchi, T.

2018-02-01

In this study, to detect the light element lithium (Li) and to detect low dosed Boron (B) in the local area at nm order, we developed an analytical electron microscope equipped with an improved serial (S)-type WDX (wavelength dispersive X-ray spectroscopy) system. In detail, to detect Li, we developed a high-conductivity multi-capillary X-ray (MCX) lens, and a diffractor with a lattice spacing (d) of 15 nm, and with a spacing variation (δ d) of 0.8 nm. Moreover, to detect low dosed light element B, we designed a high-conductivity MCX lens based on the soft X-ray reflectivity in the capillary and calculation. We developed a large-solid-angle MCX lens whose conductivity of the characteristic X-rays of B became 20 times higher than that of an MCX lens with a 30-mm focal length. Our developed analytical electron microscope was applied to a LiAl specimen and a low B-doped Si substrate specimen, and the performance of this analytical electron microscope was evaluated. As a results, this analytical electron microscope could detect the characteristic X-rays of Li with a minimum mass fraction (MMF) of 8.4 atomic % (at. %). The energy resolution was 1 eV at 55 eV. From the results of measuring the line profile of B for the unpatterned B-implantation area on a B-doped Si substrate specimen, the measured line profile data were in good agreement with secondary ion mass spectrometry data up to a depth of 100 nm with a B concentration of 0.05 at. %.

Microstructural evolution and micromechanical properties of gamma-irradiated Au ball bonds

DOE Office of Scientific and Technical Information (OSTI.GOV)

Yusoff, Wan Yusmawati Wan, E-mail: yusmawati@upnm.edu.my; Ismail, Roslina, E-mail: roslina.ismail@ukm.my; Jalar, Azman, E-mail: azmn@ukm.my

2014-07-01

The effect of gamma radiation on the mechanical and structural properties of gold ball bonds was investigated. Gold wires from thermosonic wire bonding were exposed to gamma rays from a Cobalt-60 source at a low dose (5 Gy). The load–depth curve of nanoindentation for the irradiated gold wire bond has an apparent staircase shape during loading compared to the as-received sample. The hardness of the specimens calculated from the nanoindentation shows an increase in value from 0.91 to 1.09 GPa for specimens after exposure. The reduced elastic modulus for irradiated specimens significantly increased as well, with values from 75.18 tomore » 98.55 GPa. The change in intrinsic properties due to gamma radiation was investigated using dual-focused ion beam and high-resolution transmission electron microscope analysis. The dual-focused ion beam and high-resolution transmission electron microscope images confirmed the changes in grain structure and the presence of dislocations. The scanning electron microscope micrographs of focused ion beam cross sections showed that the grain structure of the gold became elongated and smaller after exposure to gamma rays. Meanwhile, high-resolution transmission electron microscopy provided evidence that gamma radiation induced dislocation of the atomic arrangement. - Highlights: • Nanoindentation technique provides a detailed characterisation of Au ball bond. • P–h curve of irradiated Au ball bond shows an apparent pop-in event. • Hardness and reduced modulus increased after exposure. • Elongated and smaller grain structure in irradiated specimens • Prevalent presence of dislocations in the atomic arrangement.« less

Backscattered electron SEM imaging of resin sections from plant specimens: observation of histological to subcellular structure and CLEM.

PubMed

Rizzo, N W; Duncan, K E; Bourett, T M; Howard, R J

2016-08-01

We have refined methods for biological specimen preparation and low-voltage backscattered electron imaging in the scanning electron microscope that allow for observation at continuous magnifications of ca. 130-70 000 X, and documentation of tissue and subcellular ultrastructure detail. The technique, based upon early work by Ogura & Hasegawa (1980), affords use of significantly larger sections from fixed and resin-embedded specimens than is possible with transmission electron microscopy while providing similar data. After microtomy, the sections, typically ca. 750 nm thick, were dried onto the surface of glass or silicon wafer and stained with heavy metals-the use of grids avoided. The glass/wafer support was then mounted onto standard scanning electron microscopy sample stubs, carbon-coated and imaged directly at an accelerating voltage of 5 kV, using either a yttrium aluminum garnet or ExB backscattered electron detector. Alternatively, the sections could be viewed first by light microscopy, for example to document signal from a fluorescent protein, and then by scanning electron microscopy to provide correlative light/electron microscope (CLEM) data. These methods provide unobstructed access to ultrastructure in the spatial context of a section ca. 7 × 10 mm in size, significantly larger than the typical 0.2 × 0.3 mm section used for conventional transmission electron microscopy imaging. Application of this approach was especially useful when the biology of interest was rare or difficult to find, e.g. a particular cell type, developmental stage, large organ, the interface between cells of interacting organisms, when contextual information within a large tissue was obligatory, or combinations of these factors. In addition, the methods were easily adapted for immunolocalizations. © 2015 The Author. Journal of Microscopy published by John Wiley & Sons, Ltd on behalf of the Royal Microscopical Society.

DOE Office of Scientific and Technical Information (OSTI.GOV)

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

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

Cross-sectional TEM specimen preparation for W/B{sub 4}C multilayer sample using FIB

DOE Office of Scientific and Technical Information (OSTI.GOV)

Mondal, Puspen, E-mail: puspen@rrcat.gov.in; Pradhan, P. C.; Tiwari, Pragya

2016-05-23

A recent emergence of a cross-beam scanning electron microscopy (SEM)/focused-ion-beam (FIB) system have given choice to fabricate cross-sectional transmission electron microscopy (TEM) specimen of thin film multilayer sample. A 300 layer pair thin film multilayer sample of W/B{sub 4}C was used to demonstrate the specimen lift-out technique in very short time as compared to conventional cross-sectional sample preparation technique. To get large area electron transparent sample, sample prepared by FIB is followed by Ar{sup +} ion polishing at 2 kV with grazing incident. The prepared cross-sectional sample was characterized by transmission electron microscope.

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.

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.

Multi-pass transmission electron microscopy

DOE PAGES

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

2017-05-10

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

Investigation of the relations between resin and advanced composite mechanical properties. Volume 2: Appendices

NASA Technical Reports Server (NTRS)

Zimmerman, R. S.; Adams, D. F.; Walrath, D. E.

1984-01-01

One untoughened epoxy baseline resin and three toughened epoxy resin systems were evaluated. The Hercules 3502, 2220-1, and 2220-3, and Ciba-Geigy Fibredux 914 resin systems were supplied in the uncured state by NASA-Langley and cast into thin flat specimens and round dogbone specimens. Tensile and torsional shear measurements were performed at three temperatures and two moisture conditions. Coefficients of thermal expansion and moisture expansion were also measured. Extensive scanning electron microscopic examination of fracture surfaces was performed to permit the correlation of observed failure modes with the environmental conditions under which the various specimens were tested. A micromechanics analysis was used to predict the unidirectional composite response under the various test conditions, incorporating the neat resin experimental results as the required input data. The mechanical and physical test results, the scanning electron microscope observations, and the analytical predictions were then correlated.

Investigation of the relations between neat resin and advanced composite mechanical properties. Volume 1: Results

NASA Technical Reports Server (NTRS)

Zimmerman, R. S.; Adams, D. F.; Walrath, D. E.

1984-01-01

A detailed evaluation of one untoughened epoxy baseline resin and three toughened epoxy resin systems was performed. The Hercules 3502, 2220-1, and 2220-3, and Ciba-Geigy Fibredux 914 resin systems were supplied in the uncured state by NASA-Langley and cast into thin flat specimens and round dogbone specimens. Tensile and torsional shear measurements were performed at three temperatures and two moisture conditions. Coefficients of thermal expansion and moisture expansion were also measured. Extensive scanning electron microscopic examination of fracture surfaces was performed, to permit the correlation of observed failure modes with the environmental conditions under which the various specimens were tested. A micromechanics analysis was used to predict the unidirectional composite response under the various test conditions, using the neat resin experimental results as the required input data. Mechanical and physical test results, the scanning electron microscope observations, and the analytical predictions were then correlated.

Effect of Specimen Thickness on the Creep Response of a Ni-Based Single Crystal Superalloy (PREPRINT)

DTIC Science & Technology

2012-08-01

unlimited 3.1.2. Fractography Figure 5: SEM images of a 3.18mm thick sheet specimen tested at 760◦C/758MPa. (a) The region near the fracture surface... fractography using secondary electron imaging (SE) in a scanning electron microscope (SEM). No surface oxidation was observed at this temperature. The...ruptured after 210 hours. 3.2.3. Fractography The SEM image of the reconstructed creep ruptured specimen with thickness h = 3.18mm is shown in Fig. 18a

Specimen-thickness effects on transmission Kikuchi patterns in the scanning electron microscope.

PubMed

Rice, K P; Keller, R R; Stoykovich, M P

2014-06-01

We report the effects of varying specimen thickness on the generation of transmission Kikuchi patterns in the scanning electron microscope. Diffraction patterns sufficient for automated indexing were observed from films spanning nearly three orders of magnitude in thickness in several materials, from 5 nm of hafnium dioxide to 3 μm of aluminum, corresponding to a mass-thickness range of ~5 to 810 μg cm(-2) . The scattering events that are most likely to be detected in transmission are shown to be very near the exit surface of the films. The energies, spatial distribution and trajectories of the electrons that are transmitted through the film and are collected by the detector are predicted using Monte Carlo simulations. Published 2014. This article is a U.S. Government work and is in the public domain in the USA.

A scanning electron microscope technique for studying the sclerites of Cichlidogyrus.

PubMed

Fannes, Wouter; Vanhove, Maarten P M; Huyse, Tine; Paladini, Giuseppe

2015-05-01

The genus Cichlidogyrus (Monogenea: Ancyrocephalidae) includes more than 90 species, most of which are gill parasites of African cichlid fishes. Cichlidogyrus has been studied extensively in recent years, but scanning electron microscope (SEM) investigations of the isolated hard parts have not yet been undertaken. In this paper, we describe a method for isolating and scanning the sclerites of individual Cichlidogyrus worms. Twenty-year-old, formol-fixed specimens of Cichlidogyrus casuarinus were subjected to proteinase K digestion in order to release the sclerites from the surrounding soft tissues. SEM micrographs of the haptoral sclerites and the male copulatory organ are presented. The ability to digest formol-fixed specimens makes this method a useful tool for the study of historical museum collections.

Indentation-Enabled In Situ Mechanical Characterization of Micro/Nanopillars in Electron Microscopes

NASA Astrophysics Data System (ADS)

Guo, Qiang; Fu, Xidan; Guo, Xiaolei; Liu, Zhiying; Shi, Yan; Zhang, Di

2018-04-01

Indentation-enabled micro/nanomechanical characterization of small-scale specimens provides powerful new tools for probing materials properties that were once unattainable by conventional experimental methods. Recent advancement in instrumentation further allows mechanical testing to be carried out in situ in electron microscopes, with high spatial and temporal resolution. This review discusses the recent development of nanoindentation-enabled in situ mechanical testing in electron microscopes, with an emphasis on the study of micro/nanopillars. Focus is given to novel applications beyond simple compressive and tensile testing that have been developed in the past few years, and limitations and possible future research directions in this field are proposed and discussed.

The cutting of ultrathin sections with the thickness less than 20 nm from biological specimens embedded in resin blocks.

PubMed

Nebesářová, Jana; Hozák, Pavel; Frank, Luděk; Štěpan, Petr; Vancová, Marie

2016-06-01

Low voltage electron microscopes working in transmission mode, like LVEM5 (Delong Instruments, Czech Republic) working at accelerating voltage 5 kV or scanning electron microscope working in transmission mode with accelerating voltage below 1 kV, require ultrathin sections with the thickness below 20 nm. Decreasing of the primary electron energy leads to enhancement of image contrast, which is especially useful in the case of biological samples composed of elements with low atomic numbers. As a result treatments with heavy metals, like post-fixation with osmium tetroxide or ultrathin section staining, can by omitted. The disadvantage is reduced penetration ability of incident electrons influencing the usable thickness of the specimen resulting in the need of ultrathin sections of under 20 nm thickness. In this study we want to answer basic questions concerning the cutting of extremely ultrathin sections: Is it possible routinely and reproducibly to cut extremely thin sections of biological specimens embedded in commonly used resins with contemporary ultramicrotome techniques and under what conditions? Microsc. Res. Tech. 79:512-517, 2016. © 2016 Wiley Periodicals, Inc. © 2016 Wiley Periodicals, Inc.

[Electron microscopic detection rate of enteral viruses in diarrhea of dogs, cats, calves, swine and foals in the year 1988--electron microscopic study results].

PubMed

Biermann, U; Herbst, W; Krauss, H; Schliesser, T

1989-12-01

During 1988 fecal and gut samples of 641 dogs, 198 cats, 576 calves, 108 piglets and 64 foals with diarrhoea were investigated for virus infections by electron microscopy. In samples of dogs and cats parvovirus was detected at a proportion of 21.9% and 16.7%, respectively; rotavirus alone or together with coronavirus was found only in 0.3-1.5% of the specimens. In samples of calves rotavirus, as well as coronavirus dominated with a detection rate amounting to 17.4% and 26.6% respectively (including 4.5% of mixed infections); parvovirus was present in a ratio of 0.5%. Specimens of piglets mainly contained coronavirus (25.0%), and in lower percentages rotavirus (2.8%), rota- and coronaviruses (0.9%) and parvovirus (0.9%). In feces of foals rotavirus was detected in 6.3% and particles resembling picornavirus in 4.7% of cases. Not identifiable virus particles resembling corona-or picornaviruses were rarely found (between 0.6-2.5) also in specimens of the other animal species.

On the Progress of Scanning Transmission Electron Microscopy (STEM) Imaging in a Scanning Electron Microscope.

PubMed

Sun, Cheng; Müller, Erich; Meffert, Matthias; Gerthsen, Dagmar

2018-04-01

Transmission electron microscopy (TEM) with low-energy electrons has been recognized as an important addition to the family of electron microscopies as it may avoid knock-on damage and increase the contrast of weakly scattering objects. Scanning electron microscopes (SEMs) are well suited for low-energy electron microscopy with maximum electron energies of 30 keV, but they are mainly used for topography imaging of bulk samples. Implementation of a scanning transmission electron microscopy (STEM) detector and a charge-coupled-device camera for the acquisition of on-axis transmission electron diffraction (TED) patterns, in combination with recent resolution improvements, make SEMs highly interesting for structure analysis of some electron-transparent specimens which are traditionally investigated by TEM. A new aspect is correlative SEM, STEM, and TED imaging from the same specimen region in a SEM which leads to a wealth of information. Simultaneous image acquisition gives information on surface topography, inner structure including crystal defects and qualitative material contrast. Lattice-fringe resolution is obtained in bright-field STEM imaging. The benefits of correlative SEM/STEM/TED imaging in a SEM are exemplified by structure analyses from representative sample classes such as nanoparticulates and bulk materials.

Germination, growth rates, and electron microscope analysis of tomato seeds flown on the LDEF

NASA Technical Reports Server (NTRS)

Hammond, Ernest C., Jr.; Bridgers, Kevin; Brown, Cecelia Wright

1995-01-01

The tomato seeds were flown in orbit aboard the Long Duration Exposure Facility (LDEF) for nearly six years. During this time, the tomato seeds received an abundant exposure to cosmic radiation and solar wind. Upon the return of the LDEF to earth, the seeds were distributed throughout the United States and 30 foreign countries for analysis. The purpose of the experiment was to determine the long term effect of cosmic rays on living tissue. Our university analysis included germination and growth rates as well as Scanning Electron Microscopy and X-ray analysis of the control as well as Space-exposed tomato seeds. In analyzing the seeds under the Electron Microscope, usual observations were performed on the nutritional and epidermis layer of the seed. These layers appeared to be more porous in the Space-exposed seeds than on the Earth-based control seeds. This unusual characteristic may explain the increases in the space seeds growth pattern. (Several test results show that the Space-exposed seeds germinate sooner than the Earth-Based seeds. Also, the Space-exposed seeds grew at a faster rate). The porous nutritional region may allow the seeds to receive necessary nutrients and liquids more readily, thus enabling the plant to grow at a faster rate. Roots, leaves and stems were cut into small sections and mounted. After sputter coating the specimens with Argon/Gold Palladium Plasma, they were ready to be viewed under the Electron Microscope. Many micrographs were taken. The X-ray analysis displayed possible identifications of calcium, potassium, chlorine, copper, aluminum, silicon, phosphate, carbon, and sometimes sulfur and iron. The highest concentrations were shown in potassium and calcium. The Space-exposed specimens displayed a high concentration of copper and calcium in the two specimens. There was a significantly high concentration of copper in the Earth-based specimens, whereas there was no copper in the Space-exposed specimens.

Development of the field of structural physiology

PubMed Central

FUJIYOSHI, Yoshinori

2015-01-01

Electron crystallography is especially useful for studying the structure and function of membrane proteins — key molecules with important functions in neural and other cells. Electron crystallography is now an established technique for analyzing the structures of membrane proteins in lipid bilayers that closely simulate their natural biological environment. Utilizing cryo-electron microscopes with helium-cooled specimen stages that were developed through a personal motivation to understand the functions of neural systems from a structural point of view, the structures of membrane proteins can be analyzed at a higher than 3 Å resolution. This review covers four objectives. First, I introduce the new research field of structural physiology. Second, I recount some of the struggles involved in developing cryo-electron microscopes. Third, I review the structural and functional analyses of membrane proteins mainly by electron crystallography using cryo-electron microscopes. Finally, I discuss multifunctional channels named “adhennels” based on structures analyzed using electron and X-ray crystallography. PMID:26560835

Choice and maintenance of equipment for electron crystallography.

PubMed

Mills, Deryck J; Vonck, Janet

2013-01-01

The choice of equipment for an electron crystallography laboratory will ultimately be determined by the available budget; nevertheless, the ideal lab will have two electron microscopes: a dedicated 300 kV cryo-EM with a field emission gun and a smaller LaB(6) machine for screening. The high-end machine should be equipped with photographic film or a very large CCD or CMOS camera for 2D crystal data collection; the screening microscope needs a mid-size CCD for rapid evaluation of crystal samples. The microscope room installations should provide adequate space and a special environment that puts no restrictions on the collection of high-resolution data. Equipment for specimen preparation includes a carbon coater, glow discharge unit, light microscope, plunge freezer, and liquid nitrogen containers and storage dewars. When photographic film is to be used, additional requirements are a film desiccator, dark room, optical diffractometer, and a film scanner. Having the electron microscopes and ancillary equipment well maintained and always in optimum condition facilitates the production of high-quality data.

A simple cryo-holder facilitates specimen observation under a conventional scanning electron microscope.

PubMed

Tang, Chih-Yuan; Huang, Rong-Nan; Kuo-Huang, Ling-Long; Kuo, Tai-Chih; Yang, Ya-Yun; Lin, Ching-Yeh; Jane, Wann-Neng; Chen, Shiang-Jiuun

2012-02-01

A pre-cryogenic holder (cryo-holder) facilitating cryo-specimen observation under a conventional scanning electron microscope (SEM) is described. This cryo-holder includes a specimen-holding unit (the stub) and a cryogenic energy-storing unit (a composite of three cylinders assembled with a screw). After cooling, the cryo-holder can continue supplying cryogenic energy to extend the observation time for the specimen in a conventional SEM. Moreover, the cryogenic energy-storing unit could retain appropriate liquid nitrogen that can evaporate to prevent frost deposition on the surface of the specimen. This device is proved feasible for various tissues and cells, and can be applied to the fields of both biology and material science. We have employed this novel cryo-holder for observation of yeast cells, trichome, and epidermal cells in the leaf of Arabidopsis thaliana, compound eyes of insects, red blood cells, filiform papillae on the surface of rat tongue, agar medium, water molecules, penicillium, etc. All results suggested that the newly designed cryo-holder is applicable for cryo-specimen observation under a conventional SEM without cooling system. Most importantly, the design of this cryo-holder is simple and easy to operate and could adapt a conventional SEM to a plain type cryo-SEM affordable for most laboratories. Copyright © 2011 Wiley Periodicals, Inc.

Imaging and elemental mapping of biological specimens with a dual-EDS dedicated scanning transmission electron microscope

PubMed Central

Wu, J.S.; Kim, A. M.; Bleher, R.; Myers, B.D.; Marvin, R. G.; Inada, H.; Nakamura, K.; Zhang, X.F.; Roth, E.; Li, S.Y.; Woodruff, T. K.; O'Halloran, T. V.; Dravid, Vinayak P.

2013-01-01

A dedicated analytical scanning transmission electron microscope (STEM) with dual energy dispersive spectroscopy (EDS) detectors has been designed for complementary high performance imaging as well as high sensitivity elemental analysis and mapping of biological structures. The performance of this new design, based on a Hitachi HD-2300A model, was evaluated using a variety of biological specimens. With three imaging detectors, both the surface and internal structure of cells can be examined simultaneously. The whole-cell elemental mapping, especially of heavier metal species that have low cross-section for electron energy loss spectroscopy (EELS), can be faithfully obtained. Optimization of STEM imaging conditions is applied to thick sections as well as thin sections of biological cells under low-dose conditions at room- and cryogenic temperatures. Such multimodal capabilities applied to soft/biological structures usher a new era for analytical studies in biological systems. PMID:23500508

In situ sputter cleaning of thin film metal substrates for UHV-TEM corrosion studies.

NASA Technical Reports Server (NTRS)

Heinemann, K.; Poppa, H.

1973-01-01

A prerequisite for conducting valid corrosion experiments by in situ electron microscopy techniques is not only the achievement of UHV background pressure conditions at the site of the specimen but also the ability to clean the surface of the thin metal substrate specimen before initiation of the corrosive interaction. A miniaturized simple ion gun has been constructed for this purpose. The gun is small enough to be incorporated into an UHV electron microscope specimen chamber with hot stage in such a way as to permit bombardment of the substrate specimen while observing it by transmission electron microscopy TEM. It is shown that the ion beam generated is confined well enough to cause a sputtering removal of substrate material at a rate of approximately 5-10 A/min and to prevent the sputter deposition of contaminating material from the specimen holder.

Pathological Renal Findings of Chronic Renal Failure in a Patient with the E66Q Mutation in the α-galactosidase A Gene.

PubMed

Satomura, Atsushi; Fujita, Takayuki; Nakayama, Tomohiro; Kusano, Hiroyuki; Takayama, Eiichi; Hamada, Hiroaki; Maruyama, Toshiharu

2015-01-01

A 66-year-old Japanese man was diagnosed with interstitial nephritis on a renal biopsy at 45 years of age and began to receive hemodialysis at 65 years of age. He was suspected of having Fabry disease as a result of a screening study for Fabry disease performed in hemodialysis patients. He had an E66Q mutation in the α-galactosidase A gene. We conducted an electron microscopic examination of a renal biopsy specimen obtained when the patient was diagnosed with chronic renal failure at 45 years of age in order to elucidate the pathogenicity of the E66Q mutation. Interestingly, an electron microscopic examination of the renal biopsy specimen indicated no characteristic findings of Fabry disease.

Experimental Plan for the Development of Equivalent Crack Size Distributions and a Monte Carlo Model of Fatigue in Low and High-kt Specimens of Corroded AA7050-T7451

DTIC Science & Technology

2012-03-01

18 3.3.7 Fractography ... Fractography : The fracture surfaces of the fractured fatigue specimens will be examined optically and in the scanning electron microscope to measure the size...scanned and added to the report containing the results from these fatigue tests. 3.3.7 Fractography Once a specimen has been tested, and assuming

Finite element simulations of electrostatic dopant potentials in thin semiconductor specimens for electron holography.

PubMed

Somodi, P K; Twitchett-Harrison, A C; Midgley, P A; Kardynał, B E; Barnes, C H W; Dunin-Borkowski, R E

2013-11-01

Two-dimensional finite element simulations of electrostatic dopant potentials in parallel-sided semiconductor specimens that contain p-n junctions are used to assess the effect of the electrical state of the surface of a thin specimen on projected potentials measured using off-axis electron holography in the transmission electron microscope. For a specimen that is constrained to have an equipotential surface, the simulations show that the step in the projected potential across a p-n junction is always lower than would be predicted from the properties of the bulk device, but is relatively insensitive to the value of the surface state energy, especially for thicker specimens and higher dopant concentrations. The depletion width measured from the projected potential, however, has a complicated dependence on specimen thickness. The results of the simulations are of broader interest for understanding the influence of surfaces and interfaces on electrostatic potentials in nanoscale semiconductor devices. © 2013 Elsevier B.V. All rights reserved.

Whole-cell imaging of the budding yeast Saccharomyces cerevisiae by high-voltage scanning transmission electron tomography.

PubMed

Murata, Kazuyoshi; Esaki, Masatoshi; Ogura, Teru; Arai, Shigeo; Yamamoto, Yuta; Tanaka, Nobuo

2014-11-01

Electron tomography using a high-voltage electron microscope (HVEM) provides three-dimensional information about cellular components in sections thicker than 1 μm, although in bright-field mode image degradation caused by multiple inelastic scattering of transmitted electrons limit the attainable resolution. Scanning transmission electron microscopy (STEM) is believed to give enhanced contrast and resolution compared to conventional transmission electron microscopy (CTEM). Samples up to 1 μm in thickness have been analyzed with an intermediate-voltage electron microscope because inelastic scattering is not a critical limitation, and probe broadening can be minimized. Here, we employed STEM at 1 MeV high-voltage to extend the useful specimen thickness for electron tomography, which we demonstrate by a seamless tomographic reconstruction of a whole, budding Saccharomyces cerevisiae yeast cell, which is ~3 μm in thickness. High-voltage STEM tomography, especially in the bright-field mode, demonstrated sufficiently enhanced contrast and intensity, compared to CTEM tomography, to permit segmentation of major organelles in the whole cell. STEM imaging also reduced specimen shrinkage during tilt-series acquisition. The fidelity of structural preservation was limited by cytoplasmic extraction, and the spatial resolution was limited by the relatively large convergence angle of the scanning probe. However, the new technique has potential to solve longstanding problems of image blurring in biological specimens beyond 1 μm in thickness, and may facilitate new research in cellular structural biology. Copyright © 2014 Elsevier B.V. All rights reserved.

An ultrahigh vacuum multipurpose specimen chamber with sample introduction system for in situ transmission electron microscopy investigations

NASA Technical Reports Server (NTRS)

Heinemann, K.; Poppa, H.

1986-01-01

A commercial transmission electron microscope (TEM), with flat-plate upper pole piece configuration of the objective lens, and top-entry specimen introduction was modified by introducing an ultrahigh vacuum (UHV) specimen chamber for in situ TEM experimentation. The pumping and design principles and special features of this UHV chamber, which makes it possible to obtain 5 x 10 to the -10th mbar pressure at the site of the specimen, while maintaining the airlock system that allows operation in the 10 to the -10th mbar range within 15 min after specimen change, are described. Design operating pressures and image quality (resolution of metal particles smaller than 1 nm in size) were achieved. Schematic drawings and design dimensions are included.

Microstructural study of the nickel-base alloy WAZ-20 using qualitative and quantitative electron optical techniques

NASA Technical Reports Server (NTRS)

Young, S. G.

1973-01-01

The NASA nickel-base alloy WAZ-20 was analyzed by advanced metallographic techniques to qualitatively and quantitatively characterize its phases and stability. The as-cast alloy contained primary gamma-prime, a coarse gamma-gamma prime eutectic, a gamma-fine gamma prime matrix, and MC carbides. A specimen aged at 870 C for 1000 hours contained these same constituents and a few widely scattered high W particles. No detrimental phases (such as sigma or mu) were observed. Scanning electron microscope, light metallography, and replica electron microscope methods are compared. The value of quantitative electron microprobe techniques such as spot and area analysis is demonstrated.

Microscopic observations of osteoblast growth on micro-arc oxidized β titanium

NASA Astrophysics Data System (ADS)

Chen, Hsien-Te; Chung, Chi-Jen; Yang, Tsai-Ching; Tang, Chin-Hsin; He, Ju-Liang

2013-02-01

Titanium alloys are widely used in orthopedic and dental implants, owing to their excellent physical properties and biocompatibility. By using the micro-arc oxidation (MAO), we generated anatase-rich (A-TiO2) and rutile-rich (R-TiO2) titanium dioxide coatings, individually on β-Ti alloy, in which the latter achieved an enhanced in vitro and in vivo performance. Thoroughly elucidating how the osteoblasts interact with TiO2 coatings is of worthwhile interest. This study adopts the focused ion beam (FIB) to section off the TiO2 coated samples for further scanning electron microscope (SEM) and transmission electron microscope (TEM) observation. The detailed crystal structures of the TiO2 coated specimens are also characterized. Experimental results indicate osteoblasts adhered more tenaciously and grew conformably with more lamellipodia extent on the R-TiO2 specimen than on the A-TiO2 and raw β-Ti specimens. FIB/SEM cross-sectional images of the cell/TiO2 interface revealed micro gaps between the cell membrane and contact surface of A-TiO2 specimen, while it was not found on the R-TiO2 specimen. Additionally, the number of adhered and proliferated cells on the R-TiO2 specimen was visually greater than the others. Closely examining EDS line scans and elemental mappings of the FIB/TEM cross-sectional images of the cell/TiO2 interface reveals both the cell body and interior space of the TiO2 coating contain nitrogen and sulfur (the biological elements in cell). This finding supports the assumption that osteoblast can grow into the porous structure of TiO2 coatings and demonstrating that the R-TiO2 coating formed by MAO serves the best for β-Ti alloys as orthopedic and dental implants.

The effect of aging treatment on the fracture toughness and impact strength of injection molded Ni-625 superalloy parts

DOE Office of Scientific and Technical Information (OSTI.GOV)

Özgün, Özgür, E-mail: oozgun@bingol.edu.tr; Yılmaz, Ramazan; Özkan Gülsoy, H.

In this study, the effect of aging heat treatment on fracture toughness and impact strength of Ni-625 superalloy fabricated by using powder injection molding (PIM) method was examined. After a feedstock was prepared by mixing the prealloyed Ni-625 superalloy powder, which was fabricated by gas atomisation, with a polymeric binder system and then it was granulated, it was shaped through the use of injection. The molded specimens were sintered at 1300 °C for 3 h after a two-stage debinding process. Once the sintered specimens were treated in the solution at 1150 °C for 2 h, they were quenched. Aging treatmentmore » was performed by keeping specimens at 745 °C for 22 h. Fracture toughness and impact tests were performed on sintered and aged specimens. Microstructure examinations were performed by using optical microscope, scanning electron microscope, and transmission electron microscope. The results revealed that aging heat treatment led to the formation of some carbides and intermetallic phases in the microstructure. While the hardness of the aged specimens increased due to these phases, their fracture toughness and impact strength values decreased. - Highlights: • Ni-625 superalloy components were produced by means of powder injection molding. • The produced components were subjected to aging treatment. • Aging process provided approximately 50% increase in the hardness of components. • Intermetallic precipitates, carbides and TCP phases occurred within the aged parts. • Fracture toughness and impact strength values decreased due to the hard phases.« less

Ultrastructural effects of silicone oil on the clear crystalline lens of the human eye.

PubMed

Soliman, Wael; Sharaf, Mohamed; Abdelazeem, Khaled; El-Gamal, Dalia; Nafady, Allam

2018-03-01

To evaluate light and electron microscopic changes of the anterior capsule and its epithelium after clear lens extraction of vitrectomized myopic eyes with silicone oil tamponade. This prospective, controlled, non-randomized, interventional study included 20 anterior lens capsular specimens that were excised during combined clear lens extraction and silicone oil removal from previously vitrectomized highly myopic patients with silicone oil tamponade for previous retinal detachment surgeries. The specimens were examined via light microscopy and electron microscopy and compared with 20 anterior capsule specimens removed during clear lens extraction of non-vitrectomized highly myopic eyes. Light microscopic examination of clear lens anterior capsule specimens of vitrectomized myopic eyes filled with silicone oil showed relatively more flat cells with irregular outline of lens' epithelial cells with wide intercellular spaces, deeply stained nuclei, and multiple intracytoplasmic vacuoles. Scanning electron microscopy revealed collagenous surfaces filled with multiple pits, depressions, and abnormal deposits. Transmission electron microscopy revealed lens epithelial cells with apoptotic changes, many cytoplasmic vacuoles, and filopodia-like protrusions between lens epithelial cells and the capsule. Epithelial proliferation and multilayering were also observed. silicone oil may play a role in the development of apoptotic and histopathological changes in clear lens epithelial cells. Clarity of the lens at the time of silicone oil removal does not indicate an absence of cataractous changes. We found justification of combined clear lens extraction and silicone oil removal or combined phacovitrectomy when silicone oil injection is planned, but further long-term studies with larger patient groups are required.

An evaluation of the susceptibility of V-4Cr-4Ti to stress corrosion cracking in room temperature DIII-D water

DOE Office of Scientific and Technical Information (OSTI.GOV)

Kurtz, R.J.; Jones, R.H.; Johnson, W.R.

1997-04-01

Two fatigue precracked compact tension (CT) specimens of V-4Cr-4Ti were statically loaded to a stress intensity factor of about 30 MPa{radical}m in room temperature DIII-D water. The first specimen was tested for a period of about 30 days and the second specimen for about 54 days. At the conclusion of each test the specimens were fractured, and the fracture surfaces examined with a scanning electron microscope (SEM) to determine if SCC had occurred. No SCC was found in either test specimen.

ELECTRON MICROSCOPIC OBSERVATION OF SPECIMENS UNDER CONTROLLED GAS PRESSURE

PubMed Central

Heide, Hans Gunther

1962-01-01

A technique for encasing specimens in a thin gas layer during their observation in the Siemens Elmiskop I is described. All gases can be employed at pressures up to one atmosphere. Destruction of specimens can occur in the beam; all organic specimens are particularly liable to decompose. The conditions under which this can be avoided are given. A useful application of the technique allows one to prevent specimens from drying out, as they normally do in vacuum. A further application uses the controlled removal of carbon for thinning organic layers and for selective etching of organic materials. PMID:13905967

Metallurgical characterization of the fracture of several high strength aluminum alloys

NASA Technical Reports Server (NTRS)

Bhandarkar, M. D.; Lisagor, W. B.

1977-01-01

The fracture behavior for structural aluminum alloys (2024, 6061, 7075, and 7178) was examined in selected heat treatments. The investigation included tensile, shear, and precracked notch-bend specimens fractured at ambient temperature under monotonic loading. Specimens were obtained from thin sheets and thick plates and were tested in longitudinal and transverse orientations at different strain rates. Microstructures of alloys were examined using the optical microscope and the scanning electron microscope with associated energy dispersive X ray chemical analysis. Several different types of second phase particles, some not reported by other investigators, were identified in the alloys. Fracture morphology was related to microstructural variables, test variables, and type of commercial product. Specimen orientation examined in the present investigation had little effect on fracture morphology. Test strain rate changes resulted in some change in shear fracture morphology, but not in fracture morphology of tensile specimens.

What transmission electron microscopes can visualize now and in the future.

PubMed

Müller, Shirley A; Aebi, Ueli; Engel, Andreas

2008-09-01

Our review concentrates on the progress made in high-resolution transmission electron microscopy (TEM) in the past decade. This includes significant improvements in sample preparation by quick-freezing aimed at preserving the specimen in a close-to-native state in the high vacuum of the microscope. Following advances in cold stage and TEM vacuum technology systems, the observation of native, frozen hydrated specimens has become a widely used approach. It fostered the development of computer guided, fully automated low-dose data acquisition systems allowing matched pairs of images and diffraction patterns to be recorded for electron crystallography, and the collection of entire tilt-series for electron tomography. To achieve optimal information transfer to atomic resolution, field emission electron guns combined with acceleration voltages of 200-300 kV are now routinely used. The outcome of these advances is illustrated by the atomic structure of mammalian aquaporin-O and by the pore-forming bacterial cytotoxin ClyA resolved to 12 A. Further, the Yersinia injectisome needle, a bacterial pseudopilus and the binding of phalloidin to muscle actin filaments were chosen to document the advantage of the high contrast offered by dedicated scanning transmission electron microscopy (STEM) and/or the STEM's ability to measure the mass of protein complexes and directly link this to their shape. Continued progress emerging from leading research laboratories and microscope manufacturers will eventually enable us to determine the proteome of a single cell by electron tomography, and to more routinely solve the atomic structure of membrane proteins by electron crystallography.

Imaging and elemental mapping of biological specimens with a dual-EDS dedicated scanning transmission electron microscope.

PubMed

Wu, J S; Kim, A M; Bleher, R; Myers, B D; Marvin, R G; Inada, H; Nakamura, K; Zhang, X F; Roth, E; Li, S Y; Woodruff, T K; O'Halloran, T V; Dravid, Vinayak P

2013-05-01

A dedicated analytical scanning transmission electron microscope (STEM) with dual energy dispersive spectroscopy (EDS) detectors has been designed for complementary high performance imaging as well as high sensitivity elemental analysis and mapping of biological structures. The performance of this new design, based on a Hitachi HD-2300A model, was evaluated using a variety of biological specimens. With three imaging detectors, both the surface and internal structure of cells can be examined simultaneously. The whole-cell elemental mapping, especially of heavier metal species that have low cross-section for electron energy loss spectroscopy (EELS), can be faithfully obtained. Optimization of STEM imaging conditions is applied to thick sections as well as thin sections of biological cells under low-dose conditions at room and cryogenic temperatures. Such multimodal capabilities applied to soft/biological structures usher a new era for analytical studies in biological systems. Copyright © 2013 Elsevier B.V. All rights reserved.

STEMsalabim: A high-performance computing cluster friendly code for scanning transmission electron microscopy image simulations of thin specimens.

PubMed

Oelerich, Jan Oliver; Duschek, Lennart; Belz, Jürgen; Beyer, Andreas; Baranovskii, Sergei D; Volz, Kerstin

2017-06-01

We present a new multislice code for the computer simulation of scanning transmission electron microscope (STEM) images based on the frozen lattice approximation. Unlike existing software packages, the code is optimized to perform well on highly parallelized computing clusters, combining distributed and shared memory architectures. This enables efficient calculation of large lateral scanning areas of the specimen within the frozen lattice approximation and fine-grained sweeps of parameter space. Copyright © 2017 Elsevier B.V. All rights reserved.

A new electron microscope technique for the study of living materials.

PubMed

Kálmán, E

1979-07-01

In order to gain informations on the real structure of biological specimens the "wet technique" for electron microscopy has been developed. The construction and the working principle of a special microchamber are described. Applications of this technique for the investigation of blood cells, gametes and various bacteries are demonstrated by micrographs.

Description of the last instar larva and new contributions to the knowledge of the pupa of Dasyhelea mediomunda Minaya (Diptera, Culicomorpha, Ceratopogonidae).

PubMed

Díaz, Florentina; Anjos-Santos, Danielle; Funes, Amparo; Ronderos, María M

2016-01-01

The fourth instar larva of Dasyhelea mediomunda Minaya is described for the first time and a complete description of the pupa is provided, through use of phase-contrast microscope and scanning electron microscope. Studied specimens were collected in a pond connected to a small wetland "mallin" on the Patagonian steppe, Chubut province, Argentina.

ELECTRON MICROSCOPIC OBSERVATIONS OF AMOEBA PROTEUS IN GROWTH AND INANITION

PubMed Central

Cohen, Adolph I.

1957-01-01

Electron microscopic observations have been made on growing and dividing specimens of Amoeba proteus and also on starving animals. Structures presumably corresponding to the mitochondria, alpha particles, vacuoles, and Golgi material are described. A new entity, designated as a foamy particle, is noted. Descriptions are given of the cytoplasmic and nuclear membranes. During division the inner, thick nuclear membrane component is seen to vanish and the outer membrane persist. Measurements suggest a gradual reappearance of the inner component with growth. Starving animals show a loss of cytoplasmic granularity and an increase in the electron density of mitochondria, presumably due to lipide accumulation. PMID:13481020

Electron microscopic observations of amoeba proteus in growth and inanition.

PubMed

COHEN, A I

1957-11-25

Electron microscopic observations have been made on growing and dividing specimens of Amoeba proteus and also on starving animals. Structures presumably corresponding to the mitochondria, alpha particles, vacuoles, and Golgi material are described. A new entity, designated as a foamy particle, is noted. Descriptions are given of the cytoplasmic and nuclear membranes. During division the inner, thick nuclear membrane component is seen to vanish and the outer membrane persist. Measurements suggest a gradual reappearance of the inner component with growth. Starving animals show a loss of cytoplasmic granularity and an increase in the electron density of mitochondria, presumably due to lipide accumulation.

Cold-stage microscopy system for fast-frozen liquids.

PubMed

Talmon, Y; Davis, H T; Scriven, L E; Thomas, E L

1979-06-01

The least artifact-laden fixation technique for examining colloidal suspensions, microemulsions, and other microstructured liquids in the electron microscope appears to be thermal fixation, i.e., ultrafast freezing of the liquid specimen. For rapid-enough cooling and for observation in TEM/STEM a thin sample is needed. The need is met by trapping a thin layer ( approximately 100 nm) of liquid between two polyimide films ( approximately 40 nm thickness) mounted on copper grids and immersing the resulting sandwich in liquid nitrogen at its melting point. For liquids containing water, polyimides films are used since this polymer is far less susceptible to the electron beam damage observed for the commonly used polymer films such as Formvar and collodion in contact with ice. Transfer of the frozen sample into the microscope column without deleterious frost deposition and warming is accomplished with a new transfer module for the cooling stage of the JEOL JEM-100CX microscope, which makes a true cold stage out of a device originally intended for cooling specimens inside the column. Sample results obtained with the new fast-freeze, cold-stage microscopy system are given.

Method for observation of deembedded sections of fish gonad by scanning electron microscopy

NASA Astrophysics Data System (ADS)

Mao, Lian-Ju

2000-09-01

This article reports a method for examining the intracellular structure of fish gonads using a scanning electron microscope(SEM). The specimen preparation procedure is similar to that for transmission electron microscopy wherein samples cut into semi-thin sections are fixed and embedded in plastic. The embedment matrix was removed by solvents. Risen-free specimens could be observed by SEM. The morphology of matured sperms in the gonad was very clear, and the oocyte internal structures appeared in three-dimensional images. Spheroidal nucleoli and yolk vesicles and several bundles of filaments adhered on the nucleoli could be viewed by SEM for the first time.

Off-axis electron holography of bacterial cells and magnetic nanoparticles in liquid

DOE Office of Scientific and Technical Information (OSTI.GOV)

Prozorov, Tanya; Almeida, Trevor P.; Kovacs, Andras

Here, the mapping of electrostatic potentials and magnetic fields in liquids using electron holography has been considered to be unrealistic. Here, we show that hydrated cells of Magnetospirillum magneticum strain AMB-1 and assemblies of magnetic nanoparticles can be studied using off-axis electron holography in a fluid cell specimen holder within the transmission electron microscope. Considering that the holographic object and reference wave both pass through liquid, the recorded electron holograms show sufficient interference fringe contrast to permit reconstruction of the phase shift of the electron wave and mapping of the magnetic induction from bacterial magnetite nanocrystals. We assess the challengesmore » of performing in situ magnetization reversal experiments using a fluid cell specimen holder, discuss approaches for improving spatial resolution and specimen stability, and outline future perspectives for studying scientific phenomena, ranging from interparticle interactions in liquids and electrical double layers at solid–liquid interfaces to biomineralization and the mapping of electrostatic potentials associated with protein aggregation and folding.« less

Off-axis electron holography of bacterial cells and magnetic nanoparticles in liquid

DOE PAGES

Prozorov, Tanya; Almeida, Trevor P.; Kovacs, Andras; ...

2017-10-02

Here, the mapping of electrostatic potentials and magnetic fields in liquids using electron holography has been considered to be unrealistic. Here, we show that hydrated cells of Magnetospirillum magneticum strain AMB-1 and assemblies of magnetic nanoparticles can be studied using off-axis electron holography in a fluid cell specimen holder within the transmission electron microscope. Considering that the holographic object and reference wave both pass through liquid, the recorded electron holograms show sufficient interference fringe contrast to permit reconstruction of the phase shift of the electron wave and mapping of the magnetic induction from bacterial magnetite nanocrystals. We assess the challengesmore » of performing in situ magnetization reversal experiments using a fluid cell specimen holder, discuss approaches for improving spatial resolution and specimen stability, and outline future perspectives for studying scientific phenomena, ranging from interparticle interactions in liquids and electrical double layers at solid–liquid interfaces to biomineralization and the mapping of electrostatic potentials associated with protein aggregation and folding.« less

Novel perovskite coating of strontium zirconate in Inconel substrate

NASA Astrophysics Data System (ADS)

Venkatesh, G.; Blessto, B.; Rao, C. Santhosh Kumar; Subramanian, R.; Berchmans, L. John

2018-02-01

Thermal Barrier Coatings (TBC) provides a low thermal conductivity barrier to heat transfer from the hot gas in the engine to the surface of the coated alloy component. SrZrO3 powder are prepared by Sol Gel synthesis method. The synthesized powder sample is characterized by X Ray Diffraction Technique (XRD), Scanning Electron Microscope (SEM) and Transmission Electron Microscope (TEM) and the results are interpreted. The Polycrystalline nature of SrZrO3 is confirmed and lattice spacing are determined in XRD. SEM shows sub-micron sized particles and a fringed pattern is observed in TEM. The IN718 specimen is Wire Cut and Sand Blasted. A SrZrO3 double layer is coated over the Inconel specimen through a Bond Coat made of NiCoCrAlY by Plasma spraying Process and also characterized. SEM analysis of the Coating shows diffusion of Fe, Sr into the substrate.

Microscopic and ultrastructural changes of Müller's muscle in patients with simple congenital ptosis.

PubMed

Alshehri, Mohammed D; Al-Fakey, Yasser H; Alkhalidi, Hisham M; Mubark, Mohamed A; Alsuhaibani, Adel H

2014-01-01

To study microscopic and ultrastructural changes of Müller's muscle in patients with isolated congenital ptosis. In this prospective, observational case-control study, Müller's muscle specimens were collected during ptosis surgical correction for 18 consecutive patients. Each specimen was divided into 2 parts. One part was embedded in formalin for light microscopy, and the other one was fixed in 3% glutaraldehyde for electron microscopy. A neuropathologist, serving as a masked evaluator, blindly reviewed all the different features for every case and counted the number of myocytes showing distinct myofilaments in the whole grid for every case. Statistical analysis using compare means and correlation tests was conducted to investigate potential associations and/or differences within and across groups. Twelve Müller's muscle specimens from patients with simple congenital ptosis of various severities and 6 specimens from patients with aponeurotic ptosis (controls) were collected and studied. Under light microscopy, congenital ptosis slides showed a small number of dispersed myocytes in a fibrotic background, whereas acquired ptosis slides showed a greater number of well-defined myocytes. Under electron microscopy, all congenital ptosis specimens had only a very small number of myocytes with clear, distinct myofilaments. Most myocytes in the aponeurotic ptosis group showed clear, distinct myofilaments, indicating a well-preserved muscle. No relationship existed between the number of clear, distinct myofilaments observed in the congenital ptosis group by transmission electron microscopy and patient age or ptosis severity. Substantial Müller's muscle atrophy was observed in patients with different severities of isolated congenital ptosis.

Electron ptychographic phase imaging of light elements in crystalline materials using Wigner distribution deconvolution

DOE PAGES

Yang, Hao; MacLaren, Ian; Jones, Lewys; ...

2017-04-01

Recent development in fast pixelated detector technology has allowed a two dimensional diffraction pattern to be recorded at every probe position of a two dimensional raster scan in a scanning transmission electron microscope (STEM), forming an information-rich four dimensional (4D) dataset. Electron ptychography has been shown to enable efficient coherent phase imaging of weakly scattering objects from a 4D dataset recorded using a focused electron probe, which is optimised for simultaneous incoherent Z-contrast imaging and spectroscopy in STEM. Thus coherent phase contrast and incoherent Z-contrast imaging modes can be efficiently combined to provide a good sensitivity of both light andmore » heavy elements at atomic resolution. Here, we explore the application of electron ptychography for atomic resolution imaging of strongly scattering crystalline specimens, and present experiments on imaging crystalline specimens including samples containing defects, under dynamical channelling conditions using an aberration corrected microscope. A ptychographic reconstruction method called Wigner distribution deconvolution (WDD) was implemented. Our experimental results and simulation results suggest that ptychography provides a readily interpretable phase image and great sensitivity for imaging light elements at atomic resolution in relatively thin crystalline materials.« less

The qualitative f-ratio method applied to electron channelling-induced x-ray imaging with an annular silicon drift detector in a scanning electron microscope in the transmission mode.

PubMed

Brodusch, Nicolas; Gauvin, Raynald

2017-09-01

Electron channelling is known to affect the x-ray production when an accelerated electron beam is applied to a crystalline material and is highly dependent on the local crystal orientation. This effect, unless very long counting time are used, is barely noticeable on x-ray energy spectra recorded with conventional silicon drift detectors (SDD) located at a small elevation angle. However, the very high count rates provided by the new commercially available annular SDDs permit now to observe this effect routinely and may, in some circumstances, hide the true elemental x-ray variations due to the local true specimen composition. To circumvent this issue, the recently developed f-ratio method was applied to display qualitatively the true net intensity x-ray variations in a thin specimen of a Ti-6Al-4V alloy in a scanning electron microscope in transmission mode. The diffraction contrast observed in the x-ray images was successfully cancelled through the use of f-ratios and the true composition variations at the grain boundaries could be observed in relation to the dislocation alignment prior to the β-phase nucleation. The qualitative effectiveness in removing channelling effects demonstrated in this work makes the f-ratio, in its quantitative form, a possible alternative to the ZAF method in channelling conditions. © 2017 The Authors Journal of Microscopy © 2017 Royal Microscopical Society.

Densely packed beta-structure at the protein-lipid interface of porin is revealed by high-resolution cryo-electron microscopy.

PubMed

Sass, H J; Büldt, G; Beckmann, E; Zemlin, F; van Heel, M; Zeitler, E; Rosenbusch, J P; Dorset, D L; Massalski, A

1989-09-05

Porin is an integral membrane protein that forms channels across the outer membrane of Escherichia coli. Electron microscopic studies of negatively stained two-dimensional porin crystals have shown three stain accumulations per porin trimer, revealing the locations of pores spanning the membrane. In this study, reconstituted porin lattices embedded in glucose were investigated using the low-dose technique on a cryo-electron microscope equipped with a helium-cooled superconducting objective lens. The specimen temperature was maintained at 5 K to yield an improved microscopic and specimen stability. Under these conditions, we obtained for the first time electron diffraction patterns from porin lattices to a resolution of 3.2 A and images showing optical diffraction up to a resolution of 4.9 A. Applying correlation averaging techniques to the digitized micrographs, we were able to reconstruct projected images of the porin trimer to a resolution of up to 3.5 A. In the final projection maps, amplitudes from electron diffraction and phases from these images were combined. The predominant feature is a high-density narrow band (about 6 A in thickness) that delineates the outer perimeter of the trimer. Since the molecule consists of almost exclusively beta-sheet structure, as revealed by spectroscopic data, we conclude that this band is a cylindrical beta-pleated sheet crossing the membrane nearly perpendicularly to its plane. Another intriguing finding is a low-density area (about 70 A2) situated in the centre of the trimer.

The first report of new species: Trichuris landak n. sp

PubMed Central

Purwaningsih, Endang

2013-01-01

Objective To study nematode parasites morphology of Hystrix javanica (H. javanica), both through the feces and internal organs. Methods Feces were observed by direct smear method, internal organs were observed after dissecting the host. Specimens for light microscopy examination were fixed with 70% warm alcohol, cleared and mounted in lactophenol for wet mounting. Specimens for SEM examination were postfixed in cacodylate buffer and glutaraldehyde, dehydrated through a graded series of alcohol and freeze dried. The specimens were attached to stubs with double cello-tape, coated with gold and observed with a JSM5310 LV electron microscope. Figures were made with the aid of a drawing tube attached to Olympus compound microscope, other figures were photographs of scanning electron microscope images. Measurements were given in micrometers as the mean followed by the range in parentheses, unless otherwise stated. Results The nematode species found in the intestine of H. javanica are Gireterakis girardi and a new species, Trihuris landak. The new species differs with previously reported species from Hystrix because of having stylet and short cervical alae. The pattern of bacillary band is closed to Trichuris trichiurus, the species that infect human, but differs because the surface of its vulva is not covered with densely spine. Conclusions The species of nematodes found on H. javanica were Gireterakis girardi and a new species Trichuris landak n.sp. Those two species are newly recorded in Indonesia. PMID:23593584

The first report of new species: Trichuris landak n. sp.

PubMed

Purwaningsih, Endang

2013-02-01

To study nematode parasites morphology of Hystrix javanica (H. javanica), both through the feces and internal organs. Feces were observed by direct smear method, internal organs were observed after dissecting the host. Specimens for light microscopy examination were fixed with 70% warm alcohol, cleared and mounted in lactophenol for wet mounting. Specimens for SEM examination were postfixed in cacodylate buffer and glutaraldehyde, dehydrated through a graded series of alcohol and freeze dried. The specimens were attached to stubs with double cello-tape, coated with gold and observed with a JSM5310 LV electron microscope. Figures were made with the aid of a drawing tube attached to Olympus compound microscope, other figures were photographs of scanning electron microscope images. Measurements were given in micrometers as the mean followed by the range in parentheses, unless otherwise stated. The nematode species found in the intestine of H. javanica are Gireterakis girardi and a new species, Trihuris landak. The new species differs with previously reported species from Hystrix because of having stylet and short cervical alae. The pattern of bacillary band is closed to Trichuris trichiurus, the species that infect human, but differs because the surface of its vulva is not covered with densely spine. The species of nematodes found on H. javanica were Gireterakis girardi and a new species Trichuris landak n.sp. Those two species are newly recorded in Indonesia.

Optical Interferometric Micrometrology

NASA Technical Reports Server (NTRS)

Abel, Phillip B.; Lauer, James R.

1989-01-01

Resolutions in angstrom and subangstrom range sought for atomic-scale surface probes. Experimental optical micrometrological system built to demonstrate calibration of piezoelectric transducer to displacement sensitivity of few angstroms. Objective to develop relatively simple system producing and measuring translation, across surface of specimen, of stylus in atomic-force or scanning tunneling microscope. Laser interferometer used to calibrate piezoelectric transducer used in atomic-force microscope. Electronic portion of calibration system made of commercially available components.

Acid-etching technique of non-decalcified bone samples for visualizing osteocyte-lacuno-canalicular network using scanning electron microscope.

PubMed

Lampi, Tiina; Dekker, Hannah; Ten Bruggenkate, Chris M; Schulten, Engelbert A J M; Mikkonen, Jopi J W; Koistinen, Arto; Kullaa, Arja M

2018-01-01

The aim of this study was to define the acid-etching technique for bone samples embedded in polymethyl metacrylate (PMMA) in order to visualize the osteocyte lacuno-canalicular network (LCN) for scanning electron microscopy (SEM). Human jaw bone tissue samples (N = 18) were collected from the study population consisting of patients having received dental implant surgery. After collection, the bone samples were fixed in 70% ethanol and non-decalcified samples embedded routinely into polymethyl metacrylate (PMMA). The PMMA embedded specimens were acid-etched in either 9 or 37% phosphoric acid (PA) and prepared for SEM for further analysis. PMMA embedded bone specimens acid-etched by 9% PA concentration accomplishes the most informative and favorable visualization of the LCN to be observed by SEM. Etching of PMMA embedded specimens is recommendable to start with 30 s or 40 s etching duration in order to find the proper etching duration for the samples examined. Visualizing osteocytes and LCN provides a tool to study bone structure that reflects changes in bone metabolism and diseases related to bone tissue. By proper etching protocol of non-decalcified and using scanning electron microscope it is possible to visualize the morphology of osteocytes and the network supporting vitality of bone tissue.

Collection efficiency and acceptance maps of electron detectors for understanding signal detection on modern scanning electron microscopy.

PubMed

Agemura, Toshihide; Sekiguchi, Takashi

2018-02-01

Collection efficiency and acceptance maps of typical detectors in modern scanning electron microscopes (SEMs) were investigated. Secondary and backscattered electron trajectories from a specimen to through-the-lens and under-the-lens detectors placed on an electron optical axis and an Everhart-Thornley detector mounted on a specimen chamber were simulated three-dimensionally. The acceptance maps were drawn as the relationship between the energy and angle of collected electrons under different working distances. The collection efficiency considering the detector sensitivity was also estimated for the various working distances. These data indicated that the acceptance maps and collection efficiency are keys to understand the detection mechanism and image contrast for each detector in the modern SEMs. Furthermore, the working distance is the dominant parameter because electron trajectories are drastically changed with the working distance.

In-situ measurement of objective lens data of a high-resolution electron microscope.

NASA Technical Reports Server (NTRS)

Heinemann, K.

1971-01-01

Bragg-reflex images of small individual crystallites in the size range of 20-100 A diameter with known crystallographic orientation were used in a transmission electron microscope to determine in-situ: (a) the relationship between objective lens current (or accelerating voltage) changes in discrete steps and corresponding defocus, (b) the spherical aberration coefficient, and (c) the axial chromatic aberration coefficient of the objective lens. The accuracy of the described method is better than 5%. The same specimen can advantageously be used to properly aline the illuminating beam with respect to the optical axis.

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.

A Monochromatic, Aberration-Corrected, Dual-Beam Low Energy Electron Microscope

PubMed Central

Mankos, Marian; Shadman, Khashayar

2013-01-01

The monochromatic, aberration-corrected, dual-beam low energy electron microscope (MAD-LEEM) is a novel instrument aimed at imaging of nanostructures and surfaces at sub-nanometer resolution that includes a monochromator, aberration corrector and dual beam illumination. The monochromator reduces the energy spread of the illuminating electron beam, which significantly improves spectroscopic and spatial resolution. The aberration corrector utilizes an electron mirror with negative aberrations that can be used to compensate the aberrations of the LEEM objective lens for a range of electron energies. Dual flood illumination eliminates charging generated when a conventional LEEM is used to image insulating specimens. MAD-LEEM is designed for the purpose of imaging biological and insulating specimens, which are difficult to image with conventional LEEM, Low-Voltage SEM, and TEM instruments. The MAD-LEEM instrument can also be used as a general purpose LEEM with significantly improved resolution. The low impact energy of the electrons is critical for avoiding beam damage, as high energy electrons with keV kinetic energies used in SEMs and TEMs cause irreversible change to many specimens, in particular biological materials. A potential application for MAD-LEEM is in DNA sequencing, which demands imaging techniques that enable DNA sequencing at high resolution and speed, and at low cost. The key advantages of the MAD-LEEM approach for this application are the low electron impact energies, the long read lengths, and the absence of heavy-atom DNA labeling. Image contrast simulations of the detectability of individual nucleotides in a DNA strand have been developed in order to refine the optics blur and DNA base contrast requirements for this application. PMID:23582636

A monochromatic, aberration-corrected, dual-beam low energy electron microscope.

PubMed

Mankos, Marian; Shadman, Khashayar

2013-07-01

The monochromatic, aberration-corrected, dual-beam low energy electron microscope (MAD-LEEM) is a novel instrument aimed at imaging of nanostructures and surfaces at sub-nanometer resolution that includes a monochromator, aberration corrector and dual beam illumination. The monochromator reduces the energy spread of the illuminating electron beam, which significantly improves spectroscopic and spatial resolution. The aberration corrector utilizes an electron mirror with negative aberrations that can be used to compensate the aberrations of the LEEM objective lens for a range of electron energies. Dual flood illumination eliminates charging generated when a conventional LEEM is used to image insulating specimens. MAD-LEEM is designed for the purpose of imaging biological and insulating specimens, which are difficult to image with conventional LEEM, Low-Voltage SEM, and TEM instruments. The MAD-LEEM instrument can also be used as a general purpose LEEM with significantly improved resolution. The low impact energy of the electrons is critical for avoiding beam damage, as high energy electrons with keV kinetic energies used in SEMs and TEMs cause irreversible change to many specimens, in particular biological materials. A potential application for MAD-LEEM is in DNA sequencing, which demands imaging techniques that enable DNA sequencing at high resolution and speed, and at low cost. The key advantages of the MAD-LEEM approach for this application are the low electron impact energies, the long read lengths, and the absence of heavy-atom DNA labeling. Image contrast simulations of the detectability of individual nucleotides in a DNA strand have been developed in order to refine the optics blur and DNA base contrast requirements for this application. Copyright © 2013 Elsevier B.V. All rights reserved.

MORPH-II, a software package for the analysis of scanning-electron-micrograph images for the assessment of the fractal dimension of exposed stone surfaces

USGS Publications Warehouse

Mossotti, Victor G.; Eldeeb, A. Raouf

2000-01-01

Turcotte, 1997, and Barton and La Pointe, 1995, have identified many potential uses for the fractal dimension in physicochemical models of surface properties. The image-analysis program described in this report is an extension of the program set MORPH-I (Mossotti and others, 1998), which provided the fractal analysis of electron-microscope images of pore profiles (Mossotti and Eldeeb, 1992). MORPH-II, an integration of the modified kernel of the program MORPH-I with image calibration and editing facilities, was designed to measure the fractal dimension of the exposed surfaces of stone specimens as imaged in cross section in an electron microscope.

Structural analysis of ion-implanted chemical-vapor-deposited diamond by transmission electron microscope

NASA Astrophysics Data System (ADS)

Jiang, N.; Deguchi, M.; Wang, C. L.; Won, J. H.; Jeon, H. M.; Mori, Y.; Hatta, A.; Kitabatake, M.; Ito, T.; Hirao, T.; Sasaki, T.; Hiraki, A.

1997-04-01

A transmission electron microscope (TEM) study of ion-implanted chemical-vapor-deposited (CVD) diamond is presented. CVD diamond used for transmission electron microscope observation was directly deposited onto Mo TEM grids. As-deposited specimens were irradiated by C (100 keV) ions at room temperature with a wide range of implantation doses (10 12-10 17/cm 2). Transmission electron diffraction (TED) patterns indicate that there exists a critical dose ( Dc) for the onset of amorphization of CVD diamond as a result of ion induced damage and the value of critical dose is confirmed to be about 3 × 10 15/cm 2. The ion-induced transformation process is clearly revealed by high resolution electron microscope (HREM) images. For a higher dose implantation (7 × 10 15/cm 2) a large amount of diamond phase is transformed into amorphous carbon and many tiny misoriented diamond blocks are found to be left in the amorphous solid. The average size of these misoriented diamond blocks is only about 1-2 nm. Further bombardment (10 17/cm 2) almost kills all of the diamond phase within the irradiated volume and moreover leads to local formation of micropolycrystalline graphite.

Correlation of eddy current responses between fatigue cracks and electrical-discharge-machining notches

NASA Astrophysics Data System (ADS)

Seo, Sukho; Choi, Gyudong; Eom, Tae Jhoun; Lee, Bokwon; Lee, Soo Yeol

2017-07-01

The eddy current responses of Electrical Discharge Machining (EDM) notches and fatigue cracks are directly compared to verify the reliability of eddy current inspection. The fatigue crack growth tests using a constant load range control mode were conducted to obtain a variety of edge crack sizes, ranging from 0.9 to 6.6 mm for Al alloy and from 0.1 to 3 mm for Ti alloy. EDM notch specimens of Al and Ti alloys were accordingly prepared in lengths similar to that of the fatigued specimen. The crack length was determined by optical microscope and scanning electron microscope. The eddy current responses between the EDM and fatigued specimens with varying notch/crack length were examined using probe sensors at (100-500) kHz and (1-2) MHz for Al and Ti alloys, respectively. The results show a significant difference in the eddy current signal between the two specimens, based on the correlation between the eddy current response and notch/crack length. This suggests that eddy current inspection using the EDM reference specimen is inaccurate in determining the precise crack size, unless the eddy current response data base is obtained from a fatigue-cracked specimen.

New Developments in Cathodoluminescence Spectroscopy for the Study of Luminescent Materials

PubMed Central

den Engelsen, Daniel; Fern, George R.; Harris, Paul G.; Ireland, Terry G.; Silver, Jack

2017-01-01

Herein, we describe three advanced techniques for cathodoluminescence (CL) spectroscopy that have recently been developed in our laboratories. The first is a new method to accurately determine the CL-efficiency of thin layers of phosphor powders. When a wide band phosphor with a band gap (Eg > 5 eV) is bombarded with electrons, charging of the phosphor particles will occur, which eventually leads to erroneous results in the determination of the luminous efficacy. To overcome this problem of charging, a comparison method has been developed, which enables accurate measurement of the current density of the electron beam. The study of CL from phosphor specimens in a scanning electron microscope (SEM) is the second subject to be treated. A detailed description of a measuring method to determine the overall decay time of single phosphor crystals in a SEM without beam blanking is presented. The third technique is based on the unique combination of microscopy and spectrometry in the transmission electron microscope (TEM) of Brunel University London (UK). This combination enables the recording of CL-spectra of nanometre-sized specimens and determining spatial variations in CL emission across individual particles by superimposing the scanning TEM and CL-images. PMID:28772671

Ultrastructure of canine vasoformative tumors.

PubMed

Madewell, B R; Griffey, S M; Munn, R J

1992-01-01

The transmission electron microscope was used to examine 20 spontaneous canine hemangiosarcomas or hemangiopericytomas in order to define their fine ultrastructural features, and to compare those features with descriptions of human counterpart neoplasms. From specimen to specimen the neoplasms examined showed considerable structural heterogeneity but, in composite, appeared similar to the prototype human tumors. These data suggest that the canine hemangiosarcoma and hemangiopericytoma might serve as comparative models for studies of the morphogenesis of vasoformative neoplasms.

An in-plane magnetic chiral dichroism approach for measurement of intrinsic magnetic signals using transmitted electrons

PubMed Central

Song, Dongsheng; Tavabi, Amir H.; Li, Zi-An; Kovács, András; Rusz, Ján; Huang, Wenting; Richter, Gunther; Dunin-Borkowski, Rafal E.; Zhu, Jing

2017-01-01

Electron energy-loss magnetic chiral dichroism is a powerful technique that allows the local magnetic properties of materials to be measured quantitatively with close-to-atomic spatial resolution and element specificity in the transmission electron microscope. Until now, the technique has been restricted to measurements of the magnetic circular dichroism signal in the electron beam direction. However, the intrinsic magnetization directions of thin samples are often oriented in the specimen plane, especially when they are examined in magnetic-field-free conditions in the transmission electron microscope. Here, we introduce an approach that allows in-plane magnetic signals to be measured using electron magnetic chiral dichroism by selecting a specific diffraction geometry. We compare experimental results recorded from a cobalt nanoplate with simulations to demonstrate that an electron magnetic chiral dichroism signal originating from in-plane magnetization can be detected successfully. PMID:28504267

Antarctic Meteorite Newsletter, volume 9, no. 2

NASA Technical Reports Server (NTRS)

Gooding, J. L. (Editor)

1986-01-01

Preliminary description and classifications of meteorites that were completed since publication of the February issue are contained. Most large (greater than 150 g) specimens (regardless of petrologic type) and all pebble sized (less than 150 g) specimens of special petrologic type are represented by separate descriptions. However, specimens of nonspecial petrologic type are listed only as single line entries. For convenience, new specimens are also recast by petrologic type. Each macroscopic description summarizes features that were visible to the eye at the time the meteorite was first examined. Classification is based on microscopic petrography and resonnaissance-level electron-probe microanalysis. The pairing list was updated.

Technology-Enhanced Pathology Education: Nigerian Medical Students Perspectives

ERIC Educational Resources Information Center

Vhriterhire, Raymond A.; Orkuma, Joseph A.; Jegede, Olushola O.; Omotosho, Ayodele J.; Adekwu, Amali

2016-01-01

The delivery of pathology education traditionally through instructor centred didactic lectures, small group tutorials, and practical demonstrations using microscope glass slides, gross pot specimens and autopsy sessions, is paving way for electronic learner-centred methods. Successful adoption and implementation of rapidly advancing educational…

Chapter 14: Electron Microscopy on Thin Films for Solar Cells

DOE Office of Scientific and Technical Information (OSTI.GOV)

Romero, Manuel; Abou-Ras, Daniel; Nichterwitz, Melanie

2016-07-22

This chapter overviews the various techniques applied in scanning electron microscopy (SEM) and transmission electron microscopy (TEM), and highlights their possibilities and also limitations. It gives the various imaging and analysis techniques applied on a scanning electron microscope. The chapter shows that imaging is divided into that making use of secondary electrons (SEs) and of backscattered electrons (BSEs), resulting in different contrasts in the images and thus providing information on compositions, microstructures, and surface potentials. Whenever aiming for imaging and analyses at scales of down to the angstroms range, TEM and its related techniques are appropriate tools. In many cases,more » also SEM techniques provide the access to various material properties of the individual layers, not requiring specimen preparation as time consuming as TEM techniques. Finally, the chapter dedicates to cross-sectional specimen preparation for electron microscopy. The preparation decides indeed on the quality of imaging and analyses.« less

Electron microscopy study of the iron meteorite Santa Catharina

NASA Technical Reports Server (NTRS)

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

1990-01-01

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

A cylindrical specimen holder for electron cryo-tomography

PubMed Central

Palmer, Colin M.; Löwe, Jan

2014-01-01

The use of slab-like flat specimens for electron cryo-tomography restricts the range of viewing angles that can be used. This leads to the “missing wedge” problem, which causes artefacts and anisotropic resolution in reconstructed tomograms. Cylindrical specimens provide a way to eliminate the problem, since they allow imaging from a full range of viewing angles around the tilt axis. Such specimens have been used before for tomography of radiation-insensitive samples at room temperature, but never for frozen-hydrated specimens. Here, we demonstrate the use of thin-walled carbon tubes as specimen holders, allowing the preparation of cylindrical frozen-hydrated samples of ribosomes, liposomes and whole bacterial cells. Images acquired from these cylinders have equal quality at all viewing angles, and the accessible tilt range is restricted only by the physical limits of the microscope. Tomographic reconstructions of these specimens demonstrate that the effects of the missing wedge are substantially reduced, and could be completely eliminated if a full tilt range was used. The overall quality of these tomograms is still lower than that obtained by existing methods, but improvements are likely in future. PMID:24275523

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.

Minority carrier diffusion length and edge surface-recombination velocity in InP

NASA Technical Reports Server (NTRS)

Hakimzadeh, Roshanak; Bailey, Sheila G.

1993-01-01

A scanning electron microscope was used to obtain the electron-beam-induced current (EBIC) profiles in InP specimens containing a Schottky barrier perpendicular to the scanned (edge) surface. An independent technique was used to measure the edge surface-recombination velocity. These values were used in a fit of the experimental EBIC data with a theoretical expression for normalized EBIC (Donolato, 1982) to obtain the electron (minority carrier) diffusion length.

Scanning Electron Microscope Mapping System Developed for Detecting Surface Defects in Fatigue Specimens

NASA Technical Reports Server (NTRS)

Bonacuse, Peter J.; Kantzos, Peter T.

2002-01-01

An automated two-degree-of-freedom specimen positioning stage has been developed at the NASA Glenn Research Center to map and monitor defects in fatigue specimens. This system expedites the examination of the entire gauge section of fatigue specimens so that defects can be found using scanning electron microscopy (SEM). Translation and rotation stages are driven by microprocessor-based controllers that are, in turn, interfaced to a computer running custom-designed software. This system is currently being used to find and record the location of ceramic inclusions in powder metallurgy materials. The mapped inclusions are periodically examined during interrupted fatigue experiments. The number of cycles to initiate cracks from these inclusions and the rate of growth of initiated cracks can then be quantified. This information is necessary to quantify the effect of this type of defect on the durability of powder metallurgy materials. This system was developed with support of the Ultra Safe program.

Quasi-parallel precession diffraction: Alignment method for scanning transmission electron microscopes.

PubMed

Plana-Ruiz, S; Portillo, J; Estradé, S; Peiró, F; Kolb, Ute; Nicolopoulos, S

2018-06-06

A general method to set illuminating conditions for selectable beam convergence and probe size is presented in this work for Transmission Electron Microscopes (TEM) fitted with µs/pixel fast beam scanning control, (S)TEM, and an annular dark field detector. The case of interest of beam convergence and probe size, which enables diffraction pattern indexation, is then used as a starting point in this work to add 100 Hz precession to the beam while imaging the specimen at a fast rate and keeping the projector system in diffraction mode. The described systematic alignment method for the adjustment of beam precession on the specimen plane while scanning at fast rates is mainly based on the sharpness of the precessed STEM image. The complete alignment method for parallel condition and precession, Quasi-Parallel PED-STEM, is presented in block diagram scheme, as it has been tested on a variety of instruments. The immediate application of this methodology is that it renders the TEM column ready for the acquisition of Precessed Electron Diffraction Tomographies (EDT) as well as for the acquisition of slow Precessed Scanning Nanometer Electron Diffraction (SNED). Examples of the quality of the Precessed Electron Diffraction (PED) patterns and PED-STEM alignment images are presented with corresponding probe sizes and convergence angles. Copyright © 2018. Published by Elsevier B.V.

Damage analysis of CF/AF hybrid fabric reinforced plastic laminated composites with scanned image microscopy

NASA Astrophysics Data System (ADS)

Miyasaka, Chiaki; Kasano, Hideaki; Shull, Peter J.

2004-07-01

The article presents an experimental study that has been conducted to evaluate the impact loading damage within hybrid fabric laminates-carbon and Aramid fibers. The experiments have been undertaken on a series of interply hybrid specimens with different preprags stacking sequences. Impact damage was created using an air-gun like impact device propelling spherical steel balls with diameters of 5.0mm and 10.0mm and having velocities of 113m/s and 40m/s respectively. The resulting specimen surface and internal damage (e.g., micro-cracking and debonding) was visualized nondestructively by a scanning acoustic microscope (SAM) while further interrogation of specific internal damage was visualized using a scanning electron microscope (SEM) on cross-sectioned panels.

Microstructures and magnetic properties of Co-Al-O granular thin films

NASA Astrophysics Data System (ADS)

Ohnuma, M.; Hono, K.; Onodera, H.; Ohnuma, S.; Fujimori, H.; Pedersen, J. S.

2000-01-01

The microstructures of Co-Al-O thin films of wide varieties of compositions are studied by transmission electron microscopy and small angle x-ray scattering (SAXS). In the superparamagnetic specimens, high resolution electron microscope images reveal that isolated spherical Co particles are surrounded by an amorphous aluminum oxide matrix. However, in the soft ferromagnetic films, the shape of the Co particles is prolate ellipsoidal. SAXS intensities from the soft magnetic specimens decrease inversely with the wave vector, q, in a low wave-vector region, while an interparticle interference peak is observed for the superparamagnetic specimens. The scattering profiles of the soft magnetic films imply that the Co particles have a cylindrical shape and are randomly oriented. The correlation between the magnetic properties and the microstructures is discussed.

Effect of matrix resin on the impact fracture characteristics of graphite-epoxy laminates

NASA Technical Reports Server (NTRS)

Hertzberg, P. E.; Smith, B. W.; Miller, A. G.

1982-01-01

The effect of resin chemistry on basic impact energy absorbent mechanisms exibited by graphite-epoxy composites was investigated. Impact fracture modes and microscopic resin deformation characteristics were examined for 26 NASA-impacted graphite epoxy laminates with different resin chemistries. Discrete specimen fracture modes were identified through cross sectional examination after impact, and subsequently compared with measured glass transition temperatures, cure cycles, and residual impact capabilities. Microscopic resin deformation mechanisms and their overall relationship to impact loading conditions, voids, and resin content were also characterized through scanning electron microscopic examination of separated fracture surfaces.

Microscopic colitis syndrome.

PubMed Central

Veress, B; Löfberg, R; Bergman, L

1995-01-01

The colorectal biopsy specimens from 30 patients with chronic watery diarrhoea but normal endoscopic and radiographic findings were studied by light microscopy, morphometry, immunohistochemistry, and two patients with electron microscopy. The histological changes in the colorectum were originally diagnosed in six patients as lymphocytic colitis and in 24 patients as collagenous colitis. The analysis of the specimens for this study could delineate three distinct groups of microscopic colitis: lymphocytic colitis (six patients), collagenous colitis without lymphocytic attack on the surface epithelium (seven patients), and a mixed form presenting with both thickening of the collagen plate and increased number of intraepithelial lymphocytes (17 patients). No transformation was seen from one type to another during follow up of six patients for four to seven years. Increased numbers of active pericryptal myofibroblasts were found with the electron microscope in one patient with mixed microscopic colitis showing also myofibroblasts entrapped within the collagen layer. Hitherto undescribed flat mucosa of the ileum was found in one patient with lymphocytic colitis and both flat mucosa and thickening of the collagen plate in the ileum were seen in one patient with the mixed form of the disease. In another patient with mixed microscopic colitis, normalisation of the colorectal morphology occurred after temporary loop ileostomy, followed by the reappearance of both diarrhoea, inflammation, and thickening of the collagen plate after the ileostomy was reversed. No association was found between non-steroid anti-inflammatory drug (NSAID) consumption and collagenous or mixed microscopic colitis. The primary cause of microscopic colitis is probably an immunological reaction to luminal antigen/s, perhaps of ileal origin. The engagement of the pericryptal myofibroblasts in the disease process might result in the development of the various forms of microscopic colitis. An inverse relation between intraepithelial lymphocyte count and collagen thickness may indicate that microscopic colitis is a spectral disease. Images Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 PMID:7615277

PSU/WES Interlaboratory Comparative Methodology Study of an Experimental Cementitious Repository Seal Material. Report 2. Final Results.

DTIC Science & Technology

1982-03-01

meter 25 11.0 Microstructure by SEM 11.1 Introduction In order to correlate observed physical and mechanical properties in cured grout samples, a...studied at the two laboratories has proper physical properties , phase composi- tions, and microstructures for the materials used and ages covered...Scanning Electron Microscope Resolution Test Specimen ( Al -W) D. B. Ballard Research Material 100 SEM Resolution Test Specimen (AI-W)., is an alloy of

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

NASA Astrophysics Data System (ADS)

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

2011-06-01

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

Review of current progress in nanometrology with the helium ion microscope

NASA Astrophysics Data System (ADS)

Postek, Michael T.; Vladár, András; Archie, Charles; Ming, Bin

2011-02-01

Scanning electron microscopy has been employed as an imaging and measurement tool for more than 50 years and it continues as a primary tool in many research and manufacturing facilities across the world. A new challenger to this work is the helium ion microscope (HIM). The HIM is a new imaging and metrology technology. Essentially, substitution of the electron source with a helium ion source yields a tool visually similar in function to the scanning electron microscope, but very different in the fundamental imaging and measurement process. The imaged and measured signal originates differently than in the scanning electron microscope and that fact and its single atom source diameter may be able to push the obtainable resolution lower, provide greater depth-of-field and ultimately improve the metrology. Successful imaging and metrology with this instrument entails understanding and modeling of new ion beam/specimen interaction physics. As a new methodology, HIM is beginning to show promise and the abundance of potentially advantageous applications for nanometrology has yet to be fully exploited. This paper discusses some of the progress made at NIST in collaboration with IBM to understand the science behind this new technology.

The interperiosteo-dural concept applied to the perisellar compartment: a microanatomical and electron microscopic study.

PubMed

François, Patrick; Travers, Nadine; Lescanne, Emmanuel; Arbeille, Brigitte; Jan, Michel; Velut, Stéphane

2010-11-01

The dura mater has 2 dural layers: the endosteal layer (outer layer), which is firmly attached to the bone, and the meningeal layer (inner layer), which directly covers the brain. These 2 dural layers join together in the middle temporal fossa or the convexity and separate into the orbital, lateral sellar compartment (LSC), or spinal epidural space to form the extradural neural axis compartment (EDNAC). The aim of this work was to anatomically verify the concept of the EDNAC by using electron microscopy. The authors studied the cadaveric heads obtained from 13 adults. Ten of the specimens (or 20 perisellar areas) were injected with colored latex and fixed in formalin. They carefully removed each brain to allow a superior approach to the perisellar area. The 3 other specimens were studied by microscopic and ultrastructural methods to describe the EDNAC in the perisellar area. Special attention was paid to the dural layers surrounding the perisellar area. The authors studied the anatomy of the meningeal architecture of the LSC, the petroclival venous confluence, the orbit, and the trigeminal cave. After dissection, the authors took photographs of the dural layers with the aid of optical magnification. The 3 remaining heads, obtained from fresh cadavers, were prepared for electron microscopic study. The EDNAC is limited by the endosteal layer and the meningeal layer and contains fat and/or venous blood. The endosteal layer and meningeal layer were not identical on electron microscopy; this finding can be readily related to the histology of the meninges. In this study, the authors demonstrated the existence of the EDNAC concept in the perisellar area by using dissected cadaveric heads and verified the reality of the concept of the meningeal layer with electron microscopy. These findings clearly demonstrated the existence of the EDNAC, a notion that has generally been accepted but never demonstrated microscopically.

Measurement error of Young’s modulus considering the gravity and thermal expansion of thin specimens for in situ tensile testing

NASA Astrophysics Data System (ADS)

Ma, Zhichao; Zhao, Hongwei; Ren, Luquan

2016-06-01

Most miniature in situ tensile devices compatible with scanning/transmission electron microscopes or optical microscopes adopt a horizontal layout. In order to analyze and calculate the measurement error of the tensile Young’s modulus, the effects of gravity and temperature changes, which would respectively lead to and intensify the bending deformation of thin specimens, are considered as influencing factors. On the basis of a decomposition method of static indeterminacy, equations of simplified deflection curves are obtained and, accordingly, the actual gage length is confirmed. By comparing the effects of uniaxial tensile load on the change of the deflection curve with gravity, the relation between the actual and directly measured tensile Young’s modulus is obtained. Furthermore, the quantitative effects of ideal gage length l o, temperature change ΔT and the density ρ of the specimen on the modulus difference and modulus ratio are calculated. Specimens with larger l o and ρ present more obvious measurement errors for Young’s modulus, but the effect of ΔT is not significant. The calculation method of Young’s modulus is particularly suitable for thin specimens.

An historical account of the development and applications of the negative staining technique to the electron microscopy of viruses.

PubMed

Horne, R W; Wildy, P

1979-09-01

A brief historical account of the development and applications of the negative staining techniques to the study of the structure of viruses and their components as observed in the electron microscope is presented. Although the basic method of surrounding or embedding specimens in opaque dyes was used in light microscopy dating from about 1884, the equivalent preparative techniques applied to electron microscopy were comparatively recent. The combination of experiments on a sophisticated bacterial virus and the installation of a high resolution electron microscope in the Cavendish Laboratory, Cambridge, during 1954, subsequently led to the analysis of several important morphological features of animal, plant and bacterial viruses. The implications of the results from these early experiments on viruses and recent developments in negative staining methods for high resolution image analysis of electron micrographs are also discussed.

The trajectories of secondary electrons in the scanning electron microscope.

PubMed

Konvalina, Ivo; Müllerová, Ilona

2006-01-01

Three-dimensional simulations of the trajectories of secondary electrons (SE) in the scanning electron microscope have been performed for plenty of real configurations of the specimen chamber, including all its basic components. The primary purpose was to evaluate the collection efficiency of the Everhart-Thornley detector of SE and to reveal fundamental rules for tailoring the set-ups in which efficient signal acquisition can be expected. Intuitive realizations about the easiness of attracting the SEs towards the biased front grid of the detector have shown themselves likely as false, and all grounded objects in the chamber have been proven to influence the spatial distribution of the signal-extracting field. The role of the magnetic field penetrating from inside the objective lens is shown to play an ambiguous role regarding possible support for the signal collection.

Inhibition of thrombus formation on intravascular sensors by electrical polarization.

PubMed

Schmitt, J M; Baer, M; Meindl, J D; Anderson, M F; Mihm, F G

1984-09-01

Implantable biomedical sensors built on a silicon substrate capped with glass are currently being developed for intravascular applications. Electrical techniques for inhibiting thrombus formation on the surface of a proposed optical sensor in direct contact with blood have been investigated. Glass-on-silicon specimens (4 X 1.2 X 0.4 mm3) were coated with indium-tin oxide, a transparent conductor, and implanted in the vena cava and iliac veins of three dogs for 10, 20, or 33 days. The equilibrium surface-blood interface potentials of the specimens were modified by implanted current sources which supplied either direct current (8-15 microA) or 100 KHz alternating current (5 microA, root mean square). Light-microscopic and scanning electron-microscopic analyses showed each of the DC-polarized specimens to be free of thrombus, in contrast to nonpolarized (control) specimens on which varying amounts of adsorbed protein and thrombus deposits were found. Like the control specimens, the AC-polarized specimens formed thrombus, but the appearance of the deposits differed. These findings support the view that the polarity, magnitude and time dependence of the potential across conducting surface-blood interface significantly influence thrombogenicity. Further work is necessary to determine the roles of electrochemical and electrostatic factors in preventing thrombus formation on foreign materials.

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.

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 Central

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

2015-01-01

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. PMID:25392009

Diffraction contrast as a sensitive indicator of femtosecond sub-nanoscale motion in ultrafast transmission electron microscopy

NASA Astrophysics Data System (ADS)

Cremons, Daniel R.; Schliep, Karl B.; Flannigan, David J.

2013-09-01

With ultrafast transmission electron microscopy (UTEM), access can be gained to the spatiotemporal scales required to directly visualize rapid, non-equilibrium structural dynamics of materials. This is achieved by operating a transmission electron microscope (TEM) in a stroboscopic pump-probe fashion by photoelectrically generating coherent, well-timed electron packets in the gun region of the TEM. These probe photoelectrons are accelerated down the TEM column where they travel through the specimen before reaching a standard, commercially-available CCD detector. A second laser pulse is used to excite (pump) the specimen in situ. Structural changes are visualized by varying the arrival time of the pump laser pulse relative to the probe electron packet at the specimen. Here, we discuss how ultrafast nanoscale motions of crystalline materials can be visualized and precisely quantified using diffraction contrast in UTEM. Because diffraction contrast sensitively depends upon both crystal lattice orientation as well as incoming electron wavevector, minor spatial/directional variations in either will produce dynamic and often complex patterns in real-space images. This is because sections of the crystalline material that satisfy the Laue conditions may be heterogeneously distributed such that electron scattering vectors vary over nanoscale regions. Thus, minor changes in either crystal grain orientation, as occurs during specimen tilting, warping, or anisotropic expansion, or in the electron wavevector result in dramatic changes in the observed diffraction contrast. In this way, dynamic contrast patterns observed in UTEM images can be used as sensitive indicators of ultrafast specimen motion. Further, these motions can be spatiotemporally mapped such that direction and amplitude can be determined.

Modeling of electron-specimen interaction in scanning electron microscope for e-beam metrology and inspection: challenges and perspectives

NASA Astrophysics Data System (ADS)

Suzuki, Makoto; Kameda, Toshimasa; Doi, Ayumi; Borisov, Sergey; Babin, Sergey

2018-03-01

The interpretation of scanning electron microscopy (SEM) images of the latest semiconductor devices is not intuitive and requires comparison with computed images based on theoretical modeling and simulations. For quantitative image prediction and geometrical reconstruction of the specimen structure, the accuracy of the physical model is essential. In this paper, we review the current models of electron-solid interaction and discuss their accuracy. We perform the comparison of the simulated results with our experiments of SEM overlay of under-layer, grain imaging of copper interconnect, and hole bottom visualization by angular selective detectors, and show that our model well reproduces the experimental results. Remaining issues for quantitative simulation are also discussed, including the accuracy of the charge dynamics, treatment of beam skirt, and explosive increase in computing time.

Applications of the Scanning Electron Microscope to Concrete Failure (Axial, Biaxial, and Dynamic).

DTIC Science & Technology

1984-03-05

direction. If a section of the reinforced concrete specimen is viewed at a location where no reinforcement occurs the failure mech- anism is the same...blade saw. The face of each speci- men was polished using a silicon-carbide slurry on a jewelers wheel. The specimens were than squared by use of a...Fig. 14 is a photo of an actual model, with four sandstone discs, arranged in a square array, laying on a glass plate coated with mineral oil to prevent

Preparing soft-bodied arthropods for microscope examination: Aphids (Insecta: Hemiptera: Aphididae)

USDA-ARS?s Scientific Manuscript database

Proper identification of aphids (Hemiptera: Aphididae) require preparation of the specimen on a microscope slide. This training video provides visual instruction on how to prepare aphid specimens on microscope slides for examination and indentification. Steps ranging from collection, specimen clear...

Preparing soft-bodied arthropods for microscope examination: Whiteflies (Insecta: Hemiptera: Alyrodidae)

USDA-ARS?s Scientific Manuscript database

Proper identification of whiteflies (Hemiptera:Alyrodidae) requires preparation of the specimen on a microscope slide. This training video provides visual instruction on how to prepare whitefly specimens on microscope slides for examination and identification. Steps ranging from collection, specimen...

Imaia, a new truffle genus to accommodate TerJezia gigantea

Treesearch

Gabor M. Kovacs; James M. Trappe; Abulmagid Alsheikh; Karoly Boka

2008-01-01

Originally described from Japan by Sanshi Imai in 1933, the hypogeous ascomycete Terjezia gigantea was subsequently discovered in the Appalachian Mountains of the USA. Morphological, electron microscopic, and phylogenetic studies of specimens collected in both regions revealed that, despite this huge geographic disjunction, (1) the Japanese and...

Conductive resins improve charging and resolution of acquired images in electron microscopic volume imaging

PubMed Central

Nguyen, Huy Bang; Thai, Truc Quynh; Saitoh, Sei; Wu, Bao; Saitoh, Yurika; Shimo, Satoshi; Fujitani, Hiroshi; Otobe, Hirohide; Ohno, Nobuhiko

2016-01-01

Recent advances in serial block-face imaging using scanning electron microscopy (SEM) have enabled the rapid and efficient acquisition of 3-dimensional (3D) ultrastructural information from a large volume of biological specimens including brain tissues. However, volume imaging under SEM is often hampered by sample charging, and typically requires specific sample preparation to reduce charging and increase image contrast. In the present study, we introduced carbon-based conductive resins for 3D analyses of subcellular ultrastructures, using serial block-face SEM (SBF-SEM) to image samples. Conductive resins were produced by adding the carbon black filler, Ketjen black, to resins commonly used for electron microscopic observations of biological specimens. Carbon black mostly localized around tissues and did not penetrate cells, whereas the conductive resins significantly reduced the charging of samples during SBF-SEM imaging. When serial images were acquired, embedding into the conductive resins improved the resolution of images by facilitating the successful cutting of samples in SBF-SEM. These results suggest that improving the conductivities of resins with a carbon black filler is a simple and useful option for reducing charging and enhancing the resolution of images obtained for volume imaging with SEM. PMID:27020327

Design of a 300-kV gas environmental transmission electron microscope equipped with a cold field emission gun.

PubMed

Isakozawa, Shigeto; Nagaoki, Isao; Watabe, Akira; Nagakubo, Yasuhira; Saito, Nobuhiro; Matsumoto, Hiroaki; Zhang, Xiao Feng; Taniguchi, Yoshifumi; Baba, Norio

2016-08-01

A new in situ environmental transmission electron microscope (ETEM) was developed based on a 300 kV TEM with a cold field emission gun (CFEG). Particular caution was taken in the ETEM design to assure uncompromised imaging and analytical performance of the TEM. Because of the improved pumping system between the gun and column, the vacuum of CFEG was largely improved and the probe current was sufficiently stabilized to operate without tip flashing for 2-3 h or longer. A high brightness of 2.5 × 10(9) A/cm(2) sr was measured at 300 kV, verifying the high quality of the CFEG electron beam. A specially designed gas injection-heating holder was used in the in situ TEM study at elevated temperatures with or without gas around the TEM specimen. Using this holder in a 10 Pa gas atmosphere and specimen temperatures up to 1000°C, high-resolution ETEM performance and analysis were achieved. © The Author 2016. Published by Oxford University Press on behalf of The Japanese Society of Microscopy. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

Transmission Kikuchi diffraction and transmission electron forescatter imaging of electropolished and FIB manufactured TEM specimens

DOE Office of Scientific and Technical Information (OSTI.GOV)

Zieliński, W., E-mail: wiziel@inmat.pw.edu.pl; Płociński, T.; Kurzydłowski, K.J.

2015-06-15

We present a study of the efficiency of the utility of scanning electron microscope (SEM)-based transmission methods for characterizing grain structure in thinned bulk metals. Foils of type 316 stainless steel were prepared by two methods commonly used for transmission electron microscopy — double-jet electropolishing and focused ion beam milling. A customized holder allowed positioning of the foils in a configuration appropriate for both transmission electron forward scatter diffraction, and for transmission imaging by the use of a forescatter detector with two diodes. We found that both crystallographic orientation maps and dark-field transmitted images could be obtained for specimens preparedmore » by either method. However, for both methods, preparation-induced artifacts may affect the quality or accuracy of transmission SEM data, especially those acquired by the use of transmission Kikuchi diffraction. Generally, the quality of orientation data was better for specimens prepared by electropolishing, due to the absence of ion-induced damage. - Highlights: • The transmission imaging and diffraction techniques are emerging in scanning electron microscopy (SEM) as promising new field of materials characterization. • The manuscript titled: “Transmission Kikuchi Diffraction and Transmission Electron Forescatter Imaging of Electropolished and FIB Manufactured TEM Specimens” documents how different specimen thinning procedures can effect efficiency of transmission Kikuchi diffraction and transmission electron forescatter imaging. • The abilities to make precision crystallographic orientation maps and dark-field images in transmission was studied on electropolished versus focus ion beam manufactured TEM specimens. • Depending on the need, electropolished and focused ion beam technique may produce suitable specimens for transmission imaging and diffraction in SEM.« less

In situ study of live specimens in an environmental scanning electron microscope.

PubMed

Tihlaříková, Eva; Neděla, Vilém; Shiojiri, Makoto

2013-08-01

In this paper we introduce new methodology for the observation of living biological samples in an environmental scanning electron microscope (ESEM). The methodology is based on an unconventional initiation procedure for ESEM chamber pumping, free from purge-flood cycles, and on the ability to control thermodynamic processes close to the sample. The gradual and gentle change of the working environment from air to water vapor enables the study of not only living samples in dynamic in situ experiments and their manifestation of life (sample walking) but also its experimentally stimulated physiological reactions. Moreover, Monte Carlo simulations of primary electron beam energy losses in a water layer on the sample surface were studied; consequently, the influence of the water thickness on radiation, temperature, or chemical damage of the sample was considered.

A cylindrical specimen holder for electron cryo-tomography.

PubMed

Palmer, Colin M; Löwe, Jan

2014-02-01

The use of slab-like flat specimens for electron cryo-tomography restricts the range of viewing angles that can be used. This leads to the "missing wedge" problem, which causes artefacts and anisotropic resolution in reconstructed tomograms. Cylindrical specimens provide a way to eliminate the problem, since they allow imaging from a full range of viewing angles around the tilt axis. Such specimens have been used before for tomography of radiation-insensitive samples at room temperature, but never for frozen-hydrated specimens. Here, we demonstrate the use of thin-walled carbon tubes as specimen holders, allowing the preparation of cylindrical frozen-hydrated samples of ribosomes, liposomes and whole bacterial cells. Images acquired from these cylinders have equal quality at all viewing angles, and the accessible tilt range is restricted only by the physical limits of the microscope. Tomographic reconstructions of these specimens demonstrate that the effects of the missing wedge are substantially reduced, and could be completely eliminated if a full tilt range was used. The overall quality of these tomograms is still lower than that obtained by existing methods, but improvements are likely in future. © 2013 The Authors. Published by Elsevier B.V. All rights reserved.

Preparing soft-bodied arthropods for arthropods for microscope examination: Mites (Arachnida: Acari)

USDA-ARS?s Scientific Manuscript database

Proper identification of mites (Arachnida: Acari) require preparation of the specimen on a microscope slide. This training video provides visual instruction on how to prepare mite specimens on microscope slides for examination and identification. Steps ranging from collection, specimen clearing, use...

A Multiscale Material Testing System for In Situ Optical and Electron Microscopes and Its Application

PubMed Central

Ye, Xuan; Cui, Zhiguo; Fang, Huajun; Li, Xide

2017-01-01

We report a novel material testing system (MTS) that uses hierarchical designs for in-situ mechanical characterization of multiscale materials. This MTS is adaptable for use in optical microscopes (OMs) and scanning electron microscopes (SEMs). The system consists of a microscale material testing module (m-MTM) and a nanoscale material testing module (n-MTM). The MTS can measure mechanical properties of materials with characteristic lengths ranging from millimeters to tens of nanometers, while load capacity can vary from several hundred micronewtons to several nanonewtons. The m-MTM is integrated using piezoelectric motors and piezoelectric stacks/tubes to form coarse and fine testing modules, with specimen length from millimeters to several micrometers, and displacement distances of 12 mm with 0.2 µm resolution for coarse level and 8 µm with 1 nm resolution for fine level. The n-MTM is fabricated using microelectromechanical system technology to form active and passive components and realizes material testing for specimen lengths ranging from several hundred micrometers to tens of nanometers. The system’s capabilities are demonstrated by in-situ OM and SEM testing of the system’s performance and mechanical properties measurements of carbon fibers and metallic microwires. In-situ multiscale deformation tests of Bacillus subtilis filaments are also presented. PMID:28777341

Re-evaluation of differential phase contrast (DPC) in a scanning laser microscope using a split detector as an alternative to differential interference contrast (DIC) optics.

PubMed

Amos, W B; Reichelt, S; Cattermole, D M; Laufer, J

2003-05-01

In this paper, differential phase imaging (DPC) with transmitted light is implemented by adding a suitable detection system to a standard commercially available scanning confocal microscope. DPC, a long-established method in scanning optical microscopy, depends on detecting the intensity difference between opposite halves or quadrants of a split photodiode detector placed in an aperture plane. Here, DPC is compared with scanned differential interference contrast (DIC) using a variety of biological specimens and objective lenses of high numerical aperture. While DPC and DIC images are generally similar, DPC seems to have a greater depth of field. DPC has several advantages over DIC. These include low cost (no polarizing or strain-free optics are required), absence of a double scanning spot, electronically variable direction of shading and the ability to image specimens in plastic dishes where birefringence prevents the use of DIC. DPC is also here found to need 20 times less laser power at the specimen than DIC.

Interpretation of scanning electron microscope measurements of minority carrier diffusion lengths in semiconductors

NASA Technical Reports Server (NTRS)

Flat, A.; Milnes, A. G.

1978-01-01

In scanning electron microscope (SEM) injection measurements of minority carrier diffusion lengths some uncertainties of interpretation exist when the response current is nonlinear with distance. This is significant in epitaxial layers where the layer thickness is not large in relation to the diffusion length, and where there are large surface recombination velocities on the incident and contact surfaces. An image method of analysis is presented for such specimens. A method of using the results to correct the observed response in a simple convenient way is presented. The technique is illustrated with reference to measurements in epitaxial layers of GaAs. Average beam penetration depth may also be estimated from the curve shape.

Factors that Influence the Formation and Stability of Thin, Cryo-EM Specimens

DOE PAGES

Glaeser, Robert M.; Han, Bong-Gyoon; Csencsits, Roseann; ...

2015-09-17

Poor consistency of the ice thickness from one area of a cryo-electron microscope (cryo-EM) specimen grid to another, from one grid to the next, and from one type of specimen to another, motivates a reconsideration of how to best prepare suitably thin specimens. We first review the three related topics of wetting, thinning, and stability against dewetting of aqueous films spread over a hydrophilic substrate. Furthermore, we then suggest that the importance of there being a surfactant monolayer at the air-water interface of thin, cryo-EM specimens has been largely underappreciated. In fact, a surfactant layer (of uncontrolled composition and surfacemore » pressure) can hardly be avoided during standard cryo-EM specimen preparation. Thus it is suggested that better control over the composition and properties of the surfactant layer may result in more reliable production of cryo-EM specimens with the desired thickness.« less

Factors that Influence the Formation and Stability of Thin, Cryo-EM Specimens

DOE Office of Scientific and Technical Information (OSTI.GOV)

Glaeser, Robert M.; Han, Bong-Gyoon; Csencsits, Roseann

Poor consistency of the ice thickness from one area of a cryo-electron microscope (cryo-EM) specimen grid to another, from one grid to the next, and from one type of specimen to another, motivates a reconsideration of how to best prepare suitably thin specimens. We first review the three related topics of wetting, thinning, and stability against dewetting of aqueous films spread over a hydrophilic substrate. Furthermore, we then suggest that the importance of there being a surfactant monolayer at the air-water interface of thin, cryo-EM specimens has been largely underappreciated. In fact, a surfactant layer (of uncontrolled composition and surfacemore » pressure) can hardly be avoided during standard cryo-EM specimen preparation. Thus it is suggested that better control over the composition and properties of the surfactant layer may result in more reliable production of cryo-EM specimens with the desired thickness.« less

Effect of Nickel Contents on the Microstructure and Mechanical Properties for Low-Carbon Bainitic Weld Metals

NASA Astrophysics Data System (ADS)

Mao, Gaojun; Cao, Rui; Yang, Jun; Jiang, Yong; Wang, Shuai; Guo, Xili; Yuan, Junjun; Zhang, Xiaobo; Chen, Jianhong

2017-05-01

Multi-pass weld metals were deposited on Q345 base steel using metal powder-flux-cored wire with various Ni contents to investigate the effects of the Ni content on the weld microstructure and property. The types of the microstructures were identified by optical microscope, scanning electron microscope, transmission electron microscope, and micro-hardness tests. As a focusing point, the lath bainite and lath martensite were distinguished by their compositions, morphologies, and hardness. In particular, a number of black plane facets appearing between lath bainite or lath martensite packets were characterized by laser scanning confocal microscope. The results indicated that with the increase in Ni contents in the range of 0, 2, 4, and 6%, the microstructures in the weld-deposited metal were changed from the domination of the granular bainite to the majority of the lath bainite and/or the lath martensite and the micro-hardness of the weld-deposited metal increased. Meanwhile, the average width of columnar grain displays a decreasing trend and prior austenite grain size decreases while increases with higher Ni content above 4%. Yield strength and ultimate tensile strength decrease, while the reduction in fracture area increases with the decreasing Ni mass fraction and the increasing test temperature, respectively. And poor yield strength in Ni6 specimen can be attributed to elements segregation caused by weld defect. Finally, micro-hardness distribution in correspondence with specimens presents as a style of cloud-map.

Fractography of induction-hardened steel fractured in fatigue and overload

DOE Office of Scientific and Technical Information (OSTI.GOV)

Santos, C.G.; Laird, C.

1997-07-01

The fracture surfaces of induction-hardened steel specimens obtained from an auto axle were characterized, macroscopically and microscopically, after being fractured in fatigue and monotonic overload. Specimens were tested in cyclic three-point bending under load control, and the S-N curve was established for specimens that had been notched by spark machining to facilitate fractography. Scanning electron microscopy of the fractured surfaces obtained for lives spanning the range 17,000 to 418,000 cycles revealed diverse fracture morphologies, including intergranular fracture and transgranular fatigue fracture. The results are being offered to assist in the analysis of complex field failures in strongly hardened steel.

Alignment algorithms and per-particle CTF correction for single particle cryo-electron tomography.

PubMed

Galaz-Montoya, Jesús G; Hecksel, Corey W; Baldwin, Philip R; Wang, Eryu; Weaver, Scott C; Schmid, Michael F; Ludtke, Steven J; Chiu, Wah

2016-06-01

Single particle cryo-electron tomography (cryoSPT) extracts features from cryo-electron tomograms, followed by 3D classification, alignment and averaging to generate improved 3D density maps of such features. Robust methods to correct for the contrast transfer function (CTF) of the electron microscope are necessary for cryoSPT to reach its resolution potential. Many factors can make CTF correction for cryoSPT challenging, such as lack of eucentricity of the specimen stage, inherent low dose per image, specimen charging, beam-induced specimen motions, and defocus gradients resulting both from specimen tilting and from unpredictable ice thickness variations. Current CTF correction methods for cryoET make at least one of the following assumptions: that the defocus at the center of the image is the same across the images of a tiltseries, that the particles all lie at the same Z-height in the embedding ice, and/or that the specimen, the cryo-electron microscopy (cryoEM) grid and/or the carbon support are flat. These experimental conditions are not always met. We have developed a CTF correction algorithm for cryoSPT without making any of the aforementioned assumptions. We also introduce speed and accuracy improvements and a higher degree of automation to the subtomogram averaging algorithms available in EMAN2. Using motion-corrected images of isolated virus particles as a benchmark specimen, recorded with a DE20 direct detection camera, we show that our CTF correction and subtomogram alignment routines can yield subtomogram averages close to 4/5 Nyquist frequency of the detector under our experimental conditions. Copyright © 2016 Elsevier Inc. All rights reserved.

Charging/discharge events in coated spacecraft polymers during electron beam irradiation in a scanning electron microscope

NASA Astrophysics Data System (ADS)

Czeremuszkin, G.; Latrèche, M.; Wertheimer, M. R.

2001-12-01

Spacecraft, such as those operating in geosynchronous orbit (GEO), can be subjected to intense irradiation by charged particles, for example high-energy (e.g. 20 keV) electrons. The surfaces of dielectric materials (for example, polymers used as "thermal blankets") can therefore become potential sites for damaging electrostatic discharge (ESD) pulse events. We simulate these conditions by examining small specimens of three relevant polymers (polyimide, polyester and fluoropolymer), both bare and coated, in a scanning electron microscope (SEM). The coatings examined include commercial indium-tin oxide (ITO), and thin films of SiO 2 and a-Si:H deposited by plasma-enhanced chemical vapor deposition (PECVD). All coatings are found to greatly modify the observed ESD behavior, compared with that of the bare polymer counterparts. These observations are explained in terms of the model for ESD pulses proposed by Frederickson.

[An electron microscopic study on the RNA component of synaptonemal complexes in spermatocytes of Mus musculus].

PubMed

Xing, M; Jing, D Z; Hao, S

1991-01-01

The ultrastructural and cytochemical features of synaptonemal complexes (SC) in sections of spermatocytes of Mus musculus were studied under electron microscope. In specimens stained with uranyl acetate and lead citrate the SC was found consisting of three main elements. the lateral element (LE), the central element (CE) and the transverse filament (L-C filament). When stained with the Bernhard's technique, the SC was recognized as a contrasted, tripartite structure which was usually located in the bleached area occupied by the condensed chromatin and composed of highly electron-dense LEs and medium electron-dense CE and L-C filaments. The SC and the LE, stained either by uranyl acetate-lead citrate or by the Bernhard's technique, always showed diameters of about 210 nm and 60 nm, respectively. The results suggest that RNA may be an important component of the SC.

Experimental realization of the Ehrenberg-Siday thought experiment

NASA Astrophysics Data System (ADS)

Pozzi, Giulio; Boothroyd, Chris B.; Tavabi, Amir H.; Yücelen, Emrah; Dunin-Borkowski, Rafal E.; Frabboni, Stefano; Gazzadi, Gian Carlo

2016-02-01

In 1949, at the end of a paper dedicated to the concept of the refractive index in electron optics, Ehrenberg and Siday noted that wave-optical effects will arise from an isolated magnetic field even when the rays themselves travel in magnetic-field-free space. They proposed a two-slit experiment, in which a magnetic flux is enclosed between interfering electron beams. Now, through access to modern nanotechnology tools, we used a focused ion beam to open two nanosized slits in a gold-coated silicon nitride membrane and focused electron beam induced deposition to fabricate a thin magnetic bar between the two slits. We then performed Fraunhofer experiments in a transmission electron microscope equipped with a field emission gun and a Lorentz lens. By tilting the specimen in the objective lens field of the electron microscope, the magnetization of the bar could be reversed and the corresponding change in the phase of the electron wave observed directly in the form of a shift in the interference fringe pattern.

Path-separated electron interferometry in a scanning transmission electron microscope

NASA Astrophysics Data System (ADS)

Yasin, Fehmi S.; Harvey, Tyler R.; Chess, Jordan J.; Pierce, Jordan S.; McMorran, Benjamin J.

2018-05-01

We report a path-separated electron interferometer within a scanning transmission electron microscope. In this setup, we use a nanofabricated grating as an amplitude-division beamsplitter to prepare multiple spatially separated, coherent electron probe beams. We achieve path separations of 30 nm. We pass the  +1 diffraction order probe through amorphous carbon while passing the 0th and  ‑1 orders through vacuum. The probes are then made to interfere via imaging optics, and we observe an interference pattern at the CCD detector with up to 39.7% fringe visibility. We show preliminary experimental results in which the interference pattern was recorded during a 1D scan of the diffracted probes across a test phase object. These results qualitatively agree with a modeled interference predicted by an independent measurement of the specimen thickness. This experimental design can potentially be applied to phase contrast imaging and fundamental physics experiments, such as an exploration of electron wave packet coherence length.

[Pathological features and clinical manifestations in 313 children with nephropathy under 6].

PubMed

Dang, Xi-qiang; Cao, Yan; Yi, Zhu-wen; Xu, Zi-chuan; He, Xiao-jie; Huang, Dan-lin

2008-03-01

To explore the relationship between pathological features and clinical manifestations in children with nephropathy under 6 years old. Renal biopsy by rapid percutaneous puncturation was performed on 313 children under 6 who were all diagnosed clinically as kidney diseases of 14 different kinds. The specimens were divided into 3 parts for microscope, electron microscope and immuno fluorescence examination respectively and processed by HE, PAS, PASM, and Masson staining. Immunofluorescence was used to detect the deposition of IgG, IgM, IgA, C3, C4, C1q, and Fb in the renal tissues. Additional examinations were done to detect HBs-Ag, HBeAg and HBcAg deposition in some cases with positive serum HBs-Ag. Altogether 290 of the specimens (290/313, 92.65%) were examined by electron microscope. All the renal biopsy performances were successful. The clinical manifestations comprised of persistent haematuria (32.92%, 103/313), idiopathic nephritic syndrome (26.1%, 82/313), acute nephritic syndrome (20.14%, 63/313), Henoch Schonlein purpura nephritis (8.32%, 26/313), HBV-nephritis (4.79%, 15/313), and isolated proteinuria (2.56%, 8/313). The main pathological patterns of glomerular disease were identified as mesangial proliferation (51.75%, 162/313), IgM nephropathy (8.31%,26/313), minor and minimal change (7.99%, 25/313), IgA nephropathy (7.35%, 23/313), endocapillary proliferative glomerulonephritis (5.11%, 16/313), focus segmental glomerulosclerosis (4.47%, 14/313), thin basement membrane nephropathy (4.47%, 14/313), and membrane nephropathy (4.47%, 14/313). Alport syndrome, congenital nephrotic syndrome, and thin basement membrane nephropathy can be diagnosed by electron microscope, white IgA nephropathy, IgM nephropathy and C1q nephropathy by immunopathology. Similar clinical manifestations may differ in the pathology and the clinical features of one pathological diagnosis may vary greatly. Renal biopsy is of great help to the diagnosis, treatment and the prognosis evaluation for children with nephropathy under 6. Electron microscopes also play an important role in the diagnosis of nephropathy.

Cryo-FIB specimen preparation for use in a cartridge-type cryo-TEM.

PubMed

He, Jie; Hsieh, Chyongere; Wu, Yongping; Schmelzer, Thomas; Wang, Pan; Lin, Ying; Marko, Michael; Sui, Haixin

2017-08-01

Cryo-electron tomography (cryo-ET) is a well-established technique for studying 3D structural details of subcellular macromolecular complexes and organelles in their nearly native context in the cell. A primary limitation of the application of cryo-ET is the accessible specimen thickness, which is less than the diameters of almost all eukaryotic cells. It has been shown that focused ion beam (FIB) milling can be used to prepare thin, distortion-free lamellae of frozen biological material for high-resolution cryo-ET. Commercial cryosystems are available for cryo-FIB specimen preparation, however re-engineering and additional fixtures are often essential for reliable results with a particular cryo-FIB and cryo-transmission electron microscope (cryo-TEM). Here, we describe our optimized protocol and modified instrumentation for cryo-FIB milling to produce thin lamellae and subsequent damage-free cryotransfer of the lamellae into our cartridge-type cryo-TEM. Published by Elsevier Inc.

Atomically Thin Graphene Windows That Enable High Contrast Electron Microscopy without a Specimen Vacuum Chamber.

PubMed

Han, Yimo; Nguyen, Kayla X; Ogawa, Yui; Park, Jiwoong; Muller, David A

2016-12-14

Scanning electron microscopes (SEMs) require a high vacuum environment to generate and shape an electron beam for imaging; however, the vacuum conditions greatly limit the nature of specimens that can be examined. From a purely scattering physics perspective, it is not necessary to place the specimen inside the vacuum chamber-the mean free paths (MFPs) for electron scattering in air at typical SEM beam voltages are 50-100 μm. This is the idea behind the airSEM, which removes the specimen vacuum chamber from the SEM and places the sample in air. The thickness of the gas layer is less than a MFP from an electron-transparent window to preserve the shape and resolution of the incident beam, resulting in comparable imaging quality to an all-vacuum SEM. Present silicon nitride windows scatter far more strongly than the air gap and are currently the contrast and resolution limiting factor in the airSEM. Graphene windows have been used previously to wrap or seal samples in vacuum for imaging. Here we demonstrate the use of a robust bilayer graphene window for sealing the electron optics from the room environment, providing an electron transparent window with only a 2% drop in contrast. There is a 5-fold-increase in signal/noise ratio for imaging compared to multi-MFP-thick silicon nitride windows, enabling high contrast in backscattered, transmission, and surface imaging modes for the new airSEM geometry.

Diffraction effects and inelastic electron transport in angle-resolved microscopic imaging applications.

PubMed

Winkelmann, A; Nolze, G; Vespucci, S; Naresh-Kumar, G; Trager-Cowan, C; Vilalta-Clemente, A; Wilkinson, A J; Vos, M

2017-09-01

We analyse the signal formation process for scanning electron microscopic imaging applications on crystalline specimens. In accordance with previous investigations, we find nontrivial effects of incident beam diffraction on the backscattered electron distribution in energy and momentum. Specifically, incident beam diffraction causes angular changes of the backscattered electron distribution which we identify as the dominant mechanism underlying pseudocolour orientation imaging using multiple, angle-resolving detectors. Consequently, diffraction effects of the incident beam and their impact on the subsequent coherent and incoherent electron transport need to be taken into account for an in-depth theoretical modelling of the energy- and momentum distribution of electrons backscattered from crystalline sample regions. Our findings have implications for the level of theoretical detail that can be necessary for the interpretation of complex imaging modalities such as electron channelling contrast imaging (ECCI) of defects in crystals. If the solid angle of detection is limited to specific regions of the backscattered electron momentum distribution, the image contrast that is observed in ECCI and similar applications can be strongly affected by incident beam diffraction and topographic effects from the sample surface. As an application, we demonstrate characteristic changes in the resulting images if different properties of the backscattered electron distribution are used for the analysis of a GaN thin film sample containing dislocations. © 2017 The Authors. Journal of Microscopy published by JohnWiley & Sons Ltd on behalf of Royal Microscopical Society.

Effect of grinding on the fatigue life of titanium alloy (5 Al-2.5 Sn) under dry and wet conditions

NASA Technical Reports Server (NTRS)

Rangaswamy, Partha; Terutung, Hendra; Jeelani, Shaik

1989-01-01

The principal factors in the performance of aerospace materials are strength-to-weight ratio, fatigue life, fracture toughness, survivability and, of course, reliability. Machining processes and, in particular, grinding under adverse conditions have been found to cause damage to surface integrity and affect the residual stress distribution in the surface and subsurface region. These effects have a direct bearing on the fatigue life. In this investigation the effects of grinding conditions on the fatigue life of Titanium 5 Al-2.5Sn were studied. This alloy is used in ground form in the manufacturing of some critical components in the space shuttle's main engine. It is essential that materials for such applications be properly characterized for use in severe service conditions. Flat sub-size specimens 0.1 inch thick were ground on a surface grinding machine equipped with a variable speed motor at speeds of 2000 to 6000 rpm using SiC wheels of grit sizes 60 and 120. The grinding parameters used in this investigation were chosen from a separate study. The ground specimens were then fatigued at a selected stress and the resulting lives were compared with that of the virgin material. The surfaces of the specimens were examined under a scanning electron microscope, and the roughness and hardness were measured using a standard profilometer and microhardness tester, respectively. The fatigue life of the ground specimens was found to decrease with the increase in speed for both dry and wet conditions. The fatigue life of specimens ground under wet conditions showed a significant increase at the wheel speed of 2000 rpm for both the grit sizes and thereafter decreased with increase profilometry, microhardness measurements and scanning electron microscopic examination.

Microwave-accelerated cytochemical stains for the image analysis and the electron microscopic examination of light microscopy diagnostic slides.

PubMed

Hanker, J; Giammara, B

1993-01-01

Recent studies in our laboratories have shown how microwave (MW) irradiation can accelerate a number of tissue-processing techniques, especially staining, to aid in the preparation of single specimens on glass microscope slides or coverslips for examination by light microscopy (and electron microscopy, if required) for diagnostic purposes. Techniques have been developed, which give permanently stained preparations, that can be studied initially by light microscopy, their areas of interest mapped, and computer-automated image analysis performed to obtain quantitative information. This is readily performed after MW-accelerated staining with silver methenamine by the Giammara-Hanker PATS or PATS-TS reaction. This variation of the PAS reaction gives excellent markers for specific infectious agents such as lipopolysaccharides for gram-negative bacteria or mannans for fungi. It is also an excellent stain for glycogen and basement membranes and an excellent marker for type III collagen or reticulin in the endoneurium or perineurium of peripheral nerve or in the capillary walls. Our improved MW-accelerated Feulgen reaction with silver methenamine for nuclear DNA is useful to show the nuclei of bacteria and fungi as well as of cells they are infecting. Improved coating and penetration of tissue surfaces by thiocarbohydrazide bridging of ruthenium red, applied under MW-acceleration, render biologic specimens sufficiently conductive for SEM so that sputter coating with gold is unnecessary. The specimens treated with these highly visible electron-opaque stains can be screened with the light microscope after mounting in polyethylene glycol (PEG) and the structures or areas selected for EM study are mapped with a Micro-Locator slide. After removal of the water soluble PEG the specimens are remounted in the usual EM media for scanning electron microscopy (SEM) or transmission electron microscopy (TEM) study of the mapped areas. By comparing duplicate smears from areas of infection, such as two coverslips of buffy coat smears of blood from a patient with septicemia, the microorganisms responsible can occasionally be classified for antimicrobial therapy long before culture results are available; gram-negative bacteria are positive with the Giammara-Hanker PATS-TS stain, and gram-positive bacteria are positive with the SIGMA HT40 Gram stain. The gram-positive as well as gram-negative bacteria are both initially stained by the crystal violet component of the Gram stain. The crystal violet stain is readily removed from the gram-negative (but not the gram-positive) bacteria when the specimens are rinsed with alcohol/acetone. If this rinse step is omitted, the crystal violet remains attached to both gram-negative and gram-positive bacteria. It can then be rendered insoluble, electron-opaque, and conductive by treatment with silver methenamine solution under MW-irradiation. This metallized crystal violet is a more effective silver stain than the PATS-TS stain for a number of gram-negative spirochetes such as Treponema pallidum, the microbe that causes syphilis.

In-situ deformation studies of an aluminum metal-matrix composite in a scanning electron microscope

NASA Technical Reports Server (NTRS)

Manoharan, M.; Lewandowski, J. J.

1989-01-01

Tensile specimens made of a metal-matrix composite (cast and extruded aluminum alloy-based matrix reinforced with Al2O3 particulate) were tested in situ in a scanning electron microscope equipped with a deformation stage, to directly monitor the crack propagation phenomenon. The in situ SEM observations revealed the presence of microcracks both ahead of and near the crack-tip region. The microcracks were primarily associated with cracks in the alumina particles. The results suggest that a region of intense deformation exists ahead of the crack and corresponds to the region of microcracking. As the crack progresses, a region of plastically deformed material and associated microcracks remains in the wake of the crack.

Microscope basics.

PubMed

Sluder, Greenfield; Nordberg, Joshua J

2013-01-01

This chapter provides information on how microscopes work and discusses some of the microscope issues to be considered in using a video camera on the microscope. There are two types of microscopes in use today for research in cell biology-the older finite tube-length (typically 160mm mechanical tube length) microscopes and the infinity optics microscopes that are now produced. The objective lens forms a magnified, real image of the specimen at a specific distance from the objective known as the intermediate image plane. All objectives are designed to be used with the specimen at a defined distance from the front lens element of the objective (the working distance) so that the image formed is located at a specific location in the microscope. Infinity optics microscopes differ from the finite tube-length microscopes in that the objectives are designed to project the image of the specimen to infinity and do not, on their own, form a real image of the specimen. Three types of objectives are in common use today-plan achromats, plan apochromats, and plan fluorite lenses. The concept of mounting video cameras on the microscope is also presented in the chapter. Copyright © 2003 Elsevier Inc. All rights reserved.

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.

Integrated light and scanning electron microscopy of GFP-expressing cells.

PubMed

Peddie, Christopher J; Liv, Nalan; Hoogenboom, Jacob P; Collinson, Lucy M

2014-01-01

Integration of light and electron microscopes provides imaging tools in which fluorescent proteins can be localized to cellular structures with a high level of precision. However, until recently, there were few methods that could deliver specimens with sufficient fluorescent signal and electron contrast for dual imaging without intermediate staining steps. Here, we report protocols that preserve green fluorescent protein (GFP) in whole cells and in ultrathin sections of resin-embedded cells, with membrane contrast for integrated imaging. Critically, GFP is maintained in a stable and active state within the vacuum of an integrated light and scanning electron microscope. For light microscopists, additional structural information gives context to fluorescent protein expression in whole cells, illustrated here by analysis of filopodia and focal adhesions in Madin Darby canine kidney cells expressing GFP-Paxillin. For electron microscopists, GFP highlights the proteins of interest within the architectural space of the cell, illustrated here by localization of the conical lipid diacylglycerol to cellular membranes. © 2014 Elsevier Inc. All rights reserved.

Advantages of intermediate X-ray energies in Zernike phase contrast X-ray microscopy.

PubMed

Wang, Zhili; Gao, Kun; Chen, Jian; Hong, Youli; Ge, Xin; Wang, Dajiang; Pan, Zhiyun; Zhu, Peiping; Yun, Wenbing; Jacobsen, Chris; Wu, Ziyu

2013-01-01

Understanding the hierarchical organizations of molecules and organelles within the interior of large eukaryotic cells is a challenge of fundamental interest in cell biology. Light microscopy is a powerful tool for observations of the dynamics of live cells, its resolution attainable is limited and insufficient. While electron microscopy can produce images with astonishing resolution and clarity of ultra-thin (<1 μm thick) sections of biological specimens, many questions involve the three-dimensional organization of a cell or the interconnectivity of cells. X-ray microscopy offers superior imaging resolution compared to light microscopy, and unique capability of nondestructive three-dimensional imaging of hydrated unstained biological cells, complementary to existing light and electron microscopy. Until now, X-ray microscopes operating in the "water window" energy range between carbon and oxygen k-shell absorption edges have produced outstanding 3D images of cryo-preserved cells. The relatively low X-ray energy (<540 eV) of the water window imposes two important limitations: limited penetration (<10 μm) not suitable for imaging larger cells or tissues, and small depth of focus (DoF) for high resolution 3D imaging (e.g., ~1 μm DoF for 20 nm resolution). An X-ray microscope operating at intermediate energy around 2.5 keV using Zernike phase contrast can overcome the above limitations and reduces radiation dose to the specimen. Using a hydrated model cell with an average chemical composition reported in literature, we calculated the image contrast and the radiation dose for absorption and Zernike phase contrast, respectively. The results show that an X-ray microscope operating at ~2.5 keV using Zernike phase contrast offers substantial advantages in terms of specimen size, radiation dose and depth-of-focus. Copyright © 2012 Elsevier Inc. All rights reserved.

Oxidation of metals and alloys in controlled atmospheres using in situ transmission electron microscopy and Auger spectrography

NASA Technical Reports Server (NTRS)

Rao, D. B.; Heinemann, K.; Douglass, D. L.

1976-01-01

Single-crystalline thin films of copper were oxidized at an isothermal temperature of 425 C and at an oxygen partial pressure of .005 Torr in situ in a high-resolution electron microscope. The specimens were prepared by epitaxial vapor deposition onto polished 100 and 110 faces of rocksalt and mounted in a hot stage inside an ultra-high-vacuum specimen chamber of the microscope. Large amounts of sulfur, carbon, and oxygen were detected by Auger electron spectroscopy on the surface of the as-received films and were removed in situ by ion-sputter etching immediately prior to the oxidation. The nucleation and growth characteristics of Cu2O on Cu were studied. Results show that neither stacking faults nor dislocations are associated with the Cu2O nucleation sites. The growth of Cu2O nuclei is linear with time. The experimental findings, including results from oxygen dissolution experiments and from repetitive oxidation-reduction-oxidation sequences, fit well into the framework of an oxidation process involving (a) the formation of a surface-charge layer, (b) oxygen saturation in the metal and (c) nucleation, followed by surface diffusion of oxygen and bulk diffusion of copper for lateral and vertical oxide growth, respectively.

Room temperature mechanical properties of electron beam welded zircaloy-4 sheet

DOE Office of Scientific and Technical Information (OSTI.GOV)

Parga, C. J.; Rooyen, I. J.; Coryell, B. D.

Room temperature mechanical properties of electron beam welded and plain Zircaloy-4 sheet (1.6mm thick) have been measured and compared. Various welding parameters were utilized to join sheet material. Electron beam welded specimens and as-received sheet specimens show comparable mechanical properties. Zr-4 sheet displays anisotropy; tensile properties measured for transverse display higher elastic modulus, yield strength, reduction of area and slightly lower ductility than for the longitudinal (rolling direction). Higher welding power increases the alloy’s hardness, elastic modulus and yield strength, with a corresponding decrease in tensile strength and ductility. The hardness measured at weld is comparable to the parent metalmore » hardness. Hardness at heat-affected-zone is slightly higher. Electron microscopic examination shows distinct microstructure morphology and grain size at the weld zone, HAZ and parent metal. A correlation between welding parameters, mechanical properties and microstructural features was established for electron beam welded Zircaloy-4 sheet material.« less

Room temperature mechanical properties of electron beam welded zircaloy-4 sheet

DOE PAGES

Parga, C. J.; Rooyen, I. J.; Coryell, B. D.; ...

2017-11-04

Room temperature mechanical properties of electron beam welded and plain Zircaloy-4 sheet (1.6mm thick) have been measured and compared. Various welding parameters were utilized to join sheet material. Electron beam welded specimens and as-received sheet specimens show comparable mechanical properties. Zr-4 sheet displays anisotropy; tensile properties measured for transverse display higher elastic modulus, yield strength, reduction of area and slightly lower ductility than for the longitudinal (rolling direction). Higher welding power increases the alloy’s hardness, elastic modulus and yield strength, with a corresponding decrease in tensile strength and ductility. The hardness measured at weld is comparable to the parent metalmore » hardness. Hardness at heat-affected-zone is slightly higher. Electron microscopic examination shows distinct microstructure morphology and grain size at the weld zone, HAZ and parent metal. A correlation between welding parameters, mechanical properties and microstructural features was established for electron beam welded Zircaloy-4 sheet material.« less

Ionic liquid-based observation technique for nonconductive materials in the scanning electron microscope: Application to the characterization of a rare earth ore.

PubMed

Brodusch, Nicolas; Waters, Kristian; Demers, Hendrix; Gauvin, Raynald

2014-03-01

A new approach for preparing geological materials is proposed to reduce charging during their characterization in a scanning electron microscope. This technique was applied to a sample of the Nechalacho rare earth deposit, which contains a significant amount of the minerals fergusonite and zircon. Instead of covering the specimen surface with a conductive coating, the sample was immersed in a dilute solution of ionic liquid and then air dried prior to SEM analysis. Imaging at a wide range of accelerating voltages was then possible without evidence of charging when using the in-chamber secondary and backscattered electrons detectors, even at 1 kV. High resolution x-ray and electron backscatter diffraction mapping were successfully obtained at 20 and 5 kV with negligible image drifting and permitted the characterization of the microstructure of the zircon/fergusonite-Y aggregates encased in the matrix minerals. Because of the absence of a conductive layer at the surface of the specimen, the Kikuchi band contrast was improved and the backscatter electron signal increased at both 5 and 20 kV as confirmed by Monte Carlo modeling. These major developments led to an improvement of the spatial resolution and efficiency of the above characterization techniques applied to the rare earth ore and it is expected that they can be applied to other types of ores and minerals. Copyright © 2014 Wiley Periodicals, Inc.

Scanning Miniature Microscopes without Lenses

NASA Technical Reports Server (NTRS)

Wang, Yu

2009-01-01

The figure schematically depicts some alternative designs of proposed compact, lightweight optoelectronic microscopes that would contain no lenses and would generate magnified video images of specimens. Microscopes of this type were described previously in Miniature Microscope Without Lenses (NPO - 20218), NASA Tech Briefs, Vol. 22, No. 8 (August 1998), page 43 and Reflective Variants of Miniature Microscope Without Lenses (NPO 20610), NASA Tech Briefs, Vol. 26, No. 9 (September 1999), page 6a. To recapitulate: In the design and construction of a microscope of this type, the focusing optics of a conventional microscope are replaced by a combination of a microchannel filter and a charge-coupled-device (CCD) image detector. Elimination of focusing optics reduces the size and weight of the instrument and eliminates the need for the time-consuming focusing operation. The microscopes described in the cited prior articles contained two-dimensional CCDs registered with two-dimensional arrays of microchannels and, as such, were designed to produce full two-dimensional images, without need for scanning. The microscopes of the present proposal would contain one-dimensional (line image) CCDs registered with linear arrays of microchannels. In the operation of such a microscope, one would scan a specimen along a line perpendicular to the array axis (in other words, one would scan in pushbroom fashion). One could then synthesize a full two-dimensional image of the specimen from the line-image data acquired at one-pixel increments of position along the scan. In one of the proposed microscopes, a beam of unpolarized light for illuminating the specimen would enter from the side. This light would be reflected down onto the specimen by a nonpolarizing beam splitter attached to the microchannels at their lower ends. A portion of the light incident on the specimen would be reflected upward, through the beam splitter and along the microchannels, to form an image on the CCD. If the nonpolarizing beam splitter were replaced by a polarizing one, then the specimen would be illuminated by s-polarized light. Upon reflection from the specimen, some of the s-polarized light would become p-polarized. Only the p-polarized light would contribute to the image on the CCD; in other words, the image would contain information on the polarization rotating characteristic of the specimen.

Alignment Algorithms and Per-Particle CTF Correction for Single Particle Cryo-Electron Tomography

PubMed Central

Galaz-Montoya, Jesús G.; Hecksel, Corey W.; Baldwin, Philip R.; Wang, Eryu; Weaver, Scott C.; Schmid, Michael F.; Ludtke, Steven J.; Chiu, Wah

2016-01-01

Single particle cryo-electron tomography (cryoSPT) extracts features from cryo-electron tomograms, followed by 3D classification, alignment and averaging to generate improved 3D density maps of such features. Robust methods to correct for the contrast transfer function (CTF) of the electron microscope are necessary for cryoSPT to reach its resolution potential. Many factors can make CTF correction for cryoSPT challenging, such as lack of eucentricity of the specimen stage, inherent low dose per image, specimen charging, beam-induced specimen motions, and defocus gradients resulting both from specimen tilting and from unpredictable ice thickness variations. Current CTF correction methods for cryoET make at least one of the following assumptions: that the defocus at the center of the image is the same across the images of a tiltseries, that the particles all lie at the same Z-height in the embedding ice, and/or that the specimen grid and carbon support are flat. These experimental conditions are not always met. We have developed a CTF correction algorithm for cryoSPT without making any of the aforementioned assumptions. We also introduce speed and accuracy improvements and a higher degree of automation to the subtomogram averaging algorithms available in EMAN2. Using motion-corrected images of isolated virus particles as a benchmark specimen, recorded with a DE20 direct detection camera, we show that our CTF correction and subtomogram alignment routines can yield subtomogram averages close to 4/5 Nyquist frequency of the detector under our experimental conditions. PMID:27016284

SD-SEM: sparse-dense correspondence for 3D reconstruction of microscopic samples.

PubMed

Baghaie, Ahmadreza; Tafti, Ahmad P; Owen, Heather A; D'Souza, Roshan M; Yu, Zeyun

2017-06-01

Scanning electron microscopy (SEM) imaging has been a principal component of many studies in biomedical, mechanical, and materials sciences since its emergence. Despite the high resolution of captured images, they remain two-dimensional (2D). In this work, a novel framework using sparse-dense correspondence is introduced and investigated for 3D reconstruction of stereo SEM images. SEM micrographs from microscopic samples are captured by tilting the specimen stage by a known angle. The pair of SEM micrographs is then rectified using sparse scale invariant feature transform (SIFT) features/descriptors and a contrario RANSAC for matching outlier removal to ensure a gross horizontal displacement between corresponding points. This is followed by dense correspondence estimation using dense SIFT descriptors and employing a factor graph representation of the energy minimization functional and loopy belief propagation (LBP) as means of optimization. Given the pixel-by-pixel correspondence and the tilt angle of the specimen stage during the acquisition of micrographs, depth can be recovered. Extensive tests reveal the strength of the proposed method for high-quality reconstruction of microscopic samples. Copyright © 2017 Elsevier Ltd. All rights reserved.

Preparation of viral samples within biocontainment for ultrastructural analysis: Utilization of an innovative processing capsule for negative staining.

PubMed

Monninger, Mitchell K; Nguessan, Chrystal A; Blancett, Candace D; Kuehl, Kathleen A; Rossi, Cynthia A; Olschner, Scott P; Williams, Priscilla L; Goodman, Steven L; Sun, Mei G

2016-12-01

Transmission electron microscopy can be used to observe the ultrastructure of viruses and other microbial pathogens with nanometer resolution. In a transmission electron microscope (TEM), the image is created by passing an electron beam through a specimen with contrast generated by electron scattering from dense elements in the specimen. Viruses do not normally contain dense elements, so a negative stain that places dense heavy metal salts around the sample is added to create a dark border. To prepare a virus sample for a negative stain transmission electron microscopy, a virus suspension is applied to a TEM grid specimen support, which is a 3mm diameter fragile specimen screen coated with a few nanometers of plastic film. Then, deionized (dI) water rinses and a negative stain solution are applied to the grid. All infectious viruses must be handled in a biosafety cabinet (BSC) and many require a biocontainment laboratory environment. Staining viruses in biosafety levels (BSL) 3 and 4 is especially challenging because the support grids are small, fragile, and easily moved by air currents. In this study we evaluated a new device for negative staining viruses called mPrep/g capsule. It is a capsule that holds up to two TEM grids during all processing steps and for storage after staining is complete. This study reports that the mPrep/g capsule method is valid and effective to negative stain virus specimens, especially in high containment laboratory environments. Copyright © 2016 The Authors. Published by Elsevier B.V. All rights reserved.

Correlative Raman spectroscopy and focused ion beam for targeted phase boundary analysis of titania polymorphs

DOE Office of Scientific and Technical Information (OSTI.GOV)

Mangum, John S.; Chan, Lisa H.; Schmidt, Ute

Site-specific preparation of specimens using focused ion beam instruments for transmission electron microscopy is at the forefront of targeting regions of interest for nanoscale characterization. Typical methods of pinpointing desired features include electron backscatter diffraction for differentiating crystal structures and energy-dispersive X-Ray spectroscopy for probing compositional variations. Yet there are situations, notably in the titanium dioxide system, where these techniques can fail. Differentiating between the brookite and anatase polymorphs of titania is either excessively laborious or impossible with the aforementioned techniques. However, due to differences in bonding structure, Raman spectroscopy serves as an ideal candidate for polymorph differentiation. In thismore » work, a correlative approach utilizing Raman spectroscopy for targeted focused ion beam specimen preparation was employed. Dark field imaging and diffraction in the transmission electron microscope confirmed the region of interest located via Raman spectroscopy and demonstrated the validity of this new method. Correlative Raman spectroscopy, scanning electron microscopy, and focused ion beam is shown to be a promising new technique for identifying site-specific preparation of nanoscale specimens in cases where conventional approaches do not suffice.« less

Correlative Raman spectroscopy and focused ion beam for targeted phase boundary analysis of titania polymorphs.

PubMed

Mangum, John S; Chan, Lisa H; Schmidt, Ute; Garten, Lauren M; Ginley, David S; Gorman, Brian P

2018-05-01

Site-specific preparation of specimens using focused ion beam instruments for transmission electron microscopy is at the forefront of targeting regions of interest for nanoscale characterization. Typical methods of pinpointing desired features include electron backscatter diffraction for differentiating crystal structures and energy-dispersive X-Ray spectroscopy for probing compositional variations. Yet there are situations, notably in the titanium dioxide system, where these techniques can fail. Differentiating between the brookite and anatase polymorphs of titania is either excessively laborious or impossible with the aforementioned techniques. However, due to differences in bonding structure, Raman spectroscopy serves as an ideal candidate for polymorph differentiation. In this work, a correlative approach utilizing Raman spectroscopy for targeted focused ion beam specimen preparation was employed. Dark field imaging and diffraction in the transmission electron microscope confirmed the region of interest located via Raman spectroscopy and demonstrated the validity of this new method. Correlative Raman spectroscopy, scanning electron microscopy, and focused ion beam is shown to be a promising new technique for identifying site-specific preparation of nanoscale specimens in cases where conventional approaches do not suffice. Copyright © 2018 Elsevier B.V. All rights reserved.

Correlative Raman spectroscopy and focused ion beam for targeted phase boundary analysis of titania polymorphs

DOE PAGES

Mangum, John S.; Chan, Lisa H.; Schmidt, Ute; ...

2018-02-23

Site-specific preparation of specimens using focused ion beam instruments for transmission electron microscopy is at the forefront of targeting regions of interest for nanoscale characterization. Typical methods of pinpointing desired features include electron backscatter diffraction for differentiating crystal structures and energy-dispersive X-Ray spectroscopy for probing compositional variations. Yet there are situations, notably in the titanium dioxide system, where these techniques can fail. Differentiating between the brookite and anatase polymorphs of titania is either excessively laborious or impossible with the aforementioned techniques. However, due to differences in bonding structure, Raman spectroscopy serves as an ideal candidate for polymorph differentiation. In thismore » work, a correlative approach utilizing Raman spectroscopy for targeted focused ion beam specimen preparation was employed. Dark field imaging and diffraction in the transmission electron microscope confirmed the region of interest located via Raman spectroscopy and demonstrated the validity of this new method. Correlative Raman spectroscopy, scanning electron microscopy, and focused ion beam is shown to be a promising new technique for identifying site-specific preparation of nanoscale specimens in cases where conventional approaches do not suffice.« less

Dark-field imaging based on post-processed electron backscatter diffraction patterns of bulk crystalline materials in a scanning electron microscope.

PubMed

Brodusch, Nicolas; Demers, Hendrix; Gauvin, Raynald

2015-01-01

Dark-field (DF) images were acquired in the scanning electron microscope with an offline procedure based on electron backscatter diffraction (EBSD) patterns (EBSPs). These EBSD-DF images were generated by selecting a particular reflection on the electron backscatter diffraction pattern and by reporting the intensity of one or several pixels around this point at each pixel of the EBSD-DF image. Unlike previous studies, the diffraction information of the sample is the basis of the final image contrast with a pixel scale resolution at the EBSP providing DF imaging in the scanning electron microscope. The offline facility of this technique permits the selection of any diffraction condition available in the diffraction pattern and displaying the corresponding image. The high number of diffraction-based images available allows a better monitoring of deformation structures compared to electron channeling contrast imaging (ECCI) which is generally limited to a few images of the same area. This technique was applied to steel and iron specimens and showed its high capability in describing more rigorously the deformation structures around micro-hardness indents. Due to the offline relation between the reference EBSP and the EBSD-DF images, this new technique will undoubtedly greatly improve our knowledge of deformation mechanism and help to improve our understanding of the ECCI contrast mechanisms. Copyright © 2014 Elsevier B.V. All rights reserved.

ON THE FREEZING AND IDENTIFICATION OF LIPID MONOLAYER 2-D ARRAYS FOR CRYOELECTRON MICROSCOPY

PubMed Central

Taylor, Dianne W.; Kelly, Deborah F.; Cheng, Anchi; Taylor, Kenneth A.

2008-01-01

Lipid monolayers provide a convenient vehicle for the crystallization of biological macromolecules for 3-D electron microscopy. Although numerous examples of 3-D images from 2-D protein arrays have been described from negatively stained specimens, only six structures have been done from frozen hydrated specimens. We describe here a method that makes high quality frozen-hydrated specimens of lipid monolayer arrays for cryoelectron microscopy. The method uses holey carbon films with patterned holes for monolayer recovery, blotting and plunge freezing to produce thin aqueous films which cover >90% of the available grid area. With this method, even specimens with relatively infrequent crystals can be screened using automated data collection techniques. Though developed for microscopic examination of 2-D arrays, the method may have wider application to the preparation of single particle specimens for 3-D image reconstruction. PMID:17561414

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.

Iodine Vapor Staining for Atomic Number Contrast in Backscattered Electron and X-ray Imaging

PubMed Central

Boyde, Alan; Mccorkell, Fergus A; Taylor, Graham K; Bomphrey, Richard J; Doube, Michael

2014-01-01

Iodine imparts strong contrast to objects imaged with electrons and X-rays due to its high atomic number (53), and is widely used in liquid form as a microscopic stain and clinical contrast agent. We have developed a simple technique which exploits elemental iodine's sublimation-deposition state-change equilibrium to vapor stain specimens with iodine gas. Specimens are enclosed in a gas-tight container along with a small mass of solid I2. The bottle is left at ambient laboratory conditions while staining proceeds until empirically determined completion (typically days to weeks). We demonstrate the utility of iodine vapor staining by applying it to resin-embedded tissue blocks and whole locusts and imaging them with backscattered electron scanning electron microscopy (BSE SEM) or X-ray microtomography (XMT). Contrast is comparable to that achieved with liquid staining but without the consequent tissue shrinkage, stain pooling, or uneven coverage artefacts associated with immersing the specimen in iodine solutions. Unmineralized tissue histology can be read in BSE SEM images with good discrimination between tissue components. Organs within the locust head are readily distinguished in XMT images with particularly useful contrast in the chitin exoskeleton, muscle and nerves. Here, we have used iodine vapor staining for two imaging modalities in frequent use in our laboratories and on the specimen types with which we work. It is likely to be equally convenient for a wide range of specimens, and for other modalities which generate contrast from electron- and photon-sample interactions, such as transmission electron microscopy and light microscopy. Microsc. Res. Tech. 77:1044–1051, 2014. © 2014 The Authors. Microscopy Research Technique published by Wiley Periodocals, Inc. PMID:25219801

Analysis of Microcrafters in Materials Specimens after Long-Term Exposure on ISS Surface

NASA Technical Reports Server (NTRS)

Shaevich, S. K.; Aleksandrov, N. G.; Shumov, A. E.; Novikov, L. S.; Chernik, V. N.; Samokhina, M. S.; Golden, J. L.; Graves, R. F.; Kravchenko, M.; Christiansen, E. L.;

Progress toward an aberration-corrected low energy electron microscope for DNA sequencing and surface analysis.

PubMed

Mankos, Marian; Shadman, Khashayar; N'diaye, Alpha T; Schmid, Andreas K; Persson, Henrik H J; Davis, Ronald W

2012-11-01

Monochromatic, aberration-corrected, dual-beam low energy electron microscopy (MAD-LEEM) is a novel imaging technique aimed at high resolution imaging of macromolecules, nanoparticles, and surfaces. MAD-LEEM combines three innovative electron-optical concepts in a single tool: a monochromator, a mirror aberration corrector, and dual electron beam illumination. The monochromator reduces the energy spread of the illuminating electron beam, which significantly improves spectroscopic and spatial resolution. The aberration corrector is needed to achieve subnanometer resolution at landing energies of a few hundred electronvolts. The dual flood illumination approach eliminates charging effects generated when a conventional, single-beam LEEM is used to image insulating specimens. The low landing energy of electrons in the range of 0 to a few hundred electronvolts is also critical for avoiding radiation damage, as high energy electrons with kilo-electron-volt kinetic energies cause irreversible damage to many specimens, in particular biological molecules. The performance of the key electron-optical components of MAD-LEEM, the aberration corrector combined with the objective lens and a magnetic beam separator, was simulated. Initial results indicate that an electrostatic electron mirror has negative spherical and chromatic aberration coefficients that can be tuned over a large parameter range. The negative aberrations generated by the electron mirror can be used to compensate the aberrations of the LEEM objective lens for a range of electron energies and provide a path to achieving subnanometer spatial resolution. First experimental results on characterizing DNA molecules immobilized on Au substrates in a LEEM are presented. Images obtained in a spin-polarized LEEM demonstrate that high contrast is achievable at low electron energies in the range of 1-10 eV and show that small changes in landing energy have a strong impact on the achievable contrast. The MAD-LEEM approach promises to significantly improve the performance of a LEEM for a wide range of applications in the biosciences, material sciences, and nanotechnology where nanometer scale resolution and analytical capabilities are required. In particular, the microscope has the potential of delivering images of unlabeled DNA strands with nucleotide-specific contrast. This simplifies specimen preparation and significantly eases the computational complexity needed to assemble the DNA sequence from individual reads.

Effect of etching with distinct hydrofluoric acid concentrations on the flexural strength of a lithium disilicate-based glass ceramic.

PubMed

Prochnow, Catina; Venturini, Andressa B; Grasel, Rafaella; Bottino, Marco C; Valandro, Luiz Felipe

2017-05-01

This study examined the effects of distinct hydrofluoric acid concentrations on the mechanical behavior of a lithium disilicate-based glass ceramic. Bar-shaped specimens were produced from ceramic blocks (e.max CAD, Ivoclar Vivadent). The specimens were polished, chamfered, and sonically cleaned in distilled water. The specimens were randomly divided into five groups (n = 23). The HF1, HF3, HF5, and HF10 specimens were etched for 20 s with acid concentrations of 1%, 3%, 5%, and 10%, respectively, while the SC (control) sample was untreated. The etched surfaces were evaluated using a scanning electron microscope and an atomic force microscope. Finally, the roughness was measured, and 3-point bending flexural tests were performed. The data were analyzed using one-way analysis of variance (ANOVA) and Tukey's test (α = 0.05). The Weibull modulus and characteristic strength were also determined. No statistical difference in the roughness and flexural strength was determined among the groups. The structural reliabilities (Weilbull moduli) were similar for the tested groups; however, the characteristic strength of the HF1 specimen was greater than that of the HF10 specimen. Compared with the untreated ceramic, the surface roughness and flexural strength of the ceramic were unaffected upon etching, regardless of the acid concentration. © 2016 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 105B: 885-891, 2017. © 2016 Wiley Periodicals, Inc.

[Interface bond and compatibility between Mark II machining ceramic and vita VM9 veneering porcelain].

PubMed

Liu, Tian-Shuang; Li, Zhen-Chun; Chen, Xiao-Dong

2009-04-01

To investigate the interface bond and thermal compatibility between Mark II machining ceramic and Vita VM9 veneering porcelain. A bar shaped specimen (30 mm x 15 mm x 1 mm in size) of Mark II block was prepared, with 0.5 mm-deep notch (vertical to the long axis of specimen) at the middle of the bottom surface. The upper surface was veneered with 0.3 mm VM9 dentin base porcelain. Then the specimen was fractured from the notching site and the fracture surface was examined under scanning electron microscope (SEM) and electron microprobe analyzer (EMPA) with electron beam of 1 microm in diameter. Another ten specimens (30 mm x 15 mm x 1.5 mm in size) were fabricated and the temperature of thermal shock resistance were tested. SEM observation showed tight bond between these two materials and EMPA results showed penetration of Al element from Mark II block into veneering porcelain and Ca element from veneering porcelain into Mark II block occurred after sintering baking. The average temperature of thermal shock resistance for specimens in this study was (194.0+/-10.3) degrees C. Cracks were mainly distributed in veneering porcelain. Chemical bond exists between the Mark II machining ceramic and Vita VM9 veneering porcelain, and there is good thermal compatibility between them.

x-y-recording in transmission electron microscopy. A versatile and inexpensive interface to personal computers with application to stereology.

PubMed

Rickmann, M; Siklós, L; Joó, F; Wolff, J R

1990-09-01

An interface for IBM XT/AT-compatible computers is described which has been designed to read the actual specimen stage position of electron microscopes. The complete system consists of (i) optical incremental encoders attached to the x- and y-stage drivers of the microscope, (ii) two keypads for operator input, (iii) an interface card fitted to the bus of the personal computer, (iv) a standard configuration IBM XT (or compatible) personal computer optionally equipped with a (v) HP Graphic Language controllable colour plotter. The small size of the encoders and their connection to the stage drivers by simple ribbed belts allows an easy adaptation of the system to most electron microscopes. Operation of the interface card itself is supported by any high-level language available for personal computers. By the modular concept of these languages, the system can be customized to various applications, and no computer expertise is needed for actual operation. The present configuration offers an inexpensive attachment, which covers a wide range of applications from a simple notebook to high-resolution (200-nm) mapping of tissue. Since section coordinates can be processed in real-time, stereological estimations can be derived directly "on microscope". This is exemplified by an application in which particle numbers were determined by the disector method.

Minimal resin embedding of multicellular specimens for targeted FIB-SEM imaging.

PubMed

Schieber, Nicole L; Machado, Pedro; Markert, Sebastian M; Stigloher, Christian; Schwab, Yannick; Steyer, Anna M

2017-01-01

Correlative light and electron microscopy (CLEM) is a powerful tool to perform ultrastructural analysis of targeted tissues or cells. The large field of view of the light microscope (LM) enables quick and efficient surveys of the whole specimen. It is also compatible with live imaging, giving access to functional assays. CLEM protocols take advantage of the features to efficiently retrace the position of targeted sites when switching from one modality to the other. They more often rely on anatomical cues that are visible both by light and electron microscopy. We present here a simple workflow where multicellular specimens are embedded in minimal amounts of resin, exposing their surface topology that can be imaged by scanning electron microscopy (SEM). LM and SEM both benefit from a large field of view that can cover whole model organisms. As a result, targeting specific anatomic locations by focused ion beam-SEM (FIB-SEM) tomography becomes straightforward. We illustrate this application on three different model organisms, used in our laboratory: the zebrafish embryo Danio rerio, the marine worm Platynereis dumerilii, and the dauer larva of the nematode Caenorhabditis elegans. Here we focus on the experimental steps to reduce the amount of resin covering the samples and to image the specimens inside an FIB-SEM. We expect this approach to have widespread applications for volume electron microscopy on multiple model organisms. Copyright © 2017 Elsevier Inc. All rights reserved.

Trichite growth during oxidation of titanium and TA6V4 alloy by water vapor at high temperatures

NASA Technical Reports Server (NTRS)

Coddet, C.; Motte, F.; Sarrazin, P.

1982-01-01

Analysis by electron scanning microscope detected the formation of rutile trichites on the surface of specimens of titanium and titanium alloy TA6V4 oxidized in water vapor in the temperature range 650 to 950 C and the water vapor pressure range from 0.5 to 18 torr. In all specimens, two sublayers of rutile were formed: an external layer of basalt-like appearance, and a microcrystalline inner layer. Morphology of the trichites depends on temperature and the material (whether metal or alloy), but not on vapor pressure.

Diagnosis of electrocution: The application of scanning electron microscope and energy-dispersive X-ray spectroscopy in five cases.

PubMed

Visonà, S D; Chen, Y; Bernardi, P; Andrello, L; Osculati, A

2018-03-01

Deaths from electricity, generally, do not have specific findings at the autopsy. The diagnosis is commonly based on the circumstances of the death and the morphologic findings, above all the current mark. Yet, the skin injury due to an electrocution and other kinds of thermal injuries often cannot be differentiated with certainty. Therefore, there is a great interest in finding specific markers of electrocution. The search for the metallization of the skin through Scanning Electron Microscope equipped with Energy Dispersive X-Ray Spectroscopy (EDS) probe is of special importance in order to achieve a definite diagnosis in case of suspected electrocution. We selected five cases in which the electrocution was extremely likely considering the circumstances of the death. In each case a forensic autopsy was performed. Then, the skin specimens were stained with Hematoxylin Eosin and Perls. On the other hand, the skin lesions were examined with a scanning electron microscope equipped with EDS probe in order to evaluate the morphological ultrastructural features and the presence of deposits on the surface of the skin. The typical skin injury of the electrocution (current mark) were macroscopically detected in all of the cases. The microscopic examination of the skin lesions revealed the typical spherical vacuoles in the horny layer and, in the epidermis, the elongation of the cell nuclei as well as necrosis. Perls staining was negative in 4 out 6 cases. Ultrastructural morphology revealed the evident vacuolization of the horny layer, elongation of epidermic cells, coagulation of the elastic fibers. In the specimens collected from the site of contact with the conductor of case 1 and 2, the presence of the Kα peaks of iron was detected. In the corresponding specimens taken from cases 2, 4, 5 the microanalysis showed the Kα peaks of titanium. In case 3, titanium and carbon were found. In the suspicion of electrocution, the integrated use of different tools is recommended, including macroscopic observation, H&E staining, iron-specific staining, scanning electron microscopy and EDS microanalysis. Only the careful interpretation of the results provided by all these methods can allow the pathologist to correctly identify the cause of the death. Particularly, the present study suggests that the microanalysis (SEM-EDS) represents a very useful tool for the diagnosis of electrocution, allowing the detection and the identification of the metals embedded in the skin and their evaluation in the context of the ultrastructural morphology. Copyright © 2018. Published by Elsevier B.V.

Bio-reinforced self-healing concrete using magnetic iron oxide nanoparticles.

PubMed

Seifan, Mostafa; Sarmah, Ajit K; Ebrahiminezhad, Alireza; Ghasemi, Younes; Samani, Ali Khajeh; Berenjian, Aydin

2018-03-01

Immobilization has been reported as an efficient technique to address the bacterial vulnerability for application in bio self-healing concrete. In this study, for the first time, magnetic iron oxide nanoparticles (IONs) are being practically employed as the protective vehicle for bacteria to evaluate the self-healing performance in concrete environment. Magnetic IONs were successfully synthesized and characterized using different techniques. The scanning electron microscope (SEM) images show the efficient adsorption of nanoparticles to the Bacillus cells. Microscopic observation illustrates that the incorporation of the immobilized bacteria in the concrete matrix resulted in a significant crack healing behavior, while the control specimen had no healing characteristics. Analysis of bio-precipitates revealed that the induced minerals in the cracks were calcium carbonate. The effect of magnetic immobilized cells on the concrete water absorption showed that the concrete specimens supplemented with decorated bacteria with IONs had a higher resistance to water penetration. The initial and secondary water absorption rates in bio-concrete specimens were 26% and 22% lower than the control specimens. Due to the compatible behavior of IONs with the concrete compositions, the results of this study proved the potential application of IONs for developing a new generation of bio self-healing concrete.

Chemical characterization of selected LDEF polymeric materials

NASA Technical Reports Server (NTRS)

Young, Philip R.; Slemp, Wayne S.

1991-01-01

Chemical characterization of selected polymeric materials which received exposure on the Long Duration Exposure Facility (LDEF) is reported. The specimens examined include silvered fluorinated ethylene propylene Teflon thermal blanket material, polysulfone, epoxy, polyimide matrix resin/graphite fiber reinforced composites, and several high performance polymer films. These specimens came from numerous LDEF locations, and thus received different environmental exposures. The results to date show no significant change at the molecular level in the polymer that survived exposure. Scanning electron and scanning tunneling microscopes show resin loss and a texturing of some specimens which resulted in a change in optical properties. The potential effect of a silicon-containing molecular contamination on these materials is addressed. The possibility of continued post-exposure degradation of some polymeric films is also proposed.

Study of plastic strain localization mechanisms caused by nonequilibrium transitions in mesodefect ensembles under high-speed loading

DOE Office of Scientific and Technical Information (OSTI.GOV)

Sokovikov, Mikhail, E-mail: sokovikov@icmm.ru; Chudinov, Vasiliy; Bilalov, Dmitry

2015-10-27

The behavior of specimens dynamically loaded during split Hopkinson (Kolsky) bar tests in a regime close to simple shear conditions was studied. The lateral surface of the specimens was investigated in-situ using a high-speed infrared camera CEDIP Silver 450M. The temperature field distribution obtained at different time allowed one to trace the evolution of plastic strain localization. The process of target perforation involving plug formation and ejection was examined using a high-speed infrared camera and a VISAR velocity measurement system. The microstructure of tested specimens was analyzed using an optical interferometer-profiler and a scanning electron microscope. The development of plasticmore » shear instability regions has been simulated numerically.« less

Measurement of the minority carrier diffusion length and edge surface-recombination velocity in InP

NASA Technical Reports Server (NTRS)

Bailey, Sheila G.; Hakimzadeh, Roshanak

1993-01-01

A scanning electron microscope (SEM) was used to measure the electron (minority carrier) diffusion length (L(sub n)) and the edge surface-recombination velocity (V(sub s)) in zinc-doped Czochralski-grown InP wafers. Electron-beam-induced current (EBIC) profiles were obtained in specimens containing a Schottky barrier perpendicular to the scanned (edge) surface. An independent technique was used to measure V(sub s), and these values were used in a theoretical expression for normalized EBIC. A fit of the experimental data with this expression enabled us to determine L(sub n).

Preparing sternorrhynchous insects (Insecta: Hemiptera: Sternorrhyncha) for microscope examination: Hoyer’s mounting medium

USDA-ARS?s Scientific Manuscript database

Proper identification of aphids, scale insects, psyllids, and whitefles (Hemiptera: Sternorrhyncha) require preparation of the specimen on a microscope slide. This training video provides visual instruction on how to prepare sternorrhynchous specimens on microscope slides for examination and identi...

Preparing soft-bodied arthropods for microscope examination: Soft Scales (Insecta: Hemiptera: Coccidae)

USDA-ARS?s Scientific Manuscript database

Proper identification of soft scales (Hemiptera:Coccidae) requires preparation of the specimen on a microscope slide. This training video provides visual instruction on how to prepare soft scale specimens on microscope slides for examination and identification. Steps ranging from collection, speci...

Preparing soft-bodied arthropods for arthropods for microscope examination: Mealybugs (Insects: Hemiptera: Pseudococcidae)

USDA-ARS?s Scientific Manuscript database

Proper identification of mealybugs (Hemiptera: Pseudococcidae) require preparation of the specimen on a microscope slide. This training video provides visual instruction on how to prepare mealybug specimens on microscope slides for examination and identification. Steps ranging from collection, spec...

Processing of fallopian tube, ovary, and endometrial surgical pathology specimens: A survey of U.S. laboratory practices.

PubMed

Samimi, Goli; Trabert, Britton; Duggan, Máire A; Robinson, Jennifer L; Coa, Kisha I; Waibel, Elizabeth; Garcia, Edna; Minasian, Lori M; Sherman, Mark E

2018-03-01

Many high-grade serous carcinomas initiate in fallopian tubes as serous tubal intraepithelial carcinoma (STIC), a microscopic lesion identified with specimen processing according to the Sectioning and Extensive Examination of the Fimbria protocol (SEE-Fim). Given that the tubal origin of these cancers was recently recognized, we conducted a survey of pathology practices to assess processing protocols that are applied to gynecologic surgical pathology specimens in clinical contexts in which finding STIC might have different implications. We distributed a survey electronically to the American Society for Clinical Pathology list-serve to determine practice patterns and compared results between practice types by chi-square (χ2) tests for categorical variables. Free text comments were qualitatively reviewed. Survey responses were received from 159 laboratories (72 academic, 87 non-academic), which reported diverse specimen volumes and percentage of gynecologic samples. Overall, 74.1% of laboratories reported performing SEE-Fim for risk-reducing surgical specimens (82.5% academic versus 65.7% non-academic, p < 0.05). In specimens from surgery for benign indications in which initial microscopic sections showed an unanticipated suspicious finding, 75.9% of laboratories reported using SEE-Fim to process the remainder of the specimen (94.8% academic versus 76.4% non-academic, p < 0.01), and 84.6% submitted the entire fimbriae. Changes in the theories of pathogenesis of high-grade serous carcinoma have led to implementation of pathology specimen processing protocols that include detailed analysis of the fallopian tubes. These results have implications for interpreting trends in cancer incidence data and considering the feasibility of developing a bank of gynecologic tissues containing STIC or early cancer precursors. Published by Elsevier Inc.

Progress toward an aberration-corrected low energy electron microscope for DNA sequencing and surface analysis

PubMed Central

Mankos, Marian; Shadman, Khashayar; N'Diaye, Alpha T.; Schmid, Andreas K.; Persson, Henrik H. J.; Davis, Ronald W.

2012-01-01

Monochromatic, aberration-corrected, dual-beam low energy electron microscopy (MAD-LEEM) is a novel imaging technique aimed at high resolution imaging of macromolecules, nanoparticles, and surfaces. MAD-LEEM combines three innovative electron–optical concepts in a single tool: a monochromator, a mirror aberration corrector, and dual electron beam illumination. The monochromator reduces the energy spread of the illuminating electron beam, which significantly improves spectroscopic and spatial resolution. The aberration corrector is needed to achieve subnanometer resolution at landing energies of a few hundred electronvolts. The dual flood illumination approach eliminates charging effects generated when a conventional, single-beam LEEM is used to image insulating specimens. The low landing energy of electrons in the range of 0 to a few hundred electronvolts is also critical for avoiding radiation damage, as high energy electrons with kilo-electron-volt kinetic energies cause irreversible damage to many specimens, in particular biological molecules. The performance of the key electron–optical components of MAD-LEEM, the aberration corrector combined with the objective lens and a magnetic beam separator, was simulated. Initial results indicate that an electrostatic electron mirror has negative spherical and chromatic aberration coefficients that can be tuned over a large parameter range. The negative aberrations generated by the electron mirror can be used to compensate the aberrations of the LEEM objective lens for a range of electron energies and provide a path to achieving subnanometer spatial resolution. First experimental results on characterizing DNA molecules immobilized on Au substrates in a LEEM are presented. Images obtained in a spin-polarized LEEM demonstrate that high contrast is achievable at low electron energies in the range of 1–10 eV and show that small changes in landing energy have a strong impact on the achievable contrast. The MAD-LEEM approach promises to significantly improve the performance of a LEEM for a wide range of applications in the biosciences, material sciences, and nanotechnology where nanometer scale resolution and analytical capabilities are required. In particular, the microscope has the potential of delivering images of unlabeled DNA strands with nucleotide-specific contrast. This simplifies specimen preparation and significantly eases the computational complexity needed to assemble the DNA sequence from individual reads. PMID:23847748

Microscope collision protection apparatus

DOEpatents

DeNure, Charles R.

2001-10-23

A microscope collision protection apparatus for a remote control microscope which protects the optical and associated components from damage in the event of an uncontrolled collision with a specimen, regardless of the specimen size or shape. In a preferred embodiment, the apparatus includes a counterbalanced slide for mounting the microscope's optical components. This slide replaces the rigid mounts on conventional upright microscopes with a precision ball bearing slide. As the specimen contacts an optical component, the contacting force will move the slide and the optical components mounted thereon. This movement will protect the optical and associated components from damage as the movement causes a limit switch to be actuated, thereby stopping all motors responsible for the collision.

Coherence of a spin-polarized electron beam emitted from a semiconductor photocathode in a transmission electron microscope

DOE Office of Scientific and Technical Information (OSTI.GOV)

Kuwahara, Makoto, E-mail: kuwahara@esi.nagoya-u.ac.jp; Saitoh, Koh; Tanaka, Nobuo

2014-11-10

The brightness and interference fringes of a spin-polarized electron beam extracted from a semiconductor photocathode excited by laser irradiation are directly measured via its use in a transmission electron microscope. The brightness was 3.8 × 10{sup 7 }A cm{sup −2 }sr{sup −1} for a 30-keV beam energy with the polarization of 82%, which corresponds to 3.1 × 10{sup 8 }A cm{sup −2 }sr{sup −1} for a 200-keV beam energy. The resulting electron beam exhibited a long coherence length at the specimen position due to the high parallelism of (1.7 ± 0.3) × 10{sup −5 }rad, which generated interference fringes representative of a first-order correlation using an electron biprism. The beam also had amore » high degeneracy of electron wavepacket of 4 × 10{sup −6}. Due to the high polarization, the high degeneracy and the long coherence length, the spin-polarized electron beam can enhance the antibunching effect.« less

Preparing soft-bodied arthropods for arthropods for microscope examination: Armored Scales (Insects: Hemiptera: Diaspididae)

USDA-ARS?s Scientific Manuscript database

Proper identification of armored scales (Hemiptera: Diaspididae) requires preparation of the specimen on a microscope slide. This training video provides visual instruction on how to prepare armored scales specimens on microscope slides for examination and identification. Steps ranging from collect...

Micro-CT scouting for transmission electron microscopy of human tissue specimens

DOE Office of Scientific and Technical Information (OSTI.GOV)

Morales, A. G.; Stempinski, E. S.; XIAO, X.

Transmission electron microscopy (TEM) provides sub-nanometre-scale details in volumetric samples. Samples such as pathology tissue specimens are often stained with a metal element to enhance contrast, which makes them opaque to optical microscopes. As a result, it can be a lengthy procedure to find the region of interest inside a sample through sectioning. Here, we describe micro-CT scouting for TEM that allows noninvasive identification of regions of interest within a block sample to guide the sectioning step. In a tissue pathology study, a bench-top micro-CT scanner with 10 m resolution was used to determine the location of patches of themore » mucous membrane in osmium-stained human nasal scraping samples. Furthermore, once the regions of interest were located, the sample block was sectioned to expose that location, followed by ultra-thin sectioning and TEM to inspect the internal structure of the cilia of the membrane epithelial cells with nanometre resolution. This method substantially reduced the time and labour of the search process from typically 20 sections for light microscopy to three sections with no added sample preparation. Lay description Electron microscopy provides very high levels of detail in a small area, and thus the question of where to look in an opaque sample, such as a stained tissue specimen, needs to be answered by sectioning the sample in small steps and examining the sections under a light microscope, until the region of interest is found. The search process can be lengthy and labor intensive, especially for a study involving a large number of samples. Small areas of interest can be missed in the process if not enough regions are examined. We also describe a method to directly locate the region of interest within a whole sample using micro-CT imaging, bypassing the need of blindly sectioning. Micro-CT enables locating the region within 3D space; this information provides a guide for sectioning the sample to expose that precise location for high resolution electron microscopy imaging. In a human tissue specimen study, this method considerably reduced the time and labor of the search process.« less

Micro-CT scouting for transmission electron microscopy of human tissue specimens

DOE PAGES

Morales, A. G.; Stempinski, E. S.; XIAO, X.; ...

2016-02-08

Transmission electron microscopy (TEM) provides sub-nanometre-scale details in volumetric samples. Samples such as pathology tissue specimens are often stained with a metal element to enhance contrast, which makes them opaque to optical microscopes. As a result, it can be a lengthy procedure to find the region of interest inside a sample through sectioning. Here, we describe micro-CT scouting for TEM that allows noninvasive identification of regions of interest within a block sample to guide the sectioning step. In a tissue pathology study, a bench-top micro-CT scanner with 10 m resolution was used to determine the location of patches of themore » mucous membrane in osmium-stained human nasal scraping samples. Furthermore, once the regions of interest were located, the sample block was sectioned to expose that location, followed by ultra-thin sectioning and TEM to inspect the internal structure of the cilia of the membrane epithelial cells with nanometre resolution. This method substantially reduced the time and labour of the search process from typically 20 sections for light microscopy to three sections with no added sample preparation. Lay description Electron microscopy provides very high levels of detail in a small area, and thus the question of where to look in an opaque sample, such as a stained tissue specimen, needs to be answered by sectioning the sample in small steps and examining the sections under a light microscope, until the region of interest is found. The search process can be lengthy and labor intensive, especially for a study involving a large number of samples. Small areas of interest can be missed in the process if not enough regions are examined. We also describe a method to directly locate the region of interest within a whole sample using micro-CT imaging, bypassing the need of blindly sectioning. Micro-CT enables locating the region within 3D space; this information provides a guide for sectioning the sample to expose that precise location for high resolution electron microscopy imaging. In a human tissue specimen study, this method considerably reduced the time and labor of the search process.« less

Characterization of some biological specimens using TEM and SEM

NASA Astrophysics Data System (ADS)

Ghosh, Nabarun; Smith, Don W.

2009-05-01

The advent of novel techniques using the Transmission and Scanning Electron Microscopes improved observation on various biological specimens to characterize them. We studied some biological specimens using Transmission and Scanning Electron Microscopes. We followed negative staining technique with Phosphotungstic acid using bacterial culture of Bacillus subtilis. Negative staining is very convenient technique to view the structural morphology of different samples including bacteria, phage viruses and filaments in a cell. We could observe the bacterial cell wall and flagellum very well when trapped the negative stained biofilm from bacterial culture on a TEM grid. We cut ultra thin sections from the fixed root tips of Pisum sativum (Garden pea). Root tips were pre fixed with osmium tetroxide and post fixed with uranium acetate and placed in the BEEM capsule for block making. The ultrathin sections on the grid under TEM showed the granular chromatin in the nucleus. The protein bodies and large vacuoles with the storage materials were conspicuous. We followed fixation, critical point drying and sputter coating with gold to view the tissues with SEM after placing on stubs. SEM view of the leaf surface of a dangerous weed Tragia hispida showed the surface trichomes. These trichomes when break on touching releases poisonous content causing skin irritation. The cultured tissue from in vitro culture of Albizia lebbeck, a tree revealed the regenerative structures including leaf buds and stomata on the tissue surface. SEM and TEM allow investigating the minute details characteristic morphological features that can be used for classroom teaching.

Evaluation of the correctness of the feed selection based on the analysis of chip's shape

NASA Astrophysics Data System (ADS)

Chodor, Jaroslaw; Kukielka, Leon; Kaldunski, Pawel; Bohdal, Lukasz; Patyk, Radoslaw; Kulakowska, Agnieszka

2018-05-01

For the experiment needs, the own experiment plan was developed. The researches were carried out to determine the effect of variable, small feed on the chip's shape. To provide orthogonal free cutting special specimens were prepared. Obtained chips were divided according to shapes and also scanned using a scanning electron microscope (SEM). Conclusions from the experiments were given.

Histopathological and electron microscopic studies of lymphangiectasia of the small intestine in Behçet's disease

PubMed Central

Asakura, Hitoshi; Morita, Akira; Morishita, Tetsuo; Tsuchiya, Masaharu; Watanabe, Yoonosuke; Enomoto, Yasuhiro

1973-01-01

The gastrointestinal involvement and immunological findings in 16 patients with Behçet's disease are described. Four of 15 biopsy specimens of jejunal mucosa showed marked lymphangiectasia in the lamina propria similar to the appearance which was thought to be a characteristic finding in protein-losing enteropathy. None of the patients had hypoproteinaemia. Increases in serum immunoglobulin IgA were proved in six of 15 cases; in IgM, five of 15; and in IgG, one of 15. Electron microscopic studies showed that there were thousands of precipitated lymph protein bodies in the extracellular spaces of the lamina propria. Ulcers and healed ulcers of the large intestine were studied by light microscopy. ImagesFig. 1Fig. 2Fig. 3Fig. 4Fig. 7Fig. 8Fig. 9 PMID:4700433

Metal dusting behavior of 321 stainless steel: Effects of edge and corner

NASA Astrophysics Data System (ADS)

Chang, Chia-Hao; Tsai, Wen-Ta

2011-04-01

The metal dusting behavior of 321 stainless steel (SS) in a flowing mixed CO/H2/H2O gas stream at 600 °C for 500 h and 1000 h was investigated. The microstructures and chemical compositions of the reaction products at the surface and those in the substrate under the pits were examined by using a scanning electron microscope (SEM) and a transmission electron microscope (TEM), each combined with an energy dispersive spectrometer (EDS). The phenomenon of a pitting attack that occurred preferentially at the edges and corners of the specimens was the focus of this study. The carburization behavior in the steel substrate under the pits was also characterized. Matrix carbide in the form of Cr7C3 and grain boundary carbide in the form of Cr23C6 were identified by TEM.

Physical properties of concrete made with Apollo 16 lunar soil sample

NASA Technical Reports Server (NTRS)

Lin, T. D.; Love, H.; Stark, D.

1992-01-01

This paper describes the first phase of the long-term investigation for the construction of concrete lunar bases. In this phase, petrographic and scanning electron microscope examinations showed that the morphology and elemental composition of the lunar soil made it suitable for use as a fine aggregate for concrete. Based on this finding, calcium aluminate cement and distilled water were mixed with the lunar soil to fabricate test specimens. The test specimens consisted of a 1-in cube, a 1/2-in cube, and three 0.12 x 0.58 x 3.15-in beam specimens. Tests were performed on these specimens to determine compressive strength, modulus of rupture, modulus of elasticity, and thermal coefficient of expansion. Based on examination of the material and test results, it is concluded that lunar soil can be used as a fine aggregate for concrete.

Compression, bend, and tension studies on forged Al67Ti25Cr8 and Al66Ti25Mn(g) L1(2) compounds

NASA Technical Reports Server (NTRS)

Kumar, K. S.; Brown, S. A.; Whittenberger, J. D.

1991-01-01

Cast, homogenized, and isothermally forged aluminum-rich L1(2) compounds Al67Ti25Cr8 and Al66Ti25Mn(g) were tested in compression as a function of temperature and as a function of strain rate at elevated temperatures (1000 K and 1100 K). Three-point bend specimens were tested as a function of temperature in the range 300 K to 873 K. Strain gages glued on the tensile side of the ambient and 473 K specimens enabled direct strain measurements. A number of 'buttonhead' tensile specimens were electro-discharge machined, fine polished, and tested between ambient and 1073 K for yield strength and ductility as a function of temperature. Scanning electron microscope (SEM) examination of fracture surfaces from both the bend and tensile specimens revealed a gradual transition from transgranular cleavage to intergranular failure with increasing temperature.

A scanning electron microscope study and statistical analysis of adipocyte morphology in lipofilling: comparing the effects of harvesting and purification procedures with 2 different techniques.

PubMed

Rubino, Corrado; Mazzarello, Vittorio; Faenza, Mario; Montella, Andrea; Santanelli, Fabio; Farace, Francesco

2015-06-01

The aim of this study was to evaluate the effects on adipocyte morphology of 2 techniques of fat harvesting and of fat purification in lipofilling, considering that the number of viable healthy adipocytes is important in fat survival in recipient areas of lipofilling. Fat harvesting was performed in 10 female patients from flanks, on one side with a 2-mm Coleman cannula and on the other side with a 3-mm Mercedes cannula. Thirty milliliter of fat tissue from each side was collected and divided into three 10 mL syringes: A, B, and C. The fat inside syringe A was left untreated, the fat in syringe B underwent simple sedimentation, and the fat inside syringe C underwent centrifugation at 3000 rpm for 3 minutes. Each fat graft specimen was processed for examination under low-vacuum scanning electron microscope. Diameter (μ) and number of adipocytes per square millimeter and number of altered adipocytes per square millimeter were evaluated. Untreated specimens harvested with the 2 different techniques were first compared, then sedimented versus centrifuged specimens harvested with the same technique were compared. Statistical analysis was performed using Wilcoxon signed rank test. The number of adipocytes per square millimeter was statistically higher in specimens harvested with the 3-mm Mercedes cannula (P = 0.0310). The number of altered cells was statistically higher in centrifuged specimens than in sedimented ones using both methods of fat harvesting (P = 0.0080) with a 2-mm Coleman cannula and (P = 0.0050) with a 3-mm Mercedes cannula. Alterations in adipocyte morphology consisted in wrinkling of the membrane, opening of pore with leakage of oily material, reduction of cellular diameter, and total collapse of the cellular membrane. Fat harvesting by a 3-mm cannula results in a higher number of adipocytes and centrifugation of the harvested fat results in a higher number of morphologic altered cells than sedimentation.

Cryo-electron microscopy of membrane proteins.

PubMed

Goldie, Kenneth N; Abeyrathne, Priyanka; Kebbel, Fabian; Chami, Mohamed; Ringler, Philippe; Stahlberg, Henning

2014-01-01

Electron crystallography is used to study membrane proteins in the form of planar, two-dimensional (2D) crystals, or other crystalline arrays such as tubular crystals. This method has been used to determine the atomic resolution structures of bacteriorhodopsin, tubulin, aquaporins, and several other membrane proteins. In addition, a large number of membrane protein structures were studied at a slightly lower resolution, whereby at least secondary structure motifs could be identified.In order to conserve the structural details of delicate crystalline arrays, cryo-electron microscopy (cryo-EM) allows imaging and/or electron diffraction of membrane proteins in their close-to-native state within a lipid bilayer membrane.To achieve ultimate high-resolution structural information of 2D crystals, meticulous sample preparation for electron crystallography is of outmost importance. Beam-induced specimen drift and lack of specimen flatness can severely affect the attainable resolution of images for tilted samples. Sample preparations that sandwich the 2D crystals between symmetrical carbon films reduce the beam-induced specimen drift, and the flatness of the preparations can be optimized by the choice of the grid material and the preparation protocol.Data collection in the cryo-electron microscope using either the imaging or the electron diffraction mode has to be performed applying low-dose procedures. Spot-scanning further reduces the effects of beam-induced drift. Data collection using automated acquisition schemes, along with improved and user-friendlier data processing software, is increasingly being used and is likely to bring the technique to a wider user base.

Beam deceleration for block-face scanning electron microscopy of embedded biological tissue.

PubMed

Ohta, Keisuke; Sadayama, Shoji; Togo, Akinobu; Higashi, Ryuhei; Tanoue, Ryuichiro; Nakamura, Kei-ichiro

2012-04-01

The beam deceleration (BD) method for scanning electron microscopes (SEM) also referred to as "retarding" was applied to back-scattered electron (BSE) imaging of the flat block face of a resin embedded biological specimen under low accelerating voltage and low beam current conditions. BSE imaging was performed with 0-4 kV of BD on en bloc stained rat hepatocyte. BD drastically enhanced the compositional contrast of the specimen and also improved the resolution at low landing energy levels (1.5-3 keV) and a low beam current (10 pA). These effects also functioned in long working distance observation, however, stage tilting caused uncorrectable astigmatism in BD observation. Stage tilting is mechanically required for a FIB/SEM, so we designed a novel specimen holder to minimize the unfavorable tilting effect. The FIB/SEM 3D reconstruction using the new holder showed a reasonable contrast and resolution high enough to analyze individual cell organelles and also the mitochondrial cristae structures (~5 nm) of the hepatocyte. These results indicate the advantages of BD for block face imaging of biological materials such as cells and tissues under low-voltage and low beam current conditions. Copyright © 2011 Elsevier Ltd. All rights reserved.

Ultrastructure of selected struvite-containing urinary calculi from cats.

PubMed

Neumann, R D; Ruby, A L; Ling, G V; Schiffman, P S; Johnson, D L

1996-01-01

To elucidate the ultrastructural details of struvite-containing urinary calculi from cats. Specimens studied were inclusive of the range of textures visible during preliminary analysis by use of a stereoscopic dissecting microscope. Textural types, which were used to infer crystal growth conditions, were differentiated with regard to crystal habit, crystal size, growth orientation, and primary porosity. Thirty specimens were selected from a collection of approximately 1,600 feline urinary calculi: 20 of these were composed entirely of struvite, and 10 consisted of struvite and calcium phosphate (apatite). Qualitative and quantitative analyses of specimens included use of plain and polarized light microscopy, x-ray diffractometry, scanning electron microscopy with backscattered electron imagery, x-ray fluorescence scans, and electron probe microanalysis. Four textural types were recognized among struvite calculi, whereas 2 textural types of struvite-apatite calculi were described. The presence of minute, well interconnected primary pores in struvite-containing urinary calculi from cats is an important feature, which may promote possible interaction of calculi with changes in urine composition. Primary porosity, which can facilitate interaction between the calculus and changing urine composition, may explain the efficacy of dietary or medicinal manipulations to promote the dissolution of struvite-containing uroliths from this species.

High-Performance X-ray Detection in a New Analytical Electron Microscope

NASA Technical Reports Server (NTRS)

Lyman, C. E.; Goldstein, J. I.; Williams, D. B.; Ackland, D. W.; vonHarrach, S.; Nicholls, A. W.; Statham, P. J.

1994-01-01

X-ray detection by energy-dispersive spectrometry in the analytical electron microscope (AEM) is often limited by low collected X-ray intensity (P), modest peak-to-background (P/B) ratios, and limitations on total counting time (tau) due to specimen drift and contamination. A new AFM has been designed with maximization of P. P/B, and tau as the primary considerations. Maximization of P has been accomplished by employing a field-emission electron gun, X-ray detectors with high collection angles, high-speed beam blanking to allow only one photon into the detector at a time, and simultaneous collection from two detectors. P/B has been maximized by reducing extraneous background signals generated at the specimen holder, the polepieces and the detector collimator. The maximum practical tau has been increased by reducing specimen contamination and employing electronic drift correction. Performance improvments have been measured using the NIST standard Cr thin film. The 0-3 steradian solid angle of X-ray collection is the highest value available. The beam blanking scheme for X-ray detection provides 3-4 times greater throughput of X-rays at high count rates into a recorded spectrum than normal systems employing pulse-pileup rejection circuits. Simultaneous X-ray collection from two detectors allows the highest X-ray intensity yet recorded to be collected from the NIST Cr thin film. The measured P/B of 6300 is the highest level recorded for an AEM. In addition to collected X-ray intensity (cps/nA) and P/B measured on the standard Cr film, the product of these can be used as a figure-of-merit to evaluate instruments. Estimated minimum mass fraction (MMF) for Cr measured on the standard NIST Cr thin film is also proposed as a figure-of-merit for comparing X-ray detection in AEMs. Determinations here of the MMF of Cr detectable show at least a threefold improvement over previous instruments.

Experimental and numerical study of plastic shear instability under high-speed loading conditions

DOE Office of Scientific and Technical Information (OSTI.GOV)

Sokovikov, Mikhail, E-mail: sokovikov@icmm.ru, E-mail: naimark@icmm.ru; Chudinov, Vasiliy, E-mail: sokovikov@icmm.ru, E-mail: naimark@icmm.ru; Bilalov, Dmitry, E-mail: sokovikov@icmm.ru, E-mail: naimark@icmm.ru

2014-11-14

The behavior of specimens dynamically loaded during the split Hopkinson (Kolsky) bar tests in a regime close to simple shear conditions was studied. The lateral surface of the specimens was investigated in a real-time mode with the aid of a high-speed infra-red camera CEDIP Silver 450M. The temperature field distribution obtained at different time made it possible to trace the evolution of plastic strain localization. The process of target perforation involving plug formation and ejection was examined using a high-speed infra-red camera and a VISAR velocity measurement system. The microstructure of tested specimens was analyzed using an optical interferometer-profilometer andmore » a scanning electron microscope. The development of plastic shear instability regions has been simulated numerically.« less

An in-situ study in SEM of delamination in several graphite/epoxy composite material systems

NASA Technical Reports Server (NTRS)

Bradley, W. L.

1986-01-01

A three point bend fixture compatible with the current loading stage for the Scanning Electron Microscope was designed and fabricated. End-notched flexure tests were run on several materials. Work to date was on AS4/3502, T6T145/F155, and T6T145/F185. Fracture toughness was measured. Fracture of neat resin specimens was begun. The decrease in delaminatin fracture toughness, compared to neat resin toughness, due to rigid fiber filler and interlaminar failure is addressed. An experimental program was designed to try to determine the proper interpretatin for apparent microcracking in neat resin specimens.

Micro Electron MicroProbe and Sample Analyzer

NASA Technical Reports Server (NTRS)

Manohara, Harish; Bearman, Gregory; Douglas, Susanne; Bronikowski, Michael; Urgiles, Eduardo; Kowalczyk, Robert; Bryson, Charles

2009-01-01

A proposed, low-power, backpack-sized instrument, denoted the micro electron microprobe and sample analyzer (MEMSA), would serve as a means of rapidly performing high-resolution microscopy and energy-dispersive x-ray spectroscopy (EDX) of soil, dust, and rock particles in the field. The MEMSA would be similar to an environmental scanning electron microscope (ESEM) but would be much smaller and designed specifically for field use in studying effects of geological alteration at the micrometer scale. Like an ESEM, the MEMSA could be used to examine uncoated, electrically nonconductive specimens. In addition to the difference in size, other significant differences between the MEMSA and an ESEM lie in the mode of scanning and the nature of the electron source.

Effect of electric discharge machining on the fatigue life of Inconel 718

NASA Technical Reports Server (NTRS)

Jeelani, S.; Collins, M. R.

1988-01-01

The effect of electric discharge machining on the fatigue life of Inconel 718 alloy at room temperature was investigated. Data were generated in the uniaxial tension fatigue mode at ambient temperature using flat 3.175 mm thick specimens. The specimens were machined on a wire-cut electric discharge machine at cutting speeds ranging from 0.5 to 2 mm per minute. The specimens were fatigued at a selected stress, and the resulting fatigue lives compared with that of the virgin material. The surfaces of the fatigued specimens were examined under optical and scanning electron microscopes, and the roughness of the surfaces was measured using a standard profilometer. From the results of the investigation, it was concluded that the fatigue life of the specimens machined using EDM decreased slightly as compared with that of the virgin material, but remained unchanged as the cutting speed was changed. The results are explained using data produced employing microhardness measurements, profilometry, and optical and scanning microscopy.

Over-Aging Effect on Fracture Toughness of Beryllium Copper Alloy C17200

NASA Astrophysics Data System (ADS)

Jen, Kei-Peng; Xu, Liqun; Hylinski, Steven; Gildersleeve, Nate

2008-10-01

This study experimentally increased the fracture toughness of Beryllium Copper (CuBe) UNS C17200 alloy using three different age hardening processes. At the same time, the micro- and macro-fracture behavior of this alloy were comprehensively studied. ASTM E399 fracture toughness, tensile, and Charpy impact tests were conducted for all three heat-treated rods. The fracture surfaces were examined under both an optical microscope and a scanning electron microscope to investigate the failure mechanisms. Multiple test orientations were considered to explore isotropy. Increasing the temperature and duration at which age hardening was performed increased fracture toughness while decreasing ultimate tensile strength. The maximum fracture toughness was reached on the most overaged specimen, while retaining a serviceable tensile strength. The specimen test data allowed a relationship to be established among Charpy impact toughness, fracture toughness, and yield strength. Analysis of fracture behavior revealed an interesting relationship between fracture toughness and pre-cracking fatigue propagation rate.

Dynamic compressive behavior of Pr-Nd alloy at high strain rates and temperatures

DOE Office of Scientific and Technical Information (OSTI.GOV)

Wang Huanran; Cai Canyuan; Chen Danian

2012-07-01

Based on compressive tests, static on 810 material test system and dynamic on the first compressive loading in split Hopkinson pressure bar (SHPB) tests for Pr-Nd alloy cylinder specimens at high strain rates and temperatures, this study determined a J-C type [G. R. Johnson and W. H. Cook, in Proceedings of Seventh International Symposium on Ballistics (The Hague, The Netherlands, 1983), pp. 541-547] compressive constitutive equation of Pr-Nd alloy. It was recorded by a high speed camera that the Pr-Nd alloy cylinder specimens fractured during the first compressive loading in SHPB tests at high strain rates and temperatures. From highmore » speed camera images, the critical strains of the dynamic shearing instability for Pr-Nd alloy in SHPB tests were determined, which were consistent with that estimated by using Batra and Wei's dynamic shearing instability criterion [R. C. Batra and Z. G. Wei, Int. J. Impact Eng. 34, 448 (2007)] and the determined compressive constitutive equation of Pr-Nd alloy. The transmitted and reflected pulses of SHPB tests for Pr-Nd alloy cylinder specimens computed with the determined compressive constitutive equation of Pr-Nd alloy and Batra and Wei's dynamic shearing instability criterion could be consistent with the experimental data. The fractured Pr-Nd alloy cylinder specimens of compressive tests were investigated by using 3D supper depth digital microscope and scanning electron microscope.« less

Nanoscale imaging of whole cells using a liquid enclosure and a scanning transmission electron microscope.

PubMed

Peckys, Diana B; Veith, Gabriel M; Joy, David C; de Jonge, Niels

2009-12-14

Nanoscale imaging techniques are needed to investigate cellular function at the level of individual proteins and to study the interaction of nanomaterials with biological systems. We imaged whole fixed cells in liquid state with a scanning transmission electron microscope (STEM) using a micrometer-sized liquid enclosure with electron transparent windows providing a wet specimen environment. Wet-STEM images were obtained of fixed E. coli bacteria labeled with gold nanoparticles attached to surface membrane proteins. Mammalian cells (COS7) were incubated with gold-tagged epidermal growth factor and fixed. STEM imaging of these cells resulted in a resolution of 3 nm for the gold nanoparticles. The wet-STEM method has several advantages over conventional imaging techniques. Most important is the capability to image whole fixed cells in a wet environment with nanometer resolution, which can be used, e.g., to map individual protein distributions in/on whole cells. The sample preparation is compatible with that used for fluorescent microscopy on fixed cells for experiments involving nanoparticles. Thirdly, the system is rather simple and involves only minimal new equipment in an electron microscopy (EM) laboratory.

Microstructure Evolution and Mechanical Properties of Underwater Dry and Local Dry Cavity Welded Joints of 690 MPa Grade High Strength Steel

PubMed Central

Sun, Kun; Cui, Shuwan; Zeng, Min; Yi, Jianglong; Shen, Xiaoqin; Yi, Yaoyong

2018-01-01

Q690E high strength low alloy (HSLA) steel plays an important role in offshore structures. In addition, underwater local cavity welding (ULCW) technique was widely used to repair important offshore constructions. However, the high cooling rate of ULCW joints results in bad welding quality compared with underwater dry welding (UDW) joints. Q690E high strength low alloy steels were welded by multi-pass UDW and ULCW techniques, to study the microstructural evolution and mechanical properties of underwater welded joints. The microstructure and fracture morphology of welded joints were observed by scanning electron microscope and optical microscope. The elemental distribution in the microstructure was determined with an Electron Probe Microanalyzer. The results indicated that the microstructure of both two welded joints was similar. However, martensite and martensite-austenite components were significantly different with different underwater welding methods such that the micro-hardness of the HAZ and FZ in the ULCW specimen was higher than that of the corresponding regions in UDW joint. The yield strength and ultimate tensile strength of the ULCW specimen are 109 MPa lower and 77 MPa lower, respectively, than those of the UDW joint. The impact toughness of the UDW joint was superior to those of the ULCW joint. PMID:29361743

Microstructure Evolution and Mechanical Properties of Underwater Dry and Local Dry Cavity Welded Joints of 690 MPa Grade High Strength Steel.

PubMed

Shi, Yonghua; Sun, Kun; Cui, Shuwan; Zeng, Min; Yi, Jianglong; Shen, Xiaoqin; Yi, Yaoyong

2018-01-22

Q690E high strength low alloy (HSLA) steel plays an important role in offshore structures. In addition, underwater local cavity welding (ULCW) technique was widely used to repair important offshore constructions. However, the high cooling rate of ULCW joints results in bad welding quality compared with underwater dry welding (UDW) joints. Q690E high strength low alloy steels were welded by multi-pass UDW and ULCW techniques, to study the microstructural evolution and mechanical properties of underwater welded joints. The microstructure and fracture morphology of welded joints were observed by scanning electron microscope and optical microscope. The elemental distribution in the microstructure was determined with an Electron Probe Microanalyzer. The results indicated that the microstructure of both two welded joints was similar. However, martensite and martensite-austenite components were significantly different with different underwater welding methods such that the micro-hardness of the HAZ and FZ in the ULCW specimen was higher than that of the corresponding regions in UDW joint. The yield strength and ultimate tensile strength of the ULCW specimen are 109 MPa lower and 77 MPa lower, respectively, than those of the UDW joint. The impact toughness of the UDW joint was superior to those of the ULCW joint.

Scanning electron microscope analysis of gunshot defects to bone: an underutilized source of information on ballistic trauma.

PubMed

Rickman, John M; Smith, Martin J

2014-11-01

Recent years have seen increasing involvement by forensic anthropologists in the interpretation of skeletal trauma. With regard to ballistic injuries, there is now a large literature detailing gross features of such trauma; however, less attention has been given to microscopic characteristics. This article presents analysis of experimentally induced gunshot trauma in animal bone (Bos taurus scapulae) using full metal jacket (FMJ), soft point (SP), and captive bolt projectiles. The results were examined using scanning electron microscopy (SEM). Additional analysis was conducted on a purported parietal gunshot lesion in a human cranial specimen. A range of features was observed in these samples suggesting that fibrolamellar bone response to projectile impact is analogous to that observed in synthetic composite laminates. The results indicate that direction of bullet travel can be discerned microscopically even when it is ambiguous on gross examination. It was also possible to distinguish SP from FMJ lesions. SEM analysis is therefore recommended as a previously underexploited tool in the analysis of ballistic trauma. © 2014 American Academy of Forensic Sciences.

SEM-microphotogrammetry, a new take on an old method for generating high-resolution 3D models from SEM images.

PubMed

Ball, A D; Job, P A; Walker, A E L

2017-08-01

The method we present here uses a scanning electron microscope programmed via macros to automatically capture dozens of images at suitable angles to generate accurate, detailed three-dimensional (3D) surface models with micron-scale resolution. We demonstrate that it is possible to use these Scanning Electron Microscope (SEM) images in conjunction with commercially available software originally developed for photogrammetry reconstructions from Digital Single Lens Reflex (DSLR) cameras and to reconstruct 3D models of the specimen. These 3D models can then be exported as polygon meshes and eventually 3D printed. This technique offers the potential to obtain data suitable to reconstruct very tiny features (e.g. diatoms, butterfly scales and mineral fabrics) at nanometre resolution. Ultimately, we foresee this as being a useful tool for better understanding spatial relationships at very high resolution. However, our motivation is also to use it to produce 3D models to be used in public outreach events and exhibitions, especially for the blind or partially sighted. © 2017 The Authors Journal of Microscopy © 2017 Royal Microscopical Society.

Twisted ribbon structure of paired helical filaments revealed by atomic force microscopy.

PubMed Central

Pollanen, M. S.; Markiewicz, P.; Bergeron, C.; Goh, M. C.

1994-01-01

Progressive deposition of phosphorylated tau into the paired helical filaments (PHF) that compose neurofibrillary tangles, dystrophic neurites, and neuropil threads is an obligate feature of Alzheimer's disease. The standard model of PHF structure, derived from electron microscopic studies, suggests that two 8- to 10-nm filaments each composed of three to four protofilaments are wound into a helix with a maximal diameter of -20 nm and a half period of 65 to 80 nm. However, recent vertical platinum-carbon replicas of PHF more closely resemble a thin helical ribbon without constitutive protofilaments. Here we report that native PHF imaged with an atomic force microscope appear as twisted ribbons rather than the generally accepted structure derived from electron microscopic studies. These data imply that the assembly of PHF is not due to the twisting of pair-wise filaments but rather the helical winding of self-associated tau molecules arranged into a flattened structure. Future structural models of PHF should be based on quantitative data obtained from imaging techniques, such as scanning probe microscopy, which do not require harsh specimen preparation procedures. Images Figure 1 PMID:8178938

Twisted ribbon structure of paired helical filaments revealed by atomic force microscopy.

PubMed

Pollanen, M S; Markiewicz, P; Bergeron, C; Goh, M C

1994-05-01

Progressive deposition of phosphorylated tau into the paired helical filaments (PHF) that compose neurofibrillary tangles, dystrophic neurites, and neuropil threads is an obligate feature of Alzheimer's disease. The standard model of PHF structure, derived from electron microscopic studies, suggests that two 8- to 10-nm filaments each composed of three to four protofilaments are wound into a helix with a maximal diameter of -20 nm and a half period of 65 to 80 nm. However, recent vertical platinum-carbon replicas of PHF more closely resemble a thin helical ribbon without constitutive protofilaments. Here we report that native PHF imaged with an atomic force microscope appear as twisted ribbons rather than the generally accepted structure derived from electron microscopic studies. These data imply that the assembly of PHF is not due to the twisting of pair-wise filaments but rather the helical winding of self-associated tau molecules arranged into a flattened structure. Future structural models of PHF should be based on quantitative data obtained from imaging techniques, such as scanning probe microscopy, which do not require harsh specimen preparation procedures.

Phosphorus detection in vitrified bacteria by cryo-STEM annular dark-field analysis.

PubMed

Wolf, Sharon Grayer; Rez, Peter; Elbaum, Michael

2015-11-01

Bacterial cells often contain dense granules. Among these, polyphosphate bodies (PPBs) store inorganic phosphate for a variety of essential functions. Identification of PPBs has until now been accomplished by analytical methods that required drying or chemically fixing the cells. These methods entail large electron doses that are incompatible with low-dose imaging of cryogenic specimens. We show here that Scanning Transmission Electron Microscopy (STEM) of fully hydrated, intact, vitrified bacteria provides a simple means for mapping of phosphorus-containing dense granules based on quantitative sensitivity of the electron scattering to atomic number. A coarse resolution of the scattering angles distinguishes phosphorus from the abundant lighter atoms: carbon, nitrogen and oxygen. The theoretical basis is similar to Z contrast of materials science. EDX provides a positive identification of phosphorus, but importantly, the method need not involve a more severe electron dose than that required for imaging. The approach should prove useful in general for mapping of heavy elements in cryopreserved specimens when the element identity is known from the biological context. © 2015 The Authors Journal of Microscopy © 2015 Royal Microscopical Society.

Cooperative inter- and intra-layer lattice dynamics of photoexcited multi-walled carbon nanotubes studied by ultrafast electron diffraction.

PubMed

Sun, Shuaishuai; Li, Zhongwen; Li, Zi-An; Xiao, Ruijuan; Zhang, Ming; Tian, Huanfang; Yang, Huaixin; Li, Jianqi

2018-04-26

Optical tuning and probing ultrafast structural response of nanomaterials driven by electronic excitation constitute a challenging but promising approach for understanding microscopic mechanisms and applications in microelectromechanical systems and optoelectrical devices. Here we use pulsed electron diffraction in a transmission electron microscope to investigate laser-induced tubular lattice dynamics of multi-walled carbon nanotubes (MWCNTs) with varying laser fluence and initial specimen temperature. Our photoexcitation experiments demonstrate cooperative and inverse collective atomic motions in intralayer and interlayer directions, whose strengths and rates depend on pump fluence. The electron-driven and thermally driven structural responses with opposite amplitudes cause a crossover between intralayer and interlayer directions. Our ab initio calculations support these findings and reveal that electrons excited from π to π* orbitals in a carbon tube weaken the intralayer bonds while strengthening the interlayer bonds along the radial direction. Moreover, by probing the structural dynamics of MWCNTs at initial temperatures of 300 and 100 K, we uncover the concomitance of thermal and nonthermal dynamical processes and their mutual influence in MWCNTs. Our results illustrate the nature of electron-driven nonthermal process and electron-phonon thermalization in the MWCNTs, and bear implications for the intricate energy conversion and transfer in materials at the nanoscale.

The effect of immersion time to low carbon steel hardness and microstructure with hot dip galvanizing coating method

NASA Astrophysics Data System (ADS)

Hakim, A. A.; Rajagukguk, T. O.; Sumardi, S.

2018-01-01

Along with developing necessities of metal materials, these rise demands of quality improvements and material protections especially the mechanical properties of the material. This research used hot dip galvanizing coating method. The objectives of this research were to find out Rockwell hardness (HRb), layer thickness, micro structure and observation with Scanning Electron Microscope (SEM) from result of coating by using Hot Dip Galvanizing coating method with immersion time of 3, 6, 9, and 12 minutes at 460°C. The result shows that Highest Rockwell hardness test (HRb) was at 3 minutes immersion time with 76.012 HRb. Highest thickness result was 217.3 μm at 12 minutes immersion. Microstructure test result showed that coating was formed at eta, zeta, delta and gamma phases, while Scanning Electron Microscope (SEM) showed Fe, Zn, Mn, Si and S elements at the specimens after coating.

Membranous nephropathy (bubbling appearance and spike formation) without immunoglobulin deposition in a patient with systemic lupus erythematosus.

PubMed

Miura, Naoto; Mori, Yuki; Yoshino, Masabumi; Suga, Norihiro; Kitagawa, Wataru; Yamada, Harutaka; Nishikawa, Kazuhiro; Imai, Hirokazu

2008-12-01

A 53-year-old Japanese man with systemic lupus erythematosus developed proteinuria and hematuria after a urinary stone episode. A light microscopic study of a kidney biopsy specimen demonstrated a bubbling appearance and spike formation of the basement membrane. Immunofluorescent studies revealed that there were no significant depositions of immunoglobulins, such as IgG (-), IgA (-), IgM (+/-), kappa light chain (+/-), lambda light chain (+/-), or C3 (-) in the glomerular capillary wall, though C1q was present as one-plus positive staining in mesangial areas. Electron microscopic studies showed that the thickness of the basement membrane varied from thin to thick without electron dense deposits, and that the cellular components of the podocyte were irregularly present in the basement membrane. Urinary protein decreased after the usage of prednisolone and mizoribine; however, proteinuria aggravated after an episode of urinary stone during the same treatment.

The human uterotubal junction: a scanning electron microscope study during different phases of the menstrual cycle.

PubMed

Fadel, H E; Berns, D; Zaneveld, L J; Wilbanks, G D; Brueschke, E E

1976-10-01

Uterotubal junctions from surgically extirpated human uteri were examined. The specimens were obtained during different phases of the menstrual cycle. The interstitial portions of the tubes together with the cornual areas were dissected, excised, and their luminal surfaces exposed. The specimens were then processed for scanning electron microscopy. The surface epithelium of both the cornual endometrium and interstitial endosalpins. Ciliated cells were more numerous in the endosalpinx. Cyclic changes in ciliated cells were minimal, while cyclic secretory activity was demonstrated, especially in the endometrium. The transitional area between the endometrium and the endosalpinx was characterized by a marked increase in the number of ciliated cells, and a tendency of the secretory cells to assume a flattened, polygonal shape. These morphologic features suggest a possible role in the transport and/or maintenance of spermatozoa and/or ova.

High-resolution Surface Analysis by Microarea Auger Spectroscopy: Computerization and Characterization

NASA Technical Reports Server (NTRS)

Browning, R.

1986-01-01

A custom scanning Auger electron microscope (SAM) capable of introducing a 3-5 keV electron beam of several nA into a 30 nm diameter sample area was fitted with a sample introduction system and was fully computerized to be used for materials science research. The method of multispectral Auger imaging was devised and implemented. The instrument was applied to various problems in materials science, including the study of the fiber/matrix interface in a SiC reinforced titanium alloy, the study of SiC whiskers in Al alloy 2124 (in cooperation with NASA-Langley), the study of NiCrAl superalloys (in collaboration with NASA-Lewis), the study of zircalloy specimens (in collaboration with Stanford University), and the microstructure of sintered SiC specimens (in collaboration with NASA-Lewis). The report contains a number of manuscripts submitted for publication on these subjects.

Effect of prior deformation on microstructural development and Laves phase precipitation in high-chromium stainless steel.

PubMed

Hsiao, Z-W; Chen, D; Kuo, J-C; Lin, D-Y

2017-04-01

This study investigated the influence of deformation on precipitation behaviour and microstructure change during annealing. Here, the prior deformation of high-chromium stainless steel was tensile deformation of 3%, 6% and 10%, and the specimens were then annealed at 700˚C for 10 h. The specimens were subsequently analyzed using backscattered electron image and electron backscattering diffraction measurements with SEM. Compared with the deformation microstructure, the grains revealed no preferred orientation. The precipitates of TiN and NbC were formed homogenously in the grain interior and at grain boundaries after annealing. Fine Laves phase precipitates were observed in grains and along subgrain boundaries as the deformation increased. Furthermore, the volume fraction of Laves phase increased, but the average particle diameter of precipitate was reduced as the deformation increased. © 2017 The Authors Journal of Microscopy © 2017 Royal Microscopical Society.

Microstructure-Sensitive Investigation of Fracture Using Acoustic Emission Coupled With Electron Microscopy

NASA Technical Reports Server (NTRS)

Wisner, Brian; Cabal, Mike; Vanniamparambiland, Prashanth A.; Leser, William; Hochhalter, Jacob; Kontsos, Antonios

2015-01-01

A novel technique using Scanning Electron Microscopy (SEM) in conjunction with Acoustic Emission (AE) monitoring is proposed to investigate microstructure-sensitive fatigue and fracture of metals. The coupling between quasi in situ microscopy with actual in situ nondestructive evaluation falls into the ICME framework and the idea of quantitative data-driven characterization of material behavior. To validate the use of AE monitoring inside the SEM chamber, Aluminum 2024-B sharp notch specimen were tested both inside and outside the microscope using a small scale mechanical testing device. Subsequently, the same type of specimen was tested inside the SEM chamber. Load data were correlated with both AE information and observations of microcracks around grain boundaries as well as secondary cracks, voids, and slip bands. The preliminary results are in excellent agreement with similar findings at the mesoscale. Extensions of the application of this novel technique are discussed.

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.

[Evaluation of the effect of modified wet particle erosion on bond strength between 3Y-TZP zirconia framework and veneering porcelain].

PubMed

Guo, Jing; Zhu, Jia; Liu, Hon-Guang; Zhu, Hong-Shui

2017-02-01

To evaluate the effect of bond strength between 3mol% yttrium-stabilized tetragonal zirconium polycrystal (3Y-TZP) zirconia framework after modified wet particle erosion and veneering porcelain. A total of 174 [8 mm× 8 mm× 3 mm (±0.02)] specimens were prepared and then randomly divided into different groups according to different particle size, sandblasting pressure, sandblasting time through conventional sandblasting (experimental groups); specimens in the control group were not sandblasted. The bond strength between 3Y-TZP zirconia framework and veneering porcelain was measured using a universal testing machine. Statistical analysis was performed using SPSS17.0 software package. The bond strength of specimens treated by two methods-conventional sandblasting and modified wet particle erosion under the same condition (150 μm, 0.6 MPa, 30 s) were acquired, while the surface was analyzed microscopically before the specimens were veneered with the veneering ceramic under scanning electronic microscope (SEM). The bond strength of conventional sandblasting group under the condition (110 μm, 0.4 MPa,30 s) was maximal, and significantly higher than the control group (P<0.05). The bond strength of modified wet particle erosion group was significantly higher than the conventional sandblasting group and control group (P<0.01); SEM showed that the surface of specimen with modified wet particle erosion was homogeneously rough; the surface specimen with conventional sandblasting was heterogeneously rough, some microcracks were seen in some areas. Sandblasting can slightly enhance the bond strength between 3Y-TZP zirconia framework and veneering porcelain, modified wet particle erosion is recommended for 3Y-TZP zirconia framework surface treatment.

Performance of low-voltage STEM/TEM with delta corrector and cold field emission gun.

PubMed

Sasaki, Takeo; Sawada, Hidetaka; Hosokawa, Fumio; Kohno, Yuji; Tomita, Takeshi; Kaneyama, Toshikatsu; Kondo, Yukihito; Kimoto, Koji; Sato, Yuta; Suenaga, Kazu

2010-08-01

To reduce radiation damage caused by the electron beam and to obtain high-contrast images of specimens, we have developed a highly stabilized transmission electron microscope equipped with a cold field emission gun and spherical aberration correctors for image- and probe-forming systems, which operates at lower acceleration voltages than conventional transmission electron microscopes. A delta-type aberration corrector is designed to simultaneously compensate for third-order spherical aberration and fifth-order 6-fold astigmatism. Both were successfully compensated in both scanning transmission electron microscopy (STEM) and transmission electron microscopy (TEM) modes in the range 30-60 kV. The Fourier transforms of raw high-angle annular dark field (HAADF) images of a Si[110] sample revealed spots corresponding to lattice spacings of 111 and 96 pm at 30 and 60 kV, respectively, and those of raw TEM images of an amorphous Ge film with gold particles showed spots corresponding to spacings of 91 and 79 pm at 30 and 60 kV, respectively. Er@C(82)-doped single-walled carbon nanotubes, which are carbon-based samples, were successfully observed by HAADF-STEM imaging with an atomic-level resolution.

Direct observation of dopant distribution in GaAs compound semiconductors using phase-shifting electron holography and Lorentz microscopy.

PubMed

Sasaki, Hirokazu; Otomo, Shinya; Minato, Ryuichiro; Yamamoto, Kazuo; Hirayama, Tsukasa

2014-06-01

Phase-shifting electron holography and Lorentz microscopy were used to map dopant distributions in GaAs compound semiconductors with step-like dopant concentration. Transmission electron microscope specimens were prepared using a triple beam focused ion beam (FIB) system, which combines a Ga ion beam, a scanning electron microscope, and an Ar ion beam to remove the FIB damaged layers. The p-n junctions were clearly observed in both under-focused and over-focused Lorentz microscopy images. A phase image was obtained by using a phase-shifting reconstruction method to simultaneously achieve high sensitivity and high spatial resolution. Differences in dopant concentrations between 1 × 10(19) cm(-3) and 1 × 10(18) cm(-3) regions were clearly observed by using phase-shifting electron holography. We also interpreted phase profiles quantitatively by considering inactive layers induced by ion implantation during the FIB process. The thickness of an inactive layer at different dopant concentration area can be measured from the phase image. © 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.

Fatigue crack closure behavior at high stress ratios

NASA Technical Reports Server (NTRS)

Turner, C. Christopher; Carman, C. Davis; Hillberry, Ben M.

1988-01-01

Fatigue crack delay behavior at high stress ratio caused by single peak overloads was investigated in two thicknesses of 7475-T731 aluminum alloy. Closure measurements indicated no closure occurred before or throughout the overload plastic zones following the overload. This was further substantiated by comparing the specimen compliance following the overload with the compliance of a low R ratio test when the crack was fully open. Scanning electron microscope studies revealed that crack tunneling and possibly reinitiation of the crack occurred, most likely a result of crack-tip blunting. The number of delay cycles was greater for the thinner mixed mode stress state specimen than for the thicker plane strain stress state specimen, which is similar to low R ratio test results and may be due to a larger plastic zone for the mixed mode cased.

Measurement of Heavy Ion Irradiation Induced In-Plane Strain in Patterned Face-Centered-Cubic Metal Films: An in Situ Study

DOE Office of Scientific and Technical Information (OSTI.GOV)

Yu, K. Y.; Chen, Y.; Li, J.

Nanocrystalline Ag, Cu, and Ni thin films and their coarse grained counterparts are patterned in this paper using focused ion beam and then irradiated by Kr ions within an electron microscope at room temperature. Irradiation induced in-plane strain of the films is measured by tracking the location of nanosized holes. The magnitude of the strain in all specimens is linearly dose-dependent and the strain rates of nanocrystalline metals are significantly greater as compared to that of the coarse grained metals. Finally, real-time microscopic observation suggests that substantial grain boundary migration and grain rotation are responsible for the significant in-plane strain.

Measurement of Heavy Ion Irradiation Induced In-Plane Strain in Patterned Face-Centered-Cubic Metal Films: An in Situ Study

DOE PAGES

Yu, K. Y.; Chen, Y.; Li, J.; ...

2016-11-28

Nanocrystalline Ag, Cu, and Ni thin films and their coarse grained counterparts are patterned in this paper using focused ion beam and then irradiated by Kr ions within an electron microscope at room temperature. Irradiation induced in-plane strain of the films is measured by tracking the location of nanosized holes. The magnitude of the strain in all specimens is linearly dose-dependent and the strain rates of nanocrystalline metals are significantly greater as compared to that of the coarse grained metals. Finally, real-time microscopic observation suggests that substantial grain boundary migration and grain rotation are responsible for the significant in-plane strain.

Boehmite nanostructures preparation by hydrothermal method from anodic aluminium oxide membrane.

PubMed

Yang, X; Wang, J Y; Pan, H Y

2009-02-01

Boehmite nanostructures were successfully synthesized from porous anodic aluminium oxide (AAO) membrane by a simple and efficient hydro-thermal method. The experiment used high purity alumina as raw material, and the whole reaction process avoided superfluous impurities to be introduced. Thus, the purity of Boehmite products was ensured. The examinations of the morphology and structure were carried out by atomic force microscope (AFM), scanning electron microscopy (SEM) and transmission electron microscopy (TEM). Composition of the specimens was analyzed using energy dispersive X-ray spectroscope (EDX) and X-ray diffraction (XRD). Based on these observations the growth process was analyzed.

Laser shock peening without coating induced residual stress distribution, wettability characteristics and enhanced pitting corrosion resistance of austenitic stainless steel

NASA Astrophysics Data System (ADS)

Prabhakaran, S.; Kulkarni, Aniket; Vasanth, G.; Kalainathan, S.; Shukla, Pratik; Vasudevan, Vijay K.

2018-01-01

Low energy laser shock peening without coating (LSPwC) was conducted on AISI 304 austenitic stainless steel specimens with varying pulse densities or overlapping. Highest magnitude of compressive residual stress (CRS) was achieved for an optimized pulse density of 2500 pulses/cm2 (75% overlapping). The 2-D and 3-D topographical analysis were indicative of the fact that controlled roughening of the surface was achieved after the LSPwC process. After the LSPwC process, the hydrophilic unpeened surface was converted into the hydrophobic surface, thus decreasing the wettability characteristics of the surface. The X-ray diffraction (XRD) results reveal that there is a beginning of the martensite transformation and the rise in the intensity value of the peaks after LSPwC indicates the presence of compressive residual stresses induced in the specimen. The optical microscope and high-resolution transmission electron microscope results provided evidence of grain refinement and deformation induced refinement features such as multidirectional mechanical twinning, dislocations lines, micro shear cells and stacking faults in the near and sub-surface areas. The average hardness value of the LSPwC specimens was found to be increased by 28% more than the untreated specimen. The potentiodynamic polarization revealed that there was a considerable amount of increase in the pitting corrosion resistance after the LSPwC process, thus, supporting to extend the fatigue life of the specimen. The electrochemical impedance spectroscopic (EIS) analysis depicts that the LSPwC process supports the formation of the strong passivation layer in 3.5% NaCl solution.

A spherical electron-channelling pattern map for use in quartz petrofabric analysis

USGS Publications Warehouse

Lloyd, G.E.; Ferguson, C.C.

1986-01-01

Electron channelling patterns (ECP's) are formed in the scanning electron microscope (SEM) by the interaction between the incident electrons and the lattice of crystalline specimens. The patterns are unique for a particular crystallographic orientation and are therefore of considerable potential in petrofabric studies provided they can be accurately indexed. Indexing requires an ECP-map of the crystallographic stereogram or unit triangle covering all possible orientations and hence ECP patterns. Due to the presence of long-range distortions in planar ECP-maps, it is more convenient to construct the maps over a spherical surface. This also facilitates the indexing of individual ECP's. A spherical ECP-map for quartz is presented together with an example of its use in petrofabric analysis. ?? 1986.

A Survey of the Use of Iterative Reconstruction Algorithms in Electron Microscopy

PubMed Central

Otón, J.; Vilas, J. L.; Kazemi, M.; Melero, R.; del Caño, L.; Cuenca, J.; Conesa, P.; Gómez-Blanco, J.; Marabini, R.; Carazo, J. M.

2017-01-01

One of the key steps in Electron Microscopy is the tomographic reconstruction of a three-dimensional (3D) map of the specimen being studied from a set of two-dimensional (2D) projections acquired at the microscope. This tomographic reconstruction may be performed with different reconstruction algorithms that can be grouped into several large families: direct Fourier inversion methods, back-projection methods, Radon methods, or iterative algorithms. In this review, we focus on the latter family of algorithms, explaining the mathematical rationale behind the different algorithms in this family as they have been introduced in the field of Electron Microscopy. We cover their use in Single Particle Analysis (SPA) as well as in Electron Tomography (ET). PMID:29312997

The Hole-Count Test Revisited: Effects of Test Specimen Thickness

NASA Technical Reports Server (NTRS)

Lyman, C. E.; Ackland, D. W.; Williams, D. B.; Goldstein, J. I.

1989-01-01

For historical reasons the hole count, an important performance test for the Analytical Electron Microscope (AEM), is somewhat arbitrary yielding different numbers for different investigators. This was not a problem a decade ago when AEM specimens were often bathed with large fluxes of stray electrons and hard x rays. At that time the presence or absence of a thick Pt second condenser (C2) aperture could be detected by a simple comparison of the x-ray spectrum taken 'somewhere in the hole' with a spectrum collected on a 'typical thickness' of Mo or Ag foil. A high hole count of about 10-20% indicated that the electron column needed modifications; whereas a hole count of 1-2% was accepted for most AEM work. The absolute level of the hole count is a function of test specimen atomic number, overall specimen shape, and thin-foil thickness. In order that equivalent results may be obtained for any AEM in any laboratory in the world, this test must become standardized. The hole-count test we seek must be as simpl and as nonsubjective as the graphite 0.344nm lattice-line-resolution test. This lattice-resolution test spurred manufacturers to improve the image resolution of the TEM significantly in the 1970s and led to the even more stringent resolution tests of today. A similar phenomenon for AEM instruments would be welcome. The hole-count test can also indicate whether the spurious x-ray signal is generated by high-energy continuum x rays (bremsstrahlung) generated in the electron column (high K-line to L-line ratio) or uncollimated electrons passing through or around the C2 aperture (low K/L ratio).

Methods for characterizing plant fibers.

PubMed

Cruthers, Natasha; Carr, Debra; Niven, Brian; Girvan, Elizabeth; Laing, Raechel

2005-08-01

The effectiveness of different microscopy techniques for measuring the dimensions of ultimate fibers from harakeke (Phormium tenax, New Zealand flax) was investigated using a factorial experimental design. Constant variables were geographical location, location of specimens along the leaf, season (winter), individual plant, a fourth leaf from a north-facing fan, age of plant, and cultivars (two). Experimental variables were microscopy techniques and measurement axis. Measurements of width and length of harakeke ultimate fibers depended on the microscopic preparation/technique used as well as the cultivar examined. The best methods were (i) transverse sections of leaf specimens 4 microm thick, embedded in Paraplast and observed using light microscopy, and (ii) nonfixed ultimate fibers observed using scanning electron microscopy. (c) 2005 Wiley-Liss, Inc.

High-Cycle Fatigue of High-Strength Low Alloy Steel Q345 Subjected to Immersion Corrosion for Mining Wheel Applications

NASA Astrophysics Data System (ADS)

Dicecco, Sante; Altenhof, William; Hu, Henry; Banting, Richard

2017-04-01

In an effort to better understand the impact of material degradation on the fatigue life of mining wheels made of a high-strength low alloy carbon steel (Q345), this study seeks to evaluate the effect of surface corrosion on the high-cycle fatigue behavior of the Q345 alloy. The fatigue behavior of the polished and corroded alloy was investigated. Following exposure to a 3.5 wt.% NaCl saltwater solution, polished and corroded fatigue specimens were tested using an R.R. Moore rotating-bending fatigue apparatus. Microstructural analyses via both optical microscopy and scanning electron microscopy (SEM) revealed that one major phase, α-iron phase, ferrite, and one minor phase, colony pearlite, existed in the extracted Q345 alloy. The results of the fatigue testing showed that the polished and corroded specimens had an endurance strength of approximately 295 and 222 MPa, respectively, at 5,000,000 cycles. The corroded surface condition resulted in a decrease in the fatigue strength of the Q345 alloy by 24.6%. Scanning electron microscope fractography indicated that failure modes for polished and corroded fatigue specimens were consistent in the high-cycle low loading fatigue regime. Conversely, SEM fractography of low-cycle high-loading fatigue specimens found considerable differences in fracture surfaces between the corroded and polished fatigue specimens.

Apparatus and methods for controlling electron microscope stages

DOE Office of Scientific and Technical Information (OSTI.GOV)

Duden, Thomas

Methods and apparatus for generating an image of a specimen with a microscope (e.g., TEM) are disclosed. In one aspect, the microscope may generally include a beam generator, a stage, a detector, and an image generator. A plurality of crystal parameters, which describe a plurality of properties of a crystal sample, are received. In a display associated with the microscope, an interactive control sphere based at least in part on the received crystal parameters and that is rotatable by a user to different sphere orientations is presented. The sphere includes a plurality of stage coordinates that correspond to a pluralitymore » of positions of the stage and a plurality of crystallographic pole coordinates that correspond to a plurality of polar orientations of the crystal sample. Movement of the sphere causes movement of the stage, wherein the stage coordinates move in conjunction with the crystallographic coordinates represented by pole positions so as to show a relationship between stage positions and the pole positions.« less

Investigation of the aluminium-aluminium oxide reversible transformation as observed by hot stage electron microscopy.

NASA Technical Reports Server (NTRS)

Grove, C. A.; Judd, G.; Ansell, G. S.

1972-01-01

Thin foils of high purity aluminium and an Al-Al2O3 SAP type of alloy were oxidised in a specially designed hot stage specimen chamber in an electron microscope. Below 450 C, amorphous aluminium oxide formed on the foil surface and was first detectable at foil edges, holes, and pits. Islands of aluminium then nucleated in this amorphous oxide. The aluminium islands displayed either a lateral growth with eventual coalescence with other islands, or a reoxidation process which caused the islands to disappear. The aluminium island formation was determined to be related to the presence of the electron beam. A mechanism based upon electron charging due to the electron beam was proposed to explain the nucleation, growth, coalescence, disappearance, and geometry of the aluminium islands.

High resolution IVEM tomography of biological specimens

DOE Office of Scientific and Technical Information (OSTI.GOV)

Sedat, J.W.; Agard, D.A.

Electron tomography is a powerful tool for elucidating the three-dimensional architecture of large biological complexes and subcellular organelles. The introduction of intermediate voltage electron microscopes further extended the technique by providing the means to examine very large and non-symmetrical subcellular organelles, at resolutions beyond what would be possible using light microscopy. Recent studies using electron tomography on a variety of cellular organelles and assemblies such as centrosomes, kinetochores, and chromatin have clearly demonstrated the power of this technique for obtaining 3D structural information on non-symmetric cell components. When combined with biochemical and molecular observations, these 3D reconstructions have provided significantmore » new insights into biological function.« less

Understanding Imaging and Metrology with the Helium Ion Microscope

NASA Astrophysics Data System (ADS)

Postek, Michael T.; Vladár, András E.; Ming, Bin

2009-09-01

One barrier to innovation confronting all phases of nanotechnology is the lack of accurate metrology for the characterization of nanomaterials. Ultra-high resolution microscopy is a key technology needed to achieve this goal. But, current microscope technology is being pushed to its limits. The scanning and transmission electron microscopes have incrementally improved in performance and other scanned probe technologies such as atomic force microscopy, scanning tunneling microscopy and focused ion beam microscopes have all been applied to nanotechnology with various levels of success. A relatively new tool for nanotechnology is the scanning helium ion microscope (HIM). The HIM is a new complementary imaging and metrology technology for nanotechnology which may be able to push the current resolution barrier lower. But, successful imaging and metrology with this instrument entails new ion beam/specimen interaction physics which must be fully understood. As a new methodology, HIM is beginning to show promise and the abundance of potentially advantageous applications for nanotechnology have yet to be fully exploited. This presentation will discuss some of the progress made at NIST in understanding the science behind this new technique.

Correlated light and electron microscopy observations of the uterine epithelial cell actin cytoskeleton using fluorescently labeled resin-embedded sections.

PubMed

Moore, Chad L; Cheng, Delfine; Shami, Gerald J; Murphy, Christopher R

2016-05-01

In order to perform correlative light and electron microscopy (CLEM) more precisely, we have modified existing specimen preparation protocols allowing fluorescence retention within embedded and sectioned tissue, facilitating direct observation across length scales. We detail a protocol which provides a precise correlation accuracy using accessible techniques in biological specimen preparation. By combining a pre-embedding uranyl acetate staining step with the progressive lowering of temperature (PLT) technique, a methacrylate embedded tissue specimen is ultrathin sectioned and mounted onto a TEM finder grid for immediate viewing in the confocal and electron microscope. In this study, the protocol is applied to rat uterine epithelial cells in vivo during early pregnancy. Correlative overlay data was used to track changes in filamentous actin that occurs in these cells from fertilization (Day 1) to implantation on Day 6 as part of the plasma membrane transformation, a process essential in the development of uterine receptivity in the rat. CLEM confirmed that the actin cytoskeleton is disrupted as apical microvilli are progressively lost toward implantation, and revealed the thick and continuous terminal web is replaced by a thinner and irregular actin band, with individually distinguishable filaments connecting actin meshworks which correspond with remaining plasma membrane protrusions. Copyright © 2016 Elsevier Ltd. All rights reserved.

The erosive potential of soft drinks on enamel surface substrate: an in vitro scanning electron microscopy investigation.

PubMed

Owens, Barry M; Kitchens, Michael

2007-11-01

Using scanning electron and light microscopy, this study qualitatively evaluated the erosive potential of carbonated cola beverages as well as sports and high-energy drinks on enamel surface substrate. Beverages used in this study included: Coca Cola Classic, Diet Coke, Gatorade sports drink, Red Bull high-energy drink, and tap water (control). Extracted human permanent molars free of hypocalcification and/or caries were used in this study. The coronal portion of each tooth was removed and sectioned longitudinally from the buccal to the lingual surface. The crown sections were embedded in acrylic resin, leaving the enamel surfaces exposed. Following finishing and polishing of all surfaces, one side was covered with red nail varnish while the remaining side was exposed to individual beverage immersion for 14 days, 24 hours per day, at 37 degrees C. The specimens were evaluated for enamel surface changes using scanning electron and light microscopy. Enamel specimens exhibited visual surface changes following immersion in the test beverages with Red Bull and Gatorade revealing the most striking surface morphological changes. Specimens subjected to Coca Cola Classic and Diet Coke immersion also displayed irregular post-treatment surface morphology. As verified by microscopic evaluation, all test beverages displayed enamel dissolution in the following order: Red Bull>Gatorade>Coca-Cola Classic>Diet Coke.

Atom probe trajectory mapping using experimental tip shape measurements.

PubMed

Haley, D; Petersen, T; Ringer, S P; Smith, G D W

2011-11-01

Atom probe tomography is an accurate analytical and imaging technique which can reconstruct the complex structure and composition of a specimen in three dimensions. Despite providing locally high spatial resolution, atom probe tomography suffers from global distortions due to a complex projection function between the specimen and detector which is different for each experiment and can change during a single run. To aid characterization of this projection function, this work demonstrates a method for the reverse projection of ions from an arbitrary projection surface in 3D space back to an atom probe tomography specimen surface. Experimental data from transmission electron microscopy tilt tomography are combined with point cloud surface reconstruction algorithms and finite element modelling to generate a mapping back to the original tip surface in a physically and experimentally motivated manner. As a case study, aluminium tips are imaged using transmission electron microscopy before and after atom probe tomography, and the specimen profiles used as input in surface reconstruction methods. This reconstruction method is a general procedure that can be used to generate mappings between a selected surface and a known tip shape using numerical solutions to the electrostatic equation, with quantitative solutions to the projection problem readily achievable in tens of minutes on a contemporary workstation. © 2011 The Authors Journal of Microscopy © 2011 Royal Microscopical Society.

Fracture resistance of Nd:YAG laser-welded cast titanium joints with various clinical thicknesses and welding pulse energies.

PubMed

Lin, Mau-Chin; Lin, Sheng-Chieh; Wang, Yu-Tsai; Hu, Suh-Woan; Lee, Tzu-Hsin; Chen, Li-Kai; Huang, Her-Hsiung

2007-05-01

The purpose of this study was to evaluate the fracture resistance of Nd:YAG laser-welded cast titanium (Ti) joints with various clinical thicknesses and welding pulse energies. A four-point bending test was used to assess the effects of various specimen thicknesses (1-3 mm) and welding pulse energies (11-24 J) on the fracture resistance of Nd:YAG laser-welded Ti dental joints. Fracture resistance was evaluated in terms of the ratio of the number of fractured specimens to the number of tested specimens. As for the fracture frequencies, they were compared using the Cochran-Mantel-Haenszel test. Morphology of the fractured Ti joints was observed using a scanning electron microscope. Results showed that decreasing the specimen thickness and/or increasing the welding pulse energy, i.e., increasing the welded area percentage, resulted in an increase in the fracture resistance of the Ti joint. Where fracture occurred, the fracture site would be at the center of the weld metal.

Artifacts introduced by ion milling in Al-Li-Cu alloys.

PubMed

Singh, A K; Imam, M A; Sadananda, K

1988-04-01

Ion milling is commonly used to prepare specimens for observation under transmission electron microscope (TEM). This technique sometimes introduces artifacts in specimens contributing to misleading interpretation of TEM results as observed in the present investigation of Al-Li-Cu alloys. This type of alloy, in general, contains several kinds of precipitates, namely delta', T1, and theta'. It is found that ion milling even for a short time produces drastic changes in the precipitate characteristics as compared to standard electropolishing methods of specimen preparation for TEM. Careful analysis of selected area diffraction patterns and micrographs shows that after ion milling delta' precipitates are very irregular, whereas other precipitates coarsen and they are surrounded by misfit dislocations. In situ hot-stage TEM experiments were performed to relate the microstructure to that observed in the ion-milled specimen. Results and causes of ion milling effects on the microstructure are discussed in relation to standard electropolishing techniques and in situ hot-stage experiment.

Linking Regions between Helices in Bacteriorhodopsin Revealed

PubMed Central

Agard, David A.; Stroud, Robert M.

1982-01-01

Three-dimensional electron-microscopic structural analysis requires the combination of many different tilted views of the same specimen. The relative difficulty of tilting the sample to high angles >60° without introducing severe distortion due to different focal distances across the specimen entails that the observable range of electron diffraction data is often limited to this range of angles. Thus, it is generally not possible to observe the diffraction maxima that lie within the conical region of reciprocal space around the direction perpendicular to the electron microscope grid. The absence of data in this region leads to a predictable distortion in the object, and for ±60° tilting makes the resolution essentially twice as bad in the direction perpendicular to the grid as it is for the in-plane image. Constrained density map modification and refinement methods can significantly reduce these effects. A method has been developed, tested on model cases, and applied to the electron-microscopic structure determination of bacteriorhodopsin in order to visualize the location of linking regions between helices. Electron-microscopic structural analysis of bacteriorhodopsin (Henderson and Unwin. 1975 Nature [Lond.] 257:28-32.) showed that the molecule consists of seven rods of density each nearly spanning the lipid bilayer. Owing to the distortion introduced by the missing conical region of reciprocal space data, no density was visible for the polypeptide segments linking the α-helices. Density in the refined maps indicates the location of at least five of the extrahelical segments of the polypeptide. The total number of possible ways of interconnecting the helices is reduced from 7! (5,040) to the five most consistent possibilities without recourse to other considerations. In addition, the density for the helical regions is more uniform and cylindrical throughout their length, and the length of the helices increases from 35 to 45 Å, close to the membrane thickness of 49 Å obtained for membranes dried in vacuo. Only three of the five structures consistent with the location of observed linkers place the seventh helix, onto which the chromophore can be attached by reduction in the light, at a position consistent with the main peak for deuterated retinal in the structure, as derived from neutron diffraction analysis. Two of these models are also consistent with the possible location of some of the reduced chromophore on helix B, at lys 40/41 after reduction in the dark, as well as lys 216 on helix G. ImagesFIGURE 8 PMID:7074187

Stress Wave Attenuation in Aluminum Alloy and Mild Steel Specimens Under SHPB Tensile Testing

NASA Astrophysics Data System (ADS)

Pothnis, J. R.; Ravikumar, G.; Arya, H.; Yerramalli, Chandra S.; Naik, N. K.

2018-02-01

Investigations on the effect of intensity of incident pressure wave applied through the striker bar on the specimen force histories and stress wave attenuation during split Hopkinson pressure bar (SHPB) tensile testing are presented. Details of the tensile SHPB along with Lagrangian x- t diagram of the setup are included. Studies were carried out on aluminum alloy 7075 T651 and IS 2062 mild steel. While testing specimens using the tensile SHPB setup, it was observed that the force calculated from the transmitter bar strain gauge was smaller than the force obtained from the incident bar strain gauge. This mismatch between the forces in the incident bar and the transmitter bar is explained on the basis of stress wave attenuation in the specimens. A methodology to obtain force histories using the strain gauges on the specimen during SHPB tensile testing is also presented. Further, scanning electron microscope images and photomicrographs are given. Correlation between the microstructure and mechanical properties is explained. Further, uncertainty analysis was conducted to ascertain the accuracy of the results.

Three-dimensional imaging of adherent cells using FIB/SEM and STEM.

PubMed

Villinger, Clarissa; Schauflinger, Martin; Gregorius, Heiko; Kranz, Christine; Höhn, Katharina; Nafeey, Soufi; Walther, Paul

2014-01-01

In this chapter we describe three different approaches for three-dimensional imaging of electron microscopic samples: serial sectioning transmission electron microscopy (TEM), scanning transmission electron microscopy (STEM) tomography, and focused ion beam/scanning electron microscopy (FIB/SEM) tomography. With these methods, relatively large volumes of resin-embedded biological structures can be analyzed at resolutions of a few nm within a reasonable expenditure of time. The traditional method is serial sectioning and imaging the same area in all sections. Another method is TEM tomography that involves tilting a section in the electron beam and then reconstruction of the volume by back projection of the images. When the scanning transmission (STEM) mode is used, thicker sections (up to 1 μm) can be analyzed. The third approach presented here is focused ion beam/scanning electron microscopy (FIB/SEM) tomography, in which a sample is repeatedly milled with a focused ion beam (FIB) and each newly produced block face is imaged with the scanning electron microscope (SEM). This process can be repeated ad libitum in arbitrary small increments allowing 3D analysis of relatively large volumes such as eukaryotic cells. We show that resolution of this approach is considerably improved when the secondary electron signal is used. However, the most important prerequisite for three-dimensional imaging is good specimen preparation. For all three imaging methods, cryo-fixed (high-pressure frozen) and freeze-substituted samples have been used.

Interlaboratory study for nickel alloy 625 made by laser powder bed fusion to quantify mechanical property variability.

PubMed

Brown, Christopher U; Jacob, Gregor; Stoudt, Mark; Moylan, Shawn; Slotwinski, John; Donmez, Alkan

2016-08-01

Six different organizations participated in this interlaboratory study to quantify the variability in the tensile properties of Inconel 625 specimens manufactured using laser-powder-bed-fusion additive manufacturing machines. The tensile specimens were heat treated and tensile tests conducted until failure. The properties measured were yield strength, ultimate tensile strength, elastic modulus, and elongation. Statistical analysis revealed that between-participant variability for yield strength, ultimate tensile strength, and elastic modulus values were significantly higher (up to 4 times) than typical within-participant variations. Only between-participant and within-participant variability were both similar for elongation. A scanning electron microscope was used to examine one tensile specimen for fractography. The fracture surface does not have many secondary cracks or other features that would reduce the mechanical properties. In fact, the features largely consist of microvoid coalescence and are entirely consistent with ductile failure.

Recurrent herpetic keratitis: failure to detect herpes simplex virus infection using the Syva MicroTrak HSV1/HSV2 direct specimen identification/typing test.

PubMed

Kumano, Y; Yamamoto, M; Inomata, H; Sakuma, S; Hidaka, Y; Minagawa, H; Mori, R

1990-01-01

A 35-year-old man had developed recurrent herpetic keratitis characterized by dendritic keratitis at intervals of a year. We were able to culture cytopathic agents repeatedly from his lesions by inoculating Vero cells. The cultures yielded definitive evidence of a virus that caused a cytopathic effect within 3 days. However, these virus strains could not be identified as herpes simplex virus (HSV) in immunofluorescence assays using the Syva MicroTrak HSV1/HSV2 direct specimen identification/typing test. Rather they were identified as strains of HSV type 1 (HSV-1) on the basis of plaque morphology, neutralization tests, electron-microscopic examination and DNA restriction endonuclease analysis. Our results allow us to assume the existence of HSV-1 strains isolated clinically that are negative to analysis using the Syva Micro-Trak HSV1/HSV2 direct specimen identification/typing test.

Influence of irradiance on Knoop hardness, degree of conversion, and polymerization shrinkage of nanofilled and microhybrid composite resins.

PubMed

Fugolin, Ana Paula Piovezan; Correr-Sobrinho, Lourenço; Correr, Américo Bortolazzo; Sinhoreti, Mário Alexandre Coelho; Guiraldo, Ricardo Danil; Consani, Simonides

2016-01-01

The purpose of this study was to investigate the influence of the irradiance emitted by a light-curing unit on microhardness, degree of conversion (DC), and gaps resulting from shrinkage of 2 dental composite resins. Cylinders of nanofilled and microhybrid composites were fabricated and light cured. After 24 hours, the tops and bottoms of the specimens were evaluated via indentation testing and Fourier transform infrared spectroscopy to determine Knoop hardness number (KHN) and DC, respectively. Gap width (representing polymerization shrinkage) was measured under a scanning electron microscope. The nanofilled composite specimens presented significantly greater KHNs than did the microhybrid specimens (P < 0.05). The microhybrid composite resin exhibited significantly greater DC and gap width than the nanofilled material (P < 0.05). Irradiance had a mostly material-dependent influence on the hardness and DC, but not the polymerization shrinkage, of composite resins.

Every factor helps: Rapid Ptychographic Reconstruction

NASA Astrophysics Data System (ADS)

Nashed, Youssef

2015-03-01

Recent advances in microscopy, specifically higher spatial resolution and data acquisition rates, require faster and more robust phase retrieval reconstruction methods. Ptychography is a phase retrieval technique for reconstructing the complex transmission function of a specimen from a sequence of diffraction patterns in visible light, X-ray, and electron microscopes. As technical advances allow larger fields to be imaged, computational challenges arise for reconstructing the correspondingly larger data volumes. Waiting to postprocess datasets offline results in missed opportunities. Here we present a parallel method for real-time ptychographic phase retrieval. It uses a hybrid parallel strategy to divide the computation between multiple graphics processing units (GPUs). A final specimen reconstruction is then achieved by different techniques to merge sub-dataset results into a single complex phase and amplitude image. Results are shown on a simulated specimen and real datasets from X-ray experiments conducted at a synchrotron light source.

Interlaboratory study for nickel alloy 625 made by laser powder bed fusion to quantify mechanical property variability

PubMed Central

Brown, Christopher U.; Jacob, Gregor; Stoudt, Mark; Moylan, Shawn; Slotwinski, John; Donmez, Alkan

2017-01-01

Six different organizations participated in this interlaboratory study to quantify the variability in the tensile properties of Inconel 625 specimens manufactured using laser-powder-bed-fusion additive manufacturing machines. The tensile specimens were heat treated and tensile tests conducted until failure. The properties measured were yield strength, ultimate tensile strength, elastic modulus, and elongation. Statistical analysis revealed that between-participant variability for yield strength, ultimate tensile strength, and elastic modulus values were significantly higher (up to 4 times) than typical within-participant variations. Only between-participant and within-participant variability were both similar for elongation. A scanning electron microscope was used to examine one tensile specimen for fractography. The fracture surface does not have many secondary cracks or other features that would reduce the mechanical properties. In fact, the features largely consist of microvoid coalescence and are entirely consistent with ductile failure. PMID:28243032

Whole mount nuclear fluorescent imaging: convenient documentation of embryo morphology

PubMed Central

Sandell, Lisa L.; Kurosaka, Hiroshi; Trainor, Paul A.

2012-01-01

Here we describe a relatively inexpensive and easy method to produce high quality images that reveal fine topological details of vertebrate embryonic structures. The method relies on nuclear staining of whole mount embryos in combination with confocal microscopy or conventional widefield fluorescent microscopy. In cases where confocal microscopy is used in combination with whole mount nuclear staining, the resulting embryo images can rival the clarity and resolution of images of similar specimens produced by Scanning Electron Microscopy (SEM). The fluorescent nuclear staining may be performed with a variety of cell permeable nuclear dyes, enabling the technique to be performed with multiple standard microscope/illumination or confocal/laser systems. The method may be used to document morphology of embryos of a variety of organisms, as well as individual organs and tissues. Nuclear stain imaging imposes minimal impact on embryonic specimens, enabling imaged specimens to be utilized for additional assays. PMID:22930523

Surface alteration and physical properties of glass from the Cretaceous-Tertiary boundary

USGS Publications Warehouse

Barkatt, A.; Sang, J.C.; Thorpe, A.N.; Senftle, F.E.; Talmy, I.G.; Norr, M.K.; Mazer, J.J.; Izett, G.; Sigurdsson, Haraldur

1994-01-01

The scalloped surface feature on Cretaceous-Tertiary boundary glass is often explained as being due to terrestrial aqueous leaching. Leaching of man-made glass results in a reduction in density of the glass. Also, Fe, because of its relative insolubility, is concentrated by the leaching process. Thus, the Haitian glass specimens which have been heavily altered should have a thin rim of less dense glass in which the Fe is concentrated compared to the core glass. The higher Fe concentration in the rim glass should cause it to have an enhanced Curie constant and a lower density compared to the unaltered glass. The magnetic Curie constant, density, and scanning electron microscopic studies were made on altered specimens of Haitian glass and also on specimens showing a minimum of alteration. The results show that the less altered samples have the highest density and the lowest Curie constant. The data substantiate the terrestrial hypothesis. ?? 1994.

Microstructure and Properties of Zircon-Added Carbon Refractories for Blast Furnace

NASA Astrophysics Data System (ADS)

Zhu, Tianbin; Li, Yawei; Sang, Shaobai; Chen, Xilai; Zhao, Lei; Li, Yuanbing; Li, Shujing

2012-11-01

Microstructure and properties of zircon-added carbon refractory specimens for blast furnace (BF) were investigated with the aid of X-ray diffraction (XRD), a scanning electron microscope (SEM), energy-dispersive X-ray, mercury porosimetry, and a laser thermal conductivity (TC) meter. Additives could influence the matrix structures and improve the properties of specimens. With the increase of zircon powder content, the amount of SiC whiskers formed increased and their aspect ratio became larger, and the SiC whiskers tended to be distributed homogeneously. Zircon powder additions decreased the mean pore diameter and increased <1- μm pore volume by filling in pores via SiC, improved the TC and the cold crushing strength (CCS) due to the in-situ formation of the more well-developed SiC whiskers with high TC, and significantly reduced the molten iron attack to carbon specimens.

Interlaboratory Study for Nickel Alloy 625 Made by Laser Powder Bed Fusion to Quantify Mechanical Property Variability

NASA Astrophysics Data System (ADS)

Brown, Christopher U.; Jacob, Gregor; Stoudt, Mark; Moylan, Shawn; Slotwinski, John; Donmez, Alkan

2016-08-01

Six different organizations participated in this interlaboratory study to quantify the variability in the tensile properties of Inconel 625 specimens manufactured using laser powder bed fusion-additive manufacturing machines. The tensile specimens were heat treated and tensile tests were conducted until failure. The properties measured were yield strength, ultimate tensile strength, elastic modulus, and elongation. Statistical analysis revealed that between-participant variability for yield strength, ultimate tensile strength, and elastic modulus values were significantly higher (up to four times) than typical within-participant variations. Only between-participant and within-participant variability were both similar for elongation. A scanning electron microscope was used to examine one tensile specimen for fractography. The fracture surface does not have many secondary cracks or other features that would reduce the mechanical properties. In fact, the features largely consist of microvoid coalescence and are entirely consistent with ductile failure.

Automated detection of analyzable metaphase chromosome cells depicted on scanned digital microscopic images

NASA Astrophysics Data System (ADS)

Qiu, Yuchen; Wang, Xingwei; Chen, Xiaodong; Li, Yuhua; Liu, Hong; Li, Shibo; Zheng, Bin

2010-02-01

Visually searching for analyzable metaphase chromosome cells under microscopes is quite time-consuming and difficult. To improve detection efficiency, consistency, and diagnostic accuracy, an automated microscopic image scanning system was developed and tested to directly acquire digital images with sufficient spatial resolution for clinical diagnosis. A computer-aided detection (CAD) scheme was also developed and integrated into the image scanning system to search for and detect the regions of interest (ROI) that contain analyzable metaphase chromosome cells in the large volume of scanned images acquired from one specimen. Thus, the cytogeneticists only need to observe and interpret the limited number of ROIs. In this study, the high-resolution microscopic image scanning and CAD performance was investigated and evaluated using nine sets of images scanned from either bone marrow (three) or blood (six) specimens for diagnosis of leukemia. The automated CAD-selection results were compared with the visual selection. In the experiment, the cytogeneticists first visually searched for the analyzable metaphase chromosome cells from specimens under microscopes. The specimens were also automated scanned and followed by applying the CAD scheme to detect and save ROIs containing analyzable cells while deleting the others. The automated selected ROIs were then examined by a panel of three cytogeneticists. From the scanned images, CAD selected more analyzable cells than initially visual examinations of the cytogeneticists in both blood and bone marrow specimens. In general, CAD had higher performance in analyzing blood specimens. Even in three bone marrow specimens, CAD selected 50, 22, 9 ROIs, respectively. Except matching with the initially visual selection of 9, 7, and 5 analyzable cells in these three specimens, the cytogeneticists also selected 41, 15 and 4 new analyzable cells, which were missed in initially visual searching. This experiment showed the feasibility of applying this CAD-guided high-resolution microscopic image scanning system to prescreen and select ROIs that may contain analyzable metaphase chromosome cells. The success and the further improvement of this automated scanning system may have great impact on the future clinical practice in genetic laboratories to detect and diagnose diseases.

Microtensile bond strength of etch and rinse versus self-etch adhesive systems.

PubMed

Hamouda, Ibrahim M; Samra, Nagia R; Badawi, Manal F

2011-04-01

The aim of this study was to compare the microtensile bond strength of the etch and rinse adhesive versus one-component or two-component self-etch adhesives. Twelve intact human molar teeth were cleaned and the occlusal enamel of the teeth was removed. The exposed dentin surfaces were polished and rinsed, and the adhesives were applied. A microhybride composite resin was applied to form specimens of 4 mm height and 6 mm diameter. The specimens were sectioned perpendicular to the adhesive interface to produce dentin-resin composite sticks, with an adhesive area of approximately 1.4 mm(2). The sticks were subjected to tensile loading until failure occurred. The debonded areas were examined with a scanning electron microscope to determine the site of failure. The results showed that the microtensile bond strength of the etch and rinse adhesive was higher than that of one-component or two-component self-etch adhesives. The scanning electron microscope examination of the dentin surfaces revealed adhesive and mixed modes of failure. The adhesive mode of failure occurred at the adhesive/dentin interface, while the mixed mode of failure occurred partially in the composite and partially at the adhesive/dentin interface. It was concluded that the etch and rinse adhesive had higher microtensile bond strength when compared to that of the self-etch adhesives. Copyright © 2010 Elsevier Ltd. All rights reserved.

A Novel Method to Calculate the Carbides Fraction from Dilatometric Measurements During Cooling in Hot-Work Tool Steel

NASA Astrophysics Data System (ADS)

Zhao, Xiaoli; Li, Chuanwei; Han, Lizhan; Gu, Jianfeng

2018-06-01

Dilatometry is a useful technique to obtain experimental data concerning transformation. In this paper, a dilation conversional model was established to calculate carbides fraction in AISI H13 hot-work tool steel based on the measured length changes. After carbides precipitation, the alloy contents in the matrix changed. In the usual models, the content of carbon atoms after precipitation is considered as the only element that affects the lattice constant and the content of the alloy elements such as Cr, Mo, Mn, V are often ignored. In the model introduced in this paper, the alloying elements (Cr, Mo, Mn, V) changes caused by carbides precipitation are incorporated. The carbides were identified using scanning electron microscope and transmission electron microscope. The relationship between lattice constant of carbides and temperature are measured by high-temperature X-ray diffraction. The results indicate that the carbides observed in all specimens cooled at different rates are V-rich MC and Cr-rich M23C6, and most of them are V-rich MC, only very few are Cr-rich M23C6. The model including the effects of substitutional alloying elements shows a good improvement on carbides fraction predictions. In addition, lower cooling rate advances the carbides precipitation for AISI H13 specimens. The results between experiments and mathematical model agree well.

3D-profile measurement of advanced semiconductor features by using FIB as reference metrology

NASA Astrophysics Data System (ADS)

Takamasu, Kiyoshi; Iwaki, Yuuki; Takahashi, Satoru; Kawada, Hiroki; Ikota, Masami

2017-03-01

A novel method of sub-nanometer uncertainty for the 3D-profile measurement and LWR (Line Width Roughness) measurement by using FIB (Focused Ion Beam) processing, and TEM (Transmission Electron Microscope) and CD-SEM (Critical Dimension Scanning Electron Microscope) images measurement is proposed to standardize 3D-profile measurement through reference metrology. In this article, we apply the methodology to line profile measurements and roughness measurement of advanced FinFET (Fin-shaped Field-Effect Transistor) features. The FinFET features are horizontally sliced as a thin specimen by FIB micro sampling system. Horizontally images of the specimens are obtained then by a planar TEM. LWR is calculated from the edges positions on TEM images. Moreover, we already have demonstrated the novel on-wafer 3D-profile metrology as "FIB-to-CDSEM method" with FIB slope cut and CD-SEM measuring. Using the method, a few micrometers wide on a wafer is coated and cut by 45-degree slope using FIB tool. Then, the wafer is transferred to CD-SEM to measure the cross section image by top down CD-SEM measurement. We applied FIB-to-CDSEM method to a CMOS image sensor feature. The 45-degree slope cut surface is observed using AFM. The surface profile of slope cut surface and line profiles are analyzed for improving the accuracy of FIB-to-CDSEM method.

Preservation of protein fluorescence in embedded human dendritic cells for targeted 3D light and electron microscopy.

PubMed

Höhn, K; Fuchs, J; Fröber, A; Kirmse, R; Glass, B; Anders-Össwein, M; Walther, P; Kräusslich, H-G; Dietrich, C

2015-08-01

In this study, we present a correlative microscopy workflow to combine detailed 3D fluorescence light microscopy data with ultrastructural information gained by 3D focused ion beam assisted scanning electron microscopy. The workflow is based on an optimized high pressure freezing/freeze substitution protocol that preserves good ultrastructural detail along with retaining the fluorescence signal in the resin embedded specimens. Consequently, cellular structures of interest can readily be identified and imaged by state of the art 3D confocal fluorescence microscopy and are precisely referenced with respect to an imprinted coordinate system on the surface of the resin block. This allows precise guidance of the focused ion beam assisted scanning electron microscopy and limits the volume to be imaged to the structure of interest. This, in turn, minimizes the total acquisition time necessary to conduct the time consuming ultrastructural scanning electron microscope imaging while eliminating the risk to miss parts of the target structure. We illustrate the value of this workflow for targeting virus compartments, which are formed in HIV-pulsed mature human dendritic cells. © 2015 The Authors Journal of Microscopy © 2015 Royal Microscopical Society.

Nanoscale Imaging of Whole Cells Using a Liquid Enclosure and a Scanning Transmission Electron Microscope

PubMed Central

Peckys, Diana B.; Veith, Gabriel M.; Joy, David C.; de Jonge, Niels

2009-01-01

Nanoscale imaging techniques are needed to investigate cellular function at the level of individual proteins and to study the interaction of nanomaterials with biological systems. We imaged whole fixed cells in liquid state with a scanning transmission electron microscope (STEM) using a micrometer-sized liquid enclosure with electron transparent windows providing a wet specimen environment. Wet-STEM images were obtained of fixed E. coli bacteria labeled with gold nanoparticles attached to surface membrane proteins. Mammalian cells (COS7) were incubated with gold-tagged epidermal growth factor and fixed. STEM imaging of these cells resulted in a resolution of 3 nm for the gold nanoparticles. The wet-STEM method has several advantages over conventional imaging techniques. Most important is the capability to image whole fixed cells in a wet environment with nanometer resolution, which can be used, e.g., to map individual protein distributions in/on whole cells. The sample preparation is compatible with that used for fluorescent microscopy on fixed cells for experiments involving nanoparticles. Thirdly, the system is rather simple and involves only minimal new equipment in an electron microscopy (EM) laboratory. PMID:20020038

Sol-gel derived bioactive coating on zirconia: Effect on flexural strength and cell proliferation.

PubMed

Shahramian, Khalil; Leminen, Heidi; Meretoja, Ville; Linderbäck, Paula; Kangasniemi, Ilkka; Lassila, Lippo; Abdulmajeed, Aous; Närhi, Timo

2017-11-01

The purpose of this study was to evaluate the effect of sol-gel derived bioactive coatings on the biaxial flexural strength and fibroblast proliferation of zirconia, aimed to be used as an implant abutment material. Yttrium stabilized zirconia disc-shaped specimens were cut, ground, sintered, and finally cleansed ultrasonically in each of acetone and ethanol for 5 minutes. Three experimental groups (n = 15) were fabricated, zirconia with sol-gel derived titania (TiO 2 ) coating, zirconia with sol-gel derived zirconia (ZrO 2 ) coating, and non-coated zirconia as a control. The surfaces of the specimens were analyzed through images taken using a scanning electron microscope (SEM), and a non-contact tapping mode atomic force microscope (AFM) was used to record the surface topography and roughness of the coated specimens. Biaxial flexural strength values were determined using the piston-on-three ball technique. Human gingival fibroblast proliferation on the surface of the specimens was evaluated using AlamarBlue assay™. Data were analyzed using a one-way analysis of variance (ANOVA) followed by Tukey's post-hoc test. Additionally, the biaxial flexural strength data was also statistically analyzed with the Weibull distribution. The biaxial flexural strength of zirconia specimens was unaffected (p > 0.05). Weibull modulus of TiO 2 coated and ZrO 2 coated groups (5.7 and 5.4, respectively) were lower than the control (8.0). Specimens coated with ZrO 2 showed significantly lower fibroblast proliferation compared to other groups (p < 0.05). In conclusion, sol-gel derived coatings have no influence on the flexural strength of zirconia. ZrO 2 coated specimens showed significantly lower cell proliferation after 12 days than TiO 2 coated or non-coated control. © 2016 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 105B: 2401-2407, 2017. © 2016 Wiley Periodicals, Inc.

Time-of-flight atom-probe field-ion microscope for the study of defects in metals. Report No. 2357. [W--25 at. % Re

DOE Office of Scientific and Technical Information (OSTI.GOV)

Hall, T.M.; Wagner, A.; Berger, A.S.

1975-06-01

An ultra-high vacuum time-of-flight (TOF) atom-probe field ion microscope (FIM) specifically designed for the study of defects in metals is described. The variable magnification FIM image is viewed with the aid of an internal image intensification system based on a channel electron-multiplier array. The specimen is held in a liquid-helium-cooled goniometer stage, and the specimen is exchanged by means of a high-vacuum (less than 10/sup -6/ torr) specimen exchange device. This stage allows the specimen to be maintained at a tip temperature anywhere in the range from 13 to 450/sup 0/K. Specimens can also be irradiated in-situ with any low-energymore » (less than 1 keV) gas ion employing a specially constructed ion gun. The pulse-field evaporated ions are detected by a Chevron ion-detector located 2.22 m from the FIM specimen. The TOF of the ions are measured by a specially constructed eight-channel digital timer with a resolution of +-10 ns. The entire process of applying the evaporation pulse to the specimen, measuring the dc and pulse voltages, and analyzing the TOF data is controlled by a NOVA 1220 computer. The computer is also interfaced to a Tektronix graphics terminal which displays the data in the form of a histogram of the number of events versus the mass-to-charge ratio. An extensive set of computer programs to test and operate the atom-probe FIM have been developed. With this automated system we can presently record and analyze 10 TOF s/sup -1/. In the performance tests reported here the instrument has resolved the seven stable isotopes of molybdenum, the five stable isotopes of tungsten, and the two stable isotopes of rhenium in a tungsten--25 at. percent rhenium alloy. (auth)« less

Sole Dependence on Urine Testing Strips and the Ability to Identify Clinically Significant Disease: Challenging the Current Paradigm for Heme Detection in General Clinical Situations.

PubMed

Rothschild, Bruce

2016-05-01

The ability of health care professionals to provide patient care is potentially compromised when predicated on untested, although longstanding, perspectives. One such example is urinalysis testing, which has been currently simplified to use only urine testing strips for detection of microscopic hematuria. To determine whether urine testing strips are sufficient for identification of clinically significant findings in urinalysis. To determine the presence of microscopic hematuria, I examined a collection of urine specimens that had tested heme negative during the 3-month study period. Of the 342 patients from whom urine specimens were examined during this interval, 50 had microscopic hematuria, despite having tested negative for heme via urine testing strip. Also, 30% were not receiving any medication known to produce microscopic hematuria, and 18% had clinically significant pathology. Diagnosis of significant clinical pathologic manifestations would have been compromised had microscopic examination not been performed on the urine specimens from the cohort individuals. Examination of the novel approach of including microscopic examination of specimens in a specific clinical situation challenges the dominant paradigm of reliance on assaying using urine testing strips only, revealing that the current method is not only unreliable for determining microscopic hematuria but also is less than optimal in general clinical practice. The findings of this study provide evidence of the importance of microscopic evaluation as a routine component of urinalysis. © American Society for Clinical Pathology, 2016. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

The Effects of Fiber Orientation and Adhesives on Tensile Properties of Carbon Fiber Reinforced Polymer Matrix Composite with Embedded Nickel-Titanium Shape Memory Alloys

NASA Technical Reports Server (NTRS)

Quade, Derek J.; Jana, Sadhan C.; Morscher, Gregory N.; Kannan, Manigandan; McCorkle, Linda S.

2017-01-01

Nickel-titanium (NiTi) shape memory alloy (SMA) sections were embedded within carbon fiber reinforced polymer matrix composite (CFRPPMC) laminates and their tensile properties were evaluated with simultaneous monitoring of modal acoustic emissions. The test specimens were fabricated in three different layup configurations and two different thin film adhesives were applied to bond the SMA with the PMC. A trio of acoustic sensors were attached to the specimens during tensile testing to monitor the modal acoustic emission (AE) as the materials experienced mechanical failure. The values of ultimate tensile strengths, strains, and moduli were obtained. Cumulative AE energy of events and specimen failure location were determined. In conjunction, optical and scanning electron microscopy techniques were used to examine the break areas of the specimens. The analysis of AE data revealed failure locations within the specimens which were validated from the microscopic images. The placement of 90 deg plies in the outer ply gave the strongest acoustic signals during break as well as the cleanest break of the samples tested. Overlapping 0 deg ply layers surrounding the SMA was found to be the best scenario to prevent failure of the specimen itself.

Shear bond strength of indirect composite material to monolithic zirconia.

PubMed

Sari, Fatih; Secilmis, Asli; Simsek, Irfan; Ozsevik, Semih

2016-08-01

This study aimed to evaluate the effect of surface treatments on bond strength of indirect composite material (Tescera Indirect Composite System) to monolithic zirconia (inCoris TZI). Partially stabilized monolithic zirconia blocks were cut into with 2.0 mm thickness. Sintered zirconia specimens were divided into different surface treatment groups: no treatment (control), sandblasting, glaze layer & hydrofluoric acid application, and sandblasting + glaze layer & hydrofluoric acid application. The indirect composite material was applied to the surface of the monolithic zirconia specimens. Shear bond strength value of each specimen was evaluated after thermocycling. The fractured surface of each specimen was examined with a stereomicroscope and a scanning electron microscope to assess the failure types. The data were analyzed using one-way analysis of variance (ANOVA) and Tukey LSD tests (α=.05). Bond strength was significantly lower in untreated specimens than in sandblasted specimens (P<.05). No difference between the glaze layer and hydrofluoric acid application treated groups were observed. However, bond strength for these groups were significantly higher as compared with the other two groups (P<.05). Combined use of glaze layer & hydrofluoric acid application and silanization are reliable for strong and durable bonding between indirect composite material and monolithic zirconia.

Total rate imaging with x-rays (TRIX)--a simple method of forming a non-projection x-ray image in the SEM using an energy dispersive detector and its application to biological specimens.

PubMed

Ingram, P; Shelburne, J D

1980-01-01

X-ray images can be formed in a conventional scanning electron microscope equipped with a Si(Li) energy dispersive spectrometer. All the x-ray events generated in the electron beam scanning process are synchronously displayed in the same manner as for dot maps. The quasi-digital image formed using Total Rate Imaging with X-rays (TRIX) exhibits good gray scale contrast and is dependent on topography, orientation and atomic number. Although this latter dependence is complex, it has been found useful in locating several types of inclusions in lung tissue (silicosis), human alveolar macrophages and cigarette smoke condensate. This is because of the greater depth of penetration of x-rays than backscattered electrons (BSE) usually used for such localizations in a matrix, and the negligible sensitivity of the Si(Li) detector to x-rays from an organic biological matrix. The optimum procedure is to use a combination of TRIX and BSE to investigate such specimens.

Cinemicrographic specimen housing

NASA Technical Reports Server (NTRS)

Wilkins, J. R.

1979-01-01

Housing used to observe gravitation effects on specimens embedded in support media, such as agar, supports microbial specimens vertically for time-lapsed cinemicrographic studies. Procedure cannot be performed with conventional microscopes which see specimens in horizontal plane only.

Enhancement of the Microscopy Facilities at the NSLS X1A Beamline

DOE Office of Scientific and Technical Information (OSTI.GOV)

Jacobson, Chris

1999-08-31

As originally proposed, the authors constructed a new Scanning Transmission X-ray Microscope, STXM IV. The design and construction was led by Chris Jacobsen, and involved graduate students Michael Feser, Mary Carlucci-Dayton and Tobias Beetz. This microscope has the following new features: It has a new and improved high resolution scanning stage that should make it possible to perform higher resolution imaging without distortions. Preliminary results indicate that the stage performs as designed. It has an enclosure that can be evacuated and backfilled with helium. This makes it possible to perform imaging in the neighborhood of the nitrogen and oxygen edgesmore » without interference from residual air. It has a motorized detector stage for easy interchange of detectors and alignment microscope. We expect to use this to align the new segmented detector which makes it possible to perform brightfield and dark field microscopy simultaneously, and to record images in differential phase contrast as well. The microscope is located upstream of cryoSTXM, the instrument we use to examine specimens in a frozen hydrated state. The design of STXM IV is such that it makes it quick and easy to switch between STXM IV and cryo-STXM operations and vice versa. IEEE488 based control electronics provides multiple channels of data collection. The microscope is run from a LINUX PC with all new software, developed in-house. The stages for the zone plate and the order sorting aperture (OSA) have kinematic mounts. This way different sets of zone plates (optimized for different wavelengths and working distances) can be exchanged without the need for complete realignment of the instrument. The enclosure can be used as a glove-box, making it possible to examine specimens which require anaerobic handling.« less

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.

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

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

PubMed

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

2012-12-03

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

Sample holder for axial rotation of specimens in 3D microscopy.

PubMed

Bruns, T; Schickinger, S; Schneckenburger, H

2015-10-01

In common light microscopy, observation of samples is only possible from one perspective. However, especially for larger three-dimensional specimens observation from different views is desirable. Therefore, we are presenting a sample holder permitting rotation of the specimen around an axis perpendicular to the light path of the microscope. Thus, images can be put into a defined multidimensional context, enabling reliable three-dimensional reconstructions. The device can be easily adapted to a great variety of common light microscopes and is suitable for various applications in science, education and industry, where the observation of three-dimensional specimens is essential. Fluorescence z-projection images of copepods and ixodidae ticks at different rotation angles obtained by confocal laser scanning microscopy and light sheet fluorescence microscopy are reported as representative results. © 2015 The Authors Journal of Microscopy © 2015 Royal Microscopical Society.

Tilt-Pair Analysis of Images from a Range of Different Specimens in Single-Particle Electron Cryomicroscopy

PubMed Central

Henderson, Richard; Chen, Shaoxia; Chen, James Z.; Grigorieff, Nikolaus; Passmore, Lori A.; Ciccarelli, Luciano; Rubinstein, John L.; Crowther, R. Anthony; Stewart, Phoebe L.; Rosenthal, Peter B.

2011-01-01

The comparison of a pair of electron microscope images recorded at different specimen tilt angles provides a powerful approach for evaluating the quality of images, image-processing procedures, or three-dimensional structures. Here, we analyze tilt-pair images recorded from a range of specimens with different symmetries and molecular masses and show how the analysis can produce valuable information not easily obtained otherwise. We show that the accuracy of orientation determination of individual single particles depends on molecular mass, as expected theoretically since the information in each particle image increases with molecular mass. The angular uncertainty is less than 1° for particles of high molecular mass (∼ 50 MDa), several degrees for particles in the range 1–5 MDa, and tens of degrees for particles below 1 MDa. Orientational uncertainty may be the major contributor to the effective temperature factor (B-factor) describing contrast loss and therefore the maximum resolution of a structure determination. We also made two unexpected observations. Single particles that are known to be flexible showed a wider spread in orientation accuracy, and the orientations of the largest particles examined changed by several degrees during typical low-dose exposures. Smaller particles presumably also reorient during the exposure; hence, specimen movement is a second major factor that limits resolution. Tilt pairs thus enable assessment of orientation accuracy, map quality, specimen motion, and conformational heterogeneity. A convincing tilt-pair parameter plot, where 60% of the particles show a single cluster around the expected tilt axis and tilt angle, provides confidence in a structure determined using electron cryomicroscopy. PMID:21939668

Sensing mode atomic force microscope

DOEpatents

Hough, Paul V. C.; Wang, Chengpu

2003-01-01

An atomic force microscope utilizes a pulse release system and improved method of operation to minimize contact forces between a probe tip affixed to a flexible cantilever and a specimen being measured. The pulse release system includes a magnetic particle affixed proximate the probe tip and an electromagnetic coil. When energized, the electromagnetic coil generates a magnetic field which applies a driving force on the magnetic particle sufficient to overcome adhesive forces exhibited between the probe tip and specimen. The atomic force microscope includes two independently displaceable piezo elements operable along a Z-axis. A controller drives the first Z-axis piezo element to provide a controlled approach between the probe tip and specimen up to a point of contact between the probe tip and specimen. The controller then drives the first Z-axis piezo element to withdraw the cantilever from the specimen. The controller also activates the pulse release system which drives the probe tip away from the specimen during withdrawal. Following withdrawal, the controller adjusts the height of the second Z-axis piezo element to maintain a substantially constant approach distance between successive samples.

Evaluation of anterior lenticonus in alport syndrome using tracey wavefront aberrometry and transmission electron microscopy.

PubMed

Kim, Kwan Soo; Kim, Mo Sae; Kim, Joon Mo; Choi, Chul Young

2010-01-01

To evaluate the efficacy of Tracey wavefront aberrometry (Tracey Technologies, Houston, TX) and transmission electron microscopy for the detection of anterior lenticonus in Alport syndrome. Tracey wavefront aberrometry was used to treat a patient with bilateral anterior lenticonus who had a history of Alport syndrome. For transmission electron microscopic examination, anterior lens capsules were obtained during clear lens phacoemulsification and intraocular lens implantation. Spherical aberrations were the predominant higher-order aberrations in the internal optics of both eyes. The Tracey wavefront aberrometer showed that most of the irregular astigmatism originated from the lenticular portion. Transmission electron microscopy of the specimens showed anterior lens capsules with decreased thickness and multiple dehiscences. Tracey wavefront aberrometry and transmission electron microscopy are effective tools for evaluation of anterior lenticonus in Alport syndrome. Copyright 2010, SLACK Incorporated.

Evaluation of Wear Resistance of Friction Materials Prepared by Granulation.

PubMed

Ma, Yunhai; Liu, Yucheng; Menon, Carlo; Tong, Jin

2015-10-21

The tribological properties of friction materials prepared by hot-pressing pellets of different sizes were experimentally investigated. Friction and wear tests of the specimens were performed and morphological analysis was carried out by investigating images acquired with both scanning electron and confocal laser microscopes. The highest friction coefficient of friction materials was obtained with pellets having 1-5 mm size. The lowest wear rate was obtained with pellets having 8-10 mm size. Specimens processed by mixing pellets of different sizes had the highest density and the lowest roughness and were the least expensive to fabricate. The results show that granulation generally enabled increasing the friction coefficient, decreasing the wear rate, and reducing the number of defects on the surface of friction materials.

Vacuum Ultraviolet (VUV) radiation-induced degradation of Fluorinated Ethylene Propylene (FEP) Teflon aboard the Long Duration Exposure Facility (LDEF)

NASA Technical Reports Server (NTRS)

Brinza, David E.; Stiegman, A. E.; Staszak, Paul R.; Laue, Eric G.; Liang, Ranty H.

1992-01-01

Examination of fluorinated ethylene propylene (FEP) copolymer specimens recovered from the Long Duration Exposure Facility (LDEF) provides evidence for degradation attributed to extended solar vacuum ultraviolet (VUV) irradiation. Scanning electron microscope (SEM) images of sheared FEP film edges reveal the presence of a highly embrittled layer on the exposed surface of specimens obtained from the trailing edge of the LDEF. Similar images obtained for leading edge and control FEP films do not exhibit evidence for such an embrittled layer. Laboratory VUV irradiation of FEP films is found to produce a damage layer similar to that witnessed in the LDEF trailing edge films. Spectroscopic analyses of irradiated films provide data to advance a photochemical mechanism for degradation.

Surface Coating of Gypsum-Based Molds for Maxillofacial Prosthetic Silicone Elastomeric Material: The Surface Topography.

PubMed

Khalaf, Salah; Ariffin, Zaihan; Husein, Adam; Reza, Fazal

2015-07-01

This study aimed to compare the surface roughness of maxillofacial silicone elastomers fabricated in noncoated and coated gypsum materials. This study was also conducted to characterize the silicone elastomer specimens after surfaces were modified. A gypsum mold was coated with clear acrylic spray. The coated mold was then used to produce modified silicone experimental specimens (n = 35). The surface roughness of the modified silicone elastomers was compared with that of the control specimens, which were prepared by conventional flasking methods (n = 35). An atomic force microscope (AFM) was used for surface roughness measurement of silicone elastomer (unmodified and modified), and a scanning electron microscope (SEM) was used to evaluate the topographic conditions of coated and noncoated gypsum and silicone elastomer specimens (unmodified and modified) groups. After the gypsum molds were characterized, the fabricated silicone elastomers molded on noncoated and coated gypsum materials were evaluated further. Energy-dispersive X-ray spectroscopy (EDX) analysis of gypsum materials (noncoated and coated) and silicone elastomer specimens (unmodified and modified) was performed to evaluate the elemental changes after coating was conducted. Independent t test was used to analyze the differences in the surface roughness of unmodified and modified silicone at a significance level of p < 0.05. Roughness was significantly reduced in the silicone elastomers processed against coated gypsum materials (p < 0.001). The AFM and SEM analysis results showed evident differences in surface smoothness. EDX data further revealed the presence of the desired chemical components on the surface layer of unmodified and modified silicone elastomers. Silicone elastomers with lower surface roughness of maxillofacial prostheses can be obtained simply by coating a gypsum mold. © 2014 by the American College of Prosthodontists.

21 CFR 864.3600 - Microscopes and accessories.

Code of Federal Regulations, 2010 CFR

2010-04-01

... enlarge images of specimens, preparations, and cultures for medical purposes. Variations of microscopes... light. (3) Inverted stage microscopes, which permit examination of tissue cultures or other biological...

Spectral confocal reflection microscopy using a white light source

NASA Astrophysics Data System (ADS)

Booth, M.; Juškaitis, R.; Wilson, T.

2008-08-01

We present a reflection confocal microscope incorporating a white light supercontinuum source and spectral detection. The microscope provides images resolved spatially in three-dimensions, in addition to spectral resolution covering the wavelength range 450-650nm. Images and reflection spectra of artificial and natural specimens are presented, showing features that are not normally revealed in conventional microscopes or confocal microscopes using discrete line lasers. The specimens include thin film structures on semiconductor chips, iridescent structures in Papilio blumei butterfly scales, nacre from abalone shells and opal gemstones. Quantitative size and refractive index measurements of transparent beads are derived from spectral interference bands.

A hybrid scanning force and light microscope for surface imaging and three-dimensional optical sectioning in differential interference contrast.

PubMed

Stemmer, A

1995-04-01

The design of a scanned-cantilever-type force microscope is presented which is fully integrated into an inverted high-resolution video-enhanced light microscope. This set-up allows us to acquire thin optical sections in differential interference contrast (DIC) or polarization while the force microscope is in place. Such a hybrid microscope provides a unique platform to study how cell surface properties determine, or are affected by, the three-dimensional dynamic organization inside the living cell. The hybrid microscope presented in this paper has proven reliable and versatile for biological applications. It is the only instrument that can image a specimen by force microscopy and high-power DIC without having either to translate the specimen or to remove the force microscope. Adaptation of the design features could greatly enhance the suitability of other force microscopes for biological work.

Corrosion behavior of an HVOF-sprayed Fe3Al coating in a high-temperature oxidizing/sulfidizing environment

DOE Office of Scientific and Technical Information (OSTI.GOV)

Covino, Bernard S., Jr.; Bullard, Sophie J.; Cramer, Stephen D.

An iron aluminide (Fe3Al) intermetallic coating was deposited onto a F22 (2.25Cr-1Mo) steel substrate using a JP-5000 high velocity oxy-fuel (HVOF) thermal spray system. The as-sprayed coating was examined by electron microscopy and X-ray diffraction and was characterized in terms of oxidation and adhesion. Fe3Al-coated steel specimens were exposed to a mixed oxidizing/sulfidizing environment at 500, 600, 700, and 800DGC for approximately seven days. The gaseous environment consisted of N2-10%CO-5%CO2-2%H2O-0.12%H2S (by volume). All specimens gained mass after exposure to the environment and the mass gains were found to be inversely proportional to temperature increases. Representative specimens exposed at each temperaturemore » were cross-sectioned and subjected to examination under a scanning electron microscope (SEM) and X-ray mapping. Results are presented in terms of corrosion weight gain and corrosion product formation. The purpose of the research presented here was to evaluate the effectiveness of an HVOF-sprayed Fe3Al coating in protecting a steel substrate exposed to a fossil energy environment.« less

Evaluating the Use of Synthetic Replicas for SEM Identification of Bloodstains (with Emphasis on Archaeological and Ethnographic Artifacts).

PubMed

Hortolà, Policarp

2015-12-01

Some archaeological or ethnographic specimens are unavailable for direct examination using a scanning electron microscope (SEM) due to methodological obstacles or legal issues. In order to assess the feasibility of using SEM synthetic replicas for the identification of bloodstains (BSs) via morphology of red blood cells (RBCs), three fragments of different natural raw material (inorganic, stone; plant, wood; animal, shell) were smeared with peripheral human blood. Afterwards, molds and casts of the bloodstained areas were made using vinyl polysiloxane (VPS) silicone impression and polyurethane (PU) resin casting material, respectively. Then, the original samples and the resulting casts were coated with gold and examined in secondary-electron mode using a high-vacuum SEM. Results suggest that PU resin casts obtained from VPS silicone molds can preserve RBC morphology in BSs, and consequently that synthetic replicas are feasible for SEM identification of BSs on cultural heritage specimens made of natural raw materials. Although the focus of this study was on BSs, the method reported in this paper may be applicable to organic residues other than blood, as well as to the surface of other specimens when, for any reason, the original is unavailable for an SEM.

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.

Scanning electron microscopic study of the effects of Er:YAG laser on root cementum.

PubMed

Fujii, T; Baehni, P C; Kawai, O; Kawakami, T; Matsuda, K; Kowashi, Y

1998-11-01

Use of Er:YAG laser has been proposed for the removal of microbial deposits and calculus present on teeth affected by periodontal disease. However, the influence of Er:YAG laser irradiation on root surfaces has not yet been fully investigated. The aim of the present study was to evaluate the effects of Er:YAG laser irradiation on root cementum by scanning electron microscopy (SEM). Specimens were obtained from extracted human periodontally-diseased teeth using a water-cooled high-speed bur. An Er:YAG laser beam was then applied at various powers ranging from 25 to 100 mJ/ pulse/sec. The laser irradiation was performed under water irrigation, with the tip held perpendicular to the root surface in the contact mode. Following laser exposure, specimens were fixed, dehydrated, and dried at critical-point in liquid CO2. After mounting on SEM plates and sputter-coating with gold, the cementum surface was examined by SEM. Observations of the root surface showed a relatively flat surface in control specimens. In Er:YAG exposed specimens, the laser beam created a circular, notched-edge, crater-like defect on the root. The bottom of the lesion showed an irregular and sharp-pointed surface. Subsequently, the specimens were fractured with a sharp scalpel perpendicularly to the surface. SEM observations of these specimens showed a 15 microm layer of damaged tissue within the laser-irradiated cementum. The tissue presented an amorphous appearance and the Sharpey's and matrix fiber bundles were not clearly distinguishable. These observations indicate that cementum tissue could be damaged by Er:YAG laser irradiation.

Newly recognized hosts for uranium in the Hanford Site vadose zone

USGS Publications Warehouse

Stubbs, J.E.; Veblen, L.A.; Elbert, D.C.; Zachara, J.M.; Davis, J.A.; Veblen, D.R.

2009-01-01

Uranium contaminated sediments from the U.S. Department of Energy's Hanford Site have been investigated using electron microscopy. Six classes of solid hosts for uranium were identified. Preliminary sediment characterization was carried out using optical petrography, and electron microprobe analysis (EMPA) was used to locate materials that host uranium. All of the hosts are fine-grained and intergrown with other materials at spatial scales smaller than the analytical volume of the electron microprobe. A focused ion beam (FIB) was used to prepare electron-transparent specimens of each host for the transmission electron microscope (TEM). The hosts were identified as: (1) metatorbernite [Cu(UO2)2(PO4)2??8H2O]; (2) coatings on sediment clasts comprised mainly of phyllosilicates; (3) an amorphous zirconium (oxyhydr)oxide found in clast coatings; (4) amorphous and poorly crystalline materials that line voids within basalt lithic fragments; (5) amorphous palagonite surrounding fragments of basaltic glass; and (6) Fe- and Mn-oxides. These findings demonstrate the effectiveness of combining EMPA, FIB, and TEM to identify solid-phase contaminant hosts. Furthermore, they highlight the complexity of U geochemistry in the Hanford vadose zone, and illustrate the importance of microscopic transport in controlling the fate of contaminant metals in the environment. ?? 2008 Elsevier Ltd.

Simultaneous Scanning Electron Microscope Imaging of Topographical and Chemical Contrast Using In-Lens, In-Column, and Everhart-Thornley Detector Systems.

PubMed

Zhang, Xinming; Cen, Xi; Ravichandran, Rijuta; Hughes, Lauren A; van Benthem, Klaus

2016-06-01

The scanning electron microscope provides a platform for subnanometer resolution characterization of material morphology with excellent topographic and chemical contrast dependent on the used detectors. For imaging applications, the predominantly utilized signals are secondary electrons (SEs) and backscattered electrons (BSEs) that are emitted from the sample surface. Recent advances in detector technology beyond the traditional Everhart-Thornley geometry have enabled the simultaneous acquisition and discrimination of SE and BSE signals. This study demonstrates the imaging capabilities of a recently introduced new detector system that consists of the combination of two in-lens (I-L) detectors and one in-column (I-C) detector. Coupled with biasing the sample stage to reduce electron-specimen interaction volumes, this trinity of detector geometry allows simultaneous acquisition of signals to distinguish chemical contrast from topographical changes of the sample, including the identification of surface contamination. The I-C detector provides 4× improved topography, whereas the I-L detector closest to the sample offers excellent simultaneous chemical contrast imaging while not limiting the minimization of working distance to obtain optimal lateral resolution. Imaging capabilities and contrast mechanisms for all three detectors are discussed quantitatively in direct comparison to each other and the conventional Everhart-Thornley detector.

Corrosion resistance assessment of Co-Cr alloy frameworks fabricated by CAD/CAM milling, laser sintering, and casting methods.

PubMed

Tuna, Süleyman Hakan; Özçiçek Pekmez, Nuran; Kürkçüoğlu, Işin

2015-11-01

The effects of fabrication methods on the corrosion resistance of frameworks produced with Co-Cr alloys are not clear. The purpose of this in vitro study was to evaluate the electrochemical corrosion resistance of Co-Cr alloy specimens that were fabricated by conventional casting, milling, and laser sintering. The specimens fabricated with 3 different methods were investigated by potentiodynamic tests and electrochemical impedance spectroscopy in an artificial saliva. Ions released into the artificial saliva were estimated with inductively coupled plasma-mass spectrometry, and the results were statistically analyzed. The specimen surfaces were investigated with scanning electron microscopy before and after the tests. In terms of corrosion current and Rct properties, statistically significant differences were found both among the means of the methods and among the means of the material groups (P<.05). With regard to ions released, a statistically significant difference was found among the material groups (P<.05); however, no difference was found among the methods. Scanning electron microscopic imaging revealed that the specimens produced by conventional casting were affected to a greater extent by etching and electrochemical corrosion than those produced by milling and laser sintering. The corrosion resistance of a Co-Cr alloy specimens fabricated by milling or laser sintering was greater than that of the conventionally cast alloy specimens. The Co-Cr specimens produced by the same method also differed from one another in terms of corrosion resistance. These differences may be related to the variations in the alloy compositions. Copyright © 2015 Editorial Council for the Journal of Prosthetic Dentistry. Published by Elsevier Inc. All rights reserved.

Electron Microscopy Localization and Characterization of Functionalized Composite Organic-Inorganic SERS Nanoparticles on Leukemia Cells

PubMed Central

Koh, Ai Leen; Shachaf, Catherine M.; Elchuri, Sailaja; Nolan, Garry P.; Sinclair, Robert

2008-01-01

We demonstrate the use of electron microscopy as a powerful characterization tool to identify and locate antibody-conjugated composite organic-inorganic (COINs) surface enhanced Raman scattering (SERS) nanoparticles on cells. U937 leukemia cells labeled with antibody CD54-conjugated COINs were characterized in their native, hydrated state using wet Scanning Electron Microscopy (SEM) and in their dehydrated state using high-resolution SEM. In both cases, the backscattered electron detector (BSE) was used to detect and identify the silver constituents in COINs due to its high sensitivity to atomic number variations within a specimen. The imaging and analytical capabilities in the SEM were further complemented by higher resolution Transmission Electron Microscope (TEM) images and Scanning Auger Electron Spectroscopy (AES) data to give reliable and high-resolution information about nanoparticles and their binding to cell surface antigens. PMID:18995965

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

Comparative evaluation of enamel abrasivity by toothbrush and velcro: An in vitro scanning electron microscope study

PubMed Central

Ojha, Saroj Kumar; Javdekar, Sadashiv Bhaskar; Dhir, Sangeeta

2015-01-01

Context: Plaque control has been shown to be pivotal in maintaining the optimal periodontal health. Mechanical plaque control is the most popular option for establishing the optimal oral health. Toothbrushes have been the novel tool for mechanical cleansing. However, the abrasive potential of the toothbrushes on the enamel surface is an area in gray. Aims: The aim of this in vitro study is to evaluate the abrasivity of the toothbrush versus the velcro fasteners. Materials and Methods: The mounted teeth of both the groups were subjected to abrasion test, and the tooth surfaces were observed for the possible abrasions from the oscillating strokes (toothbrush) and frictional contacts (hook and loop velcro) and examined under the scanning electron microscope. Results: Comparative assessment of both velcro (hook and loop) and toothbrush bristles did not reveal any evidence of abrasion on the tooth specimens. Conclusions: Veclro fasteners are safe and qualitatively at par to the manual toothbrush for their efficacy and efficiency in teeth cleansing PMID:26229264

Nanocellulose based polymer composite for acoustical materials

NASA Astrophysics Data System (ADS)

Farid, Mohammad; Purniawan, Agung; Susanti, Diah; Priyono, Slamet; Ardhyananta, Hosta; Rahmasita, Mutia E.

2018-04-01

Natural fibers are biodegradable materials that are innovatively and widely used for composite reinforcement in automotive components. Nanocellulose derived from natural fibers oil palm empty bunches have properties that are remarkable for use as a composite reinforcement. However, there have not been many investigations related to the use of nanocellulose-based composites for wideband sound absorption materials. The specimens of nanocellulose-based polyester composite were prepared using a spray method. An impedance tube method was used to measure the sound absorption coefficient of this composite material. To reveal the characteristics of the nanocellulose-based polyester composite material, SEM (scanning electron microscope), TEM (Transmission Electron Microscope), FTIR (Fourier Transform Infra Red), TGA (Thermogravimetric Analysis), and density tests were performed. Sound absorption test results showed the average value of sound absorption coefficient of 0.36 to 0,46 for frequency between 500 and 4000 Hz indicating that this nanocellulose-based polyester composite materials had a tendency to wideband sound absorption materials and potentially used as automotive interior materials.

FluoroSim: A Visual Problem-Solving Environment for Fluorescence Microscopy

PubMed Central

Quammen, Cory W.; Richardson, Alvin C.; Haase, Julian; Harrison, Benjamin D.; Taylor, Russell M.; Bloom, Kerry S.

2010-01-01

Fluorescence microscopy provides a powerful method for localization of structures in biological specimens. However, aspects of the image formation process such as noise and blur from the microscope's point-spread function combine to produce an unintuitive image transformation on the true structure of the fluorescing molecules in the specimen, hindering qualitative and quantitative analysis of even simple structures in unprocessed images. We introduce FluoroSim, an interactive fluorescence microscope simulator that can be used to train scientists who use fluorescence microscopy to understand the artifacts that arise from the image formation process, to determine the appropriateness of fluorescence microscopy as an imaging modality in an experiment, and to test and refine hypotheses of model specimens by comparing the output of the simulator to experimental data. FluoroSim renders synthetic fluorescence images from arbitrary geometric models represented as triangle meshes. We describe three rendering algorithms on graphics processing units for computing the convolution of the specimen model with a microscope's point-spread function and report on their performance. We also discuss several cases where the microscope simulator has been used to solve real problems in biology. PMID:20431698

Measurement of specimen-induced aberrations of biological samples using phase stepping interferometry.

PubMed

Schwertner, M; Booth, M J; Neil, M A A; Wilson, T

2004-01-01

Confocal or multiphoton microscopes, which deliver optical sections and three-dimensional (3D) images of thick specimens, are widely used in biology. These techniques, however, are sensitive to aberrations that may originate from the refractive index structure of the specimen itself. The aberrations cause reduced signal intensity and the 3D resolution of the instrument is compromised. It has been suggested to correct for aberrations in confocal microscopes using adaptive optics. In order to define the design specifications for such adaptive optics systems, one has to know the amount of aberrations present for typical applications such as with biological samples. We have built a phase stepping interferometer microscope that directly measures the aberration of the wavefront. The modal content of the wavefront is extracted by employing Zernike mode decomposition. Results for typical biological specimens are presented. It was found for all samples investigated that higher order Zernike modes give only a small contribution to the overall aberration. Therefore, these higher order modes can be neglected in future adaptive optics sensing and correction schemes implemented into confocal or multiphoton microscopes, leading to more efficient designs.

Multivariate statistical analysis of low-voltage EDS spectrum images

DOE Office of Scientific and Technical Information (OSTI.GOV)

Anderson, I.M.

1998-03-01

Whereas energy-dispersive X-ray spectrometry (EDS) has been used for compositional analysis in the scanning electron microscope for 30 years, the benefits of using low operating voltages for such analyses have been explored only during the last few years. This paper couples low-voltage EDS with two other emerging areas of characterization: spectrum imaging and multivariate statistical analysis. The specimen analyzed for this study was a finished Intel Pentium processor, with the polyimide protective coating stripped off to expose the final active layers.

Manfred Girbardt and Charles Bracker: outstanding pioneers in fungal microscopy.

PubMed

Bartnicki-Garcia, Salomon

2015-01-01

Midway through the twentieth century, the availability of new and improved optical and electronic microscopes facilitated rapid advances in the elucidation of the fine structure of fungal cells. In this Essay, I pay tribute to Manfred Girbardt (1919-1991) and Charles Bracker (1938-2012)—two individuals who, despite being separated by geography and the restrictions of the Cold War, both made equally fundamental discoveries in fungal cell ultrastructure and set high standards for specimen manipulation and image processing.

Measuring nanometre-scale electric fields in scanning transmission electron microscopy using segmented detectors.

PubMed

Brown, H G; Shibata, N; Sasaki, H; Petersen, T C; Paganin, D M; Morgan, M J; Findlay, S D

2017-11-01

Electric field mapping using segmented detectors in the scanning transmission electron microscope has recently been achieved at the nanometre scale. However, converting these results to quantitative field measurements involves assumptions whose validity is unclear for thick specimens. We consider three approaches to quantitative reconstruction of the projected electric potential using segmented detectors: a segmented detector approximation to differential phase contrast and two variants on ptychographical reconstruction. Limitations to these approaches are also studied, particularly errors arising from detector segment size, inelastic scattering, and non-periodic boundary conditions. A simple calibration experiment is described which corrects the differential phase contrast reconstruction to give reliable quantitative results despite the finite detector segment size and the effects of plasmon scattering in thick specimens. A plasmon scattering correction to the segmented detector ptychography approaches is also given. Avoiding the imposition of periodic boundary conditions on the reconstructed projected electric potential leads to more realistic reconstructions. Copyright © 2017 Elsevier B.V. All rights reserved.

Fatigue and failure responses of lead zirconate titanate multilayer actuator under unipolar high-field electric cycling

NASA Astrophysics Data System (ADS)

Zeng, Fan Wen; Wang, Hong; Lin, Hua-Tay

2013-07-01

Lead zirconate titanate (PZT) multilayer actuators with an interdigital electrode design were studied under high electric fields (3 and 6 kV/mm) in a unipolar cycling mode. A 100 Hz sine wave was used in cycling. Five specimens tested under 6 kV/mm failed from 3.8 × 105 to 7 × 105 cycles, whereas three other specimens tested under 3 kV/mm were found to be still functional after 108 cycles. Variations in piezoelectric and dielectric responses of the tested specimens were observed during the fatigue test, depending on the measuring and cycling conditions. Selected fatigued and damaged actuators were characterized using an impedance analyzer or small signal measurement. Furthermore, involved fatigue and failure mechanisms were investigated using scanning acoustic microscope and scanning electron microscope. The extensive cracks and porous regions were revealed across the PZT layers on the cross sections of a failed actuator. The results from this study have demonstrated that the high-field cycling can accelerate the fatigue of PZT stacks as long as the partial discharge is controlled. The small signal measurement can also be integrated into the large signal measurement to characterize the fatigue response of PZT stacks in a more comprehensive basis. The former can further serve as an experimental method to test and monitor the behavior of PZT stacks.

Comparison of Fluorescence Microscopy and Different Growth Media Culture Methods for Acanthamoeba Keratitis Diagnosis.

PubMed

Peretz, Avi; Geffen, Yuval; Socea, Soergiu D; Pastukh, Nina; Graffi, Shmuel

2015-08-01

Acanthamoeba keratitis (AK), a potentially blinding infection of the cornea, is caused by a free-living protozoan. Culture and microscopic examination of corneal scraping tissue material is the conventional method for identifying Acanthamoeba. In this article, we compared several methods for AK diagnosis of 32 patients: microscopic examination using fluorescent dye, specific culture on growth media-non-nutrient agar (NNA), culture on liquid growth media-peptone yeast glucose (PYG), and TYI-S-33. AK was found in 14 patients. Thirteen of the specimens were found AK positive by fluorescence microscopic examination, 11 specimens were found AK positive on PYG growth media, and 9 specimens were found AK positive on TYI-S-33 growth media. Only five specimens were found AK positive on NNA growth media. Therefore, we recommend using fluorescence microscopy technique and culture method, especially PYG liquid media. © The American Society of Tropical Medicine and Hygiene.

X-ray microscopy with high-resolution zone plates: recent developments

NASA Astrophysics Data System (ADS)

Schneider, Gerd; Wilhein, Thomas; Niemann, Bastian; Guttman, P.; Schliebe, T.; Lehr, J.; Aschoff, H.; Thieme, Juergen; Rudolph, Dietbert M.; Schmahl, Guenther A.

1995-09-01

In order to expand the applications of x-ray microscopy, developments in the fields of zone plate technology, specimen preparation and imaging techniques have been made. A new cross- linked polymer chain electron beam resist allows us to record zone plate pattern down to 19 nm outermost zone width. High resolution zone plates in germanium with outermost zone widths down to 19 nm have been developed. In addition, phase zone plates in nickel down to 30 nm zone width have been made by electroplating. In order to enhance the image contrast for weak absorbing objects, the phase contrast method for x-ray microscopy was developed and implemented on the Gottingen x-ray microscope at BESSY. The effects of x ray absorption on the structure of biological specimen limits the maximum applicable radiation dose and therefore the achievable signal to noise ratio for an artifact-free x-ray image. To improve the stability especially of biological specimen, a cryogenic object chamber has been developed and tested. It turns out that at the operating temperature T less than or equal to 130 K unfixed biological specimen can be exposed to a radiation dose of 109 - 1010 Gy without any observable structural changes. A multiple-angle viewing stage allows us to take stereoscopic images with the x-ray microscope, giving a 3D-impression of the object. As an example for the applications of x-ray microscopy in biology, erythrocytes infected by malaria parasite have been examined. Studies of the aggregation of hematite by sodium sulfate gives an example for the application of x-ray microscopy in the field of colloid research.

Pre-aging time dependence of microstructure and mechanical properties in nanostructured Al-2wt%Cu alloy

NASA Astrophysics Data System (ADS)

Azad, Bahram; Borhani, Ehsan

2016-03-01

This work is focused on the effect of pre-aging time on the properties of Al-2wt%Cu alloy processed by accumulative roll bonding (ARB) process. Following aged at 190 °C for 10 or 30 min, the samples were deformed up to a strain of 4.8 by the ARB process. The microstructure evolution was investigated by transmission electron microscope and electron backscattering diffraction analyzes. The results showed that the Al2Cu precipitates were formed with different sizes due to the different pre-aging times and the finer precipitates were more effective on the formation of high angle grain boundaries during the ARB process. The grain size of Aged-10 min and Aged-30 min specimens decreased to 400 nm and 420 nm, respectively, after 6 cycles of the ARB process. Also, the final texture after 6 cycles of the ARB process, shown in the {111} pole figure, were different depending on the starting microstructures. The mechanical properties of specimens were investigated by the Vickers microhardness measurements and the tensile tests. The results showed that the mechanical properties are affected by the starting microstructure. The mechanical properties of Aged-10 min specimen were different compared to Aged-30 min specimen due to the different size of the pre-existing precipitates. Although by continuing process, the precipitates were probably dissolved due to the heavy deformation.

Tip Effect of the Tapping Mode of Atomic Force Microscope in Viscous Fluid Environments.

PubMed

Shih, Hua-Ju; Shih, Po-Jen

2015-07-28

Atomic force microscope with applicable types of operation in a liquid environment is widely used to scan the contours of biological specimens. The contact mode of operation allows a tip to touch a specimen directly but sometimes it damages the specimen; thus, a tapping mode of operation may replace the contact mode. The tapping mode triggers the cantilever of the microscope approximately at resonance frequencies, and so the tip periodically knocks the specimen. It is well known that the cantilever induces extra liquid pressure that leads to drift in the resonance frequency. Studies have noted that the heights of protein surfaces measured via the tapping mode of an atomic force microscope are ~25% smaller than those measured by other methods. This discrepancy may be attributable to the induced superficial hydrodynamic pressure, which is worth investigating. In this paper, we introduce a semi-analytical method to analyze the pressure distribution of various tip geometries. According to our analysis, the maximum hydrodynamic pressure on the specimen caused by a cone-shaped tip is ~0.5 Pa, which can, for example, pre-deform a cell by several nanometers in compression before the tip taps it. Moreover, the pressure calculated on the surface of the specimen is 20 times larger than the pressure without considering the tip effect; these results have not been motioned in other papers. Dominating factors, such as surface heights of protein surface, mechanical stiffness of protein increasing with loading velocity, and radius of tip affecting the local pressure of specimen, are also addressed in this study.

Fracture Toughness and Shear Strength of the Bonded Interface Between an Aluminium Alloy Skin and a FRP Patch

NASA Astrophysics Data System (ADS)

Kumar, Prashant; Shinde, Prakash Sonyabapu; Bhoyar, Gaurav

2018-05-01

The existing techniques to determine the fracture properties such as critical energy release rate in mode I (GIc) and mode II (GIIc) of an interface between two sheets of same material were modified to determine these properties between the sheets of dissimilar materials and thickness. In addition, the interface shear strength (ISS) was also determined. Experiments were carried out on the specimens made of a pre-cracked thin aluminium alloy skin and a Fiber reinforced polymer (FRP) patch. Two kinds of surface preparation of the aluminium skin were employed; (i) emery-paper roughened surface (ERS) and (ii) Sodium Hydroxide (NaOH) treated surface (NTS). GIc of ERS specimen was found to be 36.1 J/m2, while it was found to be much higher for NTS specimens, that is, 87.3 J/m2. GIIc was found to be 282.4 J/m2 for ERS specimens and much higher as 734.5 J/m2 for NTS specimens. ISS was determined as 32.6 MPa for ERS specimen and significantly higher for NTS specimen, that is, 44.5 MPa. The micrographs obtained from a field emission-scanning electron microscope (FE-SEM) and the surface roughness test showed that the NTS was significantly rougher than the ERS, explaining the higher values of all the three kinds of NTS specimens.

High-pressure freezing and freeze substitution of Arabidopsis for electron microscopy.

PubMed

Austin, Jotham R

2014-01-01

The objectives of electron microscopy ultrastructural studies are to examine cellular architecture and relate the cell's structural machinery to dynamic functional roles. This aspiration is difficult to achieve if specimens have not been adequately preserved in a "living state"; hence specimen preparation is of the utmost importance for the success of any electron micrographic study. High-pressure freezing (HPF)/freeze substitution (FS) has long been recognized as the primer technique for the preservation of ultrastructure in biological samples. In most cases a basic HPF/freeze substitution protocol is sufficient to obtain superior ultrastructural preservation and structural contrast, which allows one to use more advanced microscopy techniques such as 3D electron tomography. However, for plant tissues, which have a thick cell wall, large water-filled vacuoles, and air spaces (all of which are detrimental to cryopreservation), these basic HPF/FS protocols often yield undesirable results. In particular, ice crystal artifacts and the staining of membrane systems are often poorly or negatively stained, which make 3D segmentation of a tomogram difficult. To overcome these problems, various aspects of the HPF/FS protocol can be altered, including the cryo-filler(s) used, freeze substitution cocktail, and the resin infiltration process. This chapter will describe these modifications for the preparation of plant tissues for routine electron microscopic studies, immunocytochemistry, and 3D tomographic electron imaging.

Specimen Identification Errors in Breast Biopsies: Age Matters. Report of Two Near-Miss Events and Review of the Literature.

PubMed

Tozbikian, Gary; Gemignani, Mary L; Brogi, Edi

2017-09-01

The consequences of patient identification errors due to specimen mislabeling can be deleterious. We describe two near-miss events involving mislabeled breast specimens from two patients who sought treatment at our institution. In both cases, microscopic review of the slides identified inconsistencies between the histologic findings and patient age, unveiling specimen identification errors. By correlating the clinical information with the microscopic findings, we identified mistakes that had occurred at the time of specimen accessioning at the original laboratories. In both cases, thanks to a timely reassignment of the specimens, the patients suffered no harm. These cases highlight the importance of routine clinical and pathologic correlation as a critical component of quality assurance and patient safety. A review of possible specimen identification errors in the anatomic pathology setting is presented. © 2017 Wiley Periodicals, Inc.

DOE Office of Scientific and Technical Information (OSTI.GOV)

Cullen, David A; Koestner, Roland; Kukreja, Ratan

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 exposuremore » 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.« less

Undecalcified temporal bone morphology: a methodology useful for gross to fine observation and three-dimensional reconstruction.

PubMed

Fujiyoshi, T; Mogi, G; Watanabe, T; Matsushita, F

1992-01-01

Using a novel method of cutting undecalcified temporal bone specimens, quantitative structural analysis in the human and the Japanese monkey was undertaken. One millimeter thick serial slices made from unembedded temporal bones retained fine structure. Therefore, gross to fine observation could be performed systematically at the macroscopic, light, scanning, and transmission electron microscopic levels. The entire temporal bone three-dimensional reconstruction was completed from embedded sections; consequently, the volume of the tubotympanum and air cell system could be calculated. Available methods by embedding, tungsten carbide sectioning, grinding, and microwave irradiation for decalcification were also examined. These morphologic studies suggest that these novel methods offer timesaving advantages over any presently available techniques, and allow for elucidation of temporal bone morphology with only a few specimens.

The effect of inhibitor sodium nitrate on pitting corrosion of dissimilar material weldment joint of stainless steel AISI 304 and mild steel SS 400

DOE Office of Scientific and Technical Information (OSTI.GOV)

Hilca, B. R., E-mail: bangkithilca@yahoo.com; Triyono, E-mail: triyonomesin@uns.ac.id

This study experimentally evaluated the effect of Sodium Nitrate inhibitor (NaNO{sub 3}) of 0.1%, 0.3%, and 0.5% on NaCl 3.5% toward pitting corrosion of dissimilar metal welding joint between stainless steel AISI 304 and mild steel SS 400. Electrochemical corrosion was tested using potentiodynamic polarization. Further the Scanning Electron Microscope (SEM) conducted to analyze the specimen. Chemical composition analysis used Energy Dispersive X-ray Spectrometry (EDS). The highest efficiency of sodium nitrate for ER 308 attained 63.8% and 64.89%for ER 309L. The specimen surface which observed through SEM showed decrease of pitting corrosion respectively with the addition of sodium nitrate contentmore » as inhibitor.« less

Rotavirus infection in wild marsupials (Didelphis marsupialis) of the Amazon region.

PubMed

Linhares, A C; Pereira, J D; Nakauth, C M; Gabbay, Y B

1986-01-01

Rotavirus was detected by enzyme-linked immunosorbent assay in faecal specimens collected from two (1.35%) of 148 marsupials trapped in the Amazon jungle environment. The positive samples were both from the "common opossum", Didelphis marsupialis. No infections were found in the stools of 198 animals belonging to other mammalian species: the latter included small rodents, chiropterans and primates. Electron microscopic examination of one (MA 5928) rotavirus-positive specimen showed a large number of empty particles. However, both rotavirus strains grew when inoculated in MA 104 cells (foetal Rhesus monkey kidney cells) producing clear cytopathogenic effect; indirect immunofluorescence technique of these cells showed a typical granular cytoplasmic fluorescence. The electrophoretic profile of strain MA 5928 showed a high grade of homology with that of SA 11, but also showed minor differences.

Morphological, biochemical, histological, and ultrastructural protective effects of misoprostol on cisplatin induced-hepatotoxicity in adult male rats.

PubMed

Nasr, Ashraf Y

2013-12-01

To investigate the possible protective effect of misoprostol on cisplatin-induced hepatotoxicity. Four-equal sized groups (control, cisplatin-treated, misoprostol-treated, combined misoprostol, and cisplatin-treated) adult male Wistar rats (6 each) were used in this study. Body weight, liver weight, and liver weight/body weight ratio was calculated. Blood samples were obtained from the hearts of rats to determine the levels of total serum bilirubin (TSB), aspartate aminotransferase (AST), alanine aminotransferase (ALT), and albumin. Liver specimens were prepared for both light and electron microscopes. The study was carried out between June 2012 and April 2013 at the Anatomy Department, Faculty of Medicine, Zagazig University, Zagazig, Egypt, and the Department of Anatomy, Faculty of Medicine, King Abdulaziz University, Jeddah, Kingdom of Saudi Arabia. A single cisplatin dose (7.5 mg/kg intraperitoneally) resulted in significant elevation of AST, ALT, and TSB serum levels, and a significant reduction of serum albumin level, body weight, liver weight, and liver weight/body weight ratio. A combination of misoprostol (200 ug/kg/day) with cisplatin improved most of the previous parameters. Examination of specimens by both light and electron microscopes revealed pericentral hepatic necrosis, periportal fibrosis, dilatation, and congestion of central vein and blood sinusoids, diminished glycogen content, degenerated mitochondria, vesicular dilated rough endoplasmic reticulum, and nuclear changes in cisplatin-treated rats. Oral intake of misoprostol with cisplatin improved many of these changes. The results indicate that misoprostol may have a protective effect on cisplatin-induced hepatotoxicity.

Perspective: Electronic systems of knowledge in the world of virtual microscopy.

PubMed

Maybury, Terrence; Farah, Camile S

2009-09-01

Across a broad range of medical disciplines, learning how to use an optical or light microscope has been a mandatory inclusion in the undergraduate curriculum. The development of virtual microscopy (VM) technology during the past 10 years has called into question the use of the optical microscope in educational contexts. VM allows slide specimens to be digitized, which, in turn, allows the computer to mimic the workings of the light microscope. This move from analog technology (the light microscope) to digital technology (the computer as microscope) is part of the many significant changes going on in education, a singular manifestation of the broader move from print-literate traditions of knowledge (requiring literacy) to an electronics-literate, or "electrate," mode (requiring "electracy"). VM is here used as an exemplar of this broad transition from literacy to electracy, some components of which include data deluge, a multimodal structure, and modularity. Understandably, this transition is important to clarify educationally, especially in a global context mediated via digital means. A related aspect of these educational changes is the move from teacher-directed learning to student-centered learning, or "user-led education," which points to a redefinition of "pedagogy" as "andragogy." The dissemination of the specific value of VM, then, is critical to both learners and teachers and to a more coherent understanding of electracy. A practical consequence of this clarity might be a better application of this knowledge in the evolving fields of computer simulation and telemedicine, areas in which today's medical students will need future expertise.

Sensing mode atomic force microscope

DOEpatents

Hough, Paul V. C.; Wang, Chengpu

2006-08-22

An atomic force microscope is described having a cantilever comprising a base and a probe tip on an end opposite the base; a cantilever drive device connected to the base; a magnetic material coupled to the probe tip, such that when an incrementally increasing magnetic field is applied to the magnetic material an incrementally increasing force will be applied to the probe tip; a moveable specimen base; and a controller constructed to obtain a profile height of a specimen at a point based upon a contact between the probe tip and a specimen, and measure an adhesion force between the probe tip and the specimen by, under control of a program, incrementally increasing an amount of a magnetic field until a release force, sufficient to break the contact, is applied. An imaging method for atomic force microscopy involving measuring a specimen profile height and adhesion force at multiple points within an area and concurrently displaying the profile and adhesion force for each of the points is also described. A microscope controller is also described and is constructed to, for a group of points, calculate a specimen height at a point based upon a cantilever deflection, a cantilever base position and a specimen piezo position; calculate an adhesion force between a probe tip and a specimen at the point by causing an incrementally increasing force to be applied to the probe tip until the probe tip separates from a specimen; and move the probe tip to a new point in the group.

Sensing mode atomic force microscope

DOEpatents

Hough, Paul V.; Wang, Chengpu

2004-11-16

An atomic force microscope is described having a cantilever comprising a base and a probe tip on an end opposite the base; a cantilever drive device connected to the base; a magnetic material coupled to the probe tip, such that when an incrementally increasing magnetic field is applied to the magnetic material an incrementally increasing force will be applied to the probe tip; a moveable specimen base; and a controller constructed to obtain a profile height of a specimen at a point based upon a contact between the probe tip and a specimen, and measure an adhesion force between the probe tip and the specimen by, under control of a program, incrementally increasing an amount of a magnetic field until a release force, sufficient to break the contact, is applied. An imaging method for atomic force microscopy involving measuring a specimen profile height and adhesion force at multiple points within an area and concurrently displaying the profile and adhesion force for each of the points is also described. A microscope controller is also described and is constructed to, for a group of points, calculate a specimen height at a point based upon a cantilever deflection, a cantilever base position and a specimen piezo position; calculate an adhesion force between a probe tip and a specimen at the point by causing an incrementally increasing force to be applied to the probe tip until the probe tip separates from a specimen; and move the probe tip to a new point in the group.

Shear bond strength of indirect composite material to monolithic zirconia

PubMed Central

2016-01-01

PURPOSE This study aimed to evaluate the effect of surface treatments on bond strength of indirect composite material (Tescera Indirect Composite System) to monolithic zirconia (inCoris TZI). MATERIALS AND METHODS Partially stabilized monolithic zirconia blocks were cut into with 2.0 mm thickness. Sintered zirconia specimens were divided into different surface treatment groups: no treatment (control), sandblasting, glaze layer & hydrofluoric acid application, and sandblasting + glaze layer & hydrofluoric acid application. The indirect composite material was applied to the surface of the monolithic zirconia specimens. Shear bond strength value of each specimen was evaluated after thermocycling. The fractured surface of each specimen was examined with a stereomicroscope and a scanning electron microscope to assess the failure types. The data were analyzed using one-way analysis of variance (ANOVA) and Tukey LSD tests (α=.05). RESULTS Bond strength was significantly lower in untreated specimens than in sandblasted specimens (P<.05). No difference between the glaze layer and hydrofluoric acid application treated groups were observed. However, bond strength for these groups were significantly higher as compared with the other two groups (P<.05). CONCLUSION Combined use of glaze layer & hydrofluoric acid application and silanization are reliable for strong and durable bonding between indirect composite material and monolithic zirconia. PMID:27555895

Development of a multi-cycle shear-compression testing for the modeling of severe plastic deformation

NASA Astrophysics Data System (ADS)

Pesin, A.; Pustovoytov, D.; Lokotunina, N.

2017-12-01

The mechanism of severe plastic deformation comes from very significant shear strain. Shear-compression testing of materials is complicated by the fact that a state of large equivalent strain with dominant shear strain is not easily achievable. This paper presents the novel technique of laboratory simulation of severe plastic deformation by multi-cycle shear-compression testing at room temperature with equivalent strain e=1…5. The specimen consisted of a parallelepiped having an inclined gauge section created by two diametrically opposed semi-circular slots which were machined at 45°. Height of the specimen was 50 mm, section dimensions were 25×25 mm, gauge thickness was 5.0 mm and gauge width was 6.0 mm. The specimen provided dominant shear strain in an inclined gauge-section. The level of shear strain and equivalent strain was controlled through adjustment of the height reduction of the specimen, load application direction and number of cycles of shear-compression. Aluminium alloy Al-6.2Mg-0.7Mn was used as a material for specimen. FE simulation and analysis of the stress-strain state were performed. The microstructure of the specimen after multi-cycle shear-compression testing with equivalent strain e=1…5 was examined by optical and scanning electron microscope.

System design and verification of the precession electron diffraction technique

NASA Astrophysics Data System (ADS)

Own, Christopher Su-Yan

2005-07-01

Bulk structural crystallography is generally a two-part process wherein a rough starting structure model is first derived, then later refined to give an accurate model of the structure. The critical step is the determination of the initial model. As materials problems decrease in length scale, the electron microscope has proven to be a versatile and effective tool for studying many problems. However, study of complex bulk structures by electron diffraction has been hindered by the problem of dynamical diffraction. This phenomenon makes bulk electron diffraction very sensitive to specimen thickness, and expensive equipment such as aberration-corrected scanning transmission microscopes or elaborate methodology such as high resolution imaging combined with diffraction and simulation are often required to generate good starting structures. The precession electron diffraction technique (PED), which has the ability to significantly reduce dynamical effects in diffraction patterns, has shown promise as being a "philosopher's stone" for bulk electron diffraction. However, a comprehensive understanding of its abilities and limitations is necessary before it can be put into widespread use as a standalone technique. This thesis aims to bridge the gaps in understanding and utilizing precession so that practical application might be realized. Two new PED systems have been built, and optimal operating parameters have been elucidated. The role of lens aberrations is described in detail, and an alignment procedure is given that shows how to circumvent aberration in order to obtain high-quality patterns. Multislice simulation is used for investigating the errors inherent in precession, and is also used as a reference for comparison to simple models and to experimental PED data. General trends over a large sampling of parameter space are determined. In particular, we show that the primary reflection intensity errors occur near the transmitted beam and decay with increasing angle and decreasing specimen thickness. These errors, occurring at the lowest spatial frequencies, fortuitously coincide with reflections for which phases are easiest to determine via imaging methods. A general two-beam dynamical model based upon an existing approximate model is found to be fairly accurate across most experimental conditions, particularly where it is needed for providing a correction to distorted data. Finally, the practical structure solution procedure using PED is demonstrated for several model material systems. Of the experiment parameters investigated, the cone semi-angle is found to be the most important (it should be as large as possible), followed closely by specimen thickness (thinner is better). Assuming good structure projection characteristics in the specimen, the thickness tractable by PED is extended to 40-50 nm without correction, demonstrated for complex oxides. With a forward calculation based upon the two-beam dynamical model (using known structure factors), usable specimen thickness can be extended past 150 nm. For a priori correction, using the squared amplitudes approximates the two-beam model for most thicknesses if the scattering from the structure adheres to psuedo-kinematical behavior. Practically, crystals up to 60 nm in thickness can now be processed by the precession methods developed in this thesis.

The Scanning Optical Microscope.

ERIC Educational Resources Information Center

Sheppard, C. J. R.

1978-01-01

Describes the principle of the scanning optical microscope and explains its advantages over the conventional microscope in the improvement of resolution and contrast, as well as the possibility of producing a picture from optical harmonies generated within the specimen.

Microstructural studies of 35 degrees C copper Ni-Ti orthodontic wire and TEM confirmation of low-temperature martensite transformation.

PubMed

Brantley, William A; Guo, Wenhua; Clark, William A T; Iijima, Masahiro

2008-02-01

Previous temperature-modulated differential scanning calorimetry (TMDSC) study of nickel-titanium orthodontic wires revealed a large exothermic low-temperature peak that was attributed to transformation within martensitic NiTi. The purpose of this study was to use transmission electron microscopy (TEM) to verify this phase transformation in a clinically popular nickel-titanium wire, identify its mechanism and confirm other phase transformations found by TMDSC, and to provide detailed information about the microstructure of this wire. The 35 degrees C Copper nickel-titanium wire (Ormco) with cross-section dimensions of 0.016 in. x 0.022 in. used in the earlier TMDSC investigation was selected. Foils were prepared for TEM analyses by mechanical grinding, polishing, dimpling, ion milling and plasma cleaning. Standard bright-field and dark-field TEM images were obtained, along with convergent-beam electron diffraction patterns. A cryo-stage with the electron microscope (Phillips CM 200) permitted the specimen to be observed at -187, -45, and 50 degrees C, as well as at room temperature. Microstructures were also observed with an optical microscope and a scanning electron microscope. Room temperature microstructures had randomly oriented, elongated grains that were twinned. Electron diffraction patterns confirmed that phase transformations took place over temperature ranges previously found by TMDSC. TEM observations revealed a high dislocation density and fine-scale oxide particles, and that twinning is the mechanism for the low-temperature transformation in martensitic NiTi. TEM confirmed the low-temperature peak and other phase transformations observed by TMDSC, and revealed that twinning in martensite is the mechanism for the low-temperature peak. The high dislocation density and fine-scale oxide particles in the microstructure are the result of the wire manufacturing process.

Light, electron microscopic and immunohistochemical study of the effect of low-dose aspirin during the proestrus phase on rat endometrium in the preimplantation period.

PubMed

Ateş, Utku; Baka, Meral; Turgut, Mehmet; Uyanikgil, Yiğit; Ulker, Sibel; Yilmaz, Ozlem; Tavmergen, Erol; Yurtseven, Mine

2007-04-01

To evaluate structural alterations in rat endometrium at preimplantation following treatment with aspirin beginning from proestrus by light microscopy, electron microscopy and immunohistochemical techniques. Twenty rats were divided into control (n = 10) and experimental (n = 10) groups. Experimental rats were treated with low-dose aspirin daily (2 mg/kg/day) during estrus, beginning from the proestrus phase, mated at end of cycle and treated with aspirin. Untreated pregnant rats were the control group. Rats in both groups were sacrificed at the 84th pregnancy hour; the uterus was rapidly removed and dissected free of surrounding adipose tissue. Uteri specimens from nonpregnant rats were transferred into fixative solution and processed for light, electron microscopic and immunohistochemical study. Light and electron microscopy of endometrium from control rats conformed to mid-diestrus phase; endometrial histology of the aspirin-treated group conformed to late diestrus phase. The endometrial layer was significantly thicker in the aspirin-treated group compared to the untreated control group (p <0.001). No significant difference was found in vessel number between groups. Staining with alphaV integrin was more dense in the aspirin-treated group. Based on histologic findings, we suggest low-dose aspirin has positive effects on preparing endometrium before implantation.

Three-dimensional cytomorphology in fine needle aspiration biopsy of medullary thyroid carcinoma.

PubMed

Chang, T C; Lai, S M; Wen, C Y; Hsiao, Y L; Huang, S H

2001-01-01

To elucidate three-dimensional (3-D) cytomorphology in fine needle aspiration biopsy (FNAB) of medullary thyroid carcinoma (MTC). ENAB was performed on tumors from five patients with MTC. The aspirate was stained and observed under a light microscope (LM). The aspirate was also fixed, dehydrated, critical point dried, spattered with gold ions and observed with a scanning electron microscope (SEM). For transmission electron microscopy (TEM), the specimen was fixed, dehydrated, embedded in an Epon mixture, cut with an ultramicrotome, mounted on copper grids, electron doubly stained with uranium acetate and lead citrate, and observed with TEM. Findings under SEM were correlated with those under LM and TEM. Under SEM, 3-D cytomorphology of MTC displayed a disorganized cellular arrangement with indistinct cell borders in three cases. The cell surface was uneven and had granular protrusions that corresponded to secretory granules observed under TEM. In one case with multiple endocrine neoplasia type IIB, there were abundant granules on the cell surface. In one case of sporadic MTC with multinucleated tumor giant cells and small cells, granular protrusions also were noted on the cell surface. Granular protrusion was a characteristic finding in FNAB of MTC tinder SEM and might be helpful in the differential diagnosis.

Preparation and Observation of Thick Biological Samples by Scanning Transmission Electron Tomography.

PubMed

Trépout, Sylvain; Bastin, Philippe; Marco, Sergio

2017-03-12

This report describes a protocol for preparing thick biological specimens for further observation using a scanning transmission electron microscope. It also describes an imaging method for studying the 3D structure of thick biological specimens by scanning transmission electron tomography. The sample preparation protocol is based on conventional methods in which the sample is fixed using chemical agents, treated with a heavy atom salt contrasting agent, dehydrated in a series of ethanol baths, and embedded in resin. The specific imaging conditions for observing thick samples by scanning transmission electron microscopy are then described. Sections of the sample are observed using a through-focus method involving the collection of several images at various focal planes. This enables the recovery of in-focus information at various heights throughout the sample. This particular collection pattern is performed at each tilt angle during tomography data collection. A single image is then generated, merging the in-focus information from all the different focal planes. A classic tilt-series dataset is then generated. The advantage of the method is that the tilt-series alignment and reconstruction can be performed using standard tools. The collection of through-focal images allows the reconstruction of a 3D volume that contains all of the structural details of the sample in focus.

Microscopical and elemental FESEM and Phenom ProX-SEM-EDS analysis of osteocyte- and blood vessel-like microstructures obtained from fossil vertebrates of the Eocene Messel Pit, Germany.

PubMed

Cadena, Edwin

2016-01-01

The Eocene (∾48 Ma) Messel Pit in Germany is a UNESCO World Heritage Site because of its exceptionally preserved fossils, including vertebrates, invertebrates, and plants. Messel fossil vertebrates are typically characterized by their articulated state, and in some cases the skin, hair, feathers, scales and stomach contents are also preserved. Despite the exceptional macroscopic preservation of Messel fossil vertebrates, the microstructural aspect of these fossils has been poorly explored. In particular, soft tissue structures such as hair or feathers have not been chemically analyzed, nor have bone microstructures. I report here the preservation and recovery of osteocyte-like and blood vessel-like microstructures from the bone of Messel Pit specimens, including the turtles Allaeochelys crassesculpta and Neochelys franzeni, the crocodile Diplocynodon darwini, and the pangolin Eomanis krebsi. I used a Field Emission Scanning Electron Microscope (FESEM) and a Phenom ProX desktop scanning electron microscope (LOT-QuantumDesign) equipped with a thermionic CeB6 source and a high sensitivity multi-mode backscatter electron (BSE) for microscopical and elemental characterization of these bone microstructures. Osteocyte-like and blood vessel-like microstructures are constituted by a thin layer (∾50 nm thickness), external and internal mottled texture with slightly marked striations. Circular to linear marks are common on the external surface of the osteocyte-like microstructures and are interpreted as microbial troughs. Iron (Fe) is the most abundant element found in the osteocyte-like and blood vessel-like microstructures, but not in the bone matrix or collagen fibril-like microstructures. The occurrence of well-preserved soft-tissue elements (at least their physical form) establishes a promising background for future studies on preservation of biomolecules (proteins or DNA) in Messel Pit fossils.

Effect of grinding conditions on the fatigue life of titanium 5Al-2.5Sn alloy

NASA Technical Reports Server (NTRS)

Rangaswamy, P.; Terutung, H.; Jeelani, S.

1991-01-01

An investigation into the effect of grinding conditions on the fatigue life of titanium 5Al-2.5Sn is presented. Damage to surface integrity and changes in the residual stresses distribution are studied to assess changes in fatigue life. A surface grinding machine, operating at speeds ranging from 2000 to 6000 fpm and using SiC wheels of grit sizes 60 and 120, was used to grind flat subsize specimens of 0.1-in. thickness. After grinding, the specimens were fatigued at a chosen stress and compared with the unadulterated material. A standard profilometer, a microhardness tester, and a scanning electron microscope were utilized to examine surface characteristics and measure roughness and hardness. Increased grinding speed in both wet and dry applications tended to decrease the fatigue life of the specimens. Fatigue life increased markedly at 2000 fpm under wet conditions, but then decreased at higher speeds. Grit size had no effect on the fatigue life.

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.

Unsupervised segmentation of low-contrast multichannel images: discrimination of tissue components in microscopic images of unstained specimens

NASA Astrophysics Data System (ADS)

Kopriva, Ivica; Popović Hadžija, Marijana; Hadžija, Mirko; Aralica, Gorana

2015-06-01

Low-contrast images, such as color microscopic images of unstained histological specimens, are composed of objects with highly correlated spectral profiles. Such images are very hard to segment. Here, we present a method that nonlinearly maps low-contrast color image into an image with an increased number of non-physical channels and a decreased correlation between spectral profiles. The method is a proof-of-concept validated on the unsupervised segmentation of color images of unstained specimens, in which case the tissue components appear colorless when viewed under the light microscope. Specimens of human hepatocellular carcinoma, human liver with metastasis from colon and gastric cancer and mouse fatty liver were used for validation. The average correlation between the spectral profiles of the tissue components was greater than 0.9985, and the worst case correlation was greater than 0.9997. The proposed method can potentially be applied to the segmentation of low-contrast multichannel images with high spatial resolution that arise in other imaging modalities.

A Closed-Loop Proportional-Integral (PI) Control Software for Fully Mechanically Controlled Automated Electron Microscopic Tomography

DOE Office of Scientific and Technical Information (OSTI.GOV)

REN, GANG; LIU, JINXIN; LI, HONGCHANG

A closed-loop proportional-integral (PI) control software is provided for fully mechanically controlled automated electron microscopic tomography. The software is developed based on Gatan DigitalMicrograph, and is compatible with Zeiss LIBRA 120 transmission electron microscope. However, it can be expanded to other TEM instrument with modification. The software consists of a graphical user interface, a digital PI controller, an image analyzing unit, and other drive units (i.e.: image acquire unit and goniometer drive unit). During a tomography data collection process, the image analyzing unit analyzes both the accumulated shift and defocus value of the latest acquired image, and provides the resultsmore » to the digital PI controller. The digital PI control compares the results with the preset values and determines the optimum adjustments of the goniometer. The goniometer drive unit adjusts the spatial position of the specimen according to the instructions given by the digital PI controller for the next tilt angle and image acquisition. The goniometer drive unit achieves high precision positioning by using a backlash elimination method. The major benefits of the software are: 1) the goniometer drive unit keeps pre-aligned/optimized beam conditions unchanged and achieves position tracking solely through mechanical control; 2) the image analyzing unit relies on only historical data and therefore does not require additional images/exposures; 3) the PI controller enables the system to dynamically track the imaging target with extremely low system error.« less

In-situ scanning electron microscope studies of crack growth in an aluminum metal-matrix composite

NASA Technical Reports Server (NTRS)

Manoharan, M.; Lewandowski, J. J.

1990-01-01

Edge-notched specimens of a cast and extruded Al alloy-based, alumina particulate-reinforced composite in the annealed condition were tested in situ in a SEM apparatus equipped with a deformation stage permitting the direct observation of crack growth phenomena. Fracture in this composite is seen to proceed by initiation of microcracks ahead of the macrocrack; as deformation proceeds, the microcracks lengthen, and crack propagation occurs when the region of intense plastic straining becomes comparable to the macrocrack-microcrack distance. The sequence is then repeated.

Nucleation of fcc Ta when heating thin films

DOE PAGES

Janish, Matthew T.; Mook, William M.; Carter, C. Barry

2014-10-25

Thin tantalum films have been studied during in-situ heating in a transmission electron microscope. Diffraction patterns from the as-deposited films were typical of amorphous materials. Crystalline grains were observed to form when the specimen was annealed in-situ at 450°C. Particular attention was addressed to the formation and growth of grains with the face-centered cubic (fcc) crystal structure. As a result, these observations are discussed in relation to prior work on the formation of fcc Ta by deformation and during thin film deposition.

Comparative evaluation of woven graphite-epoxy composites

NASA Technical Reports Server (NTRS)

Hanagud, S.; Tayebi, A.; Clinton, R. G., Jr.; Nayak, B. M.

1979-01-01

A comparative evaluation of some of the mechanical properties of woven graphite-epoxy composites are discussed. In particular, the types of weaves and the resin contents were chosen for comparison. The types of weaves selected are plain weave, satin weave, and tridirectional weave. The composites made of the fabrics are compared to composites made from unidirectional tapes under static and fatigue loading. During static loading, acoustic emission events were monitored. Also, examinations of fracture surfaces and polished sections both away from the fracture surface, and of virgin specimens under an electron microscope are discussed.

Soft x-ray holography and microscopy of biological cells

NASA Astrophysics Data System (ADS)

Chen, Jianwen; Gao, Hongyi; Xie, Honglan; Li, Ruxin; Xu, Zhizhan

2003-10-01

Some experimental results on soft X-ray microscopy and holography imaging of biological specimens are presented in the paper. As we know, due to diffraction effects, there exists a resolution limit determined by wavelength λ and numerical aperture NA in conventional optical microscopy. In order to improve resolution, the num erical aperture should be made as large as possible and the wavelength as short as possible. Owing to the shorter wavelength, X-rays provide the potential of higher resolution in X-ray microscopy, holography image and allow for exam ination the interior structures of thicker specimens. In the experiments, we used synchrotron radiation source in Hefei as light source. Soft X-rays come from a bending magnet in 800 M eV electron storage ring with characteristic wavelength of 2.4 nm. The continuous X-ray spectrums are monochromatized by a zone-plate and a pinhole with 300 m diameter. The experimental set-up is typical contact microscopic system, its main advantage is simplicity and no special optical element is needed. The specimens used in the experiments of microscopic imaging are the colibacillus, the gingko vascular hundle and the fritillaries ovary karyon. The specimen for holographic imaging is the spider filam ents. The basic structures of plant cells such as the cell walls, the cytoplasm and the karyon especially the joint structures between the cells are observed clearly. An experimental study on a thick biological specimen that is a whole sporule w ith the thickness of about 30 μm is performed. In the holographic experiments, the experimental setup is typical Gabor in-line holography. The specimen is placed in line with X-ray source, which provides both the reference w aves and specimen illum ination. The specimen is some spider filament, which adhere to a Si3N4 film. The recording medium is PM M A, which is placed at recording distance of about 400 μm from the specimen. The hologram s were reconstructed by digital method with 300 nm resolutions. A novel method for recording in-line hologram is proposed which is called X-ray in-line holography with zone-plate magnification in this paper. The magnification factor of the micro zone plate imaging is about 103. The transverse resolution can be 48 nm in this method.

[Development of an Enterococcus faecalis periapical biofilm model for in vitro morphological study].

PubMed

Cao, Ridan; Hou, Benxiang

2014-08-01

This study aims to develop and observe a model system of the periapical biofilm structure of Enterococcus faecalis (E. faecalis). A total of 24 intact human single-rooted premolars extracted for orthodontic reasons were collected and randomly divided into eight groups (n = 3). The specimens were subjected to ultraviolet disinfection, inoculated with E. faecalis (ATCC 29212) suspension adjusted to 1 x 10(8) CFU x mL(-1), and incubated at 37 degrees C for 1, 2, and 7 d. Specimen groups were prepared for scanning electron microscope to examine the biofilm formation. The specimens in the confocal laser scanning microscope (CLSM) groups were stained with propidium iodide (PI) and ConA-fluorescein isothiocyanate (ConA-FITC) to examine the biofilm formation. The images were randomized, and biofilm coverage (%) was assessed using Photoshop CS5. The biofilm coverage (%) on the cementum increased with increasing incubation period. The biofilm coverage of the 7 d group was significantly higher than those of the 1 and 2 d groups (P < 0.05). The values of the latter two groups were not significantly different (P > 0.05). Dense aggregations composed of E. faecalis and the amorphous matrix were observed on the root cementum surfaces of the specimens in the 7 d group. The bacteria were stained red by PI, and the matrix was stained green by ConA-FITC under CLSM observation. The biofilm coverage (%) on the samples in the 7 d group was 17.23% +/- 1.52%, showing multi-level space structure and water channels. E. faecalis forms bacterial biofilms on the root cementum surface in 7 d. The biofilms were composed of E. faecalis and the amorphous matrix.

Quantitative characterization of nanoscale polycrystalline magnets with electron magnetic circular dichroism.

PubMed

Muto, Shunsuke; Rusz, Ján; Tatsumi, Kazuyoshi; Adam, Roman; Arai, Shigeo; Kocevski, Vancho; Oppeneer, Peter M; Bürgler, Daniel E; Schneider, Claus M

2014-01-01

Electron magnetic circular dichroism (EMCD) allows the quantitative, element-selective determination of spin and orbital magnetic moments, similar to its well-established X-ray counterpart, X-ray magnetic circular dichroism (XMCD). As an advantage over XMCD, EMCD measurements are made using transmission electron microscopes, which are routinely operated at sub-nanometre resolution, thereby potentially allowing nanometre magnetic characterization. However, because of the low intensity of the EMCD signal, it has not yet been possible to obtain quantitative information from EMCD signals at the nanoscale. Here we demonstrate a new approach to EMCD measurements that considerably enhances the outreach of the technique. The statistical analysis introduced here yields robust quantitative EMCD signals. Moreover, we demonstrate that quantitative magnetic information can be routinely obtained using electron beams of only a few nanometres in diameter without imposing any restriction regarding the crystalline order of the specimen.

Extraction of topographic and material contrasts on surfaces from SEM images obtained by energy filtering detection with low-energy primary electrons.

PubMed

Nagoshi, Masayasu; Aoyama, Tomohiro; Sato, Kaoru

2013-01-01

Secondary electron microscope (SEM) images have been obtained for practical materials using low primary electron energies and an in-lens type annular detector with changing negative bias voltage supplied to a grid placed in front of the detector. The kinetic-energy distribution of the detected electrons was evaluated by the gradient of the bias-energy dependence of the brightness of the images. This is divided into mainly two parts at about 500 V, high and low brightness in the low- and high-energy regions, respectively and shows difference among the surface regions having different composition and topography. The combination of the negative grid bias and the pixel-by-pixel image subtraction provides the band-pass filtered images and extracts the material and topographic information of the specimen surfaces. Copyright © 2012 Elsevier B.V. All rights reserved.

Influence of Austempering Heat Treatment on Microstructure and Mechanical Properties of Medium Carbon High Silicon Steel

NASA Astrophysics Data System (ADS)

Palaksha, P. A.; Ravishankar, K. S.

2017-08-01

In the present investigation, the influence of austempering heat treatment on the microstructure and mechanical properties of medium carbon high silicon steel was evaluated. The test specimens were machined from the as-received steel and were first austenitised at 900 °C for 45 minutes, followed by austempering heat treatment in salt bath at various temperatures 300 °C, 350 °C and 400 °C for a fixed duration of two hours, after that those specimens were air-cooled to room temperature. The characterization studies were carried out using optical microscope, scanning electron microscope (SEM) and x-ray diffractometer (XRD) and then correlated to the hardness and tensile properties. Results indicate that, the specimens austempered at lower temperature i.e. at 300 °C, which offered high hardness, tensile strength and lower ductility (1857 MPa and 13.3 %) due to the presence of acicular bainite i.e. lower bainite and also some martensite in the microstructure. At 350 °C, reduction in the tensile strength and hardness was observed, but comparatively higher ductility, which was favored by the presence of bainite laths i.e. upper bainitic structure along with higher retained austenite content. Finally at 400 °C, reduction in both ductility and tensile strength was observed, which is due to the precipitation of carbides between the banite laths, however good strain hardening response was observed at austempering temperatures of 350 °C and 400 °C.

Optical path difference microscopy with a Shack-Hartmann wavefront sensor.

PubMed

Gong, Hai; Agbana, Temitope E; Pozzi, Paolo; Soloviev, Oleg; Verhaegen, Michel; Vdovin, Gleb

2017-06-01

In this Letter, we show that a Shack-Hartmann wavefront sensor can be used for the quantitative measurement of the specimen optical path difference (OPD) in an ordinary incoherent optical microscope, if the spatial coherence of the illumination light in the plane of the specimen is larger than the microscope resolution. To satisfy this condition, the illumination numerical aperture should be smaller than the numerical aperture of the imaging lens. This principle has been successfully applied to build a high-resolution reference-free instrument for the characterization of the OPD of micro-optical components and microscopic biological samples.

Improved specimen reconstruction by Hilbert phase contrast tomography.

PubMed

Barton, Bastian; Joos, Friederike; Schröder, Rasmus R

2008-11-01

The low signal-to-noise ratio (SNR) in images of unstained specimens recorded with conventional defocus phase contrast makes it difficult to interpret 3D volumes obtained by electron tomography (ET). The high defocus applied for conventional tilt series generates some phase contrast but leads to an incomplete transfer of object information. For tomography of biological weak-phase objects, optimal image contrast and subsequently an optimized SNR are essential for the reconstruction of details such as macromolecular assemblies at molecular resolution. The problem of low contrast can be partially solved by applying a Hilbert phase plate positioned in the back focal plane (BFP) of the objective lens while recording images in Gaussian focus. Images recorded with the Hilbert phase plate provide optimized positive phase contrast at low spatial frequencies, and the contrast transfer in principle extends to the information limit of the microscope. The antisymmetric Hilbert phase contrast (HPC) can be numerically converted into isotropic contrast, which is equivalent to the contrast obtained by a Zernike phase plate. Thus, in-focus HPC provides optimal structure factor information without limiting effects of the transfer function. In this article, we present the first electron tomograms of biological specimens reconstructed from Hilbert phase plate image series. We outline the technical implementation of the phase plate and demonstrate that the technique is routinely applicable for tomography. A comparison between conventional defocus tomograms and in-focus HPC volumes shows an enhanced SNR and an improved specimen visibility for in-focus Hilbert tomography.

Monitoring damage growth in titanium matrix composites using acoustic emission

NASA Technical Reports Server (NTRS)

Bakuckas, J. G., Jr.; Prosser, W. H.; Johnson, W. S.

1993-01-01

The application of the acoustic emission (AE) technique to locate and monitor damage growth in titanium matrix composites (TMC) was investigated. Damage growth was studied using several optical techniques including a long focal length, high magnification microscope system with image acquisition capabilities. Fracture surface examinations were conducted using a scanning electron microscope (SEM). The AE technique was used to locate damage based on the arrival times of AE events between two sensors. Using model specimens exhibiting a dominant failure mechanism, correlations were established between the observed damage growth mechanisms and the AE results in terms of the events amplitude. These correlations were used to monitor the damage growth process in laminates exhibiting multiple modes of damage. Results revealed that the AE technique is a viable and effective tool to monitor damage growth in TMC.

[Clinical pathology on the verge of virtual microscopy].

PubMed

Tolonen, Teemu; Näpänkangas, Juha; Isola, Jorma

2015-01-01

For more than 100 years, examinations of pathology specimens have relied on the use of the light microscope. The technological progress of the last few years is enabling the digitizing of histologic specimen slides and application of the virtual microscope in diagnostics. Virtual microscopy will facilitate consultation possibilities, and digital image analysis serves to enhance the level of diagnostics. Organizing and monitoring clinicopathological meetings will become easier. Digital archive of histologic specimens and the virtual microscopy network are expected to benefit training and research as well, particularly what applies to the Finnish biobank network which is currently being established.

The significance of Bragg's law in electron diffraction and microscopy, and Bragg's second law.

PubMed

Humphreys, C J

2013-01-01

Bragg's second law, which deserves to be more widely known, is recounted. The significance of Bragg's law in electron diffraction and microscopy is then discussed, with particular emphasis on differences between X-ray and electron diffraction. As an example of such differences, the critical voltage effect in electron diffraction is described. It is then shown that the lattice imaging of crystals in high-resolution electron microscopy directly reveals the Bragg planes used for the imaging process, exactly as visualized by Bragg in his real-space law. Finally, it is shown how in 2012, for the first time, on the centennial anniversary of Bragg's law, single atoms have been identified in an electron microscope using X-rays emitted from the specimen. Hence atomic resolution X-ray maps of a crystal in real space can be formed which give the positions and identities of the different atoms in the crystal, or of a single impurity atom in the crystal.

Comparison of Macroscopic Pathology Measurements With Magnetic Resonance Imaging and Assessment of Microscopic Pathology Extension for Colorectal Liver Metastases

DOE Office of Scientific and Technical Information (OSTI.GOV)

Mendez Romero, Alejandra, E-mail: a.mendezromero@erasmusmc.nl; Verheij, Joanne; Dwarkasing, Roy S.

2012-01-01

Purpose: To compare pathology macroscopic tumor dimensions with magnetic resonance imaging (MRI) measurements and to establish the microscopic tumor extension of colorectal liver metastases. Methods and Materials: In a prospective pilot study we included patients with colorectal liver metastases planned for surgery and eligible for MRI. A liver MRI was performed within 48 hours before surgery. Directly after surgery, an MRI of the specimen was acquired to measure the degree of tumor shrinkage. The specimen was fixed in formalin for 48 hours, and another MRI was performed to assess the specimen/tumor shrinkage. All MRI sequences were imported into our radiotherapymore » treatment planning system, where the tumor and the specimen were delineated. For the macroscopic pathology analyses, photographs of the sliced specimens were used to delineate and reconstruct the tumor and the specimen volumes. Microscopic pathology analyses were conducted to assess the infiltration depth of tumor cell nests. Results: Between February 2009 and January 2010 we included 13 patients for analysis with 21 colorectal liver metastases. Specimen and tumor shrinkage after resection and fixation was negligible. The best tumor volume correlations between MRI and pathology were found for T1-weighted (w) echo gradient sequence (r{sub s} = 0.99, slope = 1.06), and the T2-w fast spin echo (FSE) single-shot sequence (r{sub s} = 0.99, slope = 1.08), followed by the T2-w FSE fat saturation sequence (r{sub s} = 0.99, slope = 1.23), and the T1-w gadolinium-enhanced sequence (r{sub s} = 0.98, slope = 1.24). We observed 39 tumor cell nests beyond the tumor border in 12 metastases. Microscopic extension was found between 0.2 and 10 mm from the main tumor, with 90% of the cases within 6 mm. Conclusions: MRI tumor dimensions showed a good agreement with the macroscopic pathology suggesting that MRI can be used for accurate tumor delineation. However, microscopic extensions found beyond the tumor border indicate that caution is needed in selecting appropriate tumor margins.« less

Studying dynamic processes in liquids by TEM/STEM/DTEM

NASA Astrophysics Data System (ADS)

Abellan, Patricia; Evans, James; Woehl, Taylor; Jungjohann, Katherine; Parent, Lucas; Arslan, Ilke; Ristenpart, William; Browning, Nigel; Mater. Sci. Group Team; Microsc. Group Team; Catal. Sci. Group Collaboration; Ristenpart Res. Group Collaboration

2013-03-01

In order to study dynamic phenomena such as corrosion or catalysis, extreme environmental conditions must be reproduced around the specimen - these include high-temperatures, high-pressures, specific oxidizing/reducing atmospheres or a liquid environment. The use of environmental stages specifically designed to fit in any transmission electron microscope (TEM) allows us to apply the distinct capabilities of each instrument to study dynamic processes. Localized gas/fluid conditions are created around the sample and separated from the high vacuum inside the microscope using hermetically sealed windowed-cells. Advanced capabilities of these techniques include spatial resolutions of ~1 Angstrom or better in aberration corrected instruments or temporal resolutions in the microsecond-nanosecond range in a dynamic TEM (DTEM). Here, unique qualities of the DTEM that benefit the in-situ experiments with gas/fluid environmental cells will be discussed. We also present our results with a liquid stage allowing atomic resolution imaging of nanomaterials in a colloidal suspension, core EEL spectra acquisition, continuous flow, controlled growth of nanocrystals and systematic calibration of the effect of the electron dose on silver nuclei formation.

Oxide nucleation on thin films of copper during in situ oxidation in an electron microscope

NASA Technical Reports Server (NTRS)

Heinemann, K.; Rao, D. B.; Douglass, D. L.

1975-01-01

Single-crystal copper thin films were oxidized at an isothermal temperature of 425 C and at an oxygen partial pressure of 0.005 torr. Specimens were prepared by epitaxial vapor deposition onto polished faces of rocksalt and were mounted in a hot stage inside the ultrahigh-vacuum chamber of a high-resolution electron microscope. An induction period of roughly 30 min was established which was independent of the film thickness but depended strongly on the oxygen partial pressure and to exposure to oxygen prior to oxidation. Neither stacking faults nor dislocations were found to be associated with the Cu2O nucleation sites. The experimental data, including results from oxygen dissolution experiments and from repetitive oxidation-reduction-oxidation sequences, fit well into the framework of an oxidation process involving the formation of a surface charge layer, oxygen saturation of the metal with formation of a supersaturated zone near the surface, and nucleation followed by surface diffusion of oxygen and bulk diffusion of copper for lateral and vertical oxide growth, respectively.

Ni-rich precipitates in a lead bismuth eutectic loop

NASA Astrophysics Data System (ADS)

Kikuchi, K.; Saito, S.; Hamaguchi, D.; Tezuka, M.

2010-03-01

Solidified LBE was sampled from the specimens, electro-magnetic pump, filter, drain valve and oxygen sensor at the JAEA Lead Bismuth Loop-1 (JLBL-1) where the structural material was made of SS316. The concentration of Ni, Fe and Cr in LBE were analyzed by the Inductive Coupled Plasma atomic emission spectrometer. It was concluded that the solution of Ni into LBE was not saturated although the concentration of Fe and Cr almost achieved to the values in the literature. A needle-type structure appeared on the surface of solidified LBE inside the tube specimens. It was found to be Ni-rich precipitates by X-ray analyses (Field Emission Scanning Electron Microscope, FE-SEM). LBE samples collected from a circulating loop after discharging did not show the amount of impurities equivalent to the LBE bulk property.

Infrared Pyrometry From Room Temperature To 700 Degrees C

NASA Technical Reports Server (NTRS)

Wheeler, Donald R.; Jones, William R., Jr.; Pepper, Stephen V.

1989-01-01

Consistent readings obtained when specimens prepared appropriately. New method largely overcomes limitations. Transmission of infrared increased by replacing customary metal-coated glass viewing port with quartz viewing port covered with tantalum mesh. Commercially available infrared microscope with focal distance of 53 cm focuses on spot only 1 mm wide on specimen. Microscope operated as radiometer. Output of detector varies by several orders of magnitude, processed by logarithmic amplifier before reading.

Analysis of NiAlTa precipitates in beta-NiAl + 2 at. pct Ta alloy

NASA Technical Reports Server (NTRS)

Pathare, V.; Michal, G. M.; Vedula, K.; Nathal, M. V.

1987-01-01

Results are reported from experiments performed to identify the precipitates, and their orientation in the matrix, in a beta-NiAl alloy containing 2 at. pct. Ta after undergoing creep test at 1300 K. Test specimens formed by extruding hot powders were compressed at 1300 K for about 50 hr at a strain rate averaging 6/1 million per sec. The specimens were then thinned and examined under an electron microscope and by X-ray diffractometry. An intermetallic NiAlTa compound with a hexagonal Cl4 structure appeared as second phase precipitates in the samples, exhibiting plate-like shapes and a habit plane close to (012). The prism planes of the hexagonal NiAlTa precipitates paralleled the closest packed planes in the cubic beta-NiAl matrix.

Use of a night vision intensifier for direct visualization by eye of far-red and near-infrared fluorescence through an optical microscope.

PubMed

Siddiqi, M A; Kilduff, G M; Gearhart, J D

2003-11-01

We describe the design, construction and testing of a prototype device that allows the direct visualization by eye of far-red and near-infrared (NIR) fluorescence through an optical microscope. The device incorporates a gallium arsenide (GaAs) image intensifier, typically utilized in low-light or 'night vision' applications. The intensifier converts far-red and NIR light into electrons and then into green light, which is visible to the human eye. The prototype makes possible the direct, real-time viewing by eye of normally invisible far-red and NIR fluorescence from a wide variety of fluorophores, using the full field of view of the microscope to which it is applied. The high sensitivity of the image intensifier facilitates the viewing of a wide variety of photosensitive specimens, including live cells and embryos, at vastly reduced illumination levels in both fluorescence and bright-field microscopy. Modifications to the microscope are not required in order to use the prototype, which is fully compatible with all current fluorescence techniques. Refined versions of the prototype device will have broad research and clinical applications.

The external morphology of adult female Egrasilus labracis as shown using hexamethyldisilazane treated, uncoated specimens for scanning electron microscopy.

PubMed

Murray, Harry M; Hill, Stephen J; Ang, Keng P

2016-07-01

The description and application of a modified Scanning Electron Microscope preparation technique using hexamethyldisilazane for small parasitic copepods was demonstrated though a high resolution depiction of individuals of Ergasilus labracis sampled from three spined stickleback (Gasterosteus aculeatus) in Bay D'Espoir, Newfoundland during summer 2015 and from archival samples retrieved from Atlantic salmon par (Salmo salar) stored at the Atlantic reference centre, St. Andrews, New Brunswick. The specimens were very well preserved showing high quality detail of important features and verifying those previously described using light microscopy by Hogans. Additionally the technique allowed excellent in situ demonstrations of mouth parts, swimming legs, and unusual and previously undescribed features of the second antenna including prominent striations and pore-like structures found to define the claw. It is thought that this technique will become a quick and efficient tool for describing important taxonomic features of small parasitic copepods like E. labracis or other similar small aquatic organisms. Microsc. Res. Tech. 79:657-663, 2016. © 2016 Wiley Periodicals, Inc. © 2016 Wiley Periodicals, Inc.

Diagnosis of viral agents associated with neonatal calf diarrhea.

PubMed Central

Marsolais, G; Assaf, R; Montpetit, C; Marois, P

1978-01-01

During this study, 134 samples have been examined for the detection of the viruses associated with neonatal calf diarrhea. The presence of Nebraska viruses (rotavirus and coronavirus) has been demonstrated by using the electron microscope and the fluorescent antibody techniques while the presence of other viruses has been detected by the observation of a cytopathic effect on monolayer cells of calf testis. The Nebraska viruses have been demonstrated in 107 (80%) out of 134 field case specimens. An association of rotaviruses and coronaviruses was found in 58 cases (54%) whilst the coronaviruses and the rotavirus were found singly in 34 cases (53%) and in 15 cases (14%) respectively. Four bovine virus diarrhea viruses, two infectious bovine rhinotracheitis viruses and two enteroviruses have also been isolated in the preceding 107 Nebraska positive specimens. For the detection of the Nebraska viruses, the fluorescent antibody techniques were more sensitive than the electron microscopy. However, those two techniques must be used simultaneously for a better detection of a greatest possible number of cases. Images Fig. 1. Fig. 2. PMID:208735

In situ micro-compression testing of He2+ ion irradiated titanium aluminide

NASA Astrophysics Data System (ADS)

Wei, Tao; Xu, Alan; Zhu, Hanliang; Ionescu, Mihail; Bhattacharyya, Dhriti

2017-10-01

A titanium aluminide (TiAl) alloy 45XD has been irradiated by a He ion beam with an energy of 5 MeV on a tandem accelerator at the Australian Nuclear Science and Technology Organization (ANSTO). The total fluence of He ions was 5 × 1017 ion cm-2. A 17 μm uniform damage region from the material surface with a helium concentration of about 5000 appm was achieved by using an energy degrading wheel in front of the TiAl target. The micro-size test specimens from the damage layer were fabricated using a focused ion beam & scanning electron microscope (FIB-SEM) system. The in situ SEM micromechanical compressive testing was carried out inside an SEM and the results indicated irradiation embrittlement in the helium affected region. Electron back scatter diffraction (EBSD) analysis has been applied to reveal the orientation of the lamellae in the TiAl specimens, and used to understand the deformation processes in the sample. The irradiation damage of gallium ion beam from FIB on the surface of TiAl sample was also investigated.

In situ thermomechanical testing methods for micro/nano-scale materials.

PubMed

Kang, Wonmo; Merrill, Marriner; Wheeler, Jeffrey M

2017-02-23

The advance of micro/nanotechnology in energy-harvesting, micropower, electronic devices, and transducers for automobile and aerospace applications has led to the need for accurate thermomechanical characterization of micro/nano-scale materials to ensure their reliability and performance. This persistent need has driven various efforts to develop innovative experimental techniques that overcome the critical challenges associated with precise mechanical and thermal control of micro/nano-scale specimens during material characterization. Here we review recent progress in the development of thermomechanical testing methods from miniaturized versions of conventional macroscopic test systems to the current state of the art of in situ uniaxial testing capabilities in electron microscopes utilizing either indentation-based microcompression or integrated microsystems. We discuss the major advantages/disadvantages of these methods with respect to specimen size, range of temperature control, ease of experimentation and resolution of the measurements. We also identify key challenges in each method. Finally, we summarize some of the important discoveries that have been made using in situ thermomechanical testing and the exciting research opportunities still to come in micro/nano-scale materials.

Metals handbook. Volume 12: Fractography

DOE Office of Scientific and Technical Information (OSTI.GOV)

Not Available

1987-01-01

ASM International has published this handbook in response to the growing interest in the science of fractography, the result of improved methods of preparing specimens, advances in photographic techniques and equipment, refinement of the scanning electron microscope, and the introduction of quantitative fractography. The book covers all aspects of fracture examination and interpretation, including electron and quantitative fractography. The text is accompanied by line drawings, graphs, and photographic illustrations of fracture surfaces and microstructural features. Articles explain and illustrate the principal modes of fracture and the effects of loading history, environment, and materials quality on fracture appearance. An atlas ofmore » fractographs constitutes the second half of the volume and contains more than 1300 fractographs, including a collection of ferrous and nonferrous alloy parts. Supplemental illustrations of failed metal-matrix composites, resin-matrix composites, polymers, and electronic materials are provided.« less

Exploring transmission Kikuchi diffraction using a Timepix detector

NASA Astrophysics Data System (ADS)

Vespucci, S.; Winkelmann, A.; Mingard, K.; Maneuski, D.; O'Shea, V.; Trager-Cowan, C.

2017-02-01

Electron backscatter diffraction (EBSD) is a well-established scanning electron microscope (SEM)-based technique [1]. It allows the non-destructive mapping of the crystal structure, texture, crystal phase and strain with a spatial resolution of tens of nanometers. Conventionally this is performed by placing an electron sensitive screen, typically consisting of a phosphor screen combined with a charge coupled device (CCD) camera, in front of a specimen, usually tilted 70° to the normal of the exciting electron beam. Recently, a number of authors have shown that a significant increase in spatial resolution is achievable when Kikuchi diffraction patterns are acquired in transmission geometry; that is when diffraction patterns are generated by electrons transmitted through an electron-transparent, usually thinned, specimen. The resolution of this technique, called transmission Kikuchi diffraction (TKD), has been demonstrated to be better than 10 nm [2,3]. We have recently demonstrated the advantages of a direct electron detector, Timepix [4,5], for the acquisition of standard EBSD patterns [5]. In this article we will discuss the advantages of Timepix to perform TKD and for acquiring spot diffraction patterns and more generally for acquiring scanning transmission electron microscopy micrographs in the SEM. Particularly relevant for TKD, is its very compact size, which allows much more flexibility in the positioning of the detector in the SEM chamber. We will furthermore show recent results using Timepix as a virtual forward scatter detector, and will illustrate the information derivable on producing images through processing of data acquired from different areas of the detector. We will show results from samples ranging from gold nanoparticles to nitride semiconductor nanorods.

Effect of a self-adhesive coating on the load-bearing capacity of tooth-coloured restorative materials.

PubMed

Bagheri, R; Palamara, Jea; Mese, A; Manton, D J

2017-03-01

The aim of this study was to compare the flexural strength and Vickers hardness of tooth-coloured restorative materials with and without applying a self-adhesive coating for up to 6 months. Specimens were prepared from three resin composites (RC), two resin-modified glass-ionomer cements (RM-GIC) and two conventional glass-ionomer cements (CGIC). All materials were tested both with and without applying G-Coat Plus (GCP). Specimens were conditioned in 37 °C distilled deionized water for 24 h, and 1, 3 and 6 months. The specimens were strength tested using a four-point bend test jig in a universal testing machine. The broken specimen's halves were used for Vickers hardness testing. Representative specimens were examined under an environmental scanning electron microscope. Data analysis showed that regardless of time and materials, generally the surface coating was associated with a significant increase in the flexural strength of the materials. Applying the GCP decreased the hardness of almost all materials significantly (P < 0.05) and effect of time intervals on hardness was material dependent. The load-bearing capacity of the restorative materials was affected by applying self-adhesive coating and ageing. The CGIC had significantly higher hardness but lower flexural strength than the RM-GIC and RC. © 2016 Australian Dental Association.

A sub-sampled approach to extremely low-dose STEM

DOE PAGES

Stevens, A.; Luzi, L.; Yang, H.; ...

2018-01-22

The inpainting of deliberately and randomly sub-sampled images offers a potential means to image specimens at a high resolution and under extremely low-dose conditions (≤1 e -/Å 2) using a scanning transmission electron microscope. We show that deliberate sub-sampling acquires images at least an order of magnitude faster than conventional low-dose methods for an equivalent electron dose. More importantly, when adaptive sub-sampling is implemented to acquire the images, there is a significant increase in the resolution and sensitivity which accompanies the increase in imaging speed. Lastly, we demonstrate the potential of this method for beam sensitive materials and in-situ observationsmore » by experimentally imaging the node distribution in a metal-organic framework.« less

A sub-sampled approach to extremely low-dose STEM

DOE Office of Scientific and Technical Information (OSTI.GOV)

Stevens, A.; Luzi, L.; Yang, H.

The inpainting of deliberately and randomly sub-sampled images offers a potential means to image specimens at a high resolution and under extremely low-dose conditions (≤1 e -/Å 2) using a scanning transmission electron microscope. We show that deliberate sub-sampling acquires images at least an order of magnitude faster than conventional low-dose methods for an equivalent electron dose. More importantly, when adaptive sub-sampling is implemented to acquire the images, there is a significant increase in the resolution and sensitivity which accompanies the increase in imaging speed. Lastly, we demonstrate the potential of this method for beam sensitive materials and in-situ observationsmore » by experimentally imaging the node distribution in a metal-organic framework.« less

Comparing the effectiveness of four desensitizing toothpastes on dentinal tubule occlusion: A scanning electron microscope analysis.

PubMed

Jena, Amit; Kala, Soumik; Shashirekha, Govind

2017-01-01

Dentin hypersensitivity (DH) is a sudden short sharp pain best explained by hydrodynamic theory. Several agents are available throughout the market that can treat DH either by blocking the nerves that helps in conducting pain or by blocking the open dentinal tubules. The aim of the present study was to compare the tubule occluding efficacy of four different desensitizing dentifrices under scanning electron microscope (SEM). Sixty-two dentin blocks measuring 5 mm × 5 mm × 3 mm were obtained from extracted human molar teeth and were randomly divided into five groups: Group 1 - no treatment (control, n = 2); Group 2 - Pepsodent Pro-sensitive relief and repair ( n = 15); Group 3 - Sensodyne repair and protect ( n = 15); Group 4 - Remin Pro ( n = 15); Group 5 - Test toothpaste containing 15% nano-hydroxyapatite (n-HA) crystals ( n = 15). The specimens were brushed for 2 min/day for 14 days and stored in artificial saliva. After final brushing, specimens were gold sputtered and viewed under SEM at ×2000 magnification. Results obtained were statistically analyzed using nonparametric Kruskal-Wallis test and least significant difference post hoc test. All test groups showed significant increase in dentin tubule occlusion as compared to control group. The highest percentage of tubules occluded was shown by Group 4 and Group 5 which was significantly different from other groups ( P ≤ 0.05), and there was no significant difference in tubule occlusion among them. Newer desensitizing dentifrices containing 15% n-HA and Remin Pro can provide effective tubule occlusion and thereby reduce the pain and discomfort caused by DH.

Changes in surface characteristics of two different resin composites after 1 year water storage: An SEM and AFM study.

PubMed

Tekçe, Neslihan; Pala, Kansad; Demirci, Mustafa; Tuncer, Safa

2016-11-01

To evaluate changes in surface characteristics of two different resin composites after 1 year of water storage using a profilometer, Vickers hardness, scanning electron microscopy (SEM), and atomic force microscopy (AFM). A total of 46 composite disk specimens (10 mm in diameter and 2 mm thick) were fabricated using Clearfil Majesty Esthetic and Clearfil Majesty Posterior (Kuraray Medical Co, Tokyo, Japan). Ten specimens from each composite were used for surface roughness and microhardness tests (n = 10). For each composite, scanning electron microscope (SEM, n = 2) and atomic force microscope (AFM, n = 1) images were obtained after 24 h and 1 year of water storage. The data were analyzed using two-way analysis of variance and a post-hoc Bonferroni test. Microhardness values of Clearfil Majesty Esthetic decreased significantly (78.15-63.74, p = 0.015) and surface roughness values did not change after 1 year of water storage (0.36-0.39, p = 0.464). Clearfil Majesty Posterior microhardness values were quite stable (138.74-137.25, p = 0.784), and surface roughness values increased significantly (0.39-0.48, p = 0.028) over 1 year. One year of water storage caused microhardness values for Clearfil Majesty Esthetic to decrease and the surface roughness of Clearfil Majesty Posterior increased. AFM and SEM images demonstrated surface detoration of the materials after 1 year and ensured similar results with the quantitative test methods. SCANNING 38:694-700, 2016. © 2016 Wiley Periodicals, Inc. © Wiley Periodicals, Inc.

Ethnic Distribution of Microscopic Colitis in the United States.

PubMed

Turner, Kevin; Genta, Robert M; Sonnenberg, Amnon

2015-11-01

A large electronic database of histopathology reports was used to study the ethnic distribution of microscopic colitis in the United States. Miraca Life Sciences is a nation-wide pathology laboratory that receives biopsy specimens submitted by 1500 gastroenterologists distributed throughout the United States. In a case-control study, the prevalence of microscopic colitis in 4 ethnic groups (East Asians, Indians, Hispanics, and Jews) was compared with that of all other ethnic groups (composed of American Caucasians and African Americans), serving as reference group. A total of 11,706 patients with microscopic colitis were included in the analysis. In all ethnic groups alike, microscopic colitis was more common in women than men (78% versus 22%, odds ratio = 3.40, 95% confidence interval = 3.26-3.55). In all ethnic groups, the prevalence of microscopic colitis showed a continuous age-dependent rise. Hispanic patients with microscopic colitis were on average younger than the reference group (59.4 ± 16.2 years versus 64.2 ± 13.8 years, P < 0.001). Jewish patients with microscopic colitis were slightly older than the reference group (65.6 ± 13.4 years, P = 0.015). Compared with the reference group (prevalence = 1.20%), microscopic colitis was significantly less common among patients of Indian (prevalence = 0.28%, odds ratio = 0.32, 95% confidence interval = 0.13-0.65), East Asian (0.22%, 0.19, 0.14-0.26), or Hispanic decent (0.48%, 0.40, 0.36-0.45) and significantly more common among Jewish patients (1.30%, 1.10, 1.01-1.21). Microscopic colitis shows striking variations of its occurrence among different ethnic groups. Such variations could point at differences in the exposure to environmental risk factors.

Study of the Corrosion Resistance of Austenitic Stainless Steels during Conversion of Waste to Biofuel

PubMed Central

Cabrini, Marina; Lorenzi, Sergio; Pastore, Tommaso; Pellegrini, Simone; Burattini, Mauro; Miglio, Roberta

2017-01-01

The paper deals with the corrosion behavior of stainless steels as candidate materials for biofuel production plants by liquefaction process of the sorted organic fraction of municipal solid waste. Corrosion tests were carried out on AISI 316L and AISI 304L stainless steels at 250 °C in a batch reactor during conversion of raw material to bio-oil (biofuel precursor), by exposing specimens either to water/oil phase or humid gas phase. General corrosion rate was measured by weight loss tests. The susceptibility to stress corrosion cracking was evaluated by means of U-bend specimens and slow stress rate tests at 10−6 or 10−5 s−1 strain rate. After tests, scanning electron microscope analysis was carried out to detect cracks and localized attacks. The results are discussed in relation with exposure conditions. They show very low corrosion rates strictly dependent upon time and temperature. No stress corrosion cracking was observed on U-bend specimens, under constant loading. Small cracks confined in the necking cone of specimens prove that stress corrosion cracking only occurred during slow strain rate tests at stresses exceeding the yield strength. PMID:28772682

Ductility of Advanced High-Strength Steel in the Presence of a Sheared Edge

NASA Astrophysics Data System (ADS)

Ruggles, Tim; Cluff, Stephen; Miles, Michael; Fullwood, David; Daniels, Craig; Avila, Alex; Chen, Ming

2016-07-01

The ductility of dual-phase (DP) 980 and transformation-induced plasticity (TRIP) assisted bainitic ferritic (TBF) 980 steels was studied in the presence of a sheared edge. Specimens were tested in uniaxial tension in a standard test frame as well as in situ in the scanning electron microscope (SEM). Incremental tensile straining was done in the SEM with images taken at each strain increment. Then digital image correlation (DIC) was used to compute the effective strain at the level of the individual phases in the microstructure. Shear banding across multiple phases was seen in strained TBF specimens, while the DP specimens exhibited more of a patchwork strain pattern, with high strains concentrated in ferrite and low strains observed in the martensite. Two-point statistics were applied to the strain data from the DIC work and the corresponding microstructure images to evaluate the effect of phase hardness on localization and fracture. It was observed that the DP 980 material had a greater tendency for localization around hard phases compared to the TBF 980. This at least partially explains the greater ductility of the TBF material, especially in specimens where a sheared edge was present.

Applications of Real Space Crystallography in Characterization of Dislocations in Geological Materials in a Scanning Electron Microscope (SEM)

NASA Astrophysics Data System (ADS)

Kaboli, S.; Burnley, P. C.

2017-12-01

Imaging and characterization of defects in crystalline materials is of significant importance in various disciplines including geoscience, materials science, and applied physics. Linear defects such as dislocations and planar defects such as twins and stacking faults, strongly influence many of the properties of crystalline materials and also reflect the conditions and degree of deformation. Dislocations have been conventionally imaged in thin foils in a transmission electron microscope (TEM). Since the development of field emission scanning electron microscopes (FE-SEM) with high gun brightness and small spot size, extensive efforts have been dedicated to the imaging and characterization of dislocations in semi-conductors using electron channeling contrast imaging (ECCI) in the SEM. The obvious advantages of using SEM over TEM include easier and non-destructive sample preparation and a large field of view enabling statistical examination of the density and distribution of dislocations and other defects. In this contribution, we extend this technique to geological materials and introduce the Real Space Crystallography methodology for imaging and complete characterization of dislocations based on bend contour contrast obtained by ECCI in FE-SEM. Bend contours map out the distortion in the crystal lattice across a deformed grain. The contrast of dislocations is maximum in the vicinity of bend contours where crystal planes diffract at small and positive deviations from the Bragg positions (as defined by Bragg's law of electron diffraction). Imaging is performed in a commercial FE-SEM equipped with a standard silicon photodiode backscattered (BSE) detector and an electron backscatter diffraction (EBSD) system for crystal orientation measurements. We demonstrate the practice of this technique in characterization of a number of geological materials in particular quartz, forsterite olivine and corundum, experimentally deformed at high pressure-temperature conditions. This new approach in microstructure characterization of deformed geologic materials in FE-SEM, without the use of etching or decoration techniques, has valuable applications to both experimentally deformed and naturally deformed specimens.

Markov Random Field Based Automatic Image Alignment for ElectronTomography

DOE Office of Scientific and Technical Information (OSTI.GOV)

Moussavi, Farshid; Amat, Fernando; Comolli, Luis R.

2007-11-30

Cryo electron tomography (cryo-ET) is the primary method for obtaining 3D reconstructions of intact bacteria, viruses, and complex molecular machines ([7],[2]). It first flash freezes a specimen in a thin layer of ice, and then rotates the ice sheet in a transmission electron microscope (TEM) recording images of different projections through the sample. The resulting images are aligned and then back projected to form the desired 3-D model. The typical resolution of biological electron microscope is on the order of 1 nm per pixel which means that small imprecision in the microscope's stage or lenses can cause large alignment errors.more » To enable a high precision alignment, biologists add a small number of spherical gold beads to the sample before it is frozen. These beads generate high contrast dots in the image that can be tracked across projections. Each gold bead can be seen as a marker with a fixed location in 3D, which provides the reference points to bring all the images to a common frame as in the classical structure from motion problem. A high accuracy alignment is critical to obtain a high resolution tomogram (usually on the order of 5-15nm resolution). While some methods try to automate the task of tracking markers and aligning the images ([8],[4]), they require user intervention if the SNR of the image becomes too low. Unfortunately, cryogenic electron tomography (or cryo-ET) often has poor SNR, since the samples are relatively thick (for TEM) and the restricted electron dose usually results in projections with SNR under 0 dB. This paper shows that formulating this problem as a most-likely estimation task yields an approach that is able to automatically align with high precision cryo-ET datasets using inference in graphical models. This approach has been packaged into a publicly available software called RAPTOR-Robust Alignment and Projection estimation for Tomographic Reconstruction.« less

A universal fluid cell for the imaging of biological specimens in the atomic force microscope.

PubMed

Kasas, Sandor; Radotic, Ksenja; Longo, Giovanni; Saha, Bashkar; Alonso-Sarduy, Livan; Dietler, Giovanni; Roduit, Charles

2013-04-01

Recently, atomic force microscope (AFM) manufacturers have begun producing instruments specifically designed to image biological specimens. In most instances, they are integrated with an inverted optical microscope, which permits concurrent optical and AFM imaging. An important component of the set-up is the imaging chamber, whose design determines the nature of the experiments that can be conducted. Many different imaging chamber designs are available, usually designed to optimize a single parameter, such as the dimensions of the substrate or the volume of fluid that can be used throughout the experiment. In this report, we present a universal fluid cell, which simultaneously optimizes all of the parameters that are important for the imaging of biological specimens in the AFM. This novel imaging chamber has been successfully tested using mammalian, plant, and microbial cells. Copyright © 2013 Wiley Periodicals, Inc.

Light and electron microscopic observation of regenerated fungiform taste buds in patients with recovered taste function after severing chorda tympani nerve.

PubMed

Saito, Takehisa; Ito, Tetsufumi; Narita, Norihiko; Yamada, Takechiyo; Manabe, Yasuhiro

2011-11-01

The aim of this study was to evaluate the mean number of regenerated fungiform taste buds per papilla and perform light and electron microscopic observation of taste buds in patients with recovered taste function after severing the chorda tympani nerve during middle ear surgery. We performed a biopsy on the fungiform papillae (FP) in the midlateral region of the dorsal surface of the tongue from 5 control volunteers (33 total FP) and from 7 and 5 patients with and without taste recovery (34 and 29 FP, respectively) 3 years 6 months to 18 years after surgery. The specimens were observed by light and transmission electron microscopy. The taste function was evaluated by electrogustometry. The mean number of taste buds in the FP of patients with completely recovered taste function was significantly smaller (1.9 +/- 1.4 per papilla; p < 0.01) than that of the control subjects (3.8 +/- 2.2 per papilla). By transmission electron microscopy, 4 distinct types of cell (type I, II, III, and basal cells) were identified in the regenerated taste buds. Nerve fibers and nerve terminals were also found in the taste buds. It was clarified that taste buds containing taste cells and nerve endings do regenerate in the FP of patients with recovered taste function.

Long-term effects of seven cleaning methods on light transmittance, surface roughness, and flexural modulus of polyurethane retainer material.

PubMed

Agarwal, Manika; Wible, Emily; Ramir, Tyler; Altun, Sibel; Viana, Grace; Evans, Carla; Lukic, Henry; Megremis, Spiro; Atsawasuwan, Phimon

2018-05-01

To evaluate the long-term effects of seven different cleaning methods on light transmittance, surface roughness, and flexural modulus of a polyurethane retainer material. Polyurethane retainer specimens (Vivera®, Align Technology Inc) (70 specimens, n = 10 per method, 50.8 mm × 12.7 mm × 1.0 mm) were exposed to seven cleaning methods twice a week for 6 months. Before treatment and after 6 months, light transmittance, surface roughness, and flexural modulus of the specimens were quantified. Qualitative assessment of randomly selected specimens from each solution was performed at baseline and after 6 months using a scanning electron microscope. Statistical analyses were performed at the .05 significance level. Of the three test variables, light transmittance through the specimens was the only one that changed significantly from baseline to 6 months for all cleaning solutions, with all of them causing a decrease. However, except for 0.6% sodium hypochlorite showing a change in surface roughness values and 2.5% vinegar and toothbrushing showing an increase in flexural modulus, none of the other four cleaning methods resulted in significant changes in surface roughness or flexural modulus values for the polyurethane specimens between baseline and after 6 months. Of the seven cleaning methods, Invisalign® cleaning crystals, Polident®, and Listerine® showed the least amount of change in light transmittance values for the polyurethane specimens over 6 months, and they had no effect on surface roughness and flexural modulus values.

4D multiple-cathode ultrafast electron microscopy

PubMed Central

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

2014-01-01

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

4D multiple-cathode ultrafast electron microscopy.

PubMed

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

2014-07-22

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

Biological imaging by soft X-ray diffraction microscopy

NASA Astrophysics Data System (ADS)

Shapiro, David

We have developed a microscope for soft x-ray diffraction imaging of dry or frozen hydrated biological specimens. This lensless imaging system does not suffer from the resolution or specimen thickness limitations that other short wavelength microscopes experience. The microscope, currently situated at beamline 9.0.1 of the Advanced Light Source, can collect diffraction data to 12 nm resolution with 750 eV photons and 17 nm resolution with 520 eV photons. The specimen can be rotated with a precision goniometer through an angle of 160 degrees allowing for the collection of nearly complete three-dimensional diffraction data. The microscope is fully computer controlled through a graphical user interface and a scripting language automates the collection of both two-dimensional and three-dimensional data. Diffraction data from a freeze-dried dwarf yeast cell, Saccharomyces cerevisiae carrying the CLN3-1 mutation, was collected to 12 run resolution from 8 specimen orientations spanning a total rotation of 8 degrees. The diffraction data was phased using the difference map algorithm and the reconstructions provide real space images of the cell to 30 nm resolution from each of the orientations. The agreement of the different reconstructions provides confidence in the recovered, and previously unknown, structure and indicates the three dimensionality of the cell. This work represents the first imaging of the natural complex refractive contrast from a whole unstained cell by the diffraction microscopy method and has achieved a resolution superior to lens based x-ray tomographic reconstructions of similar specimens. Studies of the effects of exposure to large radiation doses were also carried out. It was determined that the freeze-dried cell suffers from an initial collapse, which is followed by a uniform, but slow, shrinkage. This structural damage to the cell is not accompanied by a diminished ability to see small features in the specimen. Preliminary measurements on frozen-hydrated yeast indicate that the frozen specimens do not exhibit these changes even with doses as high as 5 x 109 Gray.

Development and Application of STEM for the Biological Sciences

PubMed Central

Sousa, Alioscka A.; Leapman, Richard D.

2012-01-01

The design of the scanning transmission electron microscope (STEM), as conceived originally by Crewe and coworkers, enables the highly efficient and flexible collection of different elastic and inelastic signals resulting from the interaction of a focused probe of incident electrons with a specimen. In the present paper we provide a brief review for how the STEM today can be applied towards a range of different problems in the biological sciences, emphasizing four main areas of application. (1) For three decades, the most widely used STEM technique has been the mass determination of proteins and other macromolecular assemblies. Such measurements can be performed at low electron dose by collecting the high-angle dark-field signal using an annular detector. STEM mass mapping has proven valuable for characterizing large protein assemblies such as filamentous proteins with a well-defined mass per length. (2) The annular dark-field signal can also be used to image ultrasmall, functionalized nanoparticles of heavy atoms for labeling specific aminoacid sequences in protein assemblies. (3) By acquiring electron energy loss spectra (EELS) at each pixel in a hyperspectral image, it is possible to map the distributions of specific bound elements like phosphorus, calcium and iron in isolated macromolecular assemblies or in compartments within sectioned cells. Near single atom sensitivity is feasible provided that the specimen can tolerate a very high incident electron dose. (4) Electron tomography is a new application of STEM that enables three-dimensional reconstruction of micrometer-thick sections of cells. In this technique a probe of small convergence angle gives a large depth of field throughout the thickness of the specimen while maintaining a probe diameter of < 2 nm; and the use of an on-axis bright-field detector reduces the effects of beam broadening and thus improves the spatial resolution compared to that attainable by STEM dark-field tomography. PMID:22749213

Miniature Variable Pressure Scanning Electron Microscope for In-Situ Imaging and Chemical Analysis

NASA Technical Reports Server (NTRS)

Gaskin, Jessica A.; Jerman, Gregory; Gregory, Don; Sampson, Allen R.

2012-01-01

NASA Marshall Space Flight Center (MSFC) is leading an effort to develop a Miniaturized Variable Pressure Scanning Electron Microscope (MVP-SEM) for in-situ imaging and chemical analysis of uncoated samples. This instrument development will be geared towards operation on Mars and builds on a previous MSFC design of a mini-SEM for the moon (funded through the NASA Planetary Instrument Definition and Development Program). Because Mars has a dramatically different environment than the moon, modifications to the MSFC lunar mini-SEM are necessary. Mainly, the higher atmospheric pressure calls for the use of an electron gun that can operate at High Vacuum, rather than Ultra-High Vacuum. The presence of a CO2-rich atmosphere also allows for the incorporation of a variable pressure system that enables the in-situ analysis of nonconductive geological specimens. Preliminary testing of Mars meteorites in a commercial Environmental SEM(Tradmark) (FEI) confirms the usefulness of lowcurrent/low-accelerating voltage imaging and highlights the advantages of using the Mars atmosphere for environmental imaging. The unique capabilities of the MVP-SEM make it an ideal tool for pursuing key scientific goals of NASA's Flagship Mission Max-C; to perform in-situ science and collect and cache samples in preparation for sample return from Mars.

Carbon nanotube scaffolds with controlled porosity as electromagnetic absorbing materials in the gigahertz range

NASA Astrophysics Data System (ADS)

González, M.; Crespo, M.; Baselga, J.; Pozuelo, J.

2016-05-01

Control of the microscopic structure of CNT nanocomposites allows modulation of the electromagnetic shielding in the gigahertz range. The porosity of CNT scaffolds has been controlled by two freezing protocols and a subsequent lyophilization step: fast freezing in liquid nitrogen and slow freezing at -20 °C. Mercury porosimetry shows that slowly frozen specimens present a more open pore size (100-150 μm) with a narrow distribution whereas specimens frozen rapidly show a smaller pore size and a heterogeneous distribution. 3D-scaffolds containing 3, 4, 6 and 7% CNT were infiltrated with epoxy and specimens with 2, 5 and 8 mm thicknesses were characterized in the GHz range. Samples with the highest pore size and porosity presented the lowest reflected power (about 30%) and the highest absorbed power (about 70%), which allows considering them as electromagnetic radiation absorbing materials.Control of the microscopic structure of CNT nanocomposites allows modulation of the electromagnetic shielding in the gigahertz range. The porosity of CNT scaffolds has been controlled by two freezing protocols and a subsequent lyophilization step: fast freezing in liquid nitrogen and slow freezing at -20 °C. Mercury porosimetry shows that slowly frozen specimens present a more open pore size (100-150 μm) with a narrow distribution whereas specimens frozen rapidly show a smaller pore size and a heterogeneous distribution. 3D-scaffolds containing 3, 4, 6 and 7% CNT were infiltrated with epoxy and specimens with 2, 5 and 8 mm thicknesses were characterized in the GHz range. Samples with the highest pore size and porosity presented the lowest reflected power (about 30%) and the highest absorbed power (about 70%), which allows considering them as electromagnetic radiation absorbing materials. Electronic supplementary information (ESI) available: Scheme of hydrogenated derivative of diglycidyl ether of bisphenol-A (HDGEBA) and m-xylylenediamine; X-ray diffractograms of pristine CNT and oxidized CNT; glass transition temperatures of composites; electromagnetic shielding analysis in the 1-18 GHz frequency range. See DOI: 10.1039/c6nr02133f

Electrosprayed Multi-Core Alginate Microcapsules as Novel Self-Healing Containers

NASA Astrophysics Data System (ADS)

Hia, Iee Lee; Pasbakhsh, Pooria; Chan, Eng-Seng; Chai, Siang-Piao

2016-10-01

Alginate microcapsules containing epoxy resin were developed through electrospraying method and embedded into epoxy matrix to produce a capsule-based self-healing composite system. These formaldehyde free alginate/epoxy microcapsules were characterized via light microscope, field emission scanning electron microscope, fourier transform infrared spectroscopy and thermogravimetric analysis. Results showed that epoxy resin was successfully encapsulated within alginate matrix to form porous (multi-core) microcapsules with pore size ranged from 5-100 μm. The microcapsules had an average size of 320 ± 20 μm with decomposition temperature at 220 °C. The loading capacity of these capsules was estimated to be 79%. Under in situ healing test, impact specimens showed healing efficiency as high as 86% and the ability to heal up to 3 times due to the multi-core capsule structure and the high impact energy test that triggered the released of epoxy especially in the second and third healings. TDCB specimens showed one-time healing only with the highest healing efficiency of 76%. The single healing event was attributed by the constant crack propagation rate of TDCB fracture test. For the first time, a cost effective, environmentally benign and sustainable capsule-based self-healing system with multiple healing capabilities and high healing performance was developed.

Electrosprayed Multi-Core Alginate Microcapsules as Novel Self-Healing Containers.

PubMed

Hia, Iee Lee; Pasbakhsh, Pooria; Chan, Eng-Seng; Chai, Siang-Piao

2016-10-03

Alginate microcapsules containing epoxy resin were developed through electrospraying method and embedded into epoxy matrix to produce a capsule-based self-healing composite system. These formaldehyde free alginate/epoxy microcapsules were characterized via light microscope, field emission scanning electron microscope, fourier transform infrared spectroscopy and thermogravimetric analysis. Results showed that epoxy resin was successfully encapsulated within alginate matrix to form porous (multi-core) microcapsules with pore size ranged from 5-100 μm. The microcapsules had an average size of 320 ± 20 μm with decomposition temperature at 220 °C. The loading capacity of these capsules was estimated to be 79%. Under in situ healing test, impact specimens showed healing efficiency as high as 86% and the ability to heal up to 3 times due to the multi-core capsule structure and the high impact energy test that triggered the released of epoxy especially in the second and third healings. TDCB specimens showed one-time healing only with the highest healing efficiency of 76%. The single healing event was attributed by the constant crack propagation rate of TDCB fracture test. For the first time, a cost effective, environmentally benign and sustainable capsule-based self-healing system with multiple healing capabilities and high healing performance was developed.

Electrosprayed Multi-Core Alginate Microcapsules as Novel Self-Healing Containers

PubMed Central

Hia, Iee Lee; Pasbakhsh, Pooria; Chan, Eng-Seng; Chai, Siang-Piao

2016-01-01

Alginate microcapsules containing epoxy resin were developed through electrospraying method and embedded into epoxy matrix to produce a capsule-based self-healing composite system. These formaldehyde free alginate/epoxy microcapsules were characterized via light microscope, field emission scanning electron microscope, fourier transform infrared spectroscopy and thermogravimetric analysis. Results showed that epoxy resin was successfully encapsulated within alginate matrix to form porous (multi-core) microcapsules with pore size ranged from 5–100 μm. The microcapsules had an average size of 320 ± 20 μm with decomposition temperature at 220 °C. The loading capacity of these capsules was estimated to be 79%. Under in situ healing test, impact specimens showed healing efficiency as high as 86% and the ability to heal up to 3 times due to the multi-core capsule structure and the high impact energy test that triggered the released of epoxy especially in the second and third healings. TDCB specimens showed one-time healing only with the highest healing efficiency of 76%. The single healing event was attributed by the constant crack propagation rate of TDCB fracture test. For the first time, a cost effective, environmentally benign and sustainable capsule-based self-healing system with multiple healing capabilities and high healing performance was developed. PMID:27694922

Large-scale tissue clearing (PACT): Technical evaluation and new perspectives in immunofluorescence, histology, and ultrastructure.

PubMed

Neckel, Peter H; Mattheus, Ulrich; Hirt, Bernhard; Just, Lothar; Mack, Andreas F

2016-09-29

Novel techniques, like CLARITY and PACT, render large tissue specimens transparent and thereby suitable for microscopic analysis. We used these techniques to evaluate their potential in the intestine as an exemplary organ with a complex tissue composition. Immunohistochemistry, light sheet-, and confocal scanning-microscopy enabled us to follow complex three-dimensional structures, like nerve fibers, vessels, and epithelial barriers throughout the entire organ. Moreover, in a systematic electron microscopic study, we analyzed the morphology and preservation of tissue on ultrastructural level during the clearing process. We also connect tissue clearing with classical histology and demonstrate that cleared tissues can be stained with Hematoxylin-Eosin and Heidenhain's Azan stain, suggesting potential use in histopathology. These experiments showed that a neutral pH during the clearing process results in much better preservation of tissue ultrastructure and standard stainability. Volume changes of specimens were monitored and quantified during the course of the protocol. Additionally, we employed the technique to visualize the enteric nervous system and the epithelial barrier in post mortem human gut preparations. Our data show the high potential of tissue clearing throughout different tissue types supporting its usefulness in research and diagnosis, and contribute to the technical discussion of ultrastructural tissue-retention.

Large-scale tissue clearing (PACT): Technical evaluation and new perspectives in immunofluorescence, histology, and ultrastructure

PubMed Central

Neckel, Peter H.; Mattheus, Ulrich; Hirt, Bernhard; Just, Lothar; Mack, Andreas F.

2016-01-01

Novel techniques, like CLARITY and PACT, render large tissue specimens transparent and thereby suitable for microscopic analysis. We used these techniques to evaluate their potential in the intestine as an exemplary organ with a complex tissue composition. Immunohistochemistry, light sheet-, and confocal scanning-microscopy enabled us to follow complex three-dimensional structures, like nerve fibers, vessels, and epithelial barriers throughout the entire organ. Moreover, in a systematic electron microscopic study, we analyzed the morphology and preservation of tissue on ultrastructural level during the clearing process. We also connect tissue clearing with classical histology and demonstrate that cleared tissues can be stained with Hematoxylin-Eosin and Heidenhain’s Azan stain, suggesting potential use in histopathology. These experiments showed that a neutral pH during the clearing process results in much better preservation of tissue ultrastructure and standard stainability. Volume changes of specimens were monitored and quantified during the course of the protocol. Additionally, we employed the technique to visualize the enteric nervous system and the epithelial barrier in post mortem human gut preparations. Our data show the high potential of tissue clearing throughout different tissue types supporting its usefulness in research and diagnosis, and contribute to the technical discussion of ultrastructural tissue-retention. PMID:27680942

ELECTRON MICROSCOPE STUDY OF MYCOBACTERIUM LEPRAE AND ITS ENVIRONMENT IN A VESICULAR LEPROUS LESION

PubMed Central

Imaeda, Tamotsu; Convit, Jacinto

1962-01-01

Imaeda, Tamotsu (Instituto Venezolano de Investigaciones Cientificas, Caracas, Venezuela) and Jacinto Convit. Electron microscope study of Mycobacterium leprae and its environment in a vesicular leprous lesion. J. Bacteriol. 83:43–52. 1962.—Biopsied specimens of a borderline leprosy lesion were observed with the electron microscope. In this lesion, the majority of Mycobacterium leprae were laden with cytoplasmic components. The bacilli were separated from the cytoplasm of host cells by an enclosing membrane, thus differing from the environment of well-developed lepra cells in lepromatous lesions. The cell wall is composed of a moderately dense layer. A diffuse layer is discernible outside the cell wall, separated from it by a low density space. It is suggested that the cell wall is further coated by a low density layer, although the nature of the outermost diffuse layer has not yet been determined. The plasma membrane consists of a double layer, i.e., dense inner and outer layers separated by a low density space. The outer layer is closely adjacent to the cell wall. In the region where the outer layer of the plasma membrane enters the cytoplasm and is transformed into a complex membranous structure, the inner layer encloses this membranous configuration. Together they form the intracytoplasmic membrane system. In the bacterial cytoplasm, moderately dense, presumably polyphosphate bodies are apparent. As neither these bodies nor the intracytoplasmic membrane system are visible in the degenerating bacilli, it seems probable that these two components represent indicators of the state of bacillary activity. Images PMID:16561926

Performance of surface on ultrafine grained Ti-0.2Pd in simulated body fluid

NASA Astrophysics Data System (ADS)

Wang, Xiu-Lai; Zhou, Qing; Yang, Kai; Zou, Cheng-Hong; Wang, Lei

2018-03-01

Ti-0.2 wt% Pd (Ti-0.2Pd) which has high crevice corrosion resistance is highlighted for implant applications. In this work, Ti-0.2Pd alloy is subjected to equal channel angular pressing (ECAP) for grain refinement. The effect of the microstructure on the surface performance of Ti-0.2Pd in a simulated body fluid (SBF) adding bovine serum albumin is investigated. Heat-treated specimens including furnace cooled (FC) and water quenched (WQ) specimens are also prepared for comparison. The corrosion resistance is evaluated by the tests of potentiodynamic polarization and the measurement of electrochemical impedance spectroscopy (EIS). The composition and morphology of the surface after exposing to SBF 60 days were examined by X-ray photoelectronic spectroscopy (XPS), scanning electron microscope (SEM) and energy dispersive spectrometer (EDS). The results show an ultrafine grained microstructure with average grain size of 3.6 μm is obtained after ECAP. The ultrafine grained Ti-0.2Pd has higher corrosion resistance than AR(as-received), WQ and FC specimens. The quantitative analysis of the surface shows larger numbers of precipitations formed on ECAPed Ti-0.2Pd than those formed on heat-treated. The precipitation contains more oxygen, calcium and phosphorus on ECAPed specimen than those on other specimens. The Ca:P ratio is ranged from 1:0.7 to 1:4.4, no dependent on the specimen type. A larger Warburg resistance is obtained on WQ specimen indicating a denser layer formation on WQ specimen. The precipitation formed on WQ specimens is the least among three kinds of specimens. Palladium is not found on the surfaces after exposure to SBF.

Scatter of fatigue data owing to material microscopic effects

NASA Astrophysics Data System (ADS)

Tang, XueSong

2014-01-01

A common phenomenon of fatigue test data reported in the open literature such as S-N curves exhibits the scatter of points for a group of same specimens under the same loading condition. The reason is well known that the microstructure is different from specimen to specimen even in the same group. Specifically, a fatigue failure process is a multi-scale problem so that a fatigue failure model should have the ability to take the microscopic effect into account. A physically-based trans-scale crack model is established and the analytical solution is obtained by coupling the micro- and macro-scale. Obtained is the trans-scale stress intensity factor as well as the trans-scale strain energy density (SED) factor. By taking this trans-scale SEDF as a key controlling parameter for the fatigue crack propagation from micro- to macro-scale, a trans-scale fatigue crack growth model is proposed in this work which can reflect the microscopic effect and scale transition in a fatigue process. The fatigue test data of aluminum alloy LY12 plate specimens is chosen to check the model. Two S-N experimental curves for cyclic stress ratio R=0.02 and R=0.6 are selected. The scattering test data points and two S-N curves for both R=0.02 and R=0.6 are exactly re-produced by application of the proposed model. It is demonstrated that the proposed model is able to reflect the multiscaling effect in a fatigue process. The result also shows that the microscopic effect has a pronounced influence on the fatigue life of specimens.

Friction and fretting wear characteristics of different diamond-like carbon coatings against alumina in water-lubricated fretting conditions.

PubMed

Watabe, Tsukasa; Amanov, Auezhan; Tsuboi, Ryo; Sasaki, Shinya

2013-12-01

Diamond-like carbon (DLC) coatings typically show low friction and high wear resistance. In this study, the friction and fretting wear characteristics of PVD, CVD and CVD-Si DLC coatings were investigated against an alumina (Al2O3) ball under water-lubricated fretting conditions. The objective of this study is to investigate and compare the friction and fretting wear characteristics of those DLC coatings at various fretting frequencies. The test results showed that the PVD DLC coating led to a lower friction coefficient and a higher resistance to fretting wear compared to those of the CVD and CVD-Si DLC coatings. However, the CVD DLC coating showed that the fretting wear resistance decreases with increasing frequency, while no significant difference in fretting wear resistances of the PVD and CVD-Si DLC coatings was observed. Quantitative surface analyses of the specimens were performed using an energy dispersive spectroscopy (EDS), a laser scanning microscope (LSM), a scanning electron microscope (SEM), an atomic force microscope (AFM) and the Raman spectroscopy.

An effective fixative for glucocorticoid receptors in fetal tissues

DOE Office of Scientific and Technical Information (OSTI.GOV)

Koga, T.; Kurisu, K.

1982-01-01

As a preliminary study in an autoradiographic study of glucocorticoid (GC) receptor localization in orofacial tissues of mouse fetuses, a search was made to determine the most effective fixative for preservation of the GC-receptor complex. Twelve-day-old mouse fetuses were administered tritiated triamcinolone acetonide (/sup 3/H-TAC) intraamniotically and subsequently processed by one of the following three procedures: freeze-drying, prefixation with Karnovsky's fixative, or the catechin fixative (Karnovsky's fixative containing 1% D-catechin) and postfixation with osmium tetroxide. Light microscopic autoradiography and liquid scintillation counting of the specimens revealed that the catechin fixative gave the best results for fixation of the steroid-receptor complexmore » and preservation of tissue structure. Light and electron microscopic autoradiographic studies of the time course of the localization of /sup 3/H-TAC in palatal shelves supported the catechin fixative as being the most effective in preservation of GC-receptor or ligand complexes.« less

The endocrine polypeptide cells of the human stomach, duodenum, and jejunum

PubMed Central

Pearse, A. G. E.; Coulling, I.; Weavers, B.; Friesen, S.

1970-01-01

Thirty specimens of stomach, duodenum, and jejunum, removed at operation, were examined by optical microscopical, cytochemical, and electron microscopical techniques. The overall distribution of four types of endocrine polypeptide cell in the stomach, and three in the intestine, was determined. The seven cell types are described by names and letters belonging to a scheme for nomenclature agreed upon at the 1969 Wiesbaden conference on gastrointestinal hormones. The gastrin-secreting G cell was the only cell for which firm identification with a known hormone was possible. Although there was wide variation in the distribution of the various cells, from one case to another, striking differences were nevertheless observable, with respect to the G cell, between antra from carcinoma and from ulcer cases. ImagesFig. 2Fig. 3Fig. 4Fig. 5Fig. 6Fig. 7Fig. 8Fig. 9Fig. 10Fig. 11Fig. 12Fig. 13Fig. 14Fig. 15Fig. 16Fig. 17 PMID:4919258

Bi-directional transmission of molecular information by photon or electron beams passing in the close vicinity of specific molecules, and its clinical and basic research applications: 1) Diagnosis of humans or animal patients without any direct contact; 2) Light microscopic and electron microscopic localization of neuro-transmitters, heavy metals, Oncogen C-fos (AB2), etc. of intracellular fine structures of normal and abnormal single cells using light or electro-microscopic indirect Bi-Digital O-Ring Test.

PubMed

Omura, Y; Losco, M; Omura, A K; Takeshige, C; Hisamitsu, T; Nakajima, H; Soejima, K; Yamamoto, S; Ishikawa, H; Kagoshima, T

1992-01-01

In 1985, Omura, Y. discovered that, when specific molecules were placed anywhere in the close vicinity of the path of a light beam (laser), their molecular information, as well as information on electrical & magnetic fields, is transmitted bi-directionally along the path of this light beam. Namely, this information is transmitted in the direction the light beam is projected and towards the direction from which the light beam is coming. This finding was applied to the following clinical and basic research: 1) In the past, using indirect Bi-Digital O-Ring Test, human or animal patients were diagnosed through an intermediate third person holding a good electrical conducting probe, the tip of which was touching the part of the patient to be examined. However, in order to diagnose the patient in isolation from a distance, or a dangerous or unmanagable unanesthesized animal, such as a lion or tiger, the author succeeded in making a diagnosis by replacing the metal conducting probe with a soft laser beam which is held by the one hand of the third person whose index finger is placed in close vicinity of the laser beam generated by a battery-powered penlight-type solid state laser generator. Thus, diagnosis within visible distance, without direct patient contact, became a reality. 2) Using a projection light microscope, by giving indirect Bi-Digital O-Ring Test while contacting with a fine electro-conductive probe on the magnified fine structure of normal and abnormal cells, various normal and abnormal intracellular substances were localized through a third person holding a pure reference control substance with the same hand that is holding the probe as an intermediary for the indirect Bi-Digital O-Ring Test. Instead of the photon beam in a light microscope, the author found that, using an electron beam passing through the close vicinity of specific molecules of specimens in an electron microscope, the molecular information is transmitted to the magnified fluorescent screen, and an indirect Bi-Digital O-Ring Test could be performed through a projected penlight-type solid state soft laser beam on the magnified intracellular structure through an observation glass window. Using the magnified fine structure of the cells, by either a light projection microscopic field or electron microscope, in various cancer cells of both humans and animals, Oncogen C-fos (AB2) and mercury were found inside of the nucleus. Integrin alpha 5 beta 1 was found on cell membranes and nuclear cell membranes of cancer cells. Acetylcholine was not found anywhere within cancer cells.(ABSTRACT TRUNCATED AT 400 WORDS)

Spectromicroscope for the PHotoelectron Imaging of Nanostructures with X-rays (SPHINX): performance in biology, medicine and geology.

PubMed

Frazer, Bradley H; Girasole, Marco; Wiese, Lisa M; Franz, Torsten; De Stasio, Gelsomina

2004-05-01

Several X-ray PhotoElectron Emission spectroMicroscopes (X-PEEMs) exist around the world at this time. We present recent performance and resolution tests of one of them, the Spectromicroscope for PHotoelectron Imaging of Nanostructures with X-rays (SPHINX) X-PEEM, installed at the University of Wisconsin Synchrotron Radiation Center. With this state-of-the-art instrument we demonstrate chemical analysis capabilities on conducting and insulating specimens of diverse interests, and an unprecedented lateral resolution of 10 nm with monochromatic X-rays and 7.2 nm with ultraviolet illumination.

SEM evaluation of metallization on semiconductors. [Scanning Electron Microscope

NASA Technical Reports Server (NTRS)

Fresh, D. L.; Adolphsen, J. W.

1974-01-01

A test method for the evaluation of metallization on semiconductors is presented and discussed. The method has been prepared in MIL-STD format for submittal as a proposed addition to MIL-STD-883. It is applicable to discrete devices and to integrated circuits and specifically addresses batch-process oriented defects. Quantitative accept/reject criteria are given for contact windows, other oxide steps, and general interconnecting metallization. Figures are provided that illustrate typical types of defects. Apparatus specifications, sampling plans, and specimen preparation and examination requirements are described. Procedures for glassivated devices and for multi-metal interconnection systems are included.

A method to characterize the roughness of 2-D line features: recrystallization boundaries.

PubMed

Sun, J; Zhang, Y B; Dahl, A B; Conradsen, K; Juul Jensen, D

2017-03-01

A method is presented, which allows quantification of the roughness of nonplanar boundaries of objects for which the neutral plane is not known. The method provides quantitative descriptions of both the local and global characteristics. How the method can be used to estimate the sizes of rough features and local curvatures is also presented. The potential of the method is illustrated by quantification of the roughness of two recrystallization boundaries in a pure Al specimen characterized by scanning electron microscopy. © 2016 The Authors Journal of Microscopy © 2016 Royal Microscopical Society.

Ultrastructure of the digestive system and experimental study of feeding in the monogenean skin and fin parasite Macrogyrodactylus congolensis.

PubMed

Arafa, Safaa Z; El-Naggar, Mohammed M; Kearn, Graham C

2013-12-01

In the present study, transmission electron microscopy (TEM) has been used to study the ultrastructure of the digestive system, namely the pharynx, oesophageal glands and intestine, of the monogenean skin and fin parasite Macrogyrodactylus congolensis. The pharynx consists of an anterior highly muscular region and a posterior mainly glandular syncytial region. The anterior region is provided with six pharyngeal papillae, the centre of each of which is occupied by electron dense secretory bodies, identical with those in the posterior region of the pharynx. The intestine has an uninterrupted syncytial gastrodermis and the luminal surface is provided with many unbranched lamellae. The intestine of living specimens contains large and small granules which give it a reddish brown colour. Large particles, presumed to be lipid droplets, and small granules, presumed to be melanin granules, were found in the gastrodermis and in the intestinal lumen. Parasites were induced to feed and then preserved for TEM at the following intervals: just after feeding, 30 min after feeding, 1 h 30 min after feeding and 2 h after feeding. The specimens were then processed for TEM and sections cut through the intestine of each specimen were examined with the transmission electron microscope. Three types of vacuoles (V1, V2, V3) were detected in the gastrodermis. Vacuoles V1 have thick walls and are likely to be endocytotic, enclosing luminal contents at the surface of the gastrodermis. V2 vacuoles may be lysosomes that fuse with V1 vacuoles. V3 vacuoles may serve to dispose of residual digestive material into the lumen.

Fatigue Damage Assessment Leveraging Nondestructive Evaluation Data

NASA Astrophysics Data System (ADS)

Mazur, K.; Wisner, B.; Kontsos, A.

2018-05-01

Fatigue in materials depends on several microstructural parameters. The length and time scales involved in such processes have been investigated by characterization methods that target microstructural effects or that rely on specimen-level observations. Combinations of in situ and ex situ techniques are also used to correlate microstructural changes to bulk properties. We present herein an effort to directly link local changes with specimen-level fatigue damage assessment. To achieve this goal, grain-scale observations in an aluminum alloy are linked with deformation measurements made by digital image correlation and with acoustic emission monitoring obtained from inside the scanning electron microscope. Damage assessment is attempted using a data-processing framework that involves noise removal, data reduction, and classification. The results demonstrate that nondestructive evaluation combined with small-scale testing can provide a means for fatigue damage assessment applicable to a broad range of materials and testing conditions.

[A study on the expression of anti-mitochondrial antibody in the brain of patients with MELAS syndrome].

PubMed

Qi, Xiao-Kun; Yao, Sheng; Wang, Hai-Yan; Piao, Yue-Shan; Lu, De-Hong; Yuan, Yun

2009-04-01

To investigate the pathological changes and pathogenesis of the MELAS syndrome (mitochondrial encephalopathy lactic acidosis stroke-like episodes) by using the method of immunohistochemical staining in the brain biopsy specimens with anti-mitochondrial antibody (AMA). We performed immunohistochemical staining in 3 confirmed MELAS patients' paraffin-imbued brain biopsy specimens. Small vessel proliferation and the uneven thickness of the wall were found in the 3 MELAS patients. A lot of brown deposits was shown in the wall of small vessels and also noted in neurons. The main pathological change in the MELAS brain biopsy immunohistochemical staining with AMA was the small vessel proliferation, indicating that abnormal mitochondria accumulated in the vascular smooth muscle, endothelial cell and neurons of the lesion sites. This finding was consistent with the electron microscopic discovery and valuable for the diagnosis of MELAS.

Comparative Effect of Bio-waste Ashes on Strength Properties of Cement Mortar

NASA Astrophysics Data System (ADS)

Ajay, Goyal; Hattori, Kunio; Ogata, Hidehiko; Ashraf, Muhammad; Ahmed, Mohamed Anwar

Biomass fuels produce about 400 million tonnes of ashes as waste material. This paper discusses the pozzolanic character of bio-waste ashes obtained from dry tree leaves (AML), Korai grass (KRI) and Tifton grass (TFT). Ashes were obtained by control incineration of the wastes at 600°C for 5 hours and mortar specimens were prepared by substituting cement with 10, 20 and 30% ash. Strength development of ash-blended mortar specimens was evaluated by conducting destructive tests as well as non-destructive tests till 91 days. X-ray diffraction, scanning electron microscopic and thermo-gravimetric techniques were used to analyze the influence of ash substitution on strength properties of blended-mortar. Pozzolanic reactivity of AML- and KRI-ash was confirmed, but TFT-ash did not show enough reactivity. Overall results confirmed that up to 20% substitution of cement can be made with AML- or KRI-ash with strength approaching 90% of that of control.

Whole mount nuclear fluorescent imaging: convenient documentation of embryo morphology.

PubMed

Sandell, Lisa L; Kurosaka, Hiroshi; Trainor, Paul A

2012-11-01

Here, we describe a relatively inexpensive and easy method to produce high quality images that reveal fine topological details of vertebrate embryonic structures. The method relies on nuclear staining of whole mount embryos in combination with confocal microscopy or conventional wide field fluorescent microscopy. In cases where confocal microscopy is used in combination with whole mount nuclear staining, the resulting embryo images can rival the clarity and resolution of images produced by scanning electron microscopy (SEM). The fluorescent nuclear staining may be performed with a variety of cell permeable nuclear dyes, enabling the technique to be performed with multiple standard microscope/illumination or confocal/laser systems. The method may be used to document morphology of embryos of a variety of organisms, as well as individual organs and tissues. Nuclear stain imaging imposes minimal impact on embryonic specimens, enabling imaged specimens to be utilized for additional assays. Copyright © 2012 Wiley Periodicals, Inc.

Ion Release and Galvanic Corrosion of Different Orthodontic Brackets and Wires in Artificial Saliva.

PubMed

Tahmasbi, Soodeh; Sheikh, Tahereh; Hemmati, Yasamin B

2017-03-01

To investigate the galvanic corrosion of brackets manufactured by four different companies coupled with stainless steel (SS) or nickel-titanium (NiTi) wires in an artificial saliva solution. A total of 24 mandibular central incisor Roth brackets of four different manufacturers (American Orthodontics, Dentaurum, Shinye, ORJ) were used in this experimental study. These brackets were immersed in artificial saliva along with SS or NiTi orthodontic wires (0.016'', round) for 28 days. The electric potential difference of each bracket/ wire coupled with a saturated calomel reference electrode was measured via a voltmeter and recorded constantly. Corrosion rate (CR) was calculated, and release of ions was measured with an atomic absorption spectrometer. Stereomicroscope was used to evaluate all samples. Then, samples with corrosion were further assessed by scanning electron microscope and energy-dispersive X-ray spectroscopy. Two-way analysis of variance was used to analyze data. Among ions evaluated, release of nickel ions from Shinye brackets was significantly higher than that of other brackets. The mean potential difference was significantly lower in specimens containing a couple of Shinye brackets and SS wire compared with other specimens. No significant difference was observed in the mean CR of various groups (p > 0.05). Microscopic evaluation showed corrosion in two samples only: Shinye bracket coupled with SS wire and American Orthodontics bracket coupled with NiTi wire. Shinye brackets coupled with SS wire showed more susceptibility to galvanic corrosion. There were no significant differences among specimens in terms of the CR or released ions except the release of Ni ions, which was higher in Shinye brackets.

Interface toughness of a zirconia-veneer system and the effect of a liner application.

PubMed

Wang, Gaoqi; Zhang, Song; Bian, Cuirong; Kong, Hui

2014-09-01

Chipping of veneering porcelain and delamination of a zirconia-veneer interface are 2 common clinical failure modes for zirconia-based restorations and may be partially due to weak interface bonding. The effect of liner on the bond strength of the interface has not been clearly identified. The purpose of the research was to evaluate the interface toughness between the zirconia core and veneering porcelain by means of a fracture mechanics test and to assess the effect of liner on the bond strength of the interface. Thirty bilayered beam-shape specimens were prepared and divided into 2 groups according to liner application. The specimens in each group were subdivided into 3 subgroups in accordance with 3 different veneer thicknesses. A fracture mechanics test was used on each specimen, and the energy release rate, G, and phase angle, ψ, were calculated according to the experimental results. A video microscope was used to monitor the crack propagation, and a scanning electron microscope was used to identify the fracture mode after testing. Two-way ANOVA and the Tukey honestly significant difference test were performed to analyze the experimental data (α=.05) . At each phase angle, the interfaces without a liner had higher mean G values than the interfaces with a liner. Both of the interfaces showed mixed failure mode with thin layers of a veneer or a liner that remained on the zirconia surfaces. Liner application before veneering reduced the interface toughness between zirconia and veneer. Copyright © 2014 Editorial Council for the Journal of Prosthetic Dentistry. Published by Elsevier Inc. All rights reserved.

Influence of process time on microstructure and properties of 17-4PH steel plasma nitrocarburized with rare earths addition at low temperature

NASA Astrophysics Data System (ADS)

Yan, M. F.; Liu, R. L.

2010-08-01

17-4PH stainless steel was plasma nitrocarburized at 430 °C for different time with rare earths (RE) addition. Plasma RE nitrocarburized layers were studied by optical microscope, scanning electron microscope equipped with an energy dispersive X-ray analyzer, X-ray diffraction, microhardness tests, pin-on-disc tribometer and anodic polarization tests. The results show that rare earths atoms can diffuse into the surface of 17-4PH steel. The modified layer depths increase with increasing process time and the layer growth conforms approximately to the parabolic law. The phases in the modified layer are mainly of γ'-Fe 4N, nitrogen and carbon expanded martensite (α' N) as well as some incipient CrN at short time (2 h). With increasing of process time, the phases of CrN and γ'-Fe 4N increase but α' N decomposes gradually. Interestingly, the peaks of γ'-Fe 4N display a high (2 0 0) plane preferred orientation. The hardness of the modified specimen is more than 1340 HV, which is about 3.7 times higher than that of untreated one. The friction coefficients and wear rates of specimens can be dramatically decreased by plasma RE nitrocarburizing. The surface hardness and the friction coefficients decrease gradually with increasing process time. The corrosion test shows that the 8 h treated specimen has the best corrosion resistance with the characterization of lower corrosion current density, a higher corrosion potential and a large passive region as compared with those of untreated one.

Nanoscale imaging of clinical specimens using pathology-optimized expansion microscopy

PubMed Central

Zhao, Yongxin; Bucur, Octavian; Irshad, Humayun; Chen, Fei; Weins, Astrid; Stancu, Andreea L.; Oh, Eun-Young; DiStasio, Marcello; Torous, Vanda; Glass, Benjamin; Stillman, Isaac E.; Schnitt, Stuart J.; Beck, Andrew H.; Boyden, Edward S.

2017-01-01

Expansion microscopy (ExM), a method for improving the resolution of light microscopy by physically expanding the specimen, has not been applied to clinical tissue samples. Here we report a clinically optimized form of ExM that supports nanoscale imaging of human tissue specimens that have been fixed with formalin, embedded in paraffin, stained with hematoxylin and eosin (H&E), and/or fresh frozen. The method, which we call expansion pathology (ExPath), converts clinical samples into an ExM-compatible state, then applies an ExM protocol with protein anchoring and mechanical homogenization steps optimized for clinical samples. ExPath enables ~70 nm resolution imaging of diverse biomolecules in intact tissues using conventional diffraction-limited microscopes, and standard antibody and fluorescent DNA in situ hybridization reagents. We use ExPath for optical diagnosis of kidney minimal-change disease, which previously required electron microscopy (EM), and demonstrate high-fidelity computational discrimination between early breast neoplastic lesions that to date have challenged human judgment. ExPath may enable the routine use of nanoscale imaging in pathology and clinical research. PMID:28714966

Direct observation of the residual plastic deformation caused by a single tensile overload

DOE Office of Scientific and Technical Information (OSTI.GOV)

Bichler, C.; Pippan, R.

1999-07-01

The fatigue crack growth behavior following single tensile overloads at high stress intensity ranges in a cold-rolled austenitic steel has been studied experimentally. After tensile overloads, fatigue cracks initially accelerate, followed by significant retardation, before the growth rates return to their baseline level. The initial acceleration was attributed to an immediate reduction in near-tip closure. Scanning electron micrography and stereophotogrammetric reconstruction of the fracture surface were applied to study the residual plastic deformation caused by a single tensile overload in the mid-thickness of the specimen. The measured residual opening displacement of the crack as a function of the overload ismore » presented and compared with simple estimations. Also, free specimen surface observations of the residual plastic deformation and crack growth rate were performed. In the midsection of the specimens the striation spacing-length, i.e., the microscopic growth rates, were measured before and after the applied overload. It will be shown that the measured plasticity-induced wedges from the single overload and the observed propagation behavior support the significance of the concept of crack closure.« less

Nanoscale imaging of clinical specimens using pathology-optimized expansion microscopy.

PubMed

Zhao, Yongxin; Bucur, Octavian; Irshad, Humayun; Chen, Fei; Weins, Astrid; Stancu, Andreea L; Oh, Eun-Young; DiStasio, Marcello; Torous, Vanda; Glass, Benjamin; Stillman, Isaac E; Schnitt, Stuart J; Beck, Andrew H; Boyden, Edward S

2017-08-01

Expansion microscopy (ExM), a method for improving the resolution of light microscopy by physically expanding a specimen, has not been applied to clinical tissue samples. Here we report a clinically optimized form of ExM that supports nanoscale imaging of human tissue specimens that have been fixed with formalin, embedded in paraffin, stained with hematoxylin and eosin, and/or fresh frozen. The method, which we call expansion pathology (ExPath), converts clinical samples into an ExM-compatible state, then applies an ExM protocol with protein anchoring and mechanical homogenization steps optimized for clinical samples. ExPath enables ∼70-nm-resolution imaging of diverse biomolecules in intact tissues using conventional diffraction-limited microscopes and standard antibody and fluorescent DNA in situ hybridization reagents. We use ExPath for optical diagnosis of kidney minimal-change disease, a process that previously required electron microscopy, and we demonstrate high-fidelity computational discrimination between early breast neoplastic lesions for which pathologists often disagree in classification. ExPath may enable the routine use of nanoscale imaging in pathology and clinical research.

The orthoenstatite to clinoenstatite transformation by shearing and reversion by annealing: Mechanism and potential applications

USGS Publications Warehouse

Coe, R.S.; Kirby, S.H.

1975-01-01

Clinoenstatite (CE) was produced by deforming single-crystal specimens of ortho-enstatite (OE) in several different sorta of experiments. Examination with light and trans-mission electron microscopes shows that the transformation is coherent and involves a macroscopic shear on (100) [001] through an angle of 12.8??1.3 ??, in good agreement with the theoretically expected value of 13.3 ??, and that the transformation is accomplished by glide on (100) of partial dislocations with b= 0.83[001]. Structural analysis provides further insight into the transformation mechanism. Reversion occurs in specimens annealed under a variety of conditions, and thin lamellae of CE in unconstrained, low-strain specimens recover their original shape during transformation back to OE. Our experiments and thermodynamic estimates both suggest that the equilibrium transition temperature is raised roughly 300 ?? C per kilobar of shear stress on (100) [001]. This provides the basis of a method by which it may be possible to determine the magnitude as well as the orientation of the principal stresses that produce CE in nature during deformation of enstatite-bearing rocks. ?? 1975 Springer-Verlag.

A new method to detect and correct sample tilt in scanning transmission electron microscopy bright-field imaging.

PubMed

Brown, H G; Ishikawa, R; Sánchez-Santolino, G; Lugg, N R; Ikuhara, Y; Allen, L J; Shibata, N

2017-02-01

Important properties of functional materials, such as ferroelectric shifts and octahedral distortions, are associated with displacements of the positions of lighter atoms in the unit cell. Annular bright-field scanning transmission electron microscopy is a good experimental method for investigating such phenomena due to its ability to image light and heavy atoms simultaneously. To map atomic positions at the required accuracy precise angular alignment of the sample with the microscope optical axis is necessary, since misalignment (tilt) of the specimen contributes to errors in position measurements of lighter elements in annular bright-field imaging. In this paper it is shown that it is possible to detect tilt with the aid of images recorded using a central bright-field detector placed within the inner radius of the annular bright-field detector. For a probe focus near the middle of the specimen the central bright-field image becomes especially sensitive to tilt and we demonstrate experimentally that misalignment can be detected with a precision of less than a milliradian, as we also confirm in simulation. Coma in the probe, an aberration that can be misidentified as tilt of the specimen, is also investigated and it is shown how the effects of coma and tilt can be differentiated. The effects of tilt may be offset to a large extent by shifting the diffraction plane detector an amount equivalent to the specimen tilt and we provide an experimental proof of principle of this using a segmented detector system. Copyright © 2016 Elsevier B.V. All rights reserved.

Galvanic Corrosion of and Ion Release from Various Orthodontic Brackets and Wires in a Fluoride-containing Mouthwash.

PubMed

Tahmasbi, Soodeh; Ghorbani, Mohammad; Masudrad, Mahdis

2015-01-01

Background and aims. This study compared the galvanic corrosion of orthodontic wires and brackets from various manufacturers following exposure to a fluoride mouthwash. Materials and methods. This study was conducted on 24 lower central incisor 0.022" Roth brackets of four different commercially available brands (Dentaurum, American Orthodontics, ORJ, Shinye). These brackets along with stainless steel (SS) or nickel-titanium (NiTi) orthodontic wires (0.016", round) were immersed in Oral-B mouthwash containing 0.05% sodium fluoride for 28 days. The electric potential (EP) difference of each bracket-wire couple was measured with a Saturated Calomel Reference Electrode (Ag/AgCl saturated with KCl) via a voltmeter. The ions released in the electrolyte weremeasured with an atomic absorption spectrometer. All the specimens were assessed under a stereomicroscope and specimens with corrosion were analyzed with scanning electron microscopy (SEM). Data were analyzed using ANOVA. Results. The copper ions released from specimens with NiTi wire were greater than those of samples containing SS wire. ORJ brackets released more Cu ions than other samples. The Ni ions released from Shinye brackets were significantly more than those of other specimens (P < 0.05). Corrosion rate of brackets coupled with NiTi wires was higher than that of brackets coupled with SS wires. Light and electron microscopic observations showed greater corrosion of ORJ brackets. Conclusion. In fluoride mouthwash, Shinye and ORJ brackets exhibited greater corrosion than Dentaurum and American Orthodontics brackets. Stainless steel brackets used with NiTi wires showed greater corrosion and thus caution is recommended when using them.

Dose fractionation theorem in 3-D reconstruction (tomography)

DOE Office of Scientific and Technical Information (OSTI.GOV)

Glaeser, R.M.

It is commonly assumed that the large number of projections for single-axis tomography precludes its application to most beam-labile specimens. However, Hegerl and Hoppe have pointed out that the total dose required to achieve statistical significance for each voxel of a computed 3-D reconstruction is the same as that required to obtain a single 2-D image of that isolated voxel, at the same level of statistical significance. Thus a statistically significant 3-D image can be computed from statistically insignificant projections, as along as the total dosage that is distributed among these projections is high enough that it would have resultedmore » in a statistically significant projection, if applied to only one image. We have tested this critical theorem by simulating the tomographic reconstruction of a realistic 3-D model created from an electron micrograph. The simulations verify the basic conclusions of high absorption, signal-dependent noise, varying specimen contrast and missing angular range. Furthermore, the simulations demonstrate that individual projections in the series of fractionated-dose images can be aligned by cross-correlation because they contain significant information derived from the summation of features from different depths in the structure. This latter information is generally not useful for structural interpretation prior to 3-D reconstruction, owing to the complexity of most specimens investigated by single-axis tomography. These results, in combination with dose estimates for imaging single voxels and measurements of radiation damage in the electron microscope, demonstrate that it is feasible to use single-axis tomography with soft X-ray microscopy of frozen-hydrated specimens.« less

Observations of Fabric Development in Polycrystalline Ice at Basal Pressures: Methods and Initial Results

NASA Astrophysics Data System (ADS)

Breton, D. J.; Baker, I.; Cole, D. M.

2012-12-01

Understanding and predicting the flow of polycrystalline ice is crucial to ice sheet modeling and paleoclimate reconstruction from ice cores. Ice flow rates depend strongly on the fabric (i.e. the distribution of grain sizes and crystallographic orientations) which evolves over time and enhances the flow rate in the direction of applied stress. The mechanisms for fabric evolution in ice have been extensively studied at atmospheric pressures, but little work has been done to observe these processes at the high pressures experienced deep within ice sheets where long-term changes in ice rheology are expected to have significance. We conducted compressive creep tests on a 917 kg m-3 polycrystalline ice specimen at 20 MPa hydrostatic pressure, thus simulating ~2,000 m depth. Initial specimen grain orientations were random, typical grain diameters were 1.2 mm, and the applied creep stress was 0.3 MPa. Subsequent microstructural analyses on the deformed specimen and a similarly prepared, undeformed specimen allowed characterization of crystal fabric evolution under pressure. Our microstructural analysis technique simultaneously collected grain shape and size data from Scanning Electron Microscope (SEM) micrographs and obtained crystallographic orientation data via Electron BackScatter Diffraction (EBSD). Combining these measurements allows rapid analysis of the ice fabric over large numbers of grains, yielding statistically useful numbers of grain size and full c- and a-axis grain orientation data. The combined creep and microstructural data demonstrate pressure-dependent effects on the mechanical and microstructural evolution of polycrystalline ice. We discuss possible mechanisms for the observed phenomena, and future directions for hydrostatic creep testing.

Galvanic Corrosion of and Ion Release from Various Orthodontic Brackets and Wires in a Fluoride-containing Mouthwash

PubMed Central

Tahmasbi, Soodeh; Ghorbani, Mohammad; Masudrad, Mahdis

2015-01-01

Background and aims. This study compared the galvanic corrosion of orthodontic wires and brackets from various manufacturers following exposure to a fluoride mouthwash. Materials and methods. This study was conducted on 24 lower central incisor 0.022" Roth brackets of four different commercially available brands (Dentaurum, American Orthodontics, ORJ, Shinye). These brackets along with stainless steel (SS) or nickel-titanium (NiTi) orthodontic wires (0.016", round) were immersed in Oral-B mouthwash containing 0.05% sodium fluoride for 28 days. The electric potential (EP) difference of each bracket-wire couple was measured with a Saturated Calomel Reference Electrode (Ag/AgCl saturated with KCl) via a voltmeter. The ions released in the electrolyte weremeasured with an atomic absorption spectrometer. All the specimens were assessed under a stereomicroscope and specimens with corrosion were analyzed with scanning electron microscopy (SEM). Data were analyzed using ANOVA. Results. The copper ions released from specimens with NiTi wire were greater than those of samples containing SS wire. ORJ brackets released more Cu ions than other samples. The Ni ions released from Shinye brackets were significantly more than those of other specimens (P < 0.05). Corrosion rate of brackets coupled with NiTi wires was higher than that of brackets coupled with SS wires. Light and electron microscopic observations showed greater corrosion of ORJ brackets. Conclusion. In fluoride mouthwash, Shinye and ORJ brackets exhibited greater corrosion than Dentaurum and American Orthodontics brackets. Stainless steel brackets used with NiTi wires showed greater corrosion and thus caution is recommended when using them. PMID:26697148

Understanding catalyst behavior during in situ heating through simultaneous secondary and transmitted electron imaging.

PubMed

Howe, Jane Y; Allard, Lawrence F; Bigelow, Wilbur C; Demers, Hendrix; Overbury, Steven H

2014-01-01

By coupling techniques of simultaneous secondary (SE) and transmitted electron (TE) imaging at high resolution in a modern scanning transmission electron microscope (STEM), with the ability to heat specimens using a highly stable MEMS-based heating platform, we obtained synergistic information to clarify the behavior of catalysts during in situ thermal treatments. Au/iron oxide catalyst 'leached' to remove surface Au was heated to temperatures as high as 700°C. The Fe2O3 support particle structure tended to reduce to Fe3O4 and formed surface terraces; the formation, coalescence, and mobility of 1- to 2-nm particles on the terraces were characterized in SE, STEM-ADF, and TEM-BF modes. If combined with simultaneous nanoprobe spectroscopy, this approach will open the door to a new way of studying the kinetics of nano-scaled phenomena.

Understanding catalyst behavior during in situ heating through simultaneous secondary and transmitted electron imaging

NASA Astrophysics Data System (ADS)

Howe, Jane Y.; Allard, Lawrence F.; Bigelow, Wilbur C.; Demers, Hendrix; Overbury, Steven H.

2014-11-01

By coupling techniques of simultaneous secondary (SE) and transmitted electron (TE) imaging at high resolution in a modern scanning transmission electron microscope (STEM), with the ability to heat specimens using a highly stable MEMS-based heating platform, we obtained synergistic information to clarify the behavior of catalysts during in situ thermal treatments. Au/iron oxide catalyst 'leached' to remove surface Au was heated to temperatures as high as 700°C. The Fe2O3 support particle structure tended to reduce to Fe3O4 and formed surface terraces; the formation, coalescence, and mobility of 1- to 2-nm particles on the terraces were characterized in SE, STEM-ADF, and TEM-BF modes. If combined with simultaneous nanoprobe spectroscopy, this approach will open the door to a new way of studying the kinetics of nano-scaled phenomena.

Integrating Microscopic Analysis into Existing Quality Assurance Processes

NASA Astrophysics Data System (ADS)

Frühberger, Peter; Stephan, Thomas; Beyerer, Jürgen

When technical goods, like mainboards and other electronic components, are produced, quality assurance (QA) is very important. To achieve this goal, different optical microscopes can be used to analyze a variety of specimen to gain comprehensive information by combining the acquired sensor data. In many industrial processes, cameras are used to examine these technical goods. Those cameras can analyze complete boards at once and offer a high level of accuracy when used for completeness checks. When small defects, e.g. soldered points, need to be examined in detail, those wide area cameras are limited. Microscopes with large magnification need to be used to analyze those critical areas. But microscopes alone cannot fulfill this task within a limited time schedule, because microscopic analysis of complete motherboards of a certain size is time demanding. Microscopes are limited concerning their depth of field and depth of focus, which is why additional components like XY moving tables need to be used to examine the complete surface. Yet today's industrial production quality standards require a 100 % control of the soldered components within a given time schedule. This level of quality, while keeping inspection time low, can only be achieved when combining multiple inspection devices in an optimized manner. This paper presents results and methods of combining industrial cameras with microscopy instrumenting a classificatory based approach intending to keep already deployed QA processes in place but extending them with the purpose of increasing the quality level of the produced technical goods while maintaining high throughput.

Dark-field imaging with the scanning transmission x-ray microscope

NASA Astrophysics Data System (ADS)

Morrison, G. R.; Browne, M. T.

1992-01-01

The King's College London scanning transmission x-ray microscope in use on beam line 5U2 at the SRS, SERC Daresbury Laboratory, has been modified to allow dark-field images to be formed using only the x rays scattered by the specimen. Experiments have been performed with a number of different detector geometries, and this has confirmed that the strongest scattering arises from edges or thickness gradients in the specimen. Although the dark-field signal is only a small fraction of the normal transmitted bright-field signal, features can be revealed with high contrast, and it has proved possible to detect the presence of features that are below the resolution limit of the microscope.

Effects of overnight refrigeration on the microscopic evaluation of sputum.

PubMed Central

Penn, R L; Silberman, R

1984-01-01

Microscopic evaluation of sputum permits selection of specimens suitable for culture, assessment of likely pathogens, and the best interpretation of culture results. We prospectively evaluated 50 sputum specimens which were promptly submitted to our clinical laboratory; smears and cultures were performed both immediately and after 20 h of refrigeration. Specimens were grouped according to the numbers of squamous epithelial cells and neutrophils per low-power field present on coded Gram-stained smears. The numbers of bacteria in five oil immersion fields were used to characterize smears for predominant, mixed, or scanty forms. After refrigeration, only three specimens changed group from a definite loss of squamous epithelial cells, and only two changed group from a definite loss of neutrophils. Based on cellular composition, the majority of samples would have been processed identically both before and after refrigeration. In contrast, organism forms detected on smears and their relative quantities were dramatically altered after refrigeration. A predominant smear form was gained in 11 and lost in 8 refrigerated specimens. The frequent changes on smears observed overall resulted from both increases and decreases in numbers of bacteria and yeasts. The majority of sputum culture results were insignificantly affected by the refrigeration of specimens. We conclude that 20 h of refrigeration renders sputum useless for the microscopic evaluation of potential pathogens and the subsequent interpretation of culture results. However, overnight refrigeration does not affect the determination from smears of sputum suitability for culture based on cellular composition. PMID:6699145

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

NASA Astrophysics Data System (ADS)

Karreman, M. A.

2013-03-01

Correlative microscopy is the combined use of two different forms of microscopy in the study of a specimen, allowing for the exploitation of the advantages of both imaging tools. The integrated Laser and Electron Microscope (iLEM), developed at Utrecht University, combines a fluorescence microscope (FM) and a transmission electron microscope (TEM) in a single set-up. The region of interest in the specimen is labeled or tagged with a fluorescent probe and can easily be identified within a large field of view with the FM. Next, this same area is retraced in the TEM and can be studied at high resolution. The iLEM demands samples that can be imaged with both FM and TEM. Biological specimen, typically composed of light elements, generate low image contrast in the TEM. Therefore, these samples are often ‘contrasted’ with heavy metal stains. FM, on the other hand, images fluorescent samples. Sample preparation for correlative microscopy, and iLEM in particular, is complicated by the fact that the heavy metals stains employed for TEM quench the fluorescent signal of the probe that is imaged with FM. The first part of this thesis outlines preparation procedures for biological material yielding specimen that can be imaged with the iLEM. Here, approaches for the contrasting of thin sections of cells and tissue are introduced that do not affect the fluorescence signal of the probe that marks the region of interest. Furthermore, two novel procedures, VIS2FIXH and VIS2FIX­FS are described that allow for the chemical fixation of thin sections of cryo-immobilized material. These procedures greatly expedite the sample preparation process, and open up novel possibilities for the immuno-labeling of difficult antigens, eg. proteins and lipids that are challenging to preserve. The second part of this thesis describes applications of iLEM in research in the field of life and material science. The iLEM was employed in the study of UVC induced apoptosis (programmed cell death) of human umbilical vein endothelial cells. A novel, RNA containing body was identified in the nuclei of cells going through the various stages of the apoptotic process. Furthermore, we demonstrated the potential of iLEM in the study of Facio Scapulo Humeral Dystrophy (FSHD), the third most common form of inherited muscular dystrophy. In this study, diseased cells are identified based on the immuno-labeling of proteins associated with FSHD pathology. In the field of heterogeneous catalysis, a structural and functional characterization of Fluid Catalytic Cracking (FCC) particles was performed with iLEM. FCC particles are employed in petrochemical industry, where they catalyze the breakdown of large molecules in crude oil fractions into functional products with lower molecular weight, like gasoline. The catalytic sites in the FCC particles were selectively stained with a fluorescent probe, and next their structure was investigated with TEM. The iLEM allowed for the identification and characterization of catalytically active areas in the FCC particles. Furthermore, a unique study of the deactivation processes taking place in an industrial FCC unit was performed by analyzing a sample derived from a FCC reactor

Microstructure and hot corrosion behavior of the Ni-based superalloy GH202 treated by laser shock processing

DOE Office of Scientific and Technical Information (OSTI.GOV)

Cao, Jiangdong

The effects of laser shock processing on microstructure, the residual stress, and hot corrosion behavior of the Ni-based superalloy GH202 were investigated. The microstructures of GH202 before and after laser shock processing (LSP) were characterized by electron backscattered diffraction (EBSD) and transmission electron microscope (TEM). A large number of crystal defects (twins, dislocation arrays, and high dense tangles) were generated on the surface of GH202 treated with LSP. The cross-sectional compressive residual stress and micro-hardness of specimens treated by LSP were improved significantly. The corrosion kinetics of GH202 with or without LSP treatment at 800 °C and 900 °C weremore » investigated. Analysis by X-ray diffraction (XRD) revealed that the corrosion products mainly consist of Cr{sub 2}O{sub 3}, TiO{sub 2}, Al{sub 2}O{sub 3}, NiO, CrS, Ni{sub 3}S{sub 2}, and Na{sub 2}CrO{sub 4}. The surface and cross-section morphologies were observed by scanning electron microscope (SEM) combined with energy dispersive spectroscopy (EDS). The results confirmed that the crystal defects induced by LSP promotes the creation of diffusion paths for elements (Cr, Al, and Ti), allowing the formation of tiny homogeneous oxidation films in a very short time. Additionally, the spallation of oxidation film on the treated specimens was alleviated significantly. Overall, the hot corrosion resistance of Ni-based GH202 induced by LSP was improved in Na{sub 2}SO{sub 4} and NaCl molten salt from 800 °C to 900 °C. - Highlights: • Microstructure changes of GH202 before and after LSP were observed by EBSD and TEM. • The hardness and residual compressive stress after LSP were significantly increased. • The increased diffusion paths for elements helped to form oxidation films quickly. • Hot corrosion resistance of GH202 after LSP was significantly improved.« less

Diffracting aperture based differential phase contrast for scanning X-ray microscopy.

PubMed

Kaulich, Burkhard; Polack, Francois; Neuhaeusler, Ulrich; Susini, Jean; di Fabrizio, Enzo; Wilhein, Thomas

2002-10-07

It is demonstrated that in a zone plate based scanning X-ray microscope, used to image low absorbing, heterogeneous matter at a mesoscopic scale, differential phase contrast (DPC) can be implemented without adding any additional optical component to the normal scheme of the microscope. The DPC mode is simply generated by an appropriate positioning and alignment of microscope apertures. Diffraction from the apertures produces a wave front with a non-uniform intensity. The signal recorded by a pinhole photo diode located in the intensity gradient is highly sensitive to phase changes introduced by the specimen to be recorded. The feasibility of this novel DPC technique was proven with the scanning X-ray microscope at the ID21 beamline of the European Synchrotron Radiation facility (ESRF) operated at 6 keV photon energy. We observe a differential phase contrast, similar to Nomarski's differential interference contrast for the light microscope, which results in a tremendous increase in image contrast of up to 20 % when imaging low absorbing specimen.

Mechanical properties and microstructures of glass-ionomer cements.

PubMed

Xie, D; Brantley, W A; Culbertson, B M; Wang, G

2000-03-01

The objective of this study was to determine the flexural strength (FS), compressive strength (CS), diametral tensile strength (DTS), Knoop hardness (KHN) and wear resistance of ten commercial glass-ionomer cements (GICs). The fracture surfaces of these cements were examined using scanning electron microscopic (SEM) techniques to ascertain relationships between the mechanical properties and microstructures of these cements. Specimens were fabricated according to the instructions from each manufacturer. The FS, CS, DTS, KHN and wear rate were measured after conditioning the specimens for 7 d in distilled water at 37 degrees C. One-way analysis of variance with the post hoc Tukey-Kramer multiple range test was used to determine which specimen groups were significantly different for each test. The fracture surface of one representative specimen of each GIC from the FS tests was examined using a scanning electron microscope. The resin-modified GICs (RM GICs) exhibited much higher FS and DTS, not generally higher CS, often lower Knoop hardness and generally lower wear resistance, compared to the conventional GICs (C GICs). Vitremer (3M) had the highest values of FS and DTS; Fuji II LC (GC International) and Ketac-Molar (ESPE) had the highest CS; Ketac-Fil (ESPE) had the highest KHN. Ketac-Bond (ESPE) had the lowest FS; alpha-Silver (DMG-Hamburg) had the lowest CS. Four GICs (alpha-Fil (DMG-Hamburg), alpha-Silver, Ketac-Bond and Fuji II) had the lowest values of DTS, which were not significantly different from each other; alpha-Silver and Ketac-Silver had the lowest values of KHN. The highest wear resistance was exhibited by alpha-Silver and Ketac-Fil; F2LC had the lowest wear resistance. The C GICs exhibited brittle behavior, whereas the RM GICs underwent substantial plastic deformation in compression. The more integrated the microstructure, the higher were the FS and DTS. Higher CS was correlated with smaller glass particles, and higher KHN was found where there was a combination of smaller glass particles and lower porosity. Larger glass particle sizes and a more integrated microstructure contributed to a higher wear resistance. The mechanical properties of GICs were closely related to their microstructures. Factors such as the integrity of the interface between the glass particles and the polymer matrix, the particle size, and the number and size of voids have important roles in determining the mechanical properties.

Microscopic neural image registration based on the structure of mitochondria

NASA Astrophysics Data System (ADS)

Cao, Huiwen; Han, Hua; Rao, Qiang; Xiao, Chi; Chen, Xi

2017-02-01

Microscopic image registration is a key component of the neural structure reconstruction with serial sections of neural tissue. The goal of microscopic neural image registration is to recover the 3D continuity and geometrical properties of specimen. During image registration, various distortions need to be corrected, including image rotation, translation, tissue deformation et.al, which come from the procedure of sample cutting, staining and imaging. Furthermore, there is only certain similarity between adjacent sections, and the degree of similarity depends on local structure of the tissue and the thickness of the sections. These factors make the microscopic neural image registration a challenging problem. To tackle the difficulty of corresponding landmarks extraction, we introduce a novel image registration method for Scanning Electron Microscopy (SEM) images of serial neural tissue sections based on the structure of mitochondria. The ellipsoidal shape of mitochondria ensures that the same mitochondria has similar shape between adjacent sections, and its characteristic of broad distribution in the neural tissue guarantees that landmarks based on the mitochondria distributed widely in the image. The proposed image registration method contains three parts: landmarks extraction between adjacent sections, corresponding landmarks matching and image deformation based on the correspondences. We demonstrate the performance of our method with SEM images of drosophila brain.

Influences of strain rate on yield strength aluminum alloys

NASA Astrophysics Data System (ADS)

Rizal, Samsul; Firdaus, Hamdani Teuku; Thaib, Razali; Homma, Hiroomi

2005-04-01

The simulation of aircraft has often been performing by implementing finite element code on supercomputers. The reliability an accuracy of simulation depends mainly on the material model as well as on structural model used in calculations. Consequently, an accurate knowledge of mechanical behavior of materials under impact loading is essential for safety performance evaluation of structure. Impact tension tests on specimens for aircrafts and automotive structural applications are conduct by means of the split Hopkinson bar apparatus. Small specimens having diameter 4 mm are use in the test. Tensile stress-strain relations at strain rates of 102 s-1 to over 103 s-1 are present and compared with those obtained at quasi-static strain rates. The limitations on the applicability of apparatus are also discusses. The other importance of the reference of strain, while studying void growth in elastic-viscoplastic material, is emphasized. In the present paper, a simplified plane-symmetrical two-dimensional finite element model for a SHPB with a plate specimen made of an elastic material is first established. The used of strain gage mounted at the specimens to be monitored strain during the course of impact test. Comparisons may then be made between the numerical predicted and experimentally observed of load and a specimen strain. This report also describes the apparatus and instrumentation, and also be discusses the advantages and limitations of experimental technique. Fractograph is taken by scanning electron microscope on the center of the specimens for judgment of the fracture mechanism and strain rates influences on the materials.

Science 101: How Does an Electron Microscope Work?

ERIC Educational Resources Information Center

Robertson, Bill

2013-01-01

Contrary to popular opinion, electron microscopes are not used to look at electrons. They are used to look for structure in things that are too small to observe with an optical microscope, or to obtain images that are magnified much more than is obtainable with an optical microscope. To understand how electron microscopes work, it will help to go…

Recent advances in the application of electron tomography to materials chemistry.

PubMed

Leary, Rowan; Midgley, Paul A; Thomas, John Meurig

2012-10-16

Nowadays, tomography plays a central role in pureand applied science, in medicine, and in many branches of engineering and technology. It entails reconstructing the three-dimensional (3D) structure of an object from a tilt series of two-dimensional (2D) images. Its origin goes back to 1917, when Radon showed mathematically how a series of 2D projection images could be converted to the 3D structural one. Tomographic X-ray and positron scanning for 3D medical imaging, with a resolution of ∼1 mm, is now ubiquitous in major hospitals. Electron tomography, a relatively new chemical tool, with a resolution of ∼1 nm, has been recently adopted by materials chemists as an invaluable aid for the 3D study of the morphologies, spatially-discriminating chemical compositions, and defect properties of nanostructured materials. In this Account, we review the advances that have been made in facilitating the recording of the required series of 2D electron microscopic images and the subsequent process of 3D reconstruction of specimens that are vulnerable, to a greater or lesser degree, to electron beam damage. We describe how high-fidelity 3D tomograms may be obtained from relatively few 2D images by incorporating prior structural knowledge into the reconstruction process. In particular, we highlight the vital role of compressed sensing, a recently developed procedure well-known to information theorists that exploits ideas of image compression and "sparsity" (that the important image information can be captured in a reduced data set). We also touch upon another promising approach, "discrete" tomography, which builds into the reconstruction process a prior assumption that the object can be described in discrete terms, such as the number of constituent materials and their expected densities. Other advances made recently that we outline, such as the availability of aberration-corrected electron microscopes, electron wavelength monochromators, and sophisticated specimen goniometers, have all contributed significantly to the further development of quantitative 3D studies of nanostructured materials, including nanoparticle-heterogeneous catalysts, fuel-cell components, and drug-delivery systems, as well as photovoltaic and plasmonic devices, and are likely to enhance our knowledge of many other facets of materials chemistry, such as organic-inorganic composites, solar-energy devices, bionanotechnology, biomineralization, and energy-storage systems composed of high-permittivity metal oxides.

A case of generalized argyria after ingestion of colloidal silver solution.

PubMed

Kim, Yangho; Suh, Ho Seok; Cha, Hee Jeong; Kim, Suk Hwan; Jeong, Kyoung Sook; Kim, Dong Hoon

2009-03-01

A 58-year-old woman was referred to our hospital due to progressive skin darkening, which began 5 months previously. The patient had strikingly diffuse blue-gray discoloration of the skin, most prominent in sun-exposed areas, especially her face and hands. The oral mucosa, tongue, gums, eye conjunctiva, ears, nail beds, and trunk were also involved. Bluish-gray discoloration of all nails was aggravated by cold weather. She had ingested 1 L of colloidal silver solution daily for approximately 16 months as a traditional remedy. Her serum silver concentration was 381 ng/ml which was a very high (reference level: <15 ng/ml). Light microscopic examination of a punch biopsy specimen from her nose revealed fine, minute, round, and brown-black granules deposited in the basement membrane of hair follicular epithelium. Scanning electron microscopic examination showed electron-dense granules deposited in the intercellular space of sweat glands. Energy disperse X-ray spectrometry analysis demonstrated peaks for silver and sulfur in the dense black deposits. The ingestion of colloidal silver appears to be an increasing practice among patients using alternative health practices. All silver-containing products including colloidal silver should be labeled with a clear warning to prevent argyria, especially in alternative health practices.

Asynchronous cracking with dissimilar paths in multilayer graphene.

PubMed

Jang, Bongkyun; Kim, Byungwoon; Kim, Jae-Hyun; Lee, Hak-Joo; Sumigawa, Takashi; Kitamura, Takayuki

2017-11-16

Multilayer graphene consists of a stack of single-atomic-thick monolayer graphene sheets bound with π-π interactions and is a fascinating model material opening up a new field of fracture mechanics. In this study, fracture behavior of single-crystalline multilayer graphene was investigated using an in situ mode I fracture test under a scanning electron microscope, and abnormal crack propagation in multilayer graphene was identified for the first time. The fracture toughness of graphene was determined from the measured load-displacement curves and the realistic finite element modelling of specimen geometries. Nonlinear fracture behavior of the multilayer graphene is discussed based on nonlinear elastic fracture mechanics. In situ scanning electron microscope images obtained during the fracture test showed asynchronous crack propagation along independent paths, causing interlayer shear stress and slippages. We also found that energy dissipation by interlayer slippages between the graphene layers is the reason for the enhanced fracture toughness of multilayer graphene. The asynchronous cracking with independent paths is a unique cracking and toughening mechanism for single-crystalline multilayer graphene, which is not observed for the monolayer graphene. This could provide a useful insight for the design and development of graphene-based composite materials for structural applications.

Is localized infrared spectroscopy now possible in the electron microscope?

PubMed

Rez, Peter

2014-06-01

The recently developed in-column monochromators make it possible to record energy-c spectra with resolutions better than 30 meV from nanometer-sized regions. It should therefore in principle be possible to detect localized vibrational excitations. The scattering geometry in the electron microscope means that bond stretching in the specimen plane or longitudinal optic phonons dominate the scattering. Most promising for initial studies are vibrations with energies between 300 and 400 meV from hydrogen bonded to other atoms. Estimates of the scattering cross-sections on the basis of a simple model show that they are about the same as inner shell scattering cross-sections. Cross-sections also increase with charge transfer between the atoms, and theory incorporating realistic charge distributions shows that signal/noise is the only limitation to high-resolution imaging. Given the magnitude of the scattering cross-sections, minimizing the tail of the zero-loss peak is just as important as achieving a small-width at half-maximum. Improvements in both resolution and controlling the zero-loss tail will be necessary before it is practical to detect optic phonons in solids between 40 and 60 meV.

Investigation of C3S hydration by environmental scanning electron microscope.

PubMed

Sakalli, Y; Trettin, R

2015-07-01

Tricalciumsilicate (C(3)S, Alite) is the major component of the Portland cement clinker, The hydration of the Alite is decisive for the properties of the resulting material due to the high content in cement. The mechanism of the hydration of C(3)S is very complicated and not yet fully understood. There are some models that describe the hydration of C(3)S in various ways. The Environmental Scanning Electron Microscopy (ESEM) working in gaseous atmosphere enables high-resolution dynamic observations of structure of materials, from micrometre to nanometre scale. This provides a new perspective in material research. ESEM significantly allows imaging of specimen in their natural state without the need for special preparation (coating, drying, etc.) that can alter the physical properties. This paper presents the results of our experimental studies of hydration of C(3)S using ESEM. The ESEM turned out to be an important extension of the conventional scanning microscopy. The purpose of these investigations is to gain insight of hydration mechanism to determine which hydration products are formed and to analyze if there are any differences in the composition of the hydration products. © 2015 The Authors Journal of Microscopy © 2015 Royal Microscopical Society.

Microscopic Examination of Gallbladder Stones Improves Rate of Detection of Clonorchis sinensis Infection

PubMed Central

Ma, Rui-hong; Luo, Xiao-bing; Zheng, Pei-ming; Luo, Zhen-liang; Yang, Liu-qing

2013-01-01

To improve the rate of detection of Clonorchis sinensis infection, we compared different specimens from patients with cholecystolithiasis. Feces, gallbladder bile, and gallbladder stones collected from 179 consecutive patients with cholecystolithiasis underwent microscopic examination, and according to the results, 30 egg-positive and 30 egg-negative fecal, gallbladder bile, and gallbladder stone specimens, respectively, underwent real-time fluorescent PCR. The detection rates of eggs in feces, bile, and gallbladder stones were 30.7%, 44.7%, and 69.8%, respectively, and the differences were statistically significant (P < 0.01). The PCR results confirmed that the eggs in the specimens were C. sinensis eggs. Eggs in the feces were “fresh” and in the gallbladder stones were “old.” Microscopic examination of gallbladder stones may improve the detection rates of C. sinensis infection, which is important for developing individualized treatments to prevent the recurrence of gallbladder stones and to prevent the occurrence of severe liver damage and cholangiocarcinoma. PMID:23698535

Optical properties of MgF2 nano-composite films dispersed with noble metal nanoparticles synthesized by sol-gel method

NASA Astrophysics Data System (ADS)

Wakaki, Moriaki; Soujima, Nobuaki; Shibuya, Takehisa

2015-03-01

Porous MgF2 films synthesized by a sol-gel method exhibit the lowest refractive index among the dielectric optical materials and are the most useful materials for the anti-reflection coatings. On the other hand, surface plasmon resonance (SPR) absorptions of noble metal nanoparticles in various solid matrices have been extensively studied. New functional materials like a SERS (Surface Enhanced Raman Spectroscopy) tips are expected by synthesizing composite materials between porous MgF2 films featured by the network of MgF2 nanoparticles and noble metal nanoparticles introduced within the network. In this study, fundamental physical properties including morphology and optical properties are characterized for these materials to make clear the potential of the composite system. Composite materials of MgF2 films dispersed with noble metal (Ag, Au) nanoparticles were prepared using the sol-gel technique with various annealing temperatures and densities of noble metal nanoparticles. The structural morphology was analyzed by an X-ray diffractometer (XRD) and a scanning electron microscope (SEM). The size and shape distributions of the metal nanoparticles were observed using a transmission electron microscope (TEM). The optical properties of fabricated composite films were characterized by UV-Vis-NIR and FT-IR spectrophotometers. The absorption spectra due to the surface plasmon resonance (SPR) of the metal nanoparticles were analyzed using the dielectric function considering the effective medium approximation, typically Maxwell-Garnett model. The Raman scattering spectra were also studied to check the enhancement effect of specimen dropped on the MgF2: Ag nano-composite films deposited on Si substrate. Enhancement of the Raman intensity of pyridine solution specimen was observed.

The Effects of Torsional Preloading on the Torsional Resistance of Nickel-titanium Instruments.

PubMed

Oh, Seung-Hei; Ha, Jung-Hong; Kwak, Sang Won; Ahn, Shin Wook; Lee, WooCheol; Kim, Hyeon-Cheol

2017-01-01

This study evaluated the effect of torsional preloading on the torsional resistance of nickel-titanium (NiTi) endodontic instruments. WaveOne Primary (Dentsply Maillefer, Ballaigues, Switzerland) and ProTaper Universal F2 (Dentsply Maillefer) files were used. The ultimate torsional strength until fracture was determined for each instrument. In the phase 1 experiment, the ProTaper and WaveOne files were loaded to have a maximum load from 2.0 up to 2.7 or 2.8 Ncm, respectively. In the phase 2 experiment, the number of repetitions of preloading for each file was increased from 50 to 200, whereas the preloading torque was fixed at 2.4 Ncm. Using torsionally preloaded specimens from phase 1 and 2, the torsional resistances were calculated to determine the ultimate strength, distortion angle, and toughness. The results were analyzed using 1-way analysis of variance and Duncan post hoc comparison. The fracture surfaces and longitudinal aspect of 5 specimens per group were examined under a scanning electron microscope. All preloaded groups showed significantly higher ultimate strength than the unpreloaded groups (P < .05). There was no significant difference among all groups for distortion angle and toughness. Although WaveOne had no significant difference between the repetition groups for ultimate strength, fracture angle, and toughness, ProTaper had a higher distortion angle and toughness in the 50-repetition group compared with the other repetition groups (P < .05). Scanning electron microscopic examinations of the fractured surface showed typical features of torsional fracture. Torsional preloading within the ultimate values could enhance the torsional strength of NiTi instruments. The total energy until fracture was maintained constantly, regardless of the alloy type. Copyright © 2016 American Association of Endodontists. Published by Elsevier Inc. All rights reserved.

Local electric field direct writing – Electron-beam lithography and mechanism

DOE PAGES

Jiang, Nan; Su, Dong; Spence, John C. H.

2017-08-24

Local electric field induced by a focused electron probe in silicate glass thin films is evaluated in this paper by the migration of cations. Extremely strong local electric fields can be obtained by the focused electron probe from a scanning transmission electron microscope. As a result, collective atomic displacements occur. This newly revised mechanism provides an efficient tool to write patterned nanostructures directly, and thus overcome the low efficiency of the conventional electron-beam lithography. Applying this technique to silicate glass thin films, as an example, a grid of rods of nanometer dimension can be efficiently produced by rapidly scanning amore » focused electron probe. This nanopatterning is achieved through swift phase separation in the sample, without any post-development processes. The controlled phase separation is induced by massive displacements of cations (glass modifiers) within the glass-former network, driven by the strong local electric fields. The electric field is induced by accumulated charge within the electron probed region, which is generated by the excitation of atomic electrons by the incident electron. Throughput is much improved compared to other scanning probe techniques. Finally, the half-pitch spatial resolution of nanostructure in this particular specimen is 2.5 nm.« less

Local electric field direct writing – Electron-beam lithography and mechanism

DOE Office of Scientific and Technical Information (OSTI.GOV)

Jiang, Nan; Su, Dong; Spence, John C. H.

Local electric field induced by a focused electron probe in silicate glass thin films is evaluated in this paper by the migration of cations. Extremely strong local electric fields can be obtained by the focused electron probe from a scanning transmission electron microscope. As a result, collective atomic displacements occur. This newly revised mechanism provides an efficient tool to write patterned nanostructures directly, and thus overcome the low efficiency of the conventional electron-beam lithography. Applying this technique to silicate glass thin films, as an example, a grid of rods of nanometer dimension can be efficiently produced by rapidly scanning amore » focused electron probe. This nanopatterning is achieved through swift phase separation in the sample, without any post-development processes. The controlled phase separation is induced by massive displacements of cations (glass modifiers) within the glass-former network, driven by the strong local electric fields. The electric field is induced by accumulated charge within the electron probed region, which is generated by the excitation of atomic electrons by the incident electron. Throughput is much improved compared to other scanning probe techniques. Finally, the half-pitch spatial resolution of nanostructure in this particular specimen is 2.5 nm.« less

An exploration of the accuracy of an invasive method for measuring the volume of specimens of Amoeba proteus.

PubMed

Whalen, T A; Demarco, A J

1999-10-01

A method is described for measuring the volume of individual specimens of Amoeba proteus which utilizes an easily constructed compressor to flatten the specimen to a known thickness. The microscopic image of the flattened specimen is captured on tape, digitized and analysed with the NIH Image software. The results from one specimen are given to illustrate the sources and magnitude of errors affecting these volume measurements.

High-performance serial block-face SEM of nonconductive biological samples enabled by focal gas injection-based charge compensation.

PubMed

Deerinck, T J; Shone, T M; Bushong, E A; Ramachandra, R; Peltier, S T; Ellisman, M H

2018-05-01

A longstanding limitation of imaging with serial block-face scanning electron microscopy is specimen surface charging. This charging is largely due to the difficulties in making biological specimens and the resins in which they are embedded sufficiently conductive. Local accumulation of charge on the specimen surface can result in poor image quality and distortions. Even minor charging can lead to misalignments between sequential images of the block-face due to image jitter. Typically, variable-pressure SEM is used to reduce specimen charging, but this results in a significant reduction to spatial resolution, signal-to-noise ratio and overall image quality. Here we show the development and application of a simple system that effectively mitigates specimen charging by using focal gas injection of nitrogen over the sample block-face during imaging. A standard gas injection valve is paired with a precisely positioned but retractable application nozzle, which is mechanically coupled to the reciprocating action of the serial block-face ultramicrotome. This system enables the application of nitrogen gas precisely over the block-face during imaging while allowing the specimen chamber to be maintained under high vacuum to maximise achievable SEM image resolution. The action of the ultramicrotome drives the nozzle retraction, automatically moving it away from the specimen area during the cutting cycle of the knife. The device described was added to a Gatan 3View system with minimal modifications, allowing high-resolution block-face imaging of even the most charge prone of epoxy-embedded biological samples. © 2017 The Authors Journal of Microscopy © 2017 Royal Microscopical Society.

The effect of condensation pressure on selected physical properties of mineral trioxide aggregate.

PubMed

Nekoofar, M H; Adusei, G; Sheykhrezae, M S; Hayes, S J; Bryant, S T; Dummer, P M H

2007-06-01

To examine the effect of condensation pressure on surface hardness, microstructure and compressive strength of mineral trioxide aggregate (MTA). White ProRoot MTA (Dentsply Tulsa Dental, Johnson City, TN, USA) was mixed and packed into cylindrical polycarbonate tubes. Six groups each of 10 specimens were subjected to pressures of 0.06, 0.44, 1.68, 3.22, 4.46 and 8.88 MPa respectively. The surface hardness of each specimen was measured using Vickers microhardness. Cylindrical specimens of 4 mm in diameter and 6 mm in height were prepared in polycarbonate cylindrical moulds for testing the compressive strength. Five groups of 10 specimens were prepared using pressures of 0.06, 0.44, 1.68, 3.22 or 4.46 MPa. Data were subjected to one-way anova. The microstructure was analysed using a scanning electron microscope (SEM) after sectioning specimens with a scalpel. A trend was observed for higher condensation pressures to produce lower surface hardness values. A condensation pressure of 8.88 MPa produced specimens with significantly lower values in terms of surface hardness than other groups (P<0.001). A condensation pressure of 1.68 MPa conferred the maximum compressive strength; however, it was not statistically different. Higher condensation pressures resulted in fewer voids and microchannels when analysed with SEM. In specimens prepared with lower condensation pressures distinctive crystalline structures were observed. They tended to appear around microchannels. Condensation pressure may affect the strength and hardness of MTA. Use of controlled condensation pressure in sample preparation for future studies is suggested.

Errors in quantitative backscattered electron analysis of bone standardized by energy-dispersive x-ray spectrometry.

PubMed

Vajda, E G; Skedros, J G; Bloebaum, R D

1998-10-01

Backscattered electron (BSE) imaging has proven to be a useful method for analyzing the mineral distribution in microscopic regions of bone. However, an accepted method of standardization has not been developed, limiting the utility of BSE imaging for truly quantitative analysis. Previous work has suggested that BSE images can be standardized by energy-dispersive x-ray spectrometry (EDX). Unfortunately, EDX-standardized BSE images tend to underestimate the mineral content of bone when compared with traditional ash measurements. The goal of this study is to investigate the nature of the deficit between EDX-standardized BSE images and ash measurements. A series of analytical standards, ashed bone specimens, and unembedded bone specimens were investigated to determine the source of the deficit previously reported. The primary source of error was found to be inaccurate ZAF corrections to account for the organic phase of the bone matrix. Conductive coatings, methylmethacrylate embedding media, and minor elemental constituents in bone mineral introduced negligible errors. It is suggested that the errors would remain constant and an empirical correction could be used to account for the deficit. However, extensive preliminary testing of the analysis equipment is essential.

Evaluation of breast tissue with confocal strip-mosaicking microscopy: a test approach emulating pathology-like examination

PubMed Central

Abeytunge, Sanjee; Larson, Bjorg; Peterson, Gary; Morrow, Monica; Rajadhyaksha, Milind

2017-01-01

Abstract. Confocal microscopy is an emerging technology for rapid imaging of freshly excised tissue without the need for frozen- or fixed-section processing. Initial studies have described imaging of breast tissue using fluorescence confocal microscopy with small regions of interest, typically 750×750  μm2. We present exploration with a microscope, termed confocal strip-mosaicking microscope (CSM microscope), which images an area of 2×2  cm2 of tissue with cellular-level resolution in 10 min of excision. Using the CSM microscope, we imaged 34 fresh, human, large breast tissue specimens from 18 patients, blindly analyzed by a board-certified pathologist and subsequently correlated with the corresponding standard fixed histopathology. Invasive tumors and benign tissue were clearly identified in CSM strip-mosaic images. Thirty specimens were concordant for image-to-histopathology correlation while four were discordant. PMID:28327961

A Novel Low Energy Electron Microscope for DNA Sequencing and Surface Analysis

PubMed Central

Mankos, M.; Shadman, K.; Persson, H.H.J.; N’Diaye, A.T.; Schmid, A.K.; Davis, R.W.

2014-01-01

Monochromatic, aberration-corrected, dual-beam low energy electron microscopy (MAD-LEEM) is a novel technique that is directed towards imaging nanostructures and surfaces with sub-nanometer resolution. The technique combines a monochromator, a mirror aberration corrector, an energy filter, and dual beam illumination in a single instrument. The monochromator reduces the energy spread of the illuminating electron beam, which significantly improves spectroscopic and spatial resolution. Simulation results predict that the novel aberration corrector design will eliminate the second rank chromatic and third and fifth order spherical aberrations, thereby improving the resolution into the sub-nanometer regime at landing energies as low as one hundred electron-Volts. The energy filter produces a beam that can extract detailed information about the chemical composition and local electronic states of non-periodic objects such as nanoparticles, interfaces, defects, and macromolecules. The dual flood illumination eliminates charging effects that are generated when a conventional LEEM is used to image insulating specimens. A potential application for MAD-LEEM is in DNA sequencing, which requires high resolution to distinguish the individual bases and high speed to reduce the cost. The MAD-LEEM approach images the DNA with low electron impact energies, which provides nucleobase contrast mechanisms without organometallic labels. Furthermore, the micron-size field of view when combined with imaging on the fly provides long read lengths, thereby reducing the demand on assembling the sequence. Experimental results from bulk specimens with immobilized single-base oligonucleotides demonstrate that base specific contrast is available with reflected, photo-emitted, and Auger electrons. Image contrast simulations of model rectangular features mimicking the individual nucleotides in a DNA strand have been developed to translate measurements of contrast on bulk DNA to the detectability of individual DNA bases in a sequence. PMID:24524867

A novel low energy electron microscope for DNA sequencing and surface analysis.

PubMed

Mankos, M; Shadman, K; Persson, H H J; N'Diaye, A T; Schmid, A K; Davis, R W

2014-10-01

Monochromatic, aberration-corrected, dual-beam low energy electron microscopy (MAD-LEEM) is a novel technique that is directed towards imaging nanostructures and surfaces with sub-nanometer resolution. The technique combines a monochromator, a mirror aberration corrector, an energy filter, and dual beam illumination in a single instrument. The monochromator reduces the energy spread of the illuminating electron beam, which significantly improves spectroscopic and spatial resolution. Simulation results predict that the novel aberration corrector design will eliminate the second rank chromatic and third and fifth order spherical aberrations, thereby improving the resolution into the sub-nanometer regime at landing energies as low as one hundred electron-Volts. The energy filter produces a beam that can extract detailed information about the chemical composition and local electronic states of non-periodic objects such as nanoparticles, interfaces, defects, and macromolecules. The dual flood illumination eliminates charging effects that are generated when a conventional LEEM is used to image insulating specimens. A potential application for MAD-LEEM is in DNA sequencing, which requires high resolution to distinguish the individual bases and high speed to reduce the cost. The MAD-LEEM approach images the DNA with low electron impact energies, which provides nucleobase contrast mechanisms without organometallic labels. Furthermore, the micron-size field of view when combined with imaging on the fly provides long read lengths, thereby reducing the demand on assembling the sequence. Experimental results from bulk specimens with immobilized single-base oligonucleotides demonstrate that base specific contrast is available with reflected, photo-emitted, and Auger electrons. Image contrast simulations of model rectangular features mimicking the individual nucleotides in a DNA strand have been developed to translate measurements of contrast on bulk DNA to the detectability of individual DNA bases in a sequence. Copyright © 2014 Elsevier B.V. All rights reserved.

A novel low energy electron microscope for DNA sequencing and surface analysis

DOE PAGES

Mankos, M.; Shadman, K.; Persson, H. H. J.; ...

2014-01-31

Monochromatic, aberration-corrected, dual-beam low energy electron microscopy (MAD-LEEM) is a novel technique that is directed towards imaging nanostructures and surfaces with sub-nanometer resolution. The technique combines a monochromator, a mirror aberration corrector, an energy filter, and dual beam illumination in a single instrument. The monochromator reduces the energy spread of the illuminating electron beam, which significantly improves spectroscopic and spatial resolution. Simulation results predict that the novel aberration corrector design will eliminate the second rank chromatic and third and fifth order spherical aberrations, thereby improving the resolution into the sub-nanometer regime at landing energies as low as one hundred electron-Volts.more » The energy filter produces a beam that can extract detailed information about the chemical composition and local electronic states of non-periodic objects such as nanoparticles, interfaces, defects, and macromolecules. The dual flood illumination eliminates charging effects that are generated when a conventional LEEM is used to image insulating specimens. A potential application for MAD-LEEM is in DNA sequencing, which requires high resolution to distinguish the individual bases and high speed to reduce the cost. The MAD-LEEM approach images the DNA with low electron impact energies, which provides nucleobase contrast mechanisms without organometallic labels. Furthermore, the micron-size field of view when combined with imaging on the fly provides long read lengths, thereby reducing the demand on assembling the sequence. Finally, experimental results from bulk specimens with immobilized single-base oligonucleotides demonstrate that base specific contrast is available with reflected, photo-emitted, and Auger electrons. Image contrast simulations of model rectangular features mimicking the individual nucleotides in a DNA strand have been developed to translate measurements of contrast on bulk DNA to the detectability of individual DNA bases in a sequence.« less

Analysis of the microstructure and mechanical performance of composite resins after accelerated artificial aging.

PubMed

De Oliveira Daltoé, M; Lepri, C Penazzo; Wiezel, J Guilherme G; Tornavoi, D Cremonezzi; Agnelli, J A Marcondes; Reis, A Cândido Dos

2013-03-01

Researches that assess the behavior of dental materials are important for scientific and industrial development especially when they are tested under conditions that simulate the oral environment, so this work analyzed the compressive strength and microstructure of three composite resins subjected to accelerated artificial aging (AAA). Three composites resins of 3M (P90, P60 and Z100) were analyzed and were obtained 16 specimens for each type (N.=48). Half of each type were subjected to UV-C system AAA and then were analyzed the surfaces of three aged specimens and three not aged of each type through the scanning electron microscope (SEM). After, eight specimens of each resin, aged and not aged, were subjected to compression test. After statistical analysis of compressive strength values, it was found that there was difference between groups (α <0.05). The resin specimens aged P60 presented lower values of compressive strength statistically significant when compared to the not subject to the AAA. For the other composite resins, there was no difference, regardless of aging, a fact confirmed by SEM. The results showed that the AAA influenced the compressive strength of the resin aged P60; confirmed by surface analysis by SEM, which showed greater structural disarrangement on surface material.

Development of a specimen heating holder with an evaporator and gas injector and its application for catalyst.

PubMed

Takeo, Kamino; Toshie, Yaguchi; Mitsuru, Konno; Akira, Watabe; Yasuhira, Nagakubo

2006-10-01

A specimen heating holder equipped with a gas injector and an evaporator has been developed for use with conventional transmission electron microscopes (TEMs). The developed specimen holder allows both synthesis of metal oxide support and deposition of catalyst nano-particles in situ. Since the holder is designed to be used in small gapped high-resolution objective lens pole-piece, all the procedure from the synthesis of support material to the deposition of catalyst as well as the behavior of the catalyst nano-particles on the support can be observed at near atomic resolution. The developed specimen holder was applied to the study of AuPd catalyst. First, air was injected onto heated aluminum particles via a gas injector to synthesize Al(2)O(3) support. Then, nano-particles of AuPd were deposited on the Al(2)O(3) support. After the deposition, the synthesized Al(2)O(3) support was heated and air was injected again to observe behaviors of the deposited AuPd nano-particles at elevated temperatures in the aerial environment. Behaviors of the AuPd nano-particles such as coalescence, segmentation and diffusion to the Al(2)O(3) support were dynamically observed at atomic level high resolution.

Comparative Analysis of Dentinal Erosion after Passive Ultrasonic Irrigation versus Irrigation with Reciprocating Activation: An Environmental Scanning Electron Study.

PubMed

Simezo, Ana Paula; da Silveira Bueno, Carlos Eduardo; Cunha, Rodrigo Sanches; Pelegrine, Rina Andrea; Rocha, Daniel Guimarães Pedro; de Martin, Alexandre Sigrist; Kato, Augusto Shoji

2017-01-01

The aim of this study was to assess ex vivo the erosive effects of passive ultrasonic irrigation versus irrigation with reciprocating activation on the dentinal surface of the root canal at 3 predetermined levels using environmental scanning electron microscopy. Ten roots of mandibular premolars were prepared using the ProTaper Universal system (Dentsply Maillefer, Ballaigues, Switzerland). The specimens were embedded in flasks cleaved longitudinally, and indentations were made 3.0, 6.0, and 9.0 mm from the apex. The specimens in the control group (n = 10) were cleaned in an ultrasonic bath containing 2.5% sodium hypochlorite and 17% EDTA and then dried. Then, environmental scanning electron microscopic images were obtained at magnification × 800. The specimens were then reassembled in their flasks, and the NaOCl and EDTA solutions were activated according to the conditions established for the experimental groups (ie, the passive ultrasonic irrigation group [n = 5] and the EasyClean (Easy Equipamentos Odontológicos, Belo Horizonte, MG, Brazil) group, irrigation with reciprocating activation with the EasyClean instrument [n = 5]). The specimens of both experimental groups were analyzed in the same manner as in the control group. Analysis of the dentinal surface topography was conducted using the 3D Roughness Reconstruction program (Phenom-World BV, Eindhoven, the Netherlands) as a means for assessing erosion. The data were evaluated by means of the Kruskal-Wallis, Student-Newman-Keuls, and Mann-Whitney tests. In the EasyClean group, the degree of dentinal erosion at 3.0 mm was significantly higher than at 9.0 mm. In the other comparisons, there was no statistically significant difference (P < .05). The final irrigation techniques tested were equivalent in relation to the degree of erosion caused to the dentinal surface. Copyright © 2016 American Association of Endodontists. Published by Elsevier Inc. All rights reserved.

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.

Ultrasoft magnetic films investigated with Lorentz tranmission electron microscopy and electron holography.

PubMed

De Hosson, Jeff Th M; Chechenin, Nicolai G; Alsem, Daan-Hein; Vystavel, Tomas; Kooi, Bart J; Chezan, Antoni R; Boerma, Dik O

2002-08-01

As a tribute to the scientific work of Professor Gareth Thomas in the field of structure-property relationships this paper delineates a new possibility of Lorentz transmission electron microscopy (LTEM) to study the magnetic properties of soft magnetic films. We show that in contrast to the traditional point of view, not only does the direction of the magnetization vector in nano-crystalline films make a correlated small-angle wiggling, but also the magnitude of the magnetization modulus fluctuates. This fluctuation produces a rapid modulation in the LTEM image. A novel analysis of the ripple structure in nano-crystalline Fe-Zr-N film corresponds to an amplitude of the transversal component of the magnetization deltaMy of 23 mT and a longitudinal fluctuation of the magnetization of the order of deltaMx = 30 mT. The nano-crystalline (Fe99Zr1)1-xNx films have been prepared by DC magnetron reactive sputtering with a thickness between 50 and 1000 nm. The grain size decreased monotonically with N content from typically 100 nm in the case of N-free films to less than 10 nm for films containing 8 at%. The specimens were examined with a JEOL 2010F 200 kV transmission electron microscope equipped with a post column energy filter (GIF 2000 Gatan Imaging Filter). For holography, the microscope is mounted with a biprism (JEOL biprism with a 0.6 microm diameter platinum wire).

Photodynamic therapy of urethral condylomata acuminata using topically 5-aminolevulinic acid

NASA Astrophysics Data System (ADS)

Wang, Xiuli; Wang, Hongwei; Wang, Haishan; Xu, Shizheng; Liao, Kanghuang; Hillemanns, Peter

2005-07-01

Background Electrocoagulation and laser evaporation for urethral condylomata acuminata have high recurrence rates and can be associated with urethral malformations. Objective To investigate the effect of photodynamic therapy (PDT) with topical 5-aminolevulinic acid (ALA) on urethral condylomata acuminata and to examine the histological changes in lesions of condylomata acuminata after ALA-PDT. Methods One hundred and sixty-four urethral condylomata patients were given topical ALA followed by intraurethral PDT through a cylindrical fiber. Among the cases, 16 penile and vulval condylomatous lesions in 11 patients were treated with topical ALA-PDT at same time. After the treatment, biopsy specimens were collected from the 16 penile and vulval lesions. The histological changes were then evaluated by light microscope and electron microscope. Results The complete response rate for urethral condylomata by topical ALA-PDT was 95.12% and the recurrence rate was 5.13% after 6 to 24 months follow-up. Keratinocytes in middle and upper layers of the epidermis with marked vacuolation and some necrocytosis were detected one and three hours after PDT. Necrosis in all layers of the epidermis was noted five hours after PDT by microscopy. In electron microscopy of kerationcytes, distinct ultrastructural abnormalities of mitochondrion, endoplasmic reticulum and membrane damage were observed. Apoptotic bodies were detected three hours after PDT and a large number of the keratinocytes exhibited necrosis five hours after PDT by electron microscope. Conclusions Results suggests that topical ALA-PDT is a simple, effective, relatively safe, less recurrent and comparatively well tolerated treatment for urethral condylomata acuminata. The mechanisms might be that ALA-PDT could trigger apoptotic process and necrosis in the HPV infected keratinocytes. Key words:

A Cover of Glass: First Report of Biomineralized Silicon in a Ciliate, Maryna umbrellata (Ciliophora: Colpodea)

PubMed Central

FOISSNER, WILHELM; WEISSENBACHER, BIRGIT; KRAUTGARTNER, WOLF-DIETRICH; LÜTZ-MEINDL, URSULA

2010-01-01

Using hydrofluoric acid, scanning electron microscope-assisted X-ray microanalysis, and energy-filtered transmission electron microscopy, we present the first definite proof of biomineralized silicon [(SiO2)]n in a ciliophoran protist, Maryna umbrellata, a common inhabitant of ephemeral pools. In the trophic specimen, the amorphic silicon (glass) granules are accumulated in the anterior half of the body. When entering the dormant stage, most glass granules are excreted to form the surface cover of the globular resting cyst. Most likely, the silicon granules are synthesized in vesicles of the Golgi apparatus. First, nanospheres with a size of 20–40 nm are formed in a fibrous matrix; they grow to be spongious complexes, eventually becoming amorphous glass granules with an average size of 819 nm × 630 nm. In the transmission electron microscope, the silicon granules show the characteristic fracture pattern of glass known from many other silicon-bearing organisms. A literature survey suggests that silicon is very rare in ciliates. The fine structure and genesis of silicon granules in M. umbrellata are very similar to those of other organisms, including vascular plants and animals, indicating a common mechanism. Light perception and protection against mechanical stress and predators might be functions of the silicon granules in M. umbrellata. The palaeontological significance of glass cysts in ciliates is also discussed. PMID:19883440

Adaptive optical microscope for brain imaging in vivo

NASA Astrophysics Data System (ADS)

Wang, Kai

2017-04-01

The optical heterogeneity of biological tissue imposes a major limitation to acquire detailed structural and functional information deep in the biological specimens using conventional microscopes. To restore optimal imaging performance, we developed an adaptive optical microscope based on direct wavefront sensing technique. This microscope can reliably measure and correct biological samples induced aberration. We demonstrated its performance and application in structural and functional brain imaging in various animal models, including fruit fly, zebrafish and mouse.

Impact of gastric acidic challenge on surface topography and optical properties of monolithic zirconia.

PubMed

Sulaiman, Taiseer A; Abdulmajeed, Aous A; Shahramian, Khalil; Hupa, Leena; Donovan, Terrence E; Vallittu, Pekka; Närhi, Timo O

2015-12-01

To evaluate the surface topography and optical properties of monolithic zirconia after immersion in simulated gastric acid. Four partially stabilized (PSZ) and one fully stabilized (FSZ) zirconia materials were selected for the study: Prettau (PRT, Zirkonzahn), Zenostar (ZEN, Ivoclar), Bruxzir (BRX, Glidewell), Katana (KAT, Noritake) and FSZ Prettau Anterior (PRTA, Zirkonzahn). IPS e.max (Ivoclar) was used as a control. The specimens (10×10×1.2mm, n=5 per material) were cut, sintered, polished and cleaned before immersed in 5ml of simulated gastric acid solution (Hydrochloric acid (HCl) 0.06M, 0.113% solution in deionized distal water, pH 1.2) for 96h in a 37°C incubator. Specimens were weighed and examined for morphological changes under scanning electron microscope (SEM) coupled with energy dispersive X-ray spectroscopy (EDX). Surface roughness was evaluated by a confocal microscope. Surface gloss and translucency parameter (TP) values were determined by a reflection spectrophotometer before and after acid immersion. The data was analyzed by one-way ANOVA followed by Tukey's HSD post hoc test (p<0.05). PRTA displayed the most weight loss (1.40%) among the zirconia specimens. IPS e.max showed about three times more weight loss (3.05%) than zirconia specimens as an average. SEM examination indicated areas of degradation, bead-like shapes and smoothening of the polishing scratches after acid immersion. EDX displayed ion interactions and possible ion leaching from all specimens. Sa and Sq values for PRTA, ZEN and IPS e.max were significantly lower (p<0.05) after acid immersion. TP values increased significantly for PRT, ZEN and IPS e.max (p<0.05), while the surface gloss of ZEN, PRTA and IPS e.max increased (p<0.05). Monolithic zirconia materials show some surface alterations in an acidic environment with minimum effect on their optical properties. Whether a smoother surface is in fact a sign of true corrosion resistance or is purely the result of an evenly progressive corrosive process is yet to be confirmed by further research. Copyright © 2015 Academy of Dental Materials. Published by Elsevier Ltd. All rights reserved.

Corrosive and cytotoxic properties of compact specimens and microparticles of Ni-Cr dental alloy.

PubMed

Ristic, Ljubisa; Vucevic, Dragana; Radovic, Ljubica; Djordjevic, Snezana; Nikacevic, Milutin; Colic, Miodrag

2014-04-01

Nickel-chromium (Ni-Cr) dental alloys have been widely used in prosthodontic practice, but there is a permanent concern about their biocompatibility due to the release of metal ions. This is especially important when Ni-Cr metal microparticles are incorporated into gingival tissue during prosthodontic procedures. Therefore, the aim of this study was to examine and compare the corrosion and cytotoxic properties of compact specimens and microparticles of Ni-Cr dental alloy. Ni-Cr alloy, Remanium CSe bars (4 mm diameter), were made by the standard casting method and then cut into 0.5-mm-thick disks. Metal particles were obtained by scraping the bars using a diamond instrument for crown preparation. The microstructure was observed by an optical microscope. Quantitative determination and morphological and dimensional characterization of metal particles were carried out by a scanning electron microscope and Leica Application Suite software for image analysis. Corrosion was studied by conditioning the alloy specimens in the RPMI 1640 medium, containing 10% fetal calf serum in an incubator with 5% CO2 for 72 hours at 37°C. Inductively coupled plasma-optical emission spectrometry was used to assess metal ion release. The cytotoxity of conditioning medium (CM) was investigated on L929 cells using an MTT test. One-way ANOVA was used for statistical analysis. After casting, the microstructure of the Remanium CSe compact specimen composed of Ni, Cr, Mo, Si, Fe, Al, and Co had a typical dendritic structure. Alloy microparticles had an irregular shape with a wide size range: from less than 1 μm to more than 100 μm. The release of metal ions, especially Ni and Mo from microparticles, was significantly higher, compared to the compact alloy specimen. The CM prepared from compact alloy was not cytotoxic at any tested dilutions, whereas CM from alloy microparticles showed dose-dependent cytotoxicity (90% CM and 45% CM versus control; p < 0.005). Ni-Cr microparticles showed less corrosion resistance and lower biocompatibility than compact alloy. This could affect health on long-term exposure, especially in sensitized individuals. © 2013 by the American College of Prosthodontists.

Effect of etching time and resin bond on the flexural strength of IPS e.max Press glass ceramic.

PubMed

Xiaoping, Luo; Dongfeng, Ren; Silikas, Nick

2014-12-01

To evaluate the effect of hydrofluoric acid (HFA) etching time and resin cement bond on the flexural strength of IPS e.max(®) Press glass ceramic. Two hundred and ten bars, 25mm×3mm×2mm, were made from IPS e.max(®) Press ingots through lost-wax, hot-pressed ceramic fabrication technology and randomly divided into five groups with forty-two per group after polishing. The ceramic surfaces of different groups were etched by 9.5% hydrofluoric acid gel for 0, 20, 40, 60 and 120s respectively. Two specimens of each group were selected randomly to examine the surface roughness and 3-dimensional topography with atomic force microscope (AFM), and microstructure was analyzed by the field emission scanning electron microscope (FE-SEM). Then each group were subdivided into two subgroups (n=20). One subgroup of this material was selected to receive a thin (approximately 0.1mm) layer of resin luting agent (Variolink N) whereas the other subgroup remained unaltered. Half of subgroup's specimens were thermocycled 10,000 times before a 3-point bending test in order to determine the flexural strength. Interface between resin cement and ceramic was examined with field emission scanning electronic microscope. Roughness values increased with increasing etching time. The mean flexural strength values of group 0s, 20s, 40s, 60s and 120s were 384±33, 347±43, 330±53, 327±67 and 317±41MPa respectively. Increasing HF etching times reduced the mean flexural strength (p<0.05). However, the mean flexural strength of each group, except group 0s, increased significantly to 420±31, 435±50, 400±39 and 412±58MPa after the application of dual-curing resin cement. In the present investigation, no significant differences after thermocycling on the flexural strengths were evident. Overtime HF etching could have a wakening effect on IPS e.max(®) Press glass ceramic, but resin cement bonding to appropriately etched surface would strengthen the dental ceramic. Copyright © 2014 Academy of Dental Materials. Published by Elsevier Ltd. All rights reserved.

Real imaging and size values of Saccharomyces cerevisiae cells with comparable contrast tuning to two environmental scanning electron microscopy modes.

PubMed

Misirli, Zulal; Oner, Ebru Toksoy; Kirdar, Betul

2007-01-01

The combined application of electron microscopy (EM) is frequently used for the microstructural investigation of biological specimens and plays two important roles in the quantification and in gaining an improved understanding of biological phenomena by making use of the highest resolution capability provided by EM. The possibility of imaging wet specimens in their "native" states in the environmental scanning electron microscope (ESEM) at high resolution and large depth of focus in real time is discussed in this paper. It is demonstrated here that new features can be discovered by the elimination of even the least hazardous approaches in some preparation techniques, that destroy the samples. Since the analysis conditions may influence the morphology and the extreme surface sensitivity of living biological systems, the results obtained from the same cultured cell with two different ESEM modes (Lvac mode and wet mode) were compared. This offers new opportunities compared with ESEM-wet/Lvac-mode imaging, since wet-mode imaging involves a real contrast and gives an indication of the changes in cell morphology and structure required for cell viability. In this study, wet-mode imaging was optimized using the unique ability of cell quantities for microcharacterization in situ giving very fine features of topological effects. Accordingly, the progress is reported by comparing the results of these two modes, which demonstrate interesting application details. In general, the functional comparisons have revealed that the fresh unprocessed Saccharomyces cerevisiae cells (ESEM-wet mode) were essentially unaltered with improved and minimal specimen preparation timescales, and the optimal cell viability degree was visualized and also measured quantitatively while the cell size remained unchanged with continuous images.

The collagenic architecture of human dura mater.

PubMed

Protasoni, Marina; Sangiorgi, Simone; Cividini, Andrea; Culuvaris, Gloria Tiffany; Tomei, Giustino; Dell'Orbo, Carlo; Raspanti, Mario; Balbi, Sergio; Reguzzoni, Marcella

2011-06-01

Human dura mater is the most external meningeal sheet surrounding the CNS. It provides an efficient protection to intracranial structures and represents the most important site for CSF turnover. Its intrinsic architecture is made up of fibrous tissue including collagenic and elastic fibers that guarantee the maintenance of its biophysical features. The recent technical advances in the repair of dural defects have allowed for the creation of many synthetic and biological grafts. However, no detailed studies on the 3D microscopic disposition of collagenic fibers in dura mater are available. The authors report on the collagenic 3D architecture of normal dura mater highlighting the orientation, disposition in 3 dimensions, and shape of the collagen fibers with respect to the observed layer. Thirty-two dura mater specimens were collected during cranial decompressive surgical procedures, fixed in 2.5% Karnovsky solution, and digested in 1 N NaOH solution. After a routine procedure, the specimens were observed using a scanning electron microscope. The authors distinguished the following 5 layers in the fibrous dura mater of varying thicknesses, orientation, and structures: bone surface, external median, vascular, internal median, and arachnoid layers. The description of the ultrastructural 3D organization of the different layers of dura mater will give us more information for the creation of synthetic grafts that are as similar as possible to normal dura mater. This description will be also related to the study of the neoplastic invasion.

The application of polyethylene glycol (PEG) to electron microscopy

PubMed Central

1980-01-01

The cytoplasm of cells from a variety of tissues has been viewed in sections (0.25-1 micrometers) devoid of any embedding resin. Glutaraldehyde- and osmium tetroxide-fixed tissues were infiltrated and embedded in a water-miscible wax, polyethylene glycol (PEG), and subsequently sectioned on dry glass or diamond knives. The PEG matrix was removed and the sections were placed on Formvarcarbon-polylysine- coated grids, dehydrated, dried by the critical-point method, and observed in either the high- or low-voltage electron microscope. Stereoscopic views of cells devoid of embedding resin present an image of cell utrastructure unobscured by electron-scattering resins similar to the image of whole, unembedded critical-point-dried or freeze-dried cultured cells observed by transmission electron microscopy. All organelles, including the cytoskeletal structures, are identified and appear not to have been damaged during processing, although membrane components appear somewhat less distinct. The absence of an embedding matrix eliminates the need for additional staining to increase contrast, unlike the situation with specimens embedded in standard electron-scattering resins. The PEG technique thus appears to be a valuable adjunct to conventional methods for ultrastructural analysis. PMID:7400222

The application of polyethylene glycol (PEG) to electron microscopy.

PubMed

Wolosewick, J J

1980-08-01

The cytoplasm of cells from a variety of tissues has been viewed in sections (0.25-1 micrometers) devoid of any embedding resin. Glutaraldehyde- and osmium tetroxide-fixed tissues were infiltrated and embedded in a water-miscible wax, polyethylene glycol (PEG), and subsequently sectioned on dry glass or diamond knives. The PEG matrix was removed and the sections were placed on Formvarcarbon-polylysine-coated grids, dehydrated, dried by the critical-point method, and observed in either the high- or low-voltage electron microscope. Stereoscopic views of cells devoid of embedding resin present an image of cell utrastructure unobscured by electron-scattering resins similar to the image of whole, unembedded critical-point-dried or freeze-dried cultured cells observed by transmission electron microscopy. All organelles, including the cytoskeletal structures, are identified and appear not to have been damaged during processing, although membrane components appear somewhat less distinct. The absence of an embedding matrix eliminates the need for additional staining to increase contrast, unlike the situation with specimens embedded in standard electron-scattering resins. The PEG technique thus appears to be a valuable adjunct to conventional methods for ultrastructural analysis.

Microstructures of plasma-sprayed hydroxyapatite-coated Ti-6Al-4V dental implants.

PubMed

Tufekci, E; Brantley, W A; Mitchell, J C; McGlumphy, E A

1997-01-01

The purpose of this study was to investigate the microstructure of plasma-sprayed hydroxyapatite coatings and the elemental composition near the coating-substrate interface for two commercial implants, using the scanning electron microscope. Both coating surfaces and cross-sectioned specimens were examined. The results indicated that while the surface microstructures of both implants were consistent with the plasma-spraying process, the scale of the constituents was much finer for one product. In cross-section, both coatings exhibited minimal porosity and intimate contact with the titanium alloy substrate. It was found that limited interdiffusion of titanium and calcium occurred near the interface.

Self-ion irradiation effects on mechanical properties of nanocrystalline zirconium films

DOE PAGES

Wang, Baoming; Haque, M. A.; Tomar, Vikas; ...

2017-07-13

Zirconium thin films were irradiated at room temperature with an 800 keV Zr + beam using a 6 MV HVE Tandem accelerator to 1.36 displacement per atom damage. Freestanding tensile specimens, 100 nm thick and 10 nm grain size, were tested in-situ inside a transmission electron microscope. Significant grain growth (>300%), texture evolution, and displacement damage defects were observed. Here, stress-strain profiles were mostly linear elastic below 20 nm grain size, but above this limit the samples demonstrated yielding and strain hardening. Experimental results support the hypothesis that grain boundaries in nanocrystalline metals act as very effective defect sinks.

Characterization of Polycapillary Optics in a TES Microcalorimeter EDS System Installed on an SEM

NASA Astrophysics Data System (ADS)

Takano, A.; Maehata, K.; Iyomoto, N.; Yasuda, K.; Maeno, H.; Shiiyama, K.; Tanaka, K.

2016-08-01

Energy-dispersive spectroscopic measurements are performed using a superconducting transition-edge sensor (TES) microcalorimeter mounted on a scanning electron microscope (SEM) for advanced research at Kyushu University. Because the sensitive area of the TES microcalorimeter is about 0.02~mm2, polycapillary optics is used to collect the X-rays emitted by the SEM specimen on the TES microcalorimeter. The X-ray transmission efficiency of the polycapillary optics is obtained by analyzing the X-ray energy spectra measured by the TES microcalorimeter. The obtained transmission efficiency of the polycapillary optics is reproduced by the calculated results of the simulation.

Fracture modes of high modulus graphite/epoxy angleplied laminates subjected to off-axis tensile loads

NASA Technical Reports Server (NTRS)

Sinclair, J. H.

1980-01-01

Angelplied laminates of high modulus graphite fiber/epoxy were studied in several ply configurations at various tensile loading angles to the zero ply direction in order to determine the effects of ply orientations on tensile properties, fracture modes, and fracture surface characteristics of the various plies. It was found that fracture modes in the plies of angleplied laminates can be characterized by scanning electron microscope observation. The characteristics for a given fracture mode are similar to those for the same fracture mode in unidirectional specimens. However, no simple load angle range can be associated with a given fracture mode.

Pulmonary fibrosis in a carpenter with long-lasting exposure to fiberglass.

PubMed

Takahashi, T; Munakata, M; Takekawa, H; Homma, Y; Kawakami, Y

1996-11-01

A 56-year-old male carpenter had a history of glass fiber inhalation for 41 years without any protective device. His chest radiograph showed small nodular opacities in lower lung fields and multiple cystic lesions and low attenuation areas in upper lung fields. Light and polarizing microscopic examinations of his transbronchial lung biopsy specimen revealed mild interstitial fibrosis and mononuclear cell infiltration in alveolar walls without birefringent substances. However, widespread depositions of small glass fibers (< 2.5 microns in length and 0.3 micron in diameter) were detected by analytical electron microscopy, which suggested their possible contribution to the development of his pulmonary fibrosis.

Monte Carlo Simulation to Determine Geometry Effects on Quantitative X-ray Microanalysis in Plant Cell Walls Using Gelatin Standards

NASA Astrophysics Data System (ADS)

Tylko, Grzegorz; Dubchak, Sergyi; Banach, Zuzanna; Turnau, Katarzyna

2010-04-01

Monte Carlo simulations of gelatin matrices with known elemental concentrations confirmed the suitability of protein standards to quantify elements of cellulose material in x-ray microanalysis. However, gelatin standards and cellulose plant cell walls differ in structure, what influences x-ray generation and emission in both specimens. The goal of the project was to establish the influence of gelatin structure on x-ray generation and its usefulness to calculate elemental concentrations in plant cell walls of different width. Roots of Medicago truncatula as well as gelatin standards with known elemental composition were prepared according to freeze-drying protocols. The thermanox polymer was chosen to establish background formation for flat and compact organic materials. All analyses were performed with the scanning electron microscope operated at 10 keV and probe current of 350 pA. The Monte Carlo code Casino was applied to calculate the intensities of the generated and the emitted x-rays from biological matrix of different width. No topography effects of gelatin structure were visible when the raster mode of electron impact was applied to the specimen. Monte Carlo simulations of gelatin of different width revealed that a significant decrease of the generated x-ray intensity appears at the width of the specimen around 3.5 μm. However, an increase of emission of low energy x-ray intensities (Na, Mg) was noted at 3.5 μm size with constant emission of higher energy x-rays (Cl, K) down to 2.5 μm width. It determines the minimal size of plant specimen useful for comparison to bulk gelatin standard when quantitative analysis is performed for biologically important elements.

Substantial tensile ductility in sputtered Zr-Ni-Al nano-sized metallic glass

DOE PAGES

Liontas, Rachel; Jafary-Zadeh, Mehdi; Zeng, Qiaoshi; ...

2016-08-04

We investigate the mechanical behavior and atomic-level structure of glassy Zr-Ni-Al nano-tensile specimens with widths between 75 and 215 nm. We focus our studies on two different energy states: (1) as-sputtered and (2) sputtered then annealed below the glass transition temperature (T g). In-situ tensile experiments conducted inside a scanning electron microscope (SEM) reveal substantial tensile ductility in some cases reaching >10% engineering plastic strains, >150% true plastic strains, and necking down to a point during tensile straining in specimens as wide as ~150 nm. We found the extent of ductility depends on both the specimen size and the annealingmore » conditions. Using molecular dynamics (MD) simulations, transmission electron microscopy (TEM), and synchrotron x-ray diffraction (XRD), we explain the observed mechanical behavior through changes in free volume as well as short- and medium-range atomic-level order that occur upon annealing. This work demonstrates the importance of carefully choosing the metallic glass fabrication method and post-processing conditions for achieving a certain atomic-level structure and free volume within the metallic glass, which then determine the overall mechanical response. Lastly, an important implication is that sputter deposition may be a particularly promising technique for producing thin coatings of metallic glasses with significant ductility, due to the high level of disorder and excess free volume resulting from the sputtering process and to the suitability of sputtering for producing thin coatings that may exhibit enhanced size-induced ductility.« less

Comparative study of image contrast in scanning electron microscope and helium ion microscope.

PubMed

O'Connell, R; Chen, Y; Zhang, H; Zhou, Y; Fox, D; Maguire, P; Wang, J J; Rodenburg, C

2017-12-01

Images of Ga + -implanted amorphous silicon layers in a 110 n-type silicon substrate have been collected by a range of detectors in a scanning electron microscope and a helium ion microscope. The effects of the implantation dose and imaging parameters (beam energy, dwell time, etc.) on the image contrast were investigated. We demonstrate a similar relationship for both the helium ion microscope Everhart-Thornley and scanning electron microscope Inlens detectors between the contrast of the images and the Ga + density and imaging parameters. These results also show that dynamic charging effects have a significant impact on the quantification of the helium ion microscope and scanning electron microscope contrast. © 2017 The Authors Journal of Microscopy © 2017 Royal Microscopical Society.

Experimental Characterization and Simulation of Slip Transfer at Grain Boundaries and Microstructurally-Sensitive Crack Propagation

NASA Technical Reports Server (NTRS)

Gupta, Vipul; Hochhalter, Jacob; Yamakov, Vesselin; Scott, Willard; Spear, Ashley; Smith, Stephen; Glaessgen, Edward

2013-01-01

A systematic study of crack tip interaction with grain boundaries is critical for improvement of multiscale modeling of microstructurally-sensitive fatigue crack propagation and for the computationally-assisted design of more durable materials. In this study, single, bi- and large-grain multi-crystal specimens of an aluminum-copper alloy are fabricated, characterized using electron backscattered diffraction (EBSD), and deformed under tensile loading and nano-indentation. 2D image correlation (IC) in an environmental scanning electron microscope (ESEM) is used to measure displacements near crack tips, grain boundaries and within grain interiors. The role of grain boundaries on slip transfer is examined using nano-indentation in combination with high-resolution EBSD. The use of detailed IC and EBSD-based experiments are discussed as they relate to crystal-plasticity finite element (CPFE) model calibration and validation.

Experimentally determined wear behavior of an Al2O3-SiC composite from 25 to 1200 C

NASA Technical Reports Server (NTRS)

Dellacorte, Christopher; Farmer, Serene C.; Book, Patricia O.

1990-01-01

The sliding wear behavior of a self-mated alumina-silicon carbide whisker toughened composite was studied using optical, scanning electron (SEM) and transmission electron (TEM) microscopy. Because of its excellent strength and toughness properties this composite material is under consideration for use in heat engine applications for sliding contacts which operate at elevated temperatures. The composite's wear behavior and especially its wear mechanisms are not well understood. Pin-on-disk specimens were slid in air at 2.7 m/s sliding velocity, under a 26.5-N load, at temperatures 25 to 1200 C. Pin wear increased with increasing temperature. Based upon the microscopic analyses, the wear mechanism seems to be loosening of the reinforcing whiskers due to frictional and bulk heating. This leads to whisker pullout and increased wear.

Virtual microscopy-The future of teaching histology in the medical curriculum?

PubMed

Paulsen, Friedrich P; Eichhorn, Michael; Bräuer, Lars

2010-12-20

Conventional continuing education in microscopic anatomy, histopathology, hematology and microbiology has hitherto been carried out using numerous sets of sectioned tissue specimens in a microscopy laboratory. In comparison, after digitalization of the sections it would be possible to access teaching specimens via virtual microscopy and the internet at any time and place. This would make it possible to put innumerable new learning scenarios into practice. The present article elucidates the advantages of virtual microscopy in histology instruction and presents a concept of how virtual microscopy could be introduced into the teaching of microscopic anatomy in several steps. Initially, the presently existing microscopic teaching specimens would be digitalized and made available on-line without restriction. In a second step, instruction would be shifted to an emphasis on virtual microscopy, utilizing all of the advantages offered by the technique. In a third step, the microscopic contents could be networked with other anatomical, radiological and clinical content on-line, thus opening new learning perspectives for students of human and dental medicine as well as those of medically related courses of study. The advantages and disadvantages of such a concept as well as some possibly arising consequences are discussed in the following. 2010 Elsevier GmbH. All rights reserved.

Light field creating and imaging with different order intensity derivatives

NASA Astrophysics Data System (ADS)

Wang, Yu; Jiang, Huan

2014-10-01

Microscopic image restoration and reconstruction is a challenging topic in the image processing and computer vision, which can be widely applied to life science, biology and medicine etc. A microscopic light field creating and three dimensional (3D) reconstruction method is proposed for transparent or partially transparent microscopic samples, which is based on the Taylor expansion theorem and polynomial fitting. Firstly the image stack of the specimen is divided into several groups in an overlapping or non-overlapping way along the optical axis, and the first image of every group is regarded as reference image. Then different order intensity derivatives are calculated using all the images of every group and polynomial fitting method based on the assumption that the structure of the specimen contained by the image stack in a small range along the optical axis are possessed of smooth and linear property. Subsequently, new images located any position from which to reference image the distance is Δz along the optical axis can be generated by means of Taylor expansion theorem and the calculated different order intensity derivatives. Finally, the microscopic specimen can be reconstructed in 3D form using deconvolution technology and all the images including both the observed images and the generated images. The experimental results show the effectiveness and feasibility of our method.

3DSEM++: Adaptive and intelligent 3D SEM surface reconstruction.

PubMed

Tafti, Ahmad P; Holz, Jessica D; Baghaie, Ahmadreza; Owen, Heather A; He, Max M; Yu, Zeyun

2016-08-01

Structural analysis of microscopic objects is a longstanding topic in several scientific disciplines, such as biological, mechanical, and materials sciences. The scanning electron microscope (SEM), as a promising imaging equipment has been around for decades to determine the surface properties (e.g., compositions or geometries) of specimens by achieving increased magnification, contrast, and resolution greater than one nanometer. Whereas SEM micrographs still remain two-dimensional (2D), many research and educational questions truly require knowledge and facts about their three-dimensional (3D) structures. 3D surface reconstruction from SEM images leads to remarkable understanding of microscopic surfaces, allowing informative and qualitative visualization of the samples being investigated. In this contribution, we integrate several computational technologies including machine learning, contrario methodology, and epipolar geometry to design and develop a novel and efficient method called 3DSEM++ for multi-view 3D SEM surface reconstruction in an adaptive and intelligent fashion. The experiments which have been performed on real and synthetic data assert the approach is able to reach a significant precision to both SEM extrinsic calibration and its 3D surface modeling. Copyright © 2016 Elsevier Ltd. All rights reserved.

[Microscopic extensions of head and neck squamous cell carcinomas: impact for clinical target volume definition].

PubMed

Fleury, B; Thariat, J; Barnoud, R; Buiret, G; Lebreton, F; Bancel, B; Poupart, M; Devouassoux-Shisheboran, M

2014-11-01

To assess microscopic extensions of head and neck squamous cell carcinomas aiming at a proposal for target volumes of radiation therapy. Surgical specimens were prospectively analysed macroscopically and microscopically. Tumour borders were identified per macroscopic visual examination and inked on stained slides. Then microscopic implants (perineural or lymphatic involvement, or in situ carcinomas) were looked for with an optic microscope in the macroscopic healthy tissue surrounding the tumour. The maximal length from tumour border was correlated with the maximal length of macroscopically healthy tissues assessable. Twenty-one specimens were analysed and 12 were locally advanced tumours. Mean and median maximal microscopic extensions were 2.9 and 1.0mm (0-15mm), respectively. The 90th and 95th percentiles were 5 and 11mm, respectively. The ratio between healthy tissue length and maximal microscopic tumour extension was 10%. No correlation was found with tumour grade or volume. The presence of microscopic tumour was unlikely after 5mm from macroscopic tumour (≤5% of patients in this series) but should be assessed along with other histoclinical factors and particularities of tumour behaviour by anatomic site. A rigorous terminology should authorize a relevant appreciation of local risk of recurrence, particularly in adjuvant setting or for clinical target volume definition. Larger and more homogenous confirmatory series are needed. Copyright © 2014. Published by Elsevier SAS.