WorldWideScience

Sample records for scanning acoustic microscopy

  1. Breast tissue characterization with high-frequency scanning acoustic microscopy

    Science.gov (United States)

    Kumon, R. E.; Bruno, I.; Heartwell, B.; Maeva, E.

    2004-05-01

    We have performed imaging of breast tissue using scanning acoustic microscopy (SAM) in the range of 25-50 MHz with the goal of accurately and rapidly determining the structure and composition throughout the volume of the samples. In contrast to traditional histological slides, SAM images can be obtained without special preparation, sometimes even without sectioning, but with sufficiently high spatial resolution to give information comparable to surface optical images. As a result, the use of high-frequency SAM at the time of breast lumpectomy to identify disease-free margins has the potential to reduce reoperative rates, patient anxiety, and local recurrence. However, only limited work has been performed to characterize breast tissue in the frequency range above clinical ultrasound devices. The samples are 4-cm2-thick sections (2-3 mm) taken from mastectomies and preserved in formalin. They are placed between two plates and immersed in water during imaging. Attenuation images are acquired by focusing the acoustic beam at the top and bottom of the samples, although better results were obtained for bottom focusing. For purposes of comparison and identification of histological features, acoustical images will be presented along with optical images obtained from the same samples. [Work supported by CIHR.

  2. OSTEOBLAST ADHESION OF BREAST CANCER CELLS WITH SCANNING ACOUSTIC MICROSCOPY

    Energy Technology Data Exchange (ETDEWEB)

    Chiaki Miyasaka; Robyn R. Mercer; Andrea M. Mastro; Ken L. Telschow

    2005-03-01

    Breast cancer frequently metastasizes to the bone. Upon colonizing bone tissue, the cancer cells stimulate osteoclasts (cells that break bone down), resulting in large lesions in the bone. The breast cancer cells also affect osteoblasts (cells that build new bone). Conditioned medium was collected from a bone-metastatic breast cancer cell line, MDA-MB-231, and cultured with an immature osteoblast cell line, MC3T3-E1. Under these conditions the osteoblasts acquired a changed morphology and appeared to adherer in a different way to the substrate and to each other. To characterize cell adhesion, MC3T3-E1 osteoblasts were cultured with or without MDA-MB-231 conditioned medium for two days, and then assayed with a mechanical scanning acoustic reflection microscope (SAM). The SAM indicated that in normal medium the MC3T3-E1 osteoblasts were firmly attached to their plastic substrate. However, MC3T3-E1 cells cultured with MDA-MB-231 conditioned medium displayed both an abnormal shape and poor adhesion at the substrate interface. The cells were fixed and stained to visualize cytoskeletal components using optical microscopic techniques. We were not able to observe these differences until the cells were quite confluent after 7 days of culture. However, using the SAM, we were able to detect these changes within 2 days of culture with MDA-MB-231 conditioned medium

  3. Elastic Changes of Capsule in a Rat Knee Contracture Model Assessed by Scanning Acoustic Microscopy

    Science.gov (United States)

    Hagiwara, Y.; Chimoto, E.; Ando, A.; Saijo, Y.; Itoi, E.

    Sound speed of a capsule in a rat knee contracture model was measured by scanning acoustic microscopy. There was no statistical significant difference in the anterior capsule compared with the control group. However, the sound speed of the posterior capsule was significantly greater compared with the control group after prolonged immobilization.

  4. The Observation of Martensite and Magnetic Domain Structures in Ni53Mn24Ga23 Shape Memory Alloys by Scanning Electron Acoustic Microscopy and Scanning Thermal Microscopy

    Science.gov (United States)

    Zhao, Kun-Yu; Zeng, Hua-Rong; Song, Hong-Zhang; Hui, Sen-Xing; Li, Guo-Rong; Yin, Qing-Rui

    2012-05-01

    We present observations of martensite variants and ferromagnetic domain structures of Ni53Mn24Ga23 ferromagnetic shape memory alloys with a pure tetragonal martensitic phase by using scanning electron acoustic microscopy (SEAM) and scanning thermal microscopy (SThM). Electron acoustic images show a polycrystalline morphology with martensite variants. Direct coincidence between crystallographic martensitic twin variants and magnetic domains is found. A domain-like structure, obtained by SThM, is firstly reported, and then confirmed by magnetic force microscopy (MFM). The experimental results will be helpful for investigating the local thermal properties of ferromagnets and understanding the relationship between martensite variants and magnetic domains.

  5. Acoustic microscopy

    CERN Document Server

    Briggs, Andrew

    2010-01-01

    For many years 'Acoustic Microscopy' has been the definitive book on the subject. A key development since it was first published has been the development of ultrasonic force microscopy. This edition has a major new chapter on this technique and its applications.

  6. Scanning Acoustic Microscopy for Characterization of Coatings and Near-Surface Features of Ceramics

    Energy Technology Data Exchange (ETDEWEB)

    Qu, Jun [ORNL; Blau, Peter Julian [ORNL

    2006-01-01

    Scanning Acoustic Microscopy (SAcM) has been widely used for non-destructive evaluation (NDE) in various fields such as material characterization, electronics, and biomedicine. SAcM uses high-frequency acoustic waves (60 MHz to 2.0 GHz) providing much higher resolution (up to 0.5 {micro}m) compared to conventional ultrasonic NDE, which is typically about 500 {micro}m. SAcM offers the ability to non-destructively image subsurface features and visualize the variations in elastic properties. These attributes make SAcM a valuable tool for characterizing near-surface material properties and detecting fine-scale flaws. This paper presents some recent applications of SAcM in detecting subsurface damage, assessing coatings, and visualizing residual stress for ceramic and semiconductor materials.

  7. NDE Acoustic Microscopy Research Laboratory

    Data.gov (United States)

    Federal Laboratory Consortium — The purpose is to develop advanced, more effective high-resolution micro-NDE materials characterization methods using scanning acoustic microscopy. The laboratory's...

  8. Biomechanical Changes of Collagen Cross-Linking on Human Keratoconic Corneas Using Scanning Acoustic Microscopy.

    Science.gov (United States)

    Beshtawi, Ithar M; Akhtar, Riaz; Hillarby, M Chantal; O'Donnell, Clare; Zhao, Xuegen; Brahma, Arun; Carley, Fiona; Derby, Brian; Radhakrishnan, Hema

    2016-05-01

    To assess the biomechanical changes of collagen cross-linking on keratoconic corneas in vitro. Six keratoconic corneal buttons were included in this study. Each cornea was divided into two halves, where one half was cross-linked and the other half was treated with riboflavin only and served as control. The biomechanical changes of the corneal tissue were measured across the stroma using scanning acoustic microscopy (SAM). In the cross-linked corneas, there was a steady decrease in the magnitude of speed of sound from the anterior region through to the posterior regions of the stroma. The speed of sound was found to decrease slightly across the corneal thickness in the control corneas. The increase in speed of sound between the cross-linked and control corneas in the anterior region was by a factor of 1.039×. A higher speed of sound was detected in cross-linked keratoconic corneal tissue when compared with their controls, using SAM. This in vitro model can be used to compare to the cross-linking results obtained in vivo, as well as comparing the results obtained with different protocols.

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

    Energy Technology Data Exchange (ETDEWEB)

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

    2014-09-01

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

  10. Investigation of delamination mechanisms in polymer coatings by scanning acoustic microscopy

    Energy Technology Data Exchange (ETDEWEB)

    Alig, I; Bargmann, M; Oehler, H; Lellinger, D [Deutsches Kunststoff-Institut, Schlossgartenstr. 6, 64289 Darmstadt (Germany); Wanner, M; Koch, D, E-mail: ialig@dki.tu-darmstadt.de [Fraunhofer Institut fuer Produktionstechnik und Automatisierung, Nobelstrasse 12, 70569 Stuttgart (Germany)

    2011-01-26

    Scanning acoustic microscopy (SAM) was used to investigate disbonding, delamination and blister formation in polymer coatings for different layer structures and initial defect depths during exposure to a corrosive environment. The time evolution of disbonding and blister initiation and their growth was investigated by analysing the SAM images, taken after defined times of exposure to an electrolyte solution (Harrison solution). From investigations of a model system of coatings on steel substrates, it was possible to differentiate between water and/or ion transport (i) through the coating and (ii) along the coating-steel interface. For samples without clear coat randomly distributed blisters appeared at the coating-steel interface after exposure to the electrolyte solution, irrespective of the location of initial defects. The random distribution of growing blisters is related to diffusion of water and/or ions through the coating and 'nucleation' at weak points of the substrate or within the polymer. For samples with clear coat a propagating migration front along the coating-steel interface of 3-4 {mu}m height-starting from initial defects-was detected. The linear propagation of this front cannot be explained by Fickian diffusion. Therefore, it is discussed in terms of an accelerated diffusion or crack growth kinetics. Since blistering starts only at sites, where the migration front has passed, the presence of a thin water layer at the coating-steel interface, the loss of adhesion and the following corrosive processes are prerequisites for the nucleation of blisters. The blister growth shows a square-root time dependence.

  11. Closure integrity testing of heat sealed aseptic packaging using scanning acoustic microscopy

    OpenAIRE

    Jarrosson, Bruno P.

    1992-01-01

    The objective of this study was to determine the possible application of ultrasonic inspection for non-destructive, online evaluation of the integrity of heat sealed, flexible package structure commonly used in packaging of aseptic and shelf-stable food products. A scanning acoustic microscope (SAM), Olympus UH-3, and image analysis system were used to establish the operational parameters to ultrasonically inspect the heat seal closure of various flexible packages. The...

  12. Imaging surface acoustic wave dynamics in semiconducting polymers by scanning ultrafast electron microscopy.

    Science.gov (United States)

    Najafi, Ebrahim; Liao, Bolin; Scarborough, Timothy; Zewail, Ahmed

    2018-01-01

    Understanding the mechanical properties of organic semiconductors is essential to their electronic and photovoltaic applications. Despite a large volume of research directed toward elucidating the chemical, physical and electronic properties of these materials, little attention has been directed toward understanding their thermo-mechanical behavior. Here, we report the ultrafast imaging of surface acoustic waves (SAWs) on the surface of the Poly(3-hexylthiophene-2,5-diyl) (P3HT) thin film at the picosecond and nanosecond timescales. We then use these images to measure the propagation velocity of SAWs, which we then employ to determine the Young's modulus of P3HT. We further validate our experimental observation by performing a semi-empirical transient thermoelastic finite element analysis. Our findings demonstrate the potential of ultrafast electron microscopy to not only probe charge carrier dynamics in materials as previously reported, but also to measure their mechanical properties with great accuracy. This is particularly important when in situ characterization of stiffness for thin devices and nanomaterials is required. Copyright © 2017 Elsevier B.V. All rights reserved.

  13. Scanning ultrafast electron microscopy

    OpenAIRE

    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 whic...

  14. Scanning Acoustic Microscopy-A Novel Noninvasive Method to Determine Tumor Interstitial Fluid Pressure in a Xenograft Tumor Model.

    Science.gov (United States)

    Hofmann, Matthias; Pflanzer, Ralph; Habib, Anowarul; Shelke, Amit; Bereiter-Hahn, Jürgen; Bernd, August; Kaufmann, Roland; Sader, Robert; Kippenberger, Stefan

    2016-06-01

    Elevated tumor interstitial fluid pressure (TIFP) is a prominent feature of solid tumors and hampers the transmigration of therapeutic macromolecules, for example, large monoclonal antibodies, from tumor-supplying vessels into the tumor interstitium. TIFP values of up to 40 mm Hg have been measured in experimental solid tumors using two conventional invasive techniques: the wick-in-needle and the micropuncture technique. We propose a novel noninvasive method of determining TIFP via ultrasonic investigation with scanning acoustic microscopy at 30-MHz frequency. In our experimental setup, we observed for the impedance fluctuations in the outer tumor hull of A431-vulva carcinoma-derived tumor xenograft mice. The gain dependence of signal strength was quantified, and the relaxation of tissue was calibrated with simultaneous hydrostatic pressure measurements. Signal patterns from the acoustical images were translated into TIFP curves, and a putative saturation effect was found for tumor pressures larger than 3 mm Hg. This is the first noninvasive approach to determine TIFP values in tumors. This technique can provide a potentially promising noninvasive assessment of TIFP and, therefore, can be used to determine the TIFP before treatment approach as well to measure therapeutic efficacy highlighted by lowered TFP values. Copyright © 2016 The Authors. Published by Elsevier Inc. All rights reserved.

  15. Mapping dynamical mechanical properties of osteonal bone by scanning acoustic microscopy in time-of-flight mode.

    Science.gov (United States)

    Blouin, Stéphane; Puchegger, Stephan; Roschger, Andreas; Berzlanovich, Andrea; Fratzl, Peter; Klaushofer, Klaus; Roschger, Paul

    2014-06-01

    An important determinant of mechanical properties of bone is Young's modulus and its variation in individual osteons of cortical bone tissue. Its mechanical behavior also depends on deformation rate owing to its visco- or poroelastic properties. We developed a method to measure dynamical mechanical properties of bulk bone tissue at osteonal level based on scanning acoustic microscopy (SAM) using time-of-flight (TOF) measurements in combination with quantitative backscattered electron imaging (qBEI). SAM-TOF yields local sound velocities and qBEI corresponding material densities together providing elastic properties. Osteons (n=55) were measured in three human femoral diaphyseal ground bone sections (∼ 30 µm in thickness). In addition, subchondral bone and mineralized articular cartilage were investigated. The mean mineral contents, the mean sound velocities, and the mean elastic modulus of the osteons ranged from 20 to 26 wt%, from 3,819 to 5,260 m/s, and from 21 to 44 GPa, respectively. There was a strong positive correlation between material density and sound velocity (Pearson's r=0.701; pSound velocities between cartilage and bone was similar, though material density was higher in cartilage (+4.46%, p<0.0001). These results demonstrate the power of SAM-TOF to estimate dynamic mechanical properties of the bone materials at the osteonal level.

  16. Scanning ultrafast electron microscopy.

    Science.gov (United States)

    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.

  17. Biomechanical properties of human corneas following low- and high-intensity collagen cross-linking determined with scanning acoustic microscopy.

    Science.gov (United States)

    Beshtawi, Ithar M; Akhtar, Riaz; Hillarby, M Chantal; O'Donnell, Clare; Zhao, Xuegen; Brahma, Arun; Carley, Fiona; Derby, Brian; Radhakrishnan, Hema

    2013-08-07

    To assess and compare changes in the biomechanical properties of the cornea following different corneal collagen cross-linking protocols using scanning acoustic microscopy (SAM). Ten donor human corneal pairs were divided into two groups consisting of five corneal pairs in each group. In group A, five corneas were treated with low-fluence (370 nm, 3 mW/cm(2)) cross-linking (CXL) for 30 minutes. In group B, five corneas were treated with high-fluence (370 nm, 9 mW/cm(2)) CXL for 10 minutes. The contralateral control corneas in both groups had similar treatment but without ultraviolet A. The biomechanical properties of all corneas were tested using SAM. In group A, the mean speed of sound in the treated corneas was 1677.38 ± 10.70 ms(-1) anteriorly and 1603.90 ± 9.82 ms(-1) posteriorly, while it was 1595.23 ± 9.66 ms(-1) anteriorly and 1577.13 ± 8.16 ms(-1) posteriorly in the control corneas. In group B, the mean speed of sound of the treated corneas was 1665.06 ± 9.54 ms(-1) anteriorly and 1589.89 ± 9.73 ms(-1) posteriorly, while it was 1583.55 ± 8.22 ms(-1) anteriorly and 1565.46 ± 8.13 ms(-1) posteriorly in the untreated control corneas. The increase in stiffness between the cross-linked and control corneas in both groups was by a factor of 1.051×. SAM successfully detected changes in the corneal stiffness after application of collagen cross-linking. A higher speed-of-sound value was found in the treated corneas when compared with the controls. No significant difference was found in corneal stiffness between the corneas cross-linked with low- and high-intensity protocols.

  18. Biomechanical Properties of Human Corneas Following Low- and High-Intensity Collagen Cross-Linking Determined With Scanning Acoustic Microscopy

    Science.gov (United States)

    Beshtawi, Ithar M.; Akhtar, Riaz; Hillarby, M. Chantal; O'Donnell, Clare; Zhao, Xuegen; Brahma, Arun; Carley, Fiona; Derby, Brian; Radhakrishnan, Hema

    2013-01-01

    Purpose. To assess and compare changes in the biomechanical properties of the cornea following different corneal collagen cross-linking protocols using scanning acoustic microscopy (SAM). Methods. Ten donor human corneal pairs were divided into two groups consisting of five corneal pairs in each group. In group A, five corneas were treated with low-fluence (370 nm, 3 mW/cm2) cross-linking (CXL) for 30 minutes. In group B, five corneas were treated with high-fluence (370 nm, 9 mW/cm2) CXL for 10 minutes. The contralateral control corneas in both groups had similar treatment but without ultraviolet A. The biomechanical properties of all corneas were tested using SAM. Results. In group A, the mean speed of sound in the treated corneas was 1677.38 ± 10.70 ms−1 anteriorly and 1603.90 ± 9.82 ms−1 posteriorly, while it was 1595.23 ± 9.66 ms−1 anteriorly and 1577.13 ± 8.16 ms−1 posteriorly in the control corneas. In group B, the mean speed of sound of the treated corneas was 1665.06 ± 9.54 ms−1 anteriorly and 1589.89 ± 9.73 ms−1 posteriorly, while it was 1583.55 ± 8.22 ms−1 anteriorly and 1565.46 ± 8.13 ms−1 posteriorly in the untreated control corneas. The increase in stiffness between the cross-linked and control corneas in both groups was by a factor of 1.051×. Conclusions. SAM successfully detected changes in the corneal stiffness after application of collagen cross-linking. A higher speed-of-sound value was found in the treated corneas when compared with the controls. No significant difference was found in corneal stiffness between the corneas cross-linked with low- and high-intensity protocols. PMID:23847309

  19. Multi-Layer Phase Analysis: Quantifying the Elastic Properties of Soft Tissues and Live Cells with Ultra-High Frequency Scanning Acoustic Microscopy

    Science.gov (United States)

    Zhao, Xuegen; Akhtar, Riaz; Nijenhuis, Nadja; Wilkinson, Steven J.; Murphy, Lilli; Ballestrem, Christoph; Sherratt, Michael. J.; Watson, Rachel E.B.; Derby, Brian

    2012-01-01

    Scanning acoustic microscopy is potentially a powerful tool for characterising the elastic properties of soft biological tissues and cells. In this paper, we present a method, Multi-Layer Phase Analysis (MLPA), which can be used to extract local speed of sound values, for both thin tissue sections mounted on glass slides and cultured cells grown on cell culture plastic, with a resolution close to 1 μm. The method exploits the phase information that is preserved in the interference between the acoustic wave reflected from the substrate surface and internal reflections from the acoustic lens. In practice, a stack of acoustic images are captured beginning with the acoustic focal point 4 μm above the substrate surface and moving down in 0.1 μm increments. Scanning parameters, such as acoustic wave frequency and gate position, were adjusted to obtain optimal phase and lateral resolution. The data were processed offline to extract the phase information with the contribution of any inclination in the substrate removed prior to the calculation of sound speed. Here, we apply this approach to both skin sections and fibroblast cells, and compare our data with the V(f) (voltage vs frequency) method that has previously been used for characterisation of soft tissues and cells. Compared with the V(f) method, the MPLA method not only reduces signal noise but can be implemented without making a priori assumptions with regards to tissue or cell parameters. PMID:22547273

  20. Biomechanical changes after repeated collagen cross-linking on human corneas assessed in vitro using scanning acoustic microscopy.

    Science.gov (United States)

    Beshtawi, Ithar M; Akhtar, Riaz; Hillarby, M Chantal; O'Donnell, Clare; Zhao, Xuegen; Brahma, Arun; Carley, Fiona; Derby, Brian; Radhakrishnan, Hema

    2014-03-13

    To explore the biomechanical changes induced by repeated cross-linking using scanning acoustic microscopy (SAM). Thirty human corneas were divided into three groups. In group A, five corneas were cross-linked once. In group B, five corneas were cross-linked twice, 24 hours apart. In group C, five corneas were cross-linked three times, 24 hours apart. The contralateral controls in all groups had similar treatment but without UV-A. The speed of sound, which is directly proportional to the square root of the tissue's elastic modulus, was assessed using SAM. In group A, the speed of sound of the treated corneas was 1677.38 ± 10.70 ms(-1) anteriorly and 1603.90 ± 9.82 ms(-1) posteriorly, while it was 1595.23 ± 9.66 ms(-1) anteriorly and 1577.13 ± 8.16 ms(-1) posteriorly in the controls. In group B, the speed of sound of the treated corneas was 1746.33 ± 23.37 ms(-1) anteriorly and 1631.60 ± 18.92 ms(-1) posteriorly, while it was 1637.57 ± 22.15 ms(-1) anteriorly and 1612.30 ± 22.23 ms(-1) posteriorly in the controls. In group C, the speed of sound of the treated corneas was 1717.97 ± 18.92 ms(-1) anteriorly and 1616.62 ± 17.58 ms(-1) posteriorly, while it was 1628.69 ± 9.37 ms(-1) anteriorly and 1597.68 ± 11.97 ms(-1) posteriorly in the controls. The speed of sound in the anterior (200 × 200 μm) region between the cross-linked and control corneas in groups A, B, and C was increased by a factor of 1.051 (P = 0.005), 1.066 (P = 0.010), and 1.055 (P = 0.005) respectively. However, there was no significant difference among the cross-linked corneas in all groups (P = 0.067). A significant increase in speed of sound was found in all treated groups compared with the control group; however, the difference among the treated groups is not significant, suggesting no further cross-links are induced when collagen cross-linking treatment is repeated.

  1. Analysis of residual stress in the resin of metal-resin adhesion structures by scanning acoustic microscopy.

    Science.gov (United States)

    Ohno, Hiroki; Endo, Kazuhiko; Nagano-Takebe, Futami; Ida, Yusuke; Kakino, Ken; Narita, Toshio

    2013-01-01

    The residual stress caused by polymerization shrinkage and thermal contraction of a heat-curing resin containing 4-META on a metal-resin structure was measured by a scanning acoustic microscope. The tensile residual stress in the resin occurred within 70 µm of the adhesion interface with a flat plate specimen. The maximum tensile stress was about 58 MPa at the interface. On a metal plate specimen with retention holes, ring-like cracks in the resin occurred around the retention holes with the adhesive specimen and many linear cracks occurred in the resin vertical to the longitudinal direction of the metal frame with the non-adhesive specimens. There was tensile residual stress on the resin surface at the center of the retention holes of the adhesion specimen, indicating that the stress in the specimen with surface treatment for adhesion was higher than in that without surface treatment.

  2. Confocal scanning microscopy

    DEFF Research Database (Denmark)

    Bariani, Paolo

    This report is based on a metrological investigation on confocal microscopy technique carried out by Uffe Rolf Arlø Theilade and Paolo Bariani. The purpose of the experimental activity was twofold a metrological instrument characterization and application to assessment of rough PP injection moulded...... replicated topography. Confocal microscopy is seen to be a promising technique in metrology of microstructures. Some limitations with respect to surface metrology were found during the experiments. The experiments were carried out using a Zeiss LSM 5 Pascal microscope owned by the Danish Polymer Centre...

  3. Study on Damage Evaluation and Machinability of UD-CFRP for the Orthogonal Cutting Operation Using Scanning Acoustic Microscopy and the Finite Element Method.

    Science.gov (United States)

    Wang, Dongyao; He, Xiaodong; Xu, Zhonghai; Jiao, Weicheng; Yang, Fan; Jiang, Long; Li, Linlin; Liu, Wenbo; Wang, Rongguo

    2017-02-20

    Owing to high specific strength and designability, unidirectional carbon fiber reinforced polymer (UD-CFRP) has been utilized in numerous fields to replace conventional metal materials. Post machining processes are always required for UD-CFRP to achieve dimensional tolerance and assembly specifications. Due to inhomogeneity and anisotropy, UD-CFRP differs greatly from metal materials in machining and failure mechanism. To improve the efficiency and avoid machining-induced damage, this paper undertook to study the correlations between cutting parameters, fiber orientation angle, cutting forces, and cutting-induced damage for UD-CFRP laminate. Scanning acoustic microscopy (SAM) was employed and one-/two-dimensional damage factors were then created to quantitatively characterize the damage of the laminate workpieces. According to the 3D Hashin's criteria a numerical model was further proposed in terms of the finite element method (FEM). A good agreement between simulation and experimental results was validated for the prediction and structural optimization of the UD-CFRP.

  4. Ion Acoustic Microscopy

    Science.gov (United States)

    1985-07-01

    Murphy and L. C. Aamodt , "Signal Enhancement in Photothermal Imaging Produced by Three Dimensional Heat Flow", Appl. Phys. Lett. 39, 519 (1981); L. C... Aamodt and J. C. Murphy, "Photothermal Measurements Using a Localized Excitation Source", J. Appl. Phys. 52, 4903 (1981) (9) R. L. Thomas, L. D. Favro...25 (23) J. C. Murphy, F. G. Satkiewicz and L. C. Aamodt , "Ion Acoustic Imaging of Buried Flaws in Aluminum", Review of Progress in Quantitative NDE

  5. Scanning quantum decoherence microscopy.

    Science.gov (United States)

    Cole, Jared H; Hollenberg, Lloyd C L

    2009-12-09

    The use of qubits as sensitive nanoscale magnetometers has been studied theoretically and recently demonstrated experimentally. In this paper we propose a new concept, in which a scanning two-state quantum system is used to probe a sample through the subtle effects of decoherence. Mapping both the Hamiltonian and decoherence properties of a qubit simultaneously provides a unique image of the magnetic (or electric) field properties at the nanoscale. The resulting images are sensitive to the temporal as well as spatial variation in the fields created by the sample. As examples we theoretically study two applications; one from condensed matter physics, the other biophysics. The individual components required to realize the simplest version of this device (characterization and measurement of qubits, nanoscale positioning) have already been demonstrated experimentally.

  6. Vacuum scanning capillary photoemission microscopy

    DEFF Research Database (Denmark)

    Aseyev, S.A.; Cherkun, A P; Mironov, B N

    2017-01-01

    We demonstrate the use of a conical capillary in a scanning probe microscopy for surface analysis. The probe can measure photoemission from a substrate by transmitting photoelectrons along the capillary as a function of probe position. The technique is demonstrated on a model substrate consisting...

  7. High Resolution Scanning Ion Microscopy

    NARCIS (Netherlands)

    Castaldo, V.

    2011-01-01

    The structure of the thesis is the following. The first chapter is an introduction to scanning microscopy, where the path that led to the Focused Ion Beam (FIB) is described and the main differences between electrons and ion beams are highlighted. Chapter 2 is what is normally referred to (which I

  8. Study on Damage Evaluation and Machinability of UD-CFRP for the Orthogonal Cutting Operation Using Scanning Acoustic Microscopy and the Finite Element Method

    Directory of Open Access Journals (Sweden)

    Dongyao Wang

    2017-02-01

    Full Text Available Owing to high specific strength and designability, unidirectional carbon fiber reinforced polymer (UD-CFRP has been utilized in numerous fields to replace conventional metal materials. Post machining processes are always required for UD-CFRP to achieve dimensional tolerance and assembly specifications. Due to inhomogeneity and anisotropy, UD-CFRP differs greatly from metal materials in machining and failure mechanism. To improve the efficiency and avoid machining-induced damage, this paper undertook to study the correlations between cutting parameters, fiber orientation angle, cutting forces, and cutting-induced damage for UD-CFRP laminate. Scanning acoustic microscopy (SAM was employed and one-/two-dimensional damage factors were then created to quantitatively characterize the damage of the laminate workpieces. According to the 3D Hashin’s criteria a numerical model was further proposed in terms of the finite element method (FEM. A good agreement between simulation and experimental results was validated for the prediction and structural optimization of the UD-CFRP.

  9. Study on Damage Evaluation and Machinability of UD-CFRP for the Orthogonal Cutting Operation Using Scanning Acoustic Microscopy and the Finite Element Method

    Science.gov (United States)

    Wang, Dongyao; He, Xiaodong; Xu, Zhonghai; Jiao, Weicheng; Yang, Fan; Jiang, Long; Li, Linlin; Liu, Wenbo; Wang, Rongguo

    2017-01-01

    Owing to high specific strength and designability, unidirectional carbon fiber reinforced polymer (UD-CFRP) has been utilized in numerous fields to replace conventional metal materials. Post machining processes are always required for UD-CFRP to achieve dimensional tolerance and assembly specifications. Due to inhomogeneity and anisotropy, UD-CFRP differs greatly from metal materials in machining and failure mechanism. To improve the efficiency and avoid machining-induced damage, this paper undertook to study the correlations between cutting parameters, fiber orientation angle, cutting forces, and cutting-induced damage for UD-CFRP laminate. Scanning acoustic microscopy (SAM) was employed and one-/two-dimensional damage factors were then created to quantitatively characterize the damage of the laminate workpieces. According to the 3D Hashin’s criteria a numerical model was further proposed in terms of the finite element method (FEM). A good agreement between simulation and experimental results was validated for the prediction and structural optimization of the UD-CFRP. PMID:28772565

  10. Site-matched assessment of structural and tissue properties of cortical bone using scanning acoustic microscopy and synchrotron radiation {mu}CT

    Energy Technology Data Exchange (ETDEWEB)

    Raum, K [Laboratoire d' Imagerie Parametrique, CNRS/Universite Paris 6, UMR 7623, 15, rue de l' Ecole de Medecine, 75006 Paris (France); Q-BAM Group, Department of Orthopedics, Martin Luther University of Halle-Wittenberg, Magdeburger Strasse 22, 06097 Halle (Germany); Leguerney, I [Laboratoire d' Imagerie Parametrique, CNRS/Universite Paris 6, UMR 7623, 15, rue de l' Ecole de Medecine, 75006 Paris (France); Chandelier, F [Laboratoire d' Imagerie Parametrique, CNRS/Universite Paris 6, UMR 7623, 15, rue de l' Ecole de Medecine, 75006 Paris (France); Talmant, M [Laboratoire d' Imagerie Parametrique, CNRS/Universite Paris 6, UMR 7623, 15, rue de l' Ecole de Medecine, 75006 Paris (France); Saied, A [Laboratoire d' Imagerie Parametrique, CNRS/Universite Paris 6, UMR 7623, 15, rue de l' Ecole de Medecine, 75006 Paris (France); Peyrin, F [CREATIS, CNRS, UMR 5515, INSERM, U630, INSA, 69621 Villeurbanne Cedex (France); ESRF, BP 220, 38043 Grenoble Cedex (France); Laugier, P [Laboratoire d' Imagerie Parametrique, CNRS/Universite Paris 6, UMR 7623, 15, rue de l' Ecole de Medecine, 75006 Paris (France)

    2006-02-07

    200 MHz scanning acoustic microscopy (SAM) and synchrotron radiation {mu}CT (SR-{mu}CT) were used to assess microstructural parameters and tissue properties in site-matched regions of interest in cortical bone. Anterior and postero-lateral regions of ten cross sections from human cortical radius were explored. Structural parameters, including diameter and number of Haversian canals per cortical area (Ca.Dm, N.Ca/Ar) and porosity Po were assessed with both methods using a custom-developed image fusion and analysis software. Acoustic impedance Z and degree of mineralization of bone DMB were extracted separately for osteonal and interstitial tissues from the fused images. Structural parameter estimations obtained from radiographic and acoustic images were almost identical. DMB and impedance values were in the range between 0.77 and 1.28 g cm{sup -3} and 5.13 and 12.1 Mrayl, respectively. Interindividual and regional variations were observed, whereas the strongest difference was found between osteonal and interstitial tissues (Z: 7.2 {+-} 1.1 Mrayl versus 9.3 {+-} 1.0 Mrayl, DMB: 1.06 {+-} 0.07 g cm{sup -3} versus 1.16 {+-} 0.05 g cm{sup -3}, paired t-test, p < 0.05). Weak, but significant correlations between DMB and Z were obtained for the osteonal (R{sup 2} = 0.174, p < 10{sup -4}) and for the pooled (osteonal and interstitial) data. The regression of the pooled osteonal and interstitial tissue data follows a second-order polynomial (R{sup 2} = 0.39, p < 10{sup -4}). Both modalities fulfil the requirement for a simultaneous evaluation of cortical bone microstructure and material properties at the tissue level. While SAM inspection is limited to the evaluation of carefully prepared sample surfaces, SR-{mu}CT provides volumetric information on the tissue without substantial preparation requirements. However, SAM provides a quantitative estimate of elastic properties at the tissue level that cannot be captured by SR-{mu}CT.

  11. Acoustic Microscopy at Cryogenic Temperatures.

    Science.gov (United States)

    1983-09-01

    L IIIIIrLL I~llI Illl ’.___- IImIIII...!~... 1.8 MICRO PY R[,oLUfroN uSF C HAPI NA: IN A t M I NC IA ACOUSTIC MICROSCOPY AT CRYOGENIC TEMPERATURES...ORGANIZATION NAME AND ADDRESS 10, PROGRAM ELEMENT. PROJECT, TASK Edward L. Ginzton Laboratory AREA & WORK UNfT UMBERS W.W. Hansen Laboratories of...microscope. As a follow-on to this work we are now planning to double the frequency to 8 GHz. The preliminary testing has been done and it now appears

  12. QUANTITATIVE CONFOCAL LASER SCANNING MICROSCOPY

    Directory of Open Access Journals (Sweden)

    Merete Krog Raarup

    2011-05-01

    Full Text Available This paper discusses recent advances in confocal laser scanning microscopy (CLSM for imaging of 3D structure as well as quantitative characterization of biomolecular interactions and diffusion behaviour by means of one- and two-photon excitation. The use of CLSM for improved stereological length estimation in thick (up to 0.5 mm tissue is proposed. The techniques of FRET (Fluorescence Resonance Energy Transfer, FLIM (Fluorescence Lifetime Imaging Microscopy, FCS (Fluorescence Correlation Spectroscopy and FRAP (Fluorescence Recovery After Photobleaching are introduced and their applicability for quantitative imaging of biomolecular (co-localization and trafficking in live cells described. The advantage of two-photon versus one-photon excitation in relation to these techniques is discussed.

  13. Multifunctional scanning ion conductance microscopy.

    Science.gov (United States)

    Page, Ashley; Perry, David; Unwin, Patrick R

    2017-04-01

    Scanning ion conductance microscopy (SICM) is a nanopipette-based technique that has traditionally been used to image topography or to deliver species to an interface, particularly in a biological setting. This article highlights the recent blossoming of SICM into a technique with a much greater diversity of applications and capability that can be used either standalone, with advanced control (potential-time) functions, or in tandem with other methods. SICM can be used to elucidate functional information about interfaces, such as surface charge density or electrochemical activity (ion fluxes). Using a multi-barrel probe format, SICM-related techniques can be employed to deposit nanoscale three-dimensional structures and further functionality is realized when SICM is combined with scanning electrochemical microscopy (SECM), with simultaneous measurements from a single probe opening up considerable prospects for multifunctional imaging. SICM studies are greatly enhanced by finite-element method modelling for quantitative treatment of issues such as resolution, surface charge and (tip) geometry effects. SICM is particularly applicable to the study of living systems, notably single cells, although applications extend to materials characterization and to new methods of printing and nanofabrication. A more thorough understanding of the electrochemical principles and properties of SICM provides a foundation for significant applications of SICM in electrochemistry and interfacial science.

  14. Scanning Electrochemical Microscopy in Neuroscience

    Science.gov (United States)

    Schulte, Albert; Nebel, Michaela; Schuhmann, Wolfgang

    2010-07-01

    This article reviews recent work involving the application of scanning electrochemical microscopy (SECM) to the study of individual cultured living cells, with an emphasis on topographical and functional imaging of neuronal and secretory cells of the nervous and endocrine system. The basic principles of biological SECM and associated negative amperometric-feedback and generator/collector-mode SECM imaging are discussed, and successful use of the methodology for screening soft and fragile membranous objects is outlined. The drawbacks of the constant-height mode of probe movement and the benefits of the constant-distance mode of SECM operation are described. Finally, representative examples of constant-height and constant-distance mode SECM on a variety of live cells are highlighted to demonstrate the current status of single-cell SECM in general and of SECM in neuroscience in particular.

  15. Scanning Probe Microscopy of Graphene

    Science.gov (United States)

    Tautz, Pamela

    2011-10-01

    Scanning tunneling microscopy has been used to study the unusual electronic properties of graphene. In an effort to support the graphene with minimal interaction with the substrate, we used a hexagonal boron nitride (hBN) substrate. To minimize contaminants between the CVD graphene and boron nitride, the graphene samples were cleaned with distilled water and isopropanol prior to transfer to hBN substrate. We have also examined the growth of graphene flakes by chemical vapor deposition. In particular, we examined the relationship between the orientations of the first and second layer of CVD grown graphene. We found the growth mechanism preferentially resulted in rotations of 9^o or less indicating flakes with first and second layers aligned.

  16. Acoustic microscopy of internal structure of resistance spot welds.

    Science.gov (United States)

    Chertov, Andrey M; Maev, Roman G; Severin, Fedar M

    2007-08-01

    Acoustic microscopy, although relatively new, has many advantages within the industrial quality control process. Its high degree of sensitivity, resolution, and reliability make it ideal for use in resistance spot weld analysis, aiding in visualization of small-scale nugget failures, as well as other defects, at various depths. Acoustic microscopy makes it possible to inspect fine detail of internal structures, providing reliable inspection and characterization of weld joints. Besides weld size measurements, this technique is able to provide high resolution, three-dimensional images of the weld nuggets, revealing possible imperfections within its microstructure that may affect joint quality. The high degree of accuracy allows one to consider the results of acoustic microscopy an authoritative measure of weld size, particularly in the case of high strength steels, dual phase steel, USIBOR steel, etc. Indeed, this technique is effective even when both conventional ultrasound and hammer and chisel methods are not. In this paper, the potential of scanning acoustic microscopy as a means to provide qualitative and quantitative information about the internal microstructure of the resistance spot welds is demonstrated. Thus, acoustic microscopy is shown to be a unique and effective laboratory instrument for the evaluation and calibration of weld quality.

  17. Scanning tunneling microscopy II further applications and related scanning techniques

    CERN Document Server

    Güntherodt, Hans-Joachim

    1995-01-01

    Scanning Tunneling Microscopy II, like its predecessor, presents detailed and comprehensive accounts of the basic principles and broad range of applications of STM and related scanning probe techniques. The applications discussed in this volume come predominantly from the fields of electrochemistry and biology. In contrast to those described in STM I, these studies may be performed in air and in liquids. The extensions of the basic technique to map other interactions are described in chapters on scanning force microscopy, magnetic force microscopy, and scanning near-field optical microscopy, together with a survey of other related techniques. Also described here is the use of a scanning proximal probe for surface modification. Together, the two volumes give a comprehensive account of experimental aspects of STM. They provide essential reading and reference material for all students and researchers involved in this field. In this second edition the text has been updated and new methods are discussed.

  18. Scanning tunneling microscopy II further applications and related scanning techniques

    CERN Document Server

    Güntherodt, Hans-Joachim

    1992-01-01

    Scanning Tunneling Microscopy II, like its predecessor, presents detailed and comprehensive accounts of the basic principles and broad range of applications of STM and related scanning probe techniques. The applications discussed in this volume come predominantly from the fields of electrochemistry and biology. In contrast to those described in Vol. I, these sudies may be performed in air and in liquids. The extensions of the basic technique to map other interactions are described inchapters on scanning force microscopy, magnetic force microscopy, scanning near-field optical microscopy, together with a survey of other related techniques. Also described here is the use of a scanning proximal probe for surface modification. Togehter, the two volumes give a comprehensive account of experimental aspcets of STM. They provide essentialreading and reference material for all students and researchers involvedin this field.

  19. Introduction to scanning tunneling microscopy

    CERN Document Server

    Chen, C Julian

    2008-01-01

    The scanning tunneling and the atomic force microscope, both capable of imaging individual atoms, were crowned with the Physics Nobel Prize in 1986, and are the cornerstones of nanotechnology today. This is a thoroughly updated version of this 'bible' in the field.

  20. Scanning Electron Microscopy in modern dentistry research

    OpenAIRE

    Paradella, Thaís Cachuté; Unesp-FOSJC; Bottino, Marco Antonio; Unesp-FOSJC

    2012-01-01

    The purpose of this article was to review the usage of Scanning Electron Microscopy (SEM) in dentistry research nowadays, through a careful and updated literature review. By using the key-words Scanning Electron Microscopy and one of the following areas of research in dentistry (Endodontics, Periodontics and Implant), in international database (PubMed), in the year of 2012 (from January to September), a total of 112 articles were found. This data was tabled and the articles were classified ac...

  1. Differential-concentration scanning ion conductance microscopy

    OpenAIRE

    Perry, David; Page, Ashley; Chen, Baoping; Frenguelli, Bruno G.; Unwin, Patrick R.

    2017-01-01

    Scanning ion conductance microscopy (SICM) is a nanopipette-based scanning probe microscopy technique that utilizes the ionic current flowing between an electrode inserted inside a nanopipette probe containing electrolyte solution and a second electrode placed in a bulk electrolyte bath, to provide information on a substrate of interest. For most applications to date, the composition and concentration of the electrolyte inside and outside the nanopipette is identical, but it is shown herein t...

  2. Towards high-speed scanning tunneling microscopy

    NARCIS (Netherlands)

    Tabak, Femke Chantal

    2013-01-01

    In this thesis, two routes towards high-speed scanning tunneling microscopy (STM) are described. The first possibility for high-speed scanning that is discussed is the use of MEMS (Micro-Electro Mechanical Systems) devices as high-speed add-ons in STM microscopes. The functionality of these devices

  3. Spiral scanning method for atomic force microscopy.

    Science.gov (United States)

    Hung, Shao-Kang

    2010-07-01

    A spiral scanning method is proposed for atomic force microscopy with thoroughgoing analysis and implementation. Comparing with the traditional line-by-line scanning method, the spiral scanning method demonstrates higher imaging speed, minor image distortion, and lower acceleration, which can damage the piezoelectric scanner. Employing the spiral scanning method to replace the line-by-line scanning method, the experiment shows that the time to complete an imaging cycle can be reduced from 800 s to 314 s without sacrificing the image resolution.

  4. Scanning electron microscopy of bone.

    Science.gov (United States)

    Boyde, Alan

    2012-01-01

    This chapter described methods for Scanning Electron Microscopical imaging of bone and bone cells. Backscattered electron (BSE) imaging is by far the most useful in the bone field, followed by secondary electrons (SE) and the energy dispersive X-ray (EDX) analytical modes. This chapter considers preparing and imaging samples of unembedded bone having 3D detail in a 3D surface, topography-free, polished or micromilled, resin-embedded block surfaces, and resin casts of space in bone matrix. The chapter considers methods for fixation, drying, looking at undersides of bone cells, and coating. Maceration with alkaline bacterial pronase, hypochlorite, hydrogen peroxide, and sodium or potassium hydroxide to remove cells and unmineralised matrix is described in detail. Attention is given especially to methods for 3D BSE SEM imaging of bone samples and recommendations for the types of resin embedding of bone for BSE imaging are given. Correlated confocal and SEM imaging of PMMA-embedded bone requires the use of glycerol to coverslip. Cathodoluminescence (CL) mode SEM imaging is an alternative for visualising fluorescent mineralising front labels such as calcein and tetracyclines. Making spatial casts from PMMA or other resin embedded samples is an important use of this material. Correlation with other imaging means, including microradiography and microtomography is important. Shipping wet bone samples between labs is best done in glycerol. Environmental SEM (ESEM, controlled vacuum mode) is valuable in eliminating -"charging" problems which are common with complex, cancellous bone samples.

  5. Semiconductor Surface Characterization by Scanning Probe Microscopies

    Science.gov (United States)

    2001-01-01

    potentiometry (STP)8 and ballistic electron emission microscopy (BEEM)9 which allow mapping of lateral surface potential and local subsurface Schottky...A.P.Fein. "Tunneling Spectroscopy of the Si(1 1 1)2xl Surface", Surf.Sci. 181, 295- 306, 1987. 8. P.Muralt, D.W.Pohl, "Scanning tunneling potentiometry

  6. Scanning electron microscopy study of Trichomonas gallinae.

    Science.gov (United States)

    Tasca, Tiana; De Carli, Geraldo A

    2003-12-01

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

  7. Laser scanning laser diode photoacoustic microscopy system.

    Science.gov (United States)

    Erfanzadeh, Mohsen; Kumavor, Patrick D; Zhu, Quing

    2018-03-01

    The development of low-cost and fast photoacoustic microscopy systems enhances the clinical applicability of photoacoustic imaging systems. To this end, we present a laser scanning laser diode-based photoacoustic microscopy system. In this system, a 905 nm, 325 W maximum output peak power pulsed laser diode with 50 ns pulsewidth is utilized as the light source. A combination of aspheric and cylindrical lenses is used for collimation of the laser diode beam. Two galvanometer scanning mirrors steer the beam across a focusing aspheric lens. The lateral resolution of the system was measured to be ∼21 μm using edge spread function estimation. No averaging was performed during data acquisition. The imaging speed is ∼370 A-lines per second. Photoacoustic microscopy images of human hairs, ex vivo mouse ear, and ex vivo porcine ovary are presented to demonstrate the feasibility and potentials of the proposed system.

  8. Aberration corrected Lorentz scanning transmission electron microscopy

    Energy Technology Data Exchange (ETDEWEB)

    McVitie, S., E-mail: stephen.mcvitie@glasgow.ac.uk; McGrouther, D.; McFadzean, S.; MacLaren, D.A.; O’Shea, K.J.; Benitez, M.J.

    2015-05-15

    We present results from an aberration corrected scanning transmission electron microscope which has been customised for high resolution quantitative Lorentz microscopy with the sample located in a magnetic field free or low field environment. We discuss the innovations in microscope instrumentation and additional hardware that underpin the imaging improvements in resolution and detection with a focus on developments in differential phase contrast microscopy. Examples from materials possessing nanometre scale variations in magnetisation illustrate the potential for aberration corrected Lorentz imaging as a tool to further our understanding of magnetism on this lengthscale. - Highlights: • Demonstration of nanometre scale resolution in magnetic field free environment using aberration correction in the scanning transmission electron microscope (STEM). • Implementation of differential phase contrast mode of Lorentz microscopy in aberration corrected STEM with improved sensitivity. • Quantitative imaging of magnetic induction of nanostructures in amorphous and cross-section samples.

  9. Scanning Ion Conductance Microscopy of Live Keratinocytes

    Science.gov (United States)

    Hegde, V.; Mason, A.; Saliev, T.; Smith, F. J. D.; McLean, W. H. I.; Campbell, P. A.

    2012-07-01

    Scanning ion conductance microscopy (SICM) is perhaps the least well known technique from the scanning probe microscopy (SPM) family of instruments. As with its more familiar counterpart, atomic force microscopy (AFM), the technique provides high-resolution topographic imaging, with the caveat that target structures must be immersed in a conducting solution so that a controllable ion current may be utilised as the basis for feedback. In operation, this non-contact characteristic of SICM makes it ideal for the study of delicate structures, such as live cells. Moreover, the intrinsic architecture of the instrument, incorporating as it does, a scanned micropipette, lends itself to combination approaches with complementary techniques such as patch-clamp electrophysiology: SICM therefore boasts the capability for both structural and functional imaging. For the present observations, an ICnano S system (Ionscope Ltd., Melbourn, UK) operating in 'hopping mode' was used, with the objective of assessing the instrument's utility for imaging live keratinocytes under physiological buffers. In scans employing cultured HaCaT cells (spontaneously immortalised, human keratinocytes), we compared the qualitative differences of live cells imaged with SICM and AFM, and also with their respective counterparts after chemical fixation in 4% paraformaldehyde. Characteristic surface microvilli were particularly prominent in live cell imaging by SICM. Moreover, time lapse SICM imaging on live cells revealed that changes in the pattern of microvilli could be tracked over time. By comparison, AFM imaging on live cells, even at very low contact forces (monitoring the most delicate living structures with attendant high spatial resolutions.

  10. Analysing magnetism using scanning SQUID microscopy.

    Science.gov (United States)

    Reith, P; Renshaw Wang, X; Hilgenkamp, H

    2017-12-01

    Scanning superconducting quantum interference device microscopy (SSM) is a scanning probe technique that images local magnetic flux, which allows for mapping of magnetic fields with high field and spatial accuracy. Many studies involving SSM have been published in the last few decades, using SSM to make qualitative statements about magnetism. However, quantitative analysis using SSM has received less attention. In this work, we discuss several aspects of interpreting SSM images and methods to improve quantitative analysis. First, we analyse the spatial resolution and how it depends on several factors. Second, we discuss the analysis of SSM scans and the information obtained from the SSM data. Using simulations, we show how signals evolve as a function of changing scan height, SQUID loop size, magnetization strength, and orientation. We also investigated 2-dimensional autocorrelation analysis to extract information about the size, shape, and symmetry of magnetic features. Finally, we provide an outlook on possible future applications and improvements.

  11. Scanning electron microscopy of superficial white onychomycosis*

    Science.gov (United States)

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

    2015-01-01

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

  12. Scanning electron microscopy of molluscum contagiosum*

    OpenAIRE

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

    2013-01-01

    Molluscum contagiosum is a disease caused by a poxvirus. It is more prevalent in children up to 5 years of age. There is a second peak of incidence in young adults. In order to examine its ultrastructure, three lesions were curetted without disruption, cut transversely with a scalpel, and routinely processed for scanning electron microscopy (SEM). The oval structure of molluscum contagiosum could be easily identified. In its core, there was a central umbilication and just below this depressio...

  13. Scanning electron microscopy of cold gases

    Science.gov (United States)

    Santra, Bodhaditya; Ott, Herwig

    2015-06-01

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

  14. Investigation into scanning tunnelling luminescence microscopy

    CERN Document Server

    Manson-Smith, S K

    2001-01-01

    This work reports on the development of a scanning tunnelling luminescence (STL) microscope and its application to the study of Ill-nitride semiconductor materials used in the production of light emitting devices. STL microscopy is a technique which uses the high resolution topographic imaging capabilities of the scanning tunnelling microscope (STM) to generate high resolution luminescence images. The STM tunnelling current acts as a highly localised source of electrons (or holes) which generates luminescence in certain materials. Light generated at the STM tunnelling junction is collected concurrently with the height variation of the tunnelling probe as it is scanned across a sample surface, producing simultaneous topographic and luminescence images. Due to the very localised excitation source, high resolution luminescence images can be obtained. Spectroscopic resolution can be obtained by using filters. Additionally, the variation of luminescence intensity with tunnel current and with bias voltage can provi...

  15. Interferometric Synthetic Aperture Microscopy: Computed Imaging for Scanned Coherent Microscopy

    Directory of Open Access Journals (Sweden)

    Stephen A. Boppart

    2008-06-01

    Full Text Available Three-dimensional image formation in microscopy is greatly enhanced by the use of computed imaging techniques. In particular, Interferometric Synthetic Aperture Microscopy (ISAM allows the removal of out-of-focus blur in broadband, coherent microscopy. Earlier methods, such as optical coherence tomography (OCT, utilize interferometric ranging, but do not apply computed imaging methods and therefore must scan the focal depth to acquire extended volumetric images. ISAM removes the need to scan the focus by allowing volumetric image reconstruction from data collected at a single focal depth. ISAM signal processing techniques are similar to the Fourier migration methods of seismology and the Fourier reconstruction methods of Synthetic Aperture Radar (SAR. In this article ISAM is described and the close ties between ISAM and SAR are explored. ISAM and a simple strip-map SAR system are placed in a common mathematical framework and compared to OCT and radar respectively. This article is intended to serve as a review of ISAM, and will be especially useful to readers with a background in SAR.

  16. Scanning Probe Microscopy of Organic Solar Cells

    Science.gov (United States)

    Reid, Obadiah G.

    Nanostructured composites of organic semiconductors are a promising class of materials for the manufacture of low-cost solar cells. Understanding how the nanoscale morphology of these materials affects their efficiency as solar energy harvesters is crucial to their eventual potential for large-scale deployment for primary power generation. In this thesis we describe the use of optoelectronic scanning-probe based microscopy methods to study this efficiency-structure relationship with nanoscale resolution. In particular, our objective is to make spatially resolved measurements of each step in the power conversion process from photons to an electric current, including charge generation, transport, and recombination processes, and correlate them with local device structure. We have achieved two aims in this work: first, to develop and apply novel electrically sensitive scanning probe microscopy experiments to study the optoelectronic materials and processes discussed above; and second, to deepen our understanding of the physics underpinning our experimental techniques. In the first case, we have applied conductive-, and photoconductive atomic force (cAFM & pcAFM) microscopy to measure both local photocurrent collection and dark charge transport properties in a variety of model and novel organic solar cell composites, including polymer/fullerene blends, and polymer-nanowire/fullerene blends, finding that local heterogeneity is the rule, and that improvements in the uniformity of specific beneficial nanostructures could lead to large increases in efficiency. We have used scanning Kelvin probe microscopy (SKPM) and time resolved-electrostatic force microscopy (trEFM) to characterize all-polymer blends, quantifying their sensitivity to photochemical degradation and the subsequent formation of local charge traps. We find that while trEFM provides a sensitive measure of local quantum efficiency, SKPM is generally unsuited to measurements of efficiency, less sensitive than tr

  17. Chemical Phenomena of Atomic Force Microscopy Scanning.

    Science.gov (United States)

    Ievlev, Anton V; Brown, Chance; Burch, Matthew J; Agar, Joshua C; Velarde, Gabriel A; Martin, Lane W; Maksymovych, Petro; Kalinin, Sergei V; Ovchinnikova, Olga S

    2018-02-12

    Atomic force microscopy is widely used for nanoscale characterization of materials by scientists worldwide. The long-held belief of ambient AFM is that the tip is generally chemically inert but can be functionalized with respect to the studied sample. This implies that basic imaging and scanning procedures do not affect surface and bulk chemistry of the studied sample. However, an in-depth study of the confined chemical processes taking place at the tip-surface junction and the associated chemical changes to the material surface have been missing as of now. Here, we used a hybrid system that combines time-of-flight secondary ion mass spectrometry with an atomic force microscopy to investigate the chemical interactions that take place at the tip-surface junction. Investigations showed that even basic contact mode AFM scanning is able to modify the surface of the studied sample. In particular, we found that the silicone oils deposited from the AFM tip into the scanned regions and spread to distances exceeding 15 μm from the tip. These oils were determined to come from standard gel boxes used for the storage of the tips. The explored phenomena are important for interpreting and understanding results of AFM mechanical and electrical studies relying on the state of the tip-surface junction.

  18. Hollow-tip scanning photoelectron microscopy

    Science.gov (United States)

    Cherkun, A. P.; Mironov, B. N.; Aseyev, S. A.; Chekalin, S. V.

    2014-07-01

    A new type of microscopy based on scanning in vacuum by a beam of charged particles transmitted through a hollow probe has been implemented. This approach provides controllable motion of spatially localized ion, electron, molecular (atomic), and soft X-ray beams and investigation of the surface in the shear force mode. In the photoelectron mode, in which electrons are transmitted through a 2-μm quartz capillary, a surface profile of gadolinium irradiated by 400-nm femtosecond laser pulses has been visualized with a subwave spatial resolution. The new method of microscopy opens an opportunity of investigations in the field of nanometer local photodesorption of molecular ions (one of the last ideas of V.S. Letokhov).

  19. Soft stylus probes for scanning electrochemical microscopy.

    Science.gov (United States)

    Cortés-Salazar, Fernando; Träuble, Markus; Li, Fei; Busnel, Jean-Marc; Gassner, Anne-Laure; Hojeij, Mohamad; Wittstock, Gunther; Girault, Hubert H

    2009-08-15

    A soft stylus microelectrode probe has been developed to carry out scanning electrochemical microscopy (SECM) of rough, tilted, and large substrates in contact mode. It is fabricated by first ablating a microchannel in a polyethylene terephthalate thin film and filling it with a conductive carbon ink. After curing the carbon track and lamination with a polymer film, the V-shaped stylus was cut thereby forming a probe, with the cross section of the carbon track at the tip being exposed either by UV-photoablation machining or by blade cutting followed by polishing to produce a crescent moon-shaped carbon microelectrode. The probe properties have been assessed by cyclic voltammetry, approach curves, and line scans over electrochemically active and inactive substrates of different roughness. The influence of probe bending on contact mode imaging was then characterized using simple patterns. Boundary element method simulations were employed to rationalize the distance-dependent electrochemical response of the soft stylus probes.

  20. Water-Immersible MEMS scanning mirror designed for wide-field fast-scanning photoacoustic microscopy

    Science.gov (United States)

    Yao, Junjie; Huang, Chih-Hsien; Martel, Catherine; Maslov, Konstantin I.; Wang, Lidai; Yang, Joon-Mo; Gao, Liang; Randolph, Gwendalyn; Zou, Jun; Wang, Lihong V.

    2013-03-01

    By offering images with high spatial resolution and unique optical absorption contrast, optical-resolution photoacoustic microscopy (OR-PAM) has gained increasing attention in biomedical research. Recent developments in OR-PAM have improved its imaging speed, but have sacrificed either the detection sensitivity or field of view or both. We have developed a wide-field fast-scanning OR-PAM by using a water-immersible MEMS scanning mirror (MEMS-ORPAM). Made of silicon with a gold coating, the MEMS mirror plate can reflect both optical and acoustic beams. Because it uses an electromagnetic driving force, the whole MEMS scanning system can be submerged in water. In MEMS-ORPAM, the optical and acoustic beams are confocally configured and simultaneously steered, which ensures uniform detection sensitivity. A B-scan imaging speed as high as 400 Hz can be achieved over a 3 mm scanning range. A diffraction-limited lateral resolution of 2.4 μm in water and a maximum imaging depth of 1.1 mm in soft tissue have been experimentally determined. Using the system, we imaged the flow dynamics of both red blood cells and carbon particles in a mouse ear in vivo. By using Evans blue dye as the contrast agent, we also imaged the flow dynamics of lymphatic vessels in a mouse tail in vivo. The results show that MEMS-OR-PAM could be a powerful tool for studying highly dynamic and time-sensitive biological phenomena.

  1. Differential-Concentration Scanning Ion Conductance Microscopy.

    Science.gov (United States)

    Perry, David; Page, Ashley; Chen, Baoping; Frenguelli, Bruno G; Unwin, Patrick R

    2017-11-21

    Scanning ion conductance microscopy (SICM) is a nanopipette-based scanning probe microscopy technique that utilizes the ionic current flowing between an electrode inserted inside a nanopipette probe containing electrolyte solution and a second electrode placed in a bulk electrolyte bath, to provide information on a substrate of interest. For most applications to date, the composition and concentration of the electrolyte inside and outside the nanopipette is identical, but it is shown herein that it can be very beneficial to lift this restriction. In particular, an ionic concentration gradient at the end of the nanopipette, generates an ionic current with a greatly reduced electric field strength, with particular benefits for live cell imaging. This differential concentration mode of SICM (ΔC-SICM) also enhances surface charge measurements and provides a new way to carry out reaction mapping measurements at surfaces using the tip for simultaneous delivery and sensing of the reaction rate. Comprehensive finite element method (FEM) modeling has been undertaken to enhance understanding of SICM as an electrochemical cell and to enable the interpretation and optimization of experiments. It is shown that electroosmotic flow (EOF) has much more influence on the nanopipette response in the ΔC-SICM configuration compared to standard SICM modes. The general model presented advances previous treatments, and it provides a framework for quantitative SICM studies.

  2. Phase-contrast scanning transmission electron microscopy.

    Science.gov (United States)

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

    2015-06-01

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

  3. Angular Approach Scanning Ion Conductance Microscopy.

    Science.gov (United States)

    Shevchuk, Andrew; Tokar, Sergiy; Gopal, Sahana; Sanchez-Alonso, Jose L; Tarasov, Andrei I; Vélez-Ortega, A Catalina; Chiappini, Ciro; Rorsman, Patrik; Stevens, Molly M; Gorelik, Julia; Frolenkov, Gregory I; Klenerman, David; Korchev, Yuri E

    2016-05-24

    Scanning ion conductance microscopy (SICM) is a super-resolution live imaging technique that uses a glass nanopipette as an imaging probe to produce three-dimensional (3D) images of cell surface. SICM can be used to analyze cell morphology at nanoscale, follow membrane dynamics, precisely position an imaging nanopipette close to a structure of interest, and use it to obtain ion channel recordings or locally apply stimuli or drugs. Practical implementations of these SICM advantages, however, are often complicated due to the limitations of currently available SICM systems that inherited their design from other scanning probe microscopes in which the scan assembly is placed right above the specimen. Such arrangement makes the setting of optimal illumination necessary for phase contrast or the use of high magnification upright optics difficult. Here, we describe the designs that allow mounting SICM scan head on a standard patch-clamp micromanipulator and imaging the sample at an adjustable approach angle. This angle could be as shallow as the approach angle of a patch-clamp pipette between a water immersion objective and the specimen. Using this angular approach SICM, we obtained topographical images of cells grown on nontransparent nanoneedle arrays, of islets of Langerhans, and of hippocampal neurons under upright optical microscope. We also imaged previously inaccessible areas of cells such as the side surfaces of the hair cell stereocilia and the intercalated disks of isolated cardiac myocytes, and performed targeted patch-clamp recordings from the latter. Thus, our new, to our knowledge, angular approach SICM allows imaging of living cells on nontransparent substrates and a seamless integration with most patch-clamp setups on either inverted or upright microscopes, which would facilitate research in cell biophysics and physiology. Copyright © 2016 Biophysical Society. Published by Elsevier Inc. All rights reserved.

  4. Scanning electron microscopy of molluscum contagiosum*

    Science.gov (United States)

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

    2013-01-01

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

  5. Spin-polarized scanning tunnelling microscopy

    CERN Document Server

    Bode, M

    2003-01-01

    The recent experimental progress in spin-polarized scanning tunnelling microscopy (SP-STM) - a magnetically sensitive imaging technique with ultra-high resolution - is reviewed. The basics of spin-polarized electron tunnelling are introduced as they have been investigated in planar tunnel junctions for different electrode materials, i.e. superconductors, optically excited GaAs, and ferromagnets. It is shown that ferromagnets and antiferromagnets are suitable tip materials for the realization of SP-STM. Possible tip designs and modes of operations are discussed for both classes of materials. The results of recent spatially resolved measurements as performed with different magnetic probe tips and using different modes of operation are reviewed and discussed in terms of applicability to surfaces, thin films, and nanoparticles. The limits of spatial resolution, and the impact of an external magnetic field on the imaging process.

  6. A dark mode in scanning thermal microscopy

    Science.gov (United States)

    Ramiandrisoa, Liana; Allard, Alexandre; Joumani, Youssef; Hay, Bruno; Gomés, Séverine

    2017-12-01

    The need for high lateral spatial resolution in thermal science using Scanning Thermal Microscopy (SThM) has pushed researchers to look for more and more tiny probes. SThM probes have consequently become more and more sensitive to the size effects that occur within the probe, the sample, and their interaction. Reducing the tip furthermore induces very small heat flux exchanged between the probe and the sample. The measurement of this flux, which is exploited to characterize the sample thermal properties, requires then an accurate thermal management of the probe-sample system and to reduce any phenomenon parasitic to this system. Classical experimental methodologies must then be constantly questioned to hope for relevant and interpretable results. In this paper, we demonstrate and estimate the influence of the laser of the optical force detection system used in the common SThM setup that is based on atomic-force microscopy equipment on SThM measurements. We highlight the bias induced by the overheating due to the laser illumination on the measurements performed by thermoresistive probes (palladium probe from Kelvin Nanotechnology). To face this issue, we propose a new experimental procedure based on a metrological approach of the measurement: a SThM "dark mode." The comparison with the classical procedure using the laser shows that errors between 14% and 37% can be reached on the experimental data exploited to determine the heat flux transferred from the hot probe to the sample.

  7. Comparison between optical-resolution photoacoustic microscopy and confocal laser scanning microscopy for turbid sample imaging.

    Science.gov (United States)

    U-Thainual, Paweena; Kim, Do-Hyun

    2015-12-01

    Optical-resolution photoacoustic microscopy (ORPAM) in theory provides lateral resolution equivalent to the optical diffraction limit. Scattering media, such as biological turbid media, attenuates the optical signal and also alters the diffraction-limited spot size of the focused beam. The ORPAM signal is generated only from a small voxel in scattering media with dimensions equivalent to the laser spot size after passing through scattering layers and is detected by an acoustic transducer, which is not affected by optical scattering. Thus, both ORPAM and confocal laser scanning microscopy (CLSM) reject scattered light. A multimodal optical microscopy platform that includes ORPAM and CLSM was constructed, and the lateral resolution of both modes was measured using patterned thin metal film with and without a scattering barrier. The effect of scattering media on the lateral resolution was studied using different scattering coefficients and was compared to computational results based on Monte Carlo simulations. It was found that degradation of lateral resolution due to optical scattering was not significant for either ORPAM or CLSM. The depth discrimination capability of ORPAM and CLSM was measured using microfiber embedded in a light scattering phantom material. ORPAM images demonstrated higher contrast compared to CLSM images partly due to reduced acoustic signal scattering.

  8. Scanned probe microscopy for thin film superconductor development

    Energy Technology Data Exchange (ETDEWEB)

    Moreland, J. [National Institute of Standards and Technology, Boulder, CO (United States)

    1996-12-31

    Scanned probe microscopy is a general term encompassing the science of imaging based on piezoelectric driven probes for measuring local changes in nanoscale properties of materials and devices. Techniques like scanning tunneling microscopy, atomic force microscopy, and scanning potentiometry are becoming common tools in the production and development labs in the semiconductor industry. The author presents several examples of applications specific to the development of high temperature superconducting thin films and thin-film devices.

  9. EDITORIAL: Scanning probe microscopy: a visionary development Scanning probe microscopy: a visionary development

    Science.gov (United States)

    Demming, Anna

    2013-07-01

    The development of scanning probe microscopy repositioned modern physics. When Rohrer and Binnig first used electronic tunnelling effects to image atoms and quantum states they did more than pin down theoretical hypotheses to real-world observables; the scanning tunnelling microscope fed imaginations, prompting researchers to consider new directions and possibilities [1]. As Rohrer once commented, 'We could show that you can easily manipulate or position something small in space with an accuracy of 10 pm.... When you can do that, you simply have ideas of what you can do' [2]. The development heralded a cavalry of scanning probe techniques—such as atomic force microscopy (AFM) [3-5], scanning near-field optical microscopy (SNOM) [6-8] and Kelvin probe force microscopy (KPFM) [9, 10]—that still continue to bring nanomaterials and nanoscale phenomena into fresh focus. Not long after the development of scanning tunnelling microscopy, Binnig, Quate and Gerber collaborating in California in the US published work on a new type of microscope also capable of atomic level resolution [3]. The original concept behind scanning tunnelling microscopy uses electrical conductance, which places substantial limitations on the systems that it can image. Binnig, Quate and Gerber developed the AFM to 'feel' the topology of surfaces like the needle of an old fashioned vinyl player. In this way insulators could be imaged as well. The development of a force modulation mode AFM extended the tool's reach to soft materials making images of biological samples accessible with the technique [4]. There have now been a number of demonstrations of image capture at rates that allow dynamics at the nanoscale to be tracked in real time, opening further possibilities in applications of the AFM as described in a recent review by Toshio Ando at Kanazawa University [5]. Researchers also found a way to retrieve optical information at 'super-resolution' [6, 7]. Optical microscopy provides spectral

  10. Scanning transmission electron microscopy imaging and analysis

    CERN Document Server

    Pennycook, Stephen J

    2011-01-01

    Provides the first comprehensive treatment of the physics and applications of this mainstream technique for imaging and analysis at the atomic level Presents applications of STEM in condensed matter physics, materials science, catalysis, and nanoscience Suitable for graduate students learning microscopy, researchers wishing to utilize STEM, as well as for specialists in other areas of microscopy Edited and written by leading researchers and practitioners

  11. Optimal lens design and use in laser-scanning microscopy.

    NARCIS (Netherlands)

    Negrean, A.; Mansvelder, H.D.

    2014-01-01

    In laser-scanning microscopy often an off-the-shelf achromatic doublet is used as a scan lens which can reduce the available diffraction-limited field-of-view (FOV) by a factor of 3 and introduce chromatic aberrations that are scan angle dependent. Here we present several simple lens designs of

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

    Energy Technology Data Exchange (ETDEWEB)

    Tong Yongpeng [Institute of Nuclear Techniques, Shenzhen University, Nanhai Avenue 3688, Shenzhen 518060 (China)], E-mail: yongpengt@yahoo.com.cn; Li Changming [School of Chemical and Biomedical Engineering, Nanyang Technological University, Singapore 637457 (Singapore); Liang Feng [Institute Pasteur of Shanghai, Chinese Academy of Sciences, Shanghai 200025 (China); Chen Jianmin [Shenzhen Municipal Hospital for Chronic Disease Control and Prevention, Guangdong 518020 (China); Zhang Hong; Liu Guoqing; Sun Huibin [Institute of Nuclear Techniques, Shenzhen University, Nanhai Avenue 3688, Shenzhen 518060 (China); Luong, John H.T. [Biotechnology Research Institute, National Research Council Canada, Montreal, Quebec, H4P 2R2 (Canada)

    2008-12-15

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

  13. Combined frequency modulated atomic force microscopy and scanning tunneling microscopy detection for multi-tip scanning probe microscopy applications

    Science.gov (United States)

    Morawski, Ireneusz; Spiegelberg, Richard; Korte, Stefan; Voigtländer, Bert

    2015-12-01

    A method which allows scanning tunneling microscopy (STM) tip biasing independent of the sample bias during frequency modulated atomic force microscopy (AFM) operation is presented. The AFM sensor is supplied by an electronic circuit combining both a frequency shift signal and a tunneling current signal by means of an inductive coupling. This solution enables a control of the tip potential independent of the sample potential. Individual tip biasing is specifically important in order to implement multi-tip STM/AFM applications. An extensional quartz sensor (needle sensor) with a conductive tip is applied to record simultaneously topography and conductivity of the sample. The high resonance frequency of the needle sensor (1 MHz) allows scanning of a large area of the surface being investigated in a reasonably short time. A recipe for the amplitude calibration which is based only on the frequency shift signal and does not require the tip being in contact is presented. Additionally, we show spectral measurements of the mechanical vibration noise of the scanning system used in the investigations.

  14. Plant cell wall characterization using scanning probe microscopy techniques

    Science.gov (United States)

    Yarbrough, John M; Himmel, Michael E; Ding, Shi-You

    2009-01-01

    Lignocellulosic biomass is today considered a promising renewable resource for bioenergy production. A combined chemical and biological process is currently under consideration for the conversion of polysaccharides from plant cell wall materials, mainly cellulose and hemicelluloses, to simple sugars that can be fermented to biofuels. Native plant cellulose forms nanometer-scale microfibrils that are embedded in a polymeric network of hemicelluloses, pectins, and lignins; this explains, in part, the recalcitrance of biomass to deconstruction. The chemical and structural characteristics of these plant cell wall constituents remain largely unknown today. Scanning probe microscopy techniques, particularly atomic force microscopy and its application in characterizing plant cell wall structure, are reviewed here. We also further discuss future developments based on scanning probe microscopy techniques that combine linear and nonlinear optical techniques to characterize plant cell wall nanometer-scale structures, specifically apertureless near-field scanning optical microscopy and coherent anti-Stokes Raman scattering microscopy. PMID:19703302

  15. Plant cell wall characterization using scanning probe microscopy techniques

    Directory of Open Access Journals (Sweden)

    Himmel Michael E

    2009-08-01

    Full Text Available Abstract Lignocellulosic biomass is today considered a promising renewable resource for bioenergy production. A combined chemical and biological process is currently under consideration for the conversion of polysaccharides from plant cell wall materials, mainly cellulose and hemicelluloses, to simple sugars that can be fermented to biofuels. Native plant cellulose forms nanometer-scale microfibrils that are embedded in a polymeric network of hemicelluloses, pectins, and lignins; this explains, in part, the recalcitrance of biomass to deconstruction. The chemical and structural characteristics of these plant cell wall constituents remain largely unknown today. Scanning probe microscopy techniques, particularly atomic force microscopy and its application in characterizing plant cell wall structure, are reviewed here. We also further discuss future developments based on scanning probe microscopy techniques that combine linear and nonlinear optical techniques to characterize plant cell wall nanometer-scale structures, specifically apertureless near-field scanning optical microscopy and coherent anti-Stokes Raman scattering microscopy.

  16. Integrated acoustic-resolution and optical-resolution photoacoustic microscopy using a single multifunctional acoustic lens

    Science.gov (United States)

    Guo, Heng; Xi, Lei

    2016-10-01

    With the rapid development of photoacoustic imaging, it has been widely used in various research fields such as biology, medicine and nanotechnology. Due to the huge difference among photoacoustic imaging systems, it is hard to integrate them in one platform. To solve this problem, we propose to develop a new universal photoacoustic imaging platform that integrates acoustic-resolution photoacoustic microscopy and optical-resolution photoacoustic microscopy through a multifunctional liquid lens. This lens takes advantage of an inherently low acoustic impedance and a tunable focal length that was characterized by the infusion volume of the liquid. In this paper, the liquid lens was used to realize confocal of laser illumination and acoustic detection for both acoustic-resolution and optical-resolution photoacoustic microscopy. The home-made polyvinylidene fluoride (PVDF) acoustic transducer had a center frequency of 10MHz and -6dB frequency spectrum from 4MHz to 15MHz which yielded to an axial resolution of 70 μm. The lateral resolutions of acoustic- and optical-resolution photoacoustic microscopy were evaluated to be 180 μm and 4.8 μm, respectively. The vasculature of rat ears was carried out to evaluate the performance of optical-resolution photoacoustic microscopy.

  17. Full information acquisition in scanning probe microscopy and spectroscopy

    Science.gov (United States)

    Jesse, Stephen; Belianinov, Alex; Kalinin, Sergei V.; Somnath, Suhas

    2017-04-04

    Apparatus and methods are described for scanning probe microscopy and spectroscopy based on acquisition of full probe response. The full probe response contains valuable information about the probe-sample interaction that is lost in traditional scanning probe microscopy and spectroscopy methods. The full probe response is analyzed post data acquisition using fast Fourier transform and adaptive filtering, as well as multivariate analysis. The full response data is further compressed to retain only statistically significant components before being permanently stored.

  18. Further observations on cerebellar climbing fibers. A study by means of light microscopy, confocal laser scanning microscopy and scanning and transmission electron microscopy.

    Science.gov (United States)

    Castejón, O J; Castejón, H V; Alvarado, M V

    2000-12-01

    The intracortical pathways of climbing fibers were traced in several vertebrate cerebella using light microscopy, confocal laser scanning microscopy, scanning and transmission electron microscopy. They were identified as fine fibers up to 1(micron thick, with a characteristic crossing-over bifurcation pattern. Climbing fiber collaterals were tridimensionally visualized forming thin climbing fiber glomeruli in the granular layer. Confocal laser scanning microscopy revealed three types of collateral processes at the interface between granular and Purkinje cell layers. Scanning electron microscopy showed climbing fiber retrograde collaterals in the molecular layer. Asymmetric synaptic contacts of climbing fibers with Purkinje dendritic spines and stellate neuron dendrites were characterized by transmission electron microscopy. Correlative microscopy allowed us to obtain the basic three-dimensional morphological features of climbing fibers in several vertebrates and to show with more accuracy a higher degree of lateral collateralization of these fibers within the cerebellar cortex. The correlative microscopy approach provides new views in the cerebellar cortex information processing.

  19. Optomechatronics Design and Control for Confocal Laser Scanning Microscopy

    NARCIS (Netherlands)

    Yoo, H.W.

    2015-01-01

    Confocal laser scanning microscopy (CLSM) is considered as one of the major advancements in microscopy in the last century and is widely accepted as a 3D fluorescence imaging tool for biological studies. For the emerging biological questions CLSM requires fast imaging to detect rapid biological

  20. Domain characterization of Pb(Zn1/3Nb2/3)O3-(6%-7%)PbTiO3 single crystals using scanning electron acoustic microscopy

    Science.gov (United States)

    Wong, Meng Fei; Heng, Xiangxin; Zeng, Kaiyang

    2008-10-01

    Domain structures of [001]T and [011]T-cut Pb(Zn1/3Nb2/3)O3-(6%-7%)PbTiO3 (PZN-PT) single crystals are studied using scanning electron acoustic microscope (SEAM) technique. The observation of the orientation of domain walls agree reasonably well with the trigonometric projection of rhombohedral and orthorhombic dipoles on the (001) and (011) surfaces, respectively. After mechanical loading with microindentation, domain switching is also observed to form a hyperbolic butterfly shape and extend preferentially along four diagonal directions, i.e., ⟨110⟩ on (001) surface and ⟨111¯⟩ on (011) surface. The critical shear stress to cause domain switching for PZN-PT crystal is estimated to be approximately 49 MPa for both {110} and {111¯} planes based on theoretical analysis. Generally, the SEAM technique has been successfully demonstrated to be a valid technique for observation of domain structures in single crystal PZN-PTs.

  1. Towards Automated Nanomanipulation under Scanning Electron Microscopy

    Science.gov (United States)

    Ye, Xutao

    Robotic Nanomaterial Manipulation inside scanning electron microscopes (SEM) is useful for prototyping functional devices and characterizing one-dimensional nanomaterial's properties. Conventionally, manipulation of nanowires has been performed via teleoperation, which is time-consuming and highly skill-dependent. Manual manipulation also has the limitation of low success rates and poor reproducibility. This research focuses on a robotic system capable of automated pick-place of single nanowires. Through SEM visual detection and vision-based motion control, the system transferred individual silicon nanowires from their growth substrate to a microelectromechanical systems (MEMS) device that characterized the nanowires' electromechanical properties. The performances of the nanorobotic pick-up and placement procedures were quantified by experiments. The system demonstrated automated nanowire pick-up and placement with high reliability. A software system for a load-lock-compatible nanomanipulation system is also designed and developed in this research.

  2. New Applications of Scanning Tunneling Microscopy

    Science.gov (United States)

    Smith, Douglas Philip Edward

    This dissertation describes the application of the scanning tunneling microscope (STM) technique to four new fields of study: thin organic films, phonon spectroscopy of bulk surfaces, the vibrational spectroscopy of molecules, and the tribological forces which occur between STM tip and sample. Images with atomic resolution were obtained with speeds approaching video rates. Two types of microscopes were used: one operated at room temperature in air, another at 4.2K in liquid helium. At room temperature, the STM was able to image molecules of cadmium arachidate deposited onto graphite by the Langmuir-Blodgett technique. The packing of molecules in the lipid bilayer was found to be partially ordered, with density of 1 molecule per 19.4 square angstroms. At liquid-helium temperature, inelastic electron processes were detected, and it was possible to determine within an area of a few square angstroms where the vibrational excitations occurred. On a bare graphite substrate, phonons of the sample and tip caused step increases in the tunneling conductivity at the phonon energies. Molecules of sorbic acid could be resolved when deposited onto graphite, and these molecules caused spatially localized peaks in conductivity at the energies of the bond vibrations. Although the STM is usually considered a non-contact instrument, under certain circumstances the tip and sample exerted strong forces on each other. With a tungsten tip and a graphite sample, friction and mechanical deformations on the atomic scale were observed.

  3. Visualizing subsurface defects in graphite by acoustic atomic force microscopy.

    Science.gov (United States)

    Wang, Tian; Ma, Chengfu; Hu, Wei; Chen, Yuhang; Chu, Jiaru

    2017-01-01

    We describe a versatile platform, which combines atomic force acoustic microscopy, ultrasonic atomic force microscopy and heterodyne force microscopy. The AFM system can enable in-situ switching among these operation modes flexibly and thus benefit the discrimination of differences in mechanical properties and buried subsurface nanostructures. We demonstrate the potential of this platform for visualizing the subsurface defects of graphite. Our results show that tiny topographic edges are enhanced in acoustic oscillation signals whilst embedded defects and inhomogeneous in mechanical properties are made clearly distinguishable. The possibility of detecting subsurface defects in few-layer graphene is further discussed with first-principles calculations. Microsc. Res. Tech. 80:66-74, 2017. © 2016 Wiley Periodicals, Inc. © 2016 Wiley Periodicals, Inc.

  4. Open Source Scanning Probe Microscopy Control Software Package Gxsm

    Energy Technology Data Exchange (ETDEWEB)

    Zahl P.; Wagner, T.; Moller, R.; Klust, A.

    2009-08-10

    Gxsm is a full featured and modern scanning probe microscopy (SPM) software. It can be used for powerful multidimensional image/data processing, analysis, and visualization. Connected toan instrument, it is operating many different avors of SPM, e.g., scanning tunneling microscopy(STM) and atomic force microscopy (AFM) or in general two-dimensional multi channel data acquisition instruments. The Gxsm core can handle different data types, e.g., integer and oating point numbers. An easily extendable plug-in architecture provides many image analysis and manipulation functions. A digital signal processor (DSP) subsystem runs the feedback loop, generates the scanning signals and acquires the data during SPM measurements. The programmable Gxsm vector probe engine performs virtually any thinkable spectroscopy and manipulation task, such as scanning tunneling spectroscopy (STS) or tip formation. The Gxsm software is released under the GNU general public license (GPL) and can be obtained via the Internet.

  5. Vector sensor for scanning SQUID microscopy

    Science.gov (United States)

    Dang, Vu The; Toji, Masaki; Thanh Huy, Ho; Miyajima, Shigeyuki; Shishido, Hiroaki; Hidaka, Mutsuo; Hayashi, Masahiko; Ishida, Takekazu

    2017-07-01

    We plan to build a novel 3-dimensional (3D) scanning SQUID microscope with high sensitivity and high spatial resolution. In the system, a vector sensor consists of three SQUID sensors and three pick-up coils realized on a single chip. Three pick-up coils are configured in orthogonal with each other to measure the magnetic field vector of X, Y, Z components. We fabricated some SQUID chips with one uniaxial pick-up coil or three vector pick-up coils and carried out fundamental measurements to reveal the basic characteristics. Josephson junctions (JJs) of sensors are designed to have the critical current density J c of 320 A/cm2, and the critical current I c becomes 12.5 μA for the 2.2μm × 2.2μm JJ. We carefully positioned the three pickup coils so as to keep them at the same height at the centers of all three X, Y and Z coils. This can be done by arranging them along single line parallel to a sample surface. With the aid of multilayer technology of Nb-based fabrication, we attempted to reduce an inner diameter of the pickup coils to enhance both sensitivity and spatial resolution. The method for improving a spatial resolution of a local magnetic field image is to employ an XYZ piezo-driven scanner for controlling the positions of the pick-up coils. The fundamental characteristics of our SQUID sensors confirmed the proper operation of our SQUID sensors and found a good agreement with our design parameters.

  6. Applications of orientation mapping by scanning and transmission electron microscopy

    DEFF Research Database (Denmark)

    Juul Jensen, D.

    1997-01-01

    The potentials of orientation mapping techniques (in the following referred to as OIM) for studies of thermomechanical processes are analysed. Both transmission electron microscopy (TEM) and scanning electron microscopy (SEM) based OIM techniques are considered. Among the thermomechanical processes...... information is achieved when the results of OIM and these various techniques are combined. Examples hereof are given to illustrate the potentials of OIM techniques. Finally, limitations of TEM and SEM based OIM for specific applications are discussed....

  7. System and method for compressive scanning electron microscopy

    Science.gov (United States)

    Reed, Bryan W

    2015-01-13

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

  8. Structural examination of lithium niobate ferroelectric crystals by combining scanning electron microscopy and atomic force microscopy

    Science.gov (United States)

    Efremova, P. V.; Ped'ko, B. B.; Kuznecova, Yu. V.

    2016-02-01

    The structure of lithium niobate single crystals is studied by a complex technique that combines scanning electron microscopy and atomic force microscopy. By implementing the piezoresponse force method on an atomic force microscope, the domain structure of lithium niobate crystals, which was not revealed without electron beam irradiation, is visualized

  9. Polarization contrast in photon scanning tunnelling microscopy combined with atomic force microscopy

    NARCIS (Netherlands)

    Propstra, K.; Propstra, K.; van Hulst, N.F.

    1995-01-01

    Photon scanning tunnelling microscopy combined with atomic force microscopy allows simultaneous acquisition and direct comparison of optical and topographical images, both with a lateral resolution of about 30 nm, far beyond the optical diffraction limit. The probe consists of a modified

  10. Scanning electron microscopy-energy dispersive X-ray spectrometer ...

    African Journals Online (AJOL)

    The distribution of arsenic (As) and cadmium (Cd) in himematsutake was analyzed using scanning electron microscopy-energy dispersive X-ray spectrometer (SEM-EDX). The atomic percentage of the metals was confirmed by inductively coupled plasma-mass spectrometer (ICP-MS). Results show that the accumulation of ...

  11. Challenges of scanning hall microscopy using batch fabricated probes

    NARCIS (Netherlands)

    Hatakeyama, Kodai

    2016-01-01

    Scanning Hall probe microscopy is a widely used technique for quantitative high resolution imaging of magnetic stray fields. Up to now probes with nanometer spatial resolution have only been realized by electron beam lithography, which is a slow and expensive fabrication technique. In this thesis,

  12. Nanochannel alignment analysis by scanning transmission ion microscopy

    DEFF Research Database (Denmark)

    Rajta, I.; Gál, G.A.B.; Szilasi, S.Z.

    2010-01-01

    In this paper a study on the ion transmission ratio of a nanoporous alumina sample is presented. The sample was investigated by scanning transmission ion microscopy (STIM) with different beam sizes. The hexagonally close-packed AlO nanocapillary array, realized as a suspended membrane of 15 νm...

  13. Scanning electron microscopy of Dermatobia hominis reveals cutaneous anchoring features.

    Science.gov (United States)

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

    2007-10-01

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

  14. Ultrafast terahertz scanning tunneling microscopy with atomic resolution

    DEFF Research Database (Denmark)

    Jelic, Vedran; Iwaszczuk, Krzysztof; Nguyen, Peter H.

    2016-01-01

    We demonstrate that ultrafast terahertz scanning tunneling microscopy (THz-STM) can probe single atoms on a silicon surface with simultaneous sub-nanometer and sub-picosecond spatio-temporal resolution. THz-STM is established as a new technique for exploring high-field non-equilibrium tunneling...

  15. Characterization of Polycaprolactone Films Biodeterioration by Scanning Electron Microscopy

    Czech Academy of Sciences Publication Activity Database

    Hrubanová, Kamila; Voberková, S.; Hermanová, S.; Krzyžánek, Vladislav

    2014-01-01

    Roč. 20, S3 (2014), s. 1950-1951 ISSN 1431-9276 R&D Projects: GA MŠk EE.2.3.20.0103; GA MŠk(CZ) LO1212 Institutional support: RVO:68081731 Keywords : polycaprolactone films * biodeterioration * scanning electron microscopy Subject RIV: JA - Electronics ; Optoelectronics, Electrical Engineering Impact factor: 1.877, year: 2014

  16. Scanning tunneling microscopy III theory of STM and related scanning probe methods

    CERN Document Server

    Güntherodt, Hans-Joachim

    1996-01-01

    Scanning Tunneling Microscopy III provides a unique introduction to the theoretical foundations of scanning tunneling microscopy and related scanning probe methods. The different theoretical concepts developed in the past are outlined, and the implications of the theoretical results for the interpretation of experimental data are discussed in detail. Therefore, this book serves as a most useful guide for experimentalists as well as for theoreticians working in the filed of local probe methods. In this second edition the text has been updated and new methods are discussed.

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

    Science.gov (United States)

    Hempel, Casper

    2017-07-01

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

  18. Non-linear image scanning microscopy (Conference Presentation)

    Science.gov (United States)

    Gregor, Ingo; Ros, Robert; Enderlein, Jörg

    2017-02-01

    Nowadays, multiphoton microscopy can be considered as a routine method for the observation of living cells, organs, up to whole organisms. Second-harmonics generation (SHG) imaging has evolved to a powerful qualitative and label-free method for studying fibrillar structures, like collagen networks. However, examples of super-resolution non-linear microscopy are rare. So far, such approaches require complex setups and advanced synchronization of scanning elements limiting the image acquisition rates. We describe theory and realization of a super-resolution image scanning microscope [1, 2] using two-photon excited fluorescence as well as second-harmonic generation. It requires only minor modifications compared to a classical two-photon laser-scanning microscope and allows image acquisition at the high frame rates of a resonant galvo-scanner. We achieve excellent sensitivity and high frame-rate in combination with two-times improved lateral resolution. We applied this method to fixed cells, collagen hydrogels, as well as living fly embryos. Further, we proofed the excellent image quality of our setup for deep tissue imaging. 1. Müller C.B. and Enderlein J. (2010) Image scanning microscopy. Phys. Rev. Lett. 104(19), 198101. 2. Sheppard C.J.R. (1988) Super-resolution in confocal imaging. Optik (Stuttg) 80 53-54.

  19. Investigations in optoelectronic image processing in scanning laser microscopy

    Science.gov (United States)

    Chaliha, Hiranya Kumar

    A considerable amount of work has been done on scann-ing laser microscopy since its applications were first pointed out by Roberts and Young[1], Minsky [2] and Davidovits et al [3]. The advent of laser has made it possible to focus an intense beam of laser light in a scanning optical microscope (SOM) [4, 5] and hence explore regions of microscopy[6] uncovered by conven-tional microscopy. In the simple SOM [7, 8, 9], the upper spatial frequency in amplitude transmittance or reflectance of an object for which transfer function is nonzero is same as that in a conventional optical microscope. However, in Type II SOM [7] or confocal SOM that employs a coherent or a point detector, the spatial frequency bandwidth is twice that obtained in a conventional microscope. Besides this confocal set-up is found to be very useful in optical sectioning and consequently in 3-D image processing[10, 11, 12] specially of biological specimens. Such systems are also suitable for studies of semiconductor materials [13], super-resolution [14] and various imaginative ways of image processing[15, 16, 17] including phase imaging[18]. A brief survey of related advances in scanning optical microscopy has been covered in the chapter 1 of the thesis. The performance of SOM may be investigated by concent-rating also on signal derived by one dimensional scan of the object specimen. This simplified mode may also be adapted to give wealth of information for biological and semiconductor specimens. Hence we have investigated the design of a scanning laser system suited specifically for studies of line scan image signals of microscopic specimens when probed through a focused laser spot. An electro-mechanical method of scanning of the object specimen has been designed with this aim in mind. Chapter 2, Part A of the thesis deals with the design consider-ations of such a system. For analysis of scan signals at a later instant of time so as to facilitate further processing, an arrangement of microprocessor

  20. Surface morphology of Trichinella spiralis by scanning electron microscopy

    Energy Technology Data Exchange (ETDEWEB)

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

    1980-02-01

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

  1. Scanning conductance microscopy investigations on fixed human chromosomes

    DEFF Research Database (Denmark)

    Clausen, Casper Hyttel; Lange, Jacob Moresco; Jensen, Linda Boye

    2008-01-01

    Scanning conductance microscopy investigations were carried out in air on human chromosomes fixed on pre-fabricated SiO2 surfaces with a backgate. The point of the investigation was to estimate the dielectric constant of fixed human chromosomes in order to use it for microfluidic device...... optimization. The phase shift caused by the electrostatic forces, together with geometrical measurements of the atomic force microscopy (AFM) cantilever and the chromosomes were used to estimate a value,for the dielectric constant of different human chromosomes....

  2. Ultrafast Photon Counting Applied to Resonant Scanning STED Microscopy

    Science.gov (United States)

    Wu, Xundong; Toro, Ligia; Stefani, Enrico; Wu, Yong

    2014-01-01

    Summary To take full advantage of fast resonant scanning in super-resolution STimulated Emission Depletion (STED) microscopy, we have developed an ultrafast photon counting system based on a multi-giga-sample per second analog-to-digital conversion (ADC) chip that delivers an unprecedented 450 MHz pixel clock (2.2 ns pixel dwell time in each scan). The system achieves a large field of view (~50 × 50 μm) with fast scanning that reduces photobleaching, and advances the time-gated continuous wave (CW) STED technology to the usage of resonant scanning with hardware based time-gating. The assembled system provides superb signal-to-noise ratio and highly linear quantification of light that result in superior image quality. Also, the system design allows great flexibility in processing photon signals to further improve the dynamic range. In conclusion, we have constructed a frontier photon counting image acquisition system with ultrafast readout rate, excellent counting linearity, and with the capacity of realizing resonant-scanning CW-STED microscopy with on-line time-gated detection. PMID:25227160

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

    Science.gov (United States)

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

    2015-01-01

    The growing use of nanoparticles in biomedical applications, including cancer diagnosis and treatment, demands the capability to exactly locate them within complex biological systems. In this work a correlative optical and scanning electron microscopy technique was developed to locate and observe multi-modal gold core nanoparticle accumulation in brain tumor models. Entire brain sections from mice containing orthotopic brain tumors injected intravenously with nanoparticles were imaged using both optical microscopy to identify the brain tumor, and scanning electron microscopy to identify the individual nanoparticles. Gold-based nanoparticles were readily identified in the scanning electron microscope using backscattered electron imaging as bright spots against a darker background. This information was then correlated to determine the exact location of the nanoparticles within the brain tissue. The nanoparticles were located only in areas that contained tumor cells, and not in the surrounding healthy brain tissue. This correlative technique provides a powerful method to relate the macro- and micro-scale features visible in light microscopy with the nanoscale features resolvable in scanning electron microscopy. Copyright © 2014 Elsevier Ltd. All rights reserved.

  4. [Advances of in vivo confocal scanning laser microscopy].

    Science.gov (United States)

    Tian, Ke-bin; Zhou, Guo-yu

    2006-02-01

    In vivo confocal scanning laser microscopy is being widely established as a time-saving, non-invasive, investigative methods in the study of body surfaces. Skin can be observed in its native state in vivo without the fixing, sectioning and staining that is necessary for routine histology. It is a new technology that can provide detailed images of tissue architecture and cellular morphology of living tissue. This paper reviews the fundamentals of in vivo confocal imaging and its clinical applications.

  5. Sub-Kelvin scanning tunneling microscopy on magnetic molecules

    OpenAIRE

    Zhang, Lei

    2012-01-01

    Magnetic molecules have attracted lots interest. In this work, an ultra-stable and low noise scanning tunneling microscopy operating at 400 mK using He-3 (930 mK using He-4) has been developed. The magnetic behavior of different magnetic molecules on substrates, especially the exchange interaction between the magnetic ions, the magnetic anisotropy on the surface, the magnetic excitations as well as the Kondo effect, were studied by using STM.

  6. Scanning Electron Microscopy of Cristispira Species in Chesapeake Bay Oysters

    OpenAIRE

    Tall, Ben D.; Nauman, Robert K.

    1981-01-01

    Scanning electron microscopy was employed to observe the physical interactions between Cristispira spp. and the crystalline style of the Chesapeake Bay oyster (Crassostrea virginica Gmelin 1791). Cristispira organisms were found associated with both the inner and outer layers of the posterior two-thirds of the style. The spirochetes possessed blunt-tipped ends, a cell diameter range of 0.6 to 0.8 μm, and distended spirochetal envelopes which followed the contour of the cells. Transmission ele...

  7. Playing peekaboo with graphene oxide: a scanning electrochemical microscopy investigation.

    Science.gov (United States)

    Rapino, Stefania; Treossi, Emanuele; Palermo, Vincenzo; Marcaccio, Massimo; Paolucci, Francesco; Zerbetto, Francesco

    2014-11-07

    Scanning electrochemical microscopy (SECM) can image graphene oxide (GO) flakes on insulating and conducting substrates. The contrast between GO and the substrate is controlled by the electrostatic interactions that are established between the charges of the molecular redox mediator and the charges present in the sheet/substrate. SECM also allows quantitative measurement - at the nano/microscale - of the charge transfer kinetics between single monolayer sheets and agent molecules.

  8. Scanning gate microscopy of ultra clean carbon nanotube quantum dots

    OpenAIRE

    Xue, Jiamin; Dhall, Rohan; Cronin, Stephen B.; LeRoy, Brian J.

    2015-01-01

    We perform scanning gate microscopy on individual suspended carbon nanotube quantum dots. The size and position of the quantum dots can be visually identified from the concentric high conductance rings. For the ultra clean devices used in this study, two new effects are clearly identified. Electrostatic screening creates non-overlapping multiple sets of Coulomb rings from a single quantum dot. In double quantum dots, by changing the tip voltage, the interactions between the quantum dots can b...

  9. Abrasion of 6 dentifrices measured by vertical scanning interference microscopy

    Science.gov (United States)

    PASCARETTI-GRIZON, Florence; MABILLEAU, Guillaume; CHAPPARD, Daniel

    2013-01-01

    Objectives The abrasion of dentifrices is well recognized to eliminate the dental plaque. The aims of this study were to characterize the abrasive powders of 6 dentifrices (3 toothpastes and 3 toothpowders) and to measure the abrasion on a test surface by Vertical Scanning Interference microscopy (VSI). Material and Methods Bright field and polarization microscopy were used to identify the abrasive particles on the crude dentifrices and after prolonged washes. Scanning electron microscopy and microanalysis characterized the shape and nature of the particles. Standardized and polished blocks of poly(methylmethacrylate) were brushed with a commercial electric toothbrush with the dentifrices. VSI quantified the mean roughness (Ra) and illustrated in 3D the abraded areas. Results Toothpastes induced a limited abrasion. Toothpowders induced a significantly higher roughness linked to the size of the abrasive particles. One powder (Gencix® produced a high abrasion when used with a standard testing weight. However, the powder is based on pumice particles covered by a plant homogenate that readily dissolves in water. When used in the same volume, or after dispersion in water, Ra was markedly reduced. Conclusion Light and electron microscopy characterize the abrasive particles and VSI is a new tool allowing the analysis of large surface of abraded materials. PMID:24212995

  10. Abrasion of 6 dentifrices measured by vertical scanning interference microscopy.

    Science.gov (United States)

    Pascaretti-Grizon, Florence; Mabilleau, Guillaume; Chappard, Daniel

    2013-01-01

    The abrasion of dentifrices is well recognized to eliminate the dental plaque. The aims of this study were to characterize the abrasive powders of 6 dentifrices (3 toothpastes and 3 toothpowders) and to measure the abrasion on a test surface by Vertical Scanning Interference microscopy (VSI). Bright field and polarization microscopy were used to identify the abrasive particles on the crude dentifrices and after prolonged washes. Scanning electron microscopy and microanalysis characterized the shape and nature of the particles. Standardized and polished blocks of poly(methylmethacrylate) were brushed with a commercial electric toothbrush with the dentifrices. VSI quantified the mean roughness (Ra) and illustrated in 3D the abraded areas. Toothpastes induced a limited abrasion. Toothpowders induced a significantly higher roughness linked to the size of the abrasive particles. One powder (Gencix® produced a high abrasion when used with a standard testing weight. However, the powder is based on pumice particles covered by a plant homogenate that readily dissolves in water. When used in the same volume, or after dispersion in water, Ra was markedly reduced. Light and electron microscopy characterize the abrasive particles and VSI is a new tool allowing the analysis of large surface of abraded materials.

  11. Analysis of leaf surfaces using scanning ion conductance microscopy.

    Science.gov (United States)

    Walker, Shaun C; Allen, Stephanie; Bell, Gordon; Roberts, Clive J

    2015-05-01

    Leaf surfaces are highly complex functional systems with well defined chemistry and structure dictating the barrier and transport properties of the leaf cuticle. It is a significant imaging challenge to analyse the very thin and often complex wax-like leaf cuticle morphology in their natural state. Scanning electron microscopy (SEM) and to a lesser extent Atomic force microscopy are techniques that have been used to study the leaf surface but their remains information that is difficult to obtain via these approaches. SEM is able to produce highly detailed and high-resolution images needed to study leaf structures at the submicron level. It typically operates in a vacuum or low pressure environment and as a consequence is generally unable to deal with the in situ analysis of dynamic surface events at submicron scales. Atomic force microscopy also possess the high-resolution imaging required and can follow dynamic events in ambient and liquid environments, but can over exaggerate small features and cannot image most leaf surfaces due to their inherent roughness at the micron scale. Scanning ion conductance microscopy (SICM), which operates in a liquid environment, provides a potential complementary analytical approach able to address these issues and which is yet to be explored for studying leaf surfaces. Here we illustrate the potential of SICM on various leaf surfaces and compare the data to SEM and atomic force microscopy images on the same samples. In achieving successful imaging we also show that SICM can be used to study the wetting of hydrophobic surfaces in situ. This has potentially wider implications than the study of leaves alone as surface wetting phenomena are important in a range of fundamental and applied studies. © 2015 The Authors Journal of Microscopy © 2015 Royal Microscopical Society.

  12. Microvascular quantification based on contour-scanning photoacoustic microscopy

    Science.gov (United States)

    Yeh, Chenghung; Soetikno, Brian; Hu, Song; Maslov, Konstantin I.; Wang, Lihong V.

    2014-09-01

    Accurate quantification of microvasculature remains of interest in fundamental pathophysiological studies and clinical trials. Current photoacoustic microscopy can noninvasively quantify properties of the microvasculature, including vessel density and diameter, with a high spatial resolution. However, the depth range of focus (i.e., focal zone) of optical-resolution photoacoustic microscopy (OR-PAM) is often insufficient to encompass the depth variations of features of interest-such as blood vessels-due to uneven tissue surfaces. Thus, time-consuming image acquisitions at multiple different focal planes are required to maintain the region of interest in the focal zone. We have developed continuous three-dimensional motorized contour-scanning OR-PAM, which enables real-time adjustment of the focal plane to track the vessels' profile. We have experimentally demonstrated that contour scanning improves the signal-to-noise ratio of conventional OR-PAM by as much as 41% and shortens the image acquisition time by 3.2 times. Moreover, contour-scanning OR-PAM more accurately quantifies vessel density and diameter, and has been applied to studying tumors with uneven surfaces.

  13. Integrated Confocal and Scanning Probe Microscopy for Biomedical Research

    Directory of Open Access Journals (Sweden)

    B.J. Haupt

    2006-01-01

    Full Text Available Atomic force microscopy (AFM continues to be developed, not only in design, but also in application. The new focus of using AFM is changing from pure material to biomedical studies. More frequently, it is being used in combination with other optical imaging methods, such as confocal laser scanning microscopy (CLSM and fluorescent imaging, to provide a more comprehensive understanding of biological systems. To date, AFM has been used increasingly as a precise micromanipulator, probing and altering the mechanobiological characteristics of living cells and tissues, in order to examine specific, receptor-ligand interactions, material properties, and cell behavior. In this review, we discuss the development of this new hybrid AFM, current research, and potential applications in diagnosis and the detection of disease.

  14. High Resolution Helium Ion Scanning Microscopy of the Rat Kidney

    Science.gov (United States)

    Rice, William L.; Van Hoek, Alfred N.; Păunescu, Teodor G.; Huynh, Chuong; Goetze, Bernhard; Singh, Bipin; Scipioni, Larry; Stern, Lewis A.; Brown, Dennis

    2013-01-01

    Helium ion scanning microscopy is a novel imaging technology with the potential to provide sub-nanometer resolution images of uncoated biological tissues. So far, however, it has been used mainly in materials science applications. Here, we took advantage of helium ion microscopy to explore the epithelium of the rat kidney with unsurpassed image quality and detail. In addition, we evaluated different tissue preparation methods for their ability to preserve tissue architecture. We found that high contrast, high resolution imaging of the renal tubule surface is possible with a relatively simple processing procedure that consists of transcardial perfusion with aldehyde fixatives, vibratome tissue sectioning, tissue dehydration with graded methanol solutions and careful critical point drying. Coupled with the helium ion system, fine details such as membrane texture and membranous nanoprojections on the glomerular podocytes were visualized, and pores within the filtration slit diaphragm could be seen in much greater detail than in previous scanning EM studies. In the collecting duct, the extensive and striking apical microplicae of the intercalated cells were imaged without the shrunken or distorted appearance that is typical with conventional sample processing and scanning electron microscopy. Membrane depressions visible on principal cells suggest possible endo- or exocytotic events, and central cilia on these cells were imaged with remarkable preservation and clarity. We also demonstrate the use of colloidal gold probes for highlighting specific cell-surface proteins and find that 15 nm gold labels are practical and easily distinguishable, indicating that external labels of various sizes can be used to detect multiple targets in the same tissue. We conclude that this technology represents a technical breakthrough in imaging the topographical ultrastructure of animal tissues. Its use in future studies should allow the study of fine cellular details and provide

  15. High resolution helium ion scanning microscopy of the rat kidney.

    Science.gov (United States)

    Rice, William L; Van Hoek, Alfred N; Păunescu, Teodor G; Huynh, Chuong; Goetze, Bernhard; Singh, Bipin; Scipioni, Larry; Stern, Lewis A; Brown, Dennis

    2013-01-01

    Helium ion scanning microscopy is a novel imaging technology with the potential to provide sub-nanometer resolution images of uncoated biological tissues. So far, however, it has been used mainly in materials science applications. Here, we took advantage of helium ion microscopy to explore the epithelium of the rat kidney with unsurpassed image quality and detail. In addition, we evaluated different tissue preparation methods for their ability to preserve tissue architecture. We found that high contrast, high resolution imaging of the renal tubule surface is possible with a relatively simple processing procedure that consists of transcardial perfusion with aldehyde fixatives, vibratome tissue sectioning, tissue dehydration with graded methanol solutions and careful critical point drying. Coupled with the helium ion system, fine details such as membrane texture and membranous nanoprojections on the glomerular podocytes were visualized, and pores within the filtration slit diaphragm could be seen in much greater detail than in previous scanning EM studies. In the collecting duct, the extensive and striking apical microplicae of the intercalated cells were imaged without the shrunken or distorted appearance that is typical with conventional sample processing and scanning electron microscopy. Membrane depressions visible on principal cells suggest possible endo- or exocytotic events, and central cilia on these cells were imaged with remarkable preservation and clarity. We also demonstrate the use of colloidal gold probes for highlighting specific cell-surface proteins and find that 15 nm gold labels are practical and easily distinguishable, indicating that external labels of various sizes can be used to detect multiple targets in the same tissue. We conclude that this technology represents a technical breakthrough in imaging the topographical ultrastructure of animal tissues. Its use in future studies should allow the study of fine cellular details and provide

  16. High resolution helium ion scanning microscopy of the rat kidney.

    Directory of Open Access Journals (Sweden)

    William L Rice

    Full Text Available Helium ion scanning microscopy is a novel imaging technology with the potential to provide sub-nanometer resolution images of uncoated biological tissues. So far, however, it has been used mainly in materials science applications. Here, we took advantage of helium ion microscopy to explore the epithelium of the rat kidney with unsurpassed image quality and detail. In addition, we evaluated different tissue preparation methods for their ability to preserve tissue architecture. We found that high contrast, high resolution imaging of the renal tubule surface is possible with a relatively simple processing procedure that consists of transcardial perfusion with aldehyde fixatives, vibratome tissue sectioning, tissue dehydration with graded methanol solutions and careful critical point drying. Coupled with the helium ion system, fine details such as membrane texture and membranous nanoprojections on the glomerular podocytes were visualized, and pores within the filtration slit diaphragm could be seen in much greater detail than in previous scanning EM studies. In the collecting duct, the extensive and striking apical microplicae of the intercalated cells were imaged without the shrunken or distorted appearance that is typical with conventional sample processing and scanning electron microscopy. Membrane depressions visible on principal cells suggest possible endo- or exocytotic events, and central cilia on these cells were imaged with remarkable preservation and clarity. We also demonstrate the use of colloidal gold probes for highlighting specific cell-surface proteins and find that 15 nm gold labels are practical and easily distinguishable, indicating that external labels of various sizes can be used to detect multiple targets in the same tissue. We conclude that this technology represents a technical breakthrough in imaging the topographical ultrastructure of animal tissues. Its use in future studies should allow the study of fine cellular details

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

    Science.gov (United States)

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

    2010-01-01

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

  18. Evaluation of the bleached human enamel by Scanning Electron Microscopy

    DEFF Research Database (Denmark)

    Miranda, Carolina Baptista; Pagani, Clovis; Benetti, Ana Raquel

    2005-01-01

    Since bleaching has become a popular procedure, the effect of peroxides on dental hard tissues is of great interest in research. Purpose: The aim of this in vitro study was to perform a qualitative analysis of the human enamel after the application of in-office bleaching agents, using Scanning...... Electron Microscopy (SEM). Materials and Methods: Twenty intact human third molars extracted for orthodontic reasons were randomly divided into four groups (n=5) treated as follows: G1- storage in artificial saliva (control group); G2- four 30-minute applications of 35% carbamide peroxide (total exposure...

  19. [Pulmonary hydatidosis. Comparison of cytology and scanning electron microscopy].

    Science.gov (United States)

    Lavaud, F; Nou, J M; Sadrin, R; de Montreynaud, J M; Adnet, J J

    1986-01-01

    The puncture of a hydatid cyst with a fine needle is not generally recommended as a procedure and may even be contra-indicated in the first instance. Sometimes, however, the cytologist will be surprised to discover some scolices in the aspirate when the radiology is misleading, or not suggestive, and the serology is negative. We report two cases where the diagnosis was made by the cytological examination of the aspirate. The cytological study of the liquids was compared with electron microscopy scanning, enabling the stages of development of the parasite in the tissue of the pulmonary parenchyma to be assessed.

  20. Advanced Scanning Electron Microscopy and X Ray Microanalysis

    Science.gov (United States)

    Krinsley, David

    This text is the third in a group that evolved from a short course taught annually at Lehigh University, Bethlehem, Pa., since 1972. Chapters on magnetic contrast a nd electron channeling, dropped from the second volume for reasons of space, are included here along with new topics such as image processing. The first seven chapters should be oT value to those geologists interested in scanning electron microscopy (SEM) and microanalysis. Chapters 8 and 9, concerned with specimen preparation for biological SEM a nd cryomicroscopy, make up about one third of the text.

  1. Transfer functions in collection scanning near-field optical microscopy

    DEFF Research Database (Denmark)

    Bozhevolnyi, Sergey I.; Vohnsen, Brian; Bozhevolnaya, Elena A.

    1999-01-01

    are considered with respect to the relation between near-field optical images and the corresponding intensity distributions. Our conclusions are supported with numerical simulations and experimental results obtained by using a photon scanning tunneling microscope with an uncoated fiber tip.......It is generally accepted that, if in collection near-field optical microscopy the probe-sample coupling can be disregarded, a fiber probe can be considered as a detector of the near-field intensity whose size can be accounted for via an intensity transfer function. We show that, in general...

  2. High-speed Lissajous-scan atomic force microscopy: Scan pattern planning and control design issues

    Science.gov (United States)

    Bazaei, A.; Yong, Yuen K.; Moheimani, S. O. Reza

    2012-06-01

    Tracking of triangular or sawtooth waveforms is a major difficulty for achieving high-speed operation in many scanning applications such as scanning probe microscopy. Such non-smooth waveforms contain high order harmonics of the scan frequency that can excite mechanical resonant modes of the positioning system, limiting the scan range and bandwidth. Hence, fast raster scanning often leads to image distortion. This paper proposes analysis and design methodologies for a nonlinear and smooth closed curve, known as Lissajous pattern, which allows much faster operations compared to the ordinary scan patterns. A simple closed-form measure is formulated for the image resolution of the Lissajous pattern. This enables us to systematically determine the scan parameters. Using internal model controllers (IMC), this non-raster scan method is implemented on a commercial atomic force microscope driven by a low resonance frequency positioning stage. To reduce the tracking errors due to actuator nonlinearities, higher order harmonic oscillators are included in the IMC controllers. This results in significant improvement compared to the traditional IMC method. It is shown that the proposed IMC controller achieves much better tracking performances compared to integral controllers when the noise rejection performances is a concern.

  3. Nanometrology using a through-focus scanning optical microscopy method

    Science.gov (United States)

    Attota, Ravikiran; Silver, Richard

    2011-02-01

    We present an initial review of a novel through-focus scanning optical microscopy (TSOM pronounced as 'tee-som') imaging method that produces nanometer-dimensional measurement sensitivity using a conventional bright-field optical microscope. In the TSOM method a target is scanned through the focus of an optical microscope, acquiring conventional optical images at different focal positions. The TSOM images are constructed using the through-focus optical images. A TSOM image is unique under given experimental conditions and is sensitive to changes in the dimensions of a target in a distinct way. We use this characteristic for nanoscale-dimensional metrology. This technique can be used to identify the dimension which is changing between two nanosized targets and to determine the dimensions using a library-matching method. This methodology has potential utility for a wide range of target geometries and application areas, including nanometrology, nanomanufacturing, defect analysis, inspection, process control and biotechnology.

  4. Quantitative phase imaging with scanning holographic microscopy: an experimental assessment.

    Science.gov (United States)

    Indebetouw, Guy; Tada, Yoshitaka; Leacock, John

    2006-11-28

    This paper demonstrates experimentally how quantitative phase information can be obtained in scanning holographic microscopy. Scanning holography can operate in both coherent and incoherent modes, simultaneously if desired, with different detector geometries. A spatially integrating detector provides an incoherent hologram of the object's intensity distribution (absorption and/or fluorescence, for example), while a point detector in a conjugate plane of the pupil provides a coherent hologram of the object's complex amplitude, from which a quantitative measure of its phase distribution can be extracted. The possibility of capturing simultaneously holograms of three-dimensional specimens, leading to three-dimensional reconstructions with absorption contrast, reflectance contrast, fluorescence contrast, as was previously demonstrated, and quantitative phase contrast, as shown here for the first time, opens up new avenues for multimodal imaging in biological studies.

  5. Quantitative single-molecule imaging by confocal laser scanning microscopy.

    Science.gov (United States)

    Vukojevic, Vladana; Heidkamp, Marcus; Ming, Yu; Johansson, Björn; Terenius, Lars; Rigler, Rudolf

    2008-11-25

    A new approach to quantitative single-molecule imaging by confocal laser scanning microscopy (CLSM) is presented. It relies on fluorescence intensity distribution to analyze the molecular occurrence statistics captured by digital imaging and enables direct determination of the number of fluorescent molecules and their diffusion rates without resorting to temporal or spatial autocorrelation analyses. Digital images of fluorescent molecules were recorded by using fast scanning and avalanche photodiode detectors. In this way the signal-to-background ratio was significantly improved, enabling direct quantitative imaging by CLSM. The potential of the proposed approach is demonstrated by using standard solutions of fluorescent dyes, fluorescently labeled DNA molecules, quantum dots, and the Enhanced Green Fluorescent Protein in solution and in live cells. The method was verified by using fluorescence correlation spectroscopy. The relevance for biological applications, in particular, for live cell imaging, is discussed.

  6. Elimination of periodic damped artifacts in scanning probe microscopy images

    Science.gov (United States)

    Chen, Yuhang; Huang, Wenhao

    2010-04-01

    When scanning probe microscopy (SPM) is operated at high scan rates, stripe-like artifacts will appear frequently in the SPM images. The removal of the image artifacts is highly demanded because they will distort the results in precise measurements. In this work, a method based on Prony analysis has been introduced to erase such periodic damped artifacts. Results demonstrate that this method prevails against the conventional fast Fourier transformation (FFT) method. Clean eliminations of the image artifacts are obtained, with almost no sacrifice of the detailed surface information. Even for arbitrary rough surfaces, the image artifacts can also be reduced by more than one order of magnitude. However, small amounts of stripes may still remain in the images. In these cases, the Prony analysis combined with locally weighted smoothing will provide better image quality. The artifacts reduction can have a meaning in the SPM-based visualization of dynamic phenomena with a nanoscale resolution.

  7. Quantitative phase imaging with scanning holographic microscopy: an experimental assesment

    Directory of Open Access Journals (Sweden)

    Tada Yoshitaka

    2006-11-01

    Full Text Available Abstract This paper demonstrates experimentally how quantitative phase information can be obtained in scanning holographic microscopy. Scanning holography can operate in both coherent and incoherent modes, simultaneously if desired, with different detector geometries. A spatially integrating detector provides an incoherent hologram of the object's intensity distribution (absorption and/or fluorescence, for example, while a point detector in a conjugate plane of the pupil provides a coherent hologram of the object's complex amplitude, from which a quantitative measure of its phase distribution can be extracted. The possibility of capturing simultaneously holograms of three-dimensional specimens, leading to three-dimensional reconstructions with absorption contrast, reflectance contrast, fluorescence contrast, as was previously demonstrated, and quantitative phase contrast, as shown here for the first time, opens up new avenues for multimodal imaging in biological studies.

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

    Science.gov (United States)

    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.

  9. Acoustic and photoacoustic microscopy imaging of single leukocytes

    Science.gov (United States)

    Strohm, Eric M.; Moore, Michael J.; Kolios, Michael C.

    2016-03-01

    An acoustic/photoacoustic microscope was used to create micrometer resolution images of stained cells from a blood smear. Pulse echo ultrasound images were made using a 1000 MHz transducer with 1 μm resolution. Photoacoustic images were made using a fiber coupled 532 nm laser, where energy losses through stimulated Raman scattering enabled output wavelengths from 532 nm to 620 nm. The laser was focused onto the sample using a 20x objective, and the laser spot co-aligned with the 1000 MHz transducer opposite the laser. The blood smear was stained with Wright-Giemsa, a common metachromatic dye that differentially stains the cellular components for visual identification. A neutrophil, lymphocyte and a monocyte were imaged using acoustic and photoacoustic microscopy at two different wavelengths, 532 nm and 600 nm. Unique features in each imaging modality enabled identification of the different cell types. This imaging method provides a new way of imaging stained leukocytes, with applications towards identifying and differentiating cell types, and detecting disease at the single cell level.

  10. Re-scan confocal microscopy (RCM) improves the resolution of confocal microscopy and increases the sensitivity

    Science.gov (United States)

    De Luca, Giulia; Breedijk, Ronald; Hoebe, Ron; Stallinga, Sjoerd; Manders, Erik

    2017-03-01

    Re-scan confocal microscopy (RCM) is a new super-resolution technique based on a standard confocal microscope extended with a re-scan unit in the detection path that projects the emitted light onto a sensitive camera. In this paper the fundamental properties of RCM, lateral resolution, axial resolution and signal-to-noise ratio, are characterized and compared with properties of standard confocal microscopy. The results show that the lateral resolution of RCM is ~170 nm compared to ~240 nm of confocal microscopy for 488 nm excitation and 1.49 NA. As the theory predicts, this improved lateral resolution is independent of the pinhole diameter. In standard confocal microscopy, the same lateral resolution can only be achieved with an almost closed pinhole and, consequently, with a major loss of signal. We show that the sectioning capabilities of the standard confocal microscope are preserved in RCM and that the axial resolution of RCM is slightly better (~15%) than the standard confocal microscope. Furthermore, the signal-to-noise ratio in RCM is a factor of 2 higher than in standard confocal microscopy, also due to the use of highly sensitive modern cameras. In case the pinhole of a confocal microscope is adjusted in such way that the lateral resolution is comparable to that of RCM, the signal-to-noise ratio in RCM is 4 times higher than standard confocal microscopy. Therefore, RCM offers a good alternative to standard confocal microscopy for higher lateral resolution with the main advantage of strongly improved sensitivity.

  11. Simultaneous Scanning Ion Conductance Microscopy and Atomic Force Microscopy with Microchanneled Cantilevers.

    Science.gov (United States)

    Ossola, Dario; Dorwling-Carter, Livie; Dermutz, Harald; Behr, Pascal; Vörös, János; Zambelli, Tomaso

    2015-12-04

    We combined scanning ion conductance microscopy (SICM) and atomic force microscopy (AFM) into a single tool using AFM cantilevers with an embedded microchannel flowing into the nanosized aperture at the apex of the hollow pyramid. An electrode was positioned in the AFM fluidic circuit connected to a second electrode in the bath. We could thus simultaneously measure the ionic current and the cantilever bending (in optical beam deflection mode). First, we quantitatively compared the SICM and AFM contact points on the approach curves. Second, we estimated where the probe in SICM mode touches the sample during scanning on a calibration grid and applied the finding to image a network of neurites on a Petri dish. Finally, we assessed the feasibility of a double controller using both the ionic current and the deflection as input signals of the piezofeedback. The experimental data were rationalized in the framework of finite elements simulations.

  12. Humidity effects on scanning polarization force microscopy imaging

    Energy Technology Data Exchange (ETDEWEB)

    Shen, Yue, E-mail: shenyue@isl.ac.cn [Key Laboratory of Comprehensive and Highly Efficient Utilization of Salt Lake Resources, Key Laboratory of Salt Lake Resources Chemistry of Qinghai Province, Qinghai Institute of Salt Lakes, Chinese Academy of Sciences, Xining, Qinghai 810008 (China); Key Laboratory of Interfacial Physics and Technology of Chinese Academy of Sciences, Shanghai Institute of Applied Physics, Chinese Academy of Sciences, Shanghai 201800 (China); Zhou, Yuan, E-mail: zhouy@isl.ac.cn [Key Laboratory of Comprehensive and Highly Efficient Utilization of Salt Lake Resources, Key Laboratory of Salt Lake Resources Chemistry of Qinghai Province, Qinghai Institute of Salt Lakes, Chinese Academy of Sciences, Xining, Qinghai 810008 (China); Sun, Yanxia; Zhang, Lijuan [Key Laboratory of Comprehensive and Highly Efficient Utilization of Salt Lake Resources, Key Laboratory of Salt Lake Resources Chemistry of Qinghai Province, Qinghai Institute of Salt Lakes, Chinese Academy of Sciences, Xining, Qinghai 810008 (China); University of Chinese Academy of Sciences, Beijing 100049 (China); Wang, Ying; Hu, Jun; Zhang, Yi [Key Laboratory of Interfacial Physics and Technology of Chinese Academy of Sciences, Shanghai Institute of Applied Physics, Chinese Academy of Sciences, Shanghai 201800 (China)

    2017-08-01

    Highlights: • The humidity dramatically affects the contrast of scanning polarization force microscopy (SPFM) imaging on mica surface. • This influence roots in the sensitive dielectric constant of mica surface to the humidity change. • A strategy of controllable and repeatable imaging the local dielectric properties of nanomaterials with SPFM is proposed. - Abstract: Scanning polarization force microscopy (SPFM) is a useful surface characterization technique to visually characterize and distinguish nanomaterial with different local dielectric properties at nanometer scale. In this paper, taking the individual one-atom-thick graphene oxide (GO) and reduced graphene oxide (rGO) sheets on mica as examples, we described the influences of environmental humidity on SPFM imaging. We found that the apparent heights (AHs) or contrast of SPFM imaging was influenced significantly by relative humidity (RH) at a response time of a few seconds. And this influence rooted in the sensitive dielectric constant of mica surface to the RH change. While dielectric properties of GO and rGO sheets were almost immune to the humidity change. In addition, we gave the method to determine the critical humidity at which the contrast conversion happened under different conditions. And this is important to the contrast control and repeatable imaging of SPFM through RH adjusting. These findings suggest a strategy of controllable and repeatable imaging the local dielectric properties of nanomaterials with SPFM, which is critically important for further distinguishment, manipulation, electronic applications, etc.

  13. Two-color two-photon fluorescence laser scanning microscopy.

    Science.gov (United States)

    Quentmeier, S; Denicke, S; Gericke, K-H

    2009-11-01

    We present the first realization of a Two-Color Two-Photon Laser-Scanning Microscope (2c2pLSM) and UV fluorescence images of cells acquired with this technique. Fluorescence is induced by two-color two-photon absorption using the fundamental and the second harmonic of a Ti:Sa femtosecond laser. Simultaneous absorption of an 800 nm photon and a 400 nm photon energetically corresponds to one-photon absorption at 266 nm. This technique for Laser-Scanning Microscopy extends the excitation wavelength range of a Ti:Sa powered fluorescence microscope to the UV. In addition to the known advantages of multi-photon microscopy like intrinsic 3D resolution, reduced photo damage and high penetration depth 2c2pLSM offers the possibility of using standard high numeric aperture objectives for UV fluorescence imaging. The effective excitation wavelength of 266 nm corresponds especially well to the excitation spectrum of tryptophan. Hence, it is an ideal tool for label free fluorescence studies and imaging of intrinsic protein fluorescence which originates mainly from tryptophan. Thus a very sensitive natural lifetime probe can be used for monitoring protein reactions or changes in conformation. First measurements of living MIN-6 cells reveal differences between the UV fluorescence lifetimes of the nucleus and cytoplasm. The significance of this method was further demonstrated by monitoring the binding of biotin to avidin.

  14. Gold nanocone near-field scanning optical microscopy probes.

    Science.gov (United States)

    Fleischer, Monika; Weber-Bargioni, Alexander; Altoe, M Virginia P; Schwartzberg, Adam M; Schuck, P James; Cabrini, Stefano; Kern, Dieter P

    2011-04-26

    Near-field scanning optical microscopy enables the simultaneous topographical and subdiffraction limited optical imaging of surfaces. A process is presented for the implementation of single individually engineered gold cones at the tips of atomic force microscopy cantilevers. These cantilevers act as novel high-performance optical near-field probes. In the fabrication, thin-film metallization, electron beam induced deposition of etch masks, and Ar ion milling are combined. The cone constitutes a well-defined highly efficient optical antenna with a tip radius on the order of 10 nm and an adjustable plasmon resonance frequency. The sharp tip enables high resolution topographical imaging. By controllably varying the cone size, the resonance frequency can be adapted to the application of choice. Structural properties of these sharp-tipped probes are presented together with topographical images recorded with a cone probe. The antenna functionality is demonstrated by gathering the near-field enhanced Raman signature of individual carbon nanotubes with a gold cone scanning probe.

  15. Scanning ion microscopy with low energy lithium ions

    Energy Technology Data Exchange (ETDEWEB)

    Twedt, Kevin A. [Center for Nanoscale Science and Technology, National Institute of Standards and Technology, Gaithersburg, MD 20899 (United States); Maryland NanoCenter, University of Maryland, College Park, MD 20742 (United States); Chen, Lei [Center for Nanoscale Science and Technology, National Institute of Standards and Technology, Gaithersburg, MD 20899 (United States); McClelland, Jabez J., E-mail: jabez.mcclelland@nist.gov [Center for Nanoscale Science and Technology, National Institute of Standards and Technology, Gaithersburg, MD 20899 (United States)

    2014-07-01

    Using an ion source based on photoionization of laser-cooled lithium atoms, we have developed a scanning ion microscope with probe sizes of a few tens of nanometers and beam energies from 500 eV to 5 keV. These beam energies are much lower than the typical operating energies of the helium ion microscope or gallium focused ion beam systems. We demonstrate how low energy can be advantageous in ion microscopy when detecting backscattered ions, due to a decreased interaction volume and the potential for surface sensitive composition analysis. As an example application that demonstrates these advantages, we non-destructively image the removal of a thin residual resist layer during plasma etching in a nano-imprint lithography process. - Highlights: • We use an ion source based on photoionization of laser-cooled lithium atoms. • The ion source makes possible a low energy (500 eV to 5 keV) scanning ion microscope. • Low energy is preferred for ion microscopy with backscattered ions. • We use the microscope to image a thin resist used in nano-imprint lithography.

  16. Confocal laser scanning microscopy in study of bone calcification

    Science.gov (United States)

    Nishikawa, Tetsunari; Kokubu, Mayu; Kato, Hirohito; Imai, Koichi; Tanaka, Akio

    2012-12-01

    Bone regeneration in mandible and maxillae after extraction of teeth or tumor resection and the use of rough surface implants in bone induction must be investigated to elucidate the mechanism of calcification. The calcified tissues are subjected to chemical decalcification or physical grinding to observe their microscopic features with light microscopy and transmission electron microscopy where the microscopic tissue morphology is significantly altered. We investigated the usefulness of confocal laser scanning microscopy (CLSM) for this purpose. After staggering the time of administration of calcein and alizarin red to experimental rats and dogs, rat alveolar bone and dog femur grafted with coral as scaffold or dental implants were observed with CLSM. In rat alveolar bone, the calcification of newly-formed bone and net-like canaliculi was observed at the mesial bone from the roots progressed at the rate of 15 μm/day. In dog femur grafted with coral, newly-formed bones along the space of coral were observed in an orderly manner. In dog femur with dental implants, after 8 weeks, newly-formed bone proceeded along the rough surface of the implants. CLSM produced high-magnification images of newly-formed bone and thin sections were not needed.

  17. Immunolabeling for scanning electron microscopy (SEM) and field emission SEM.

    Science.gov (United States)

    Goldberg, Martin W

    2008-01-01

    Scanning electron microscopy (SEM) is a high resolution surface imaging technique. Many biological process and structures occur at surfaces and if antibodies are available, their components can be located within the surface structure. This is usually done in a similar way to immuno-fluorescence, using an unconjugated primary antibody followed by a tagged secondary antibody against the primary. In this case the tag is usually a colloidal gold particle instead of a fluorophore. Therefore it is quite straightforward to adapt an immuno-fluorescence procedure for SEM, as long as certain precautions are followed, as discussed here. Progressing from immuno-fluorescence, which essentially only indicates the position of a protein within the volume of a cell, to immuno-SEM, puts the labeling into the context of cellular structures. The principles and practices of sample preparation, labeling and imaging are described here.

  18. Ultramicrosensors based on transition metal hexacyanoferrates for scanning electrochemical microscopy

    Directory of Open Access Journals (Sweden)

    Maria A. Komkova

    2013-10-01

    Full Text Available We report here a way for improving the stability of ultramicroelectrodes (UME based on hexacyanoferrate-modified metals for the detection of hydrogen peroxide. The most stable sensors were obtained by electrochemical deposition of six layers of hexacyanoferrates (HCF, more specifically, an alternating pattern of three layers of Prussian Blue and three layers of Ni–HCF. The microelectrodes modified with mixed layers were continuously monitored in 1 mM hydrogen peroxide and proved to be stable for more than 5 h under these conditions. The mixed layer microelectrodes exhibited a stability which is five times as high as the stability of conventional Prussian Blue-modified UMEs. The sensitivity of the mixed layer sensor was 0.32 A·M−1·cm−2, and the detection limit was 10 µM. The mixed layer-based UMEs were used as sensors in scanning electrochemical microscopy (SECM experiments for imaging of hydrogen peroxide evolution.

  19. Confocal laser scanning microscopy-guided surgery for neurofibroma.

    Science.gov (United States)

    Koller, S; Horn, M; Weger, W; Massone, C; Smolle, J; Gerger, A

    2009-12-01

    The neurofibromatoses comprise at least two separate genetic disorders with variable clinical features and an unpredictable course. The most common type, neurofibromatosis 1, is characterized by > or = 6 café-au-lait spots and the occurrence of neurofibromas, which may present as cutaneous, subcutaneous or plexiform lesions. Normally, excision of neurofibromas is only indicated in the presence of neurological symptoms, suspicion of malignancy or for exceptional cosmetic reasons. For a good functional and aesthetic result with the least danger of recurrence, the surgeon's goal is to excise as much tissue as necessary and as little tissue as possible. One of the main issues during the surgical procedure is to distinguish between neurofibroma and surrounding tissue. We report for the first time the use of confocal laser scanning microscopy to differentiate between neurofibroma and healthy skin.

  20. Cryo-Scanning Electron Microscopy of Captured Cirrus Ice Particles

    Science.gov (United States)

    Magee, N. B.; Boaggio, K.; Bandamede, M.; Bancroft, L.; Hurler, K.

    2016-12-01

    We present the latest collection of high-resolution cryo-scanning electron microscopy images and microanalysis of cirrus ice particles captured by high-altitude balloon (ICE-Ball, see abstracts by K. Boaggio and M. Bandamede). Ice particle images and sublimation-residues are derived from particles captured during approximately 15 balloon flights conducted in Pennsylvania and New Jersey over the past 12 months. Measurements include 3D digital elevation model reconstructions of ice particles, and associated statistical analyses of entire particles and particle sub-facets and surfaces. This 3D analysis reveals that morphologies of most ice particles captured deviate significantly from ideal habits, and display geometric complexity and surface roughness at multiple measureable scales, ranging from 100's nanometers to 100's of microns. The presentation suggests potential a path forward for representing scattering from a realistically complex array of ice particle shapes and surfaces.

  1. Local deposition of anisotropic nanoparticles using scanning electrochemical microscopy (SECM).

    Science.gov (United States)

    Fedorov, Roman G; Mandler, Daniel

    2013-02-28

    We demonstrate localized electrodeposition of anisotropic metal nanoobjects, namely Au nanorods (GNR), on indium tin oxide (ITO) using scanning electrochemical microscopy (SECM). A gold microelectrode was the source of the gold ions whereby double pulse chronoamperometry was employed to generate initially Au seeds which were further grown under controlled conditions. The distance between the microelectrode and the ITO surface as well as the different experimental parameters (electrodeposition regime, solution composition and temperature) were optimized to produce faceted gold seeds with the required characteristics (size and distribution). Colloidal chemical synthesis was successfully exploited for better understanding the role of the surfactant and different additives in breaking the crystallographic symmetry and anisotropic growth of GNR. Experiments performed in a conventional three-electrode cell revealed the most appropriate electrochemical conditions allowing high yield synthesis of nanorods with well-defined shape as well as nanocubes and bipyramids.

  2. Preparation of platinum/iridium scanning probe microscopy tips

    DEFF Research Database (Denmark)

    Sørensen, Alexis Hammer; Hvid, U.; Mortensen, M.W.

    1999-01-01

    material being etched is platinum/iridium (10%) the influence of the stop phase of the ac current terminating each pulse in the second etching is found to be negligible, while in the case of second etching of tungsten wires it is important to break the pulse in a certain phase to avoid formation of a thick...... of platinum from the wire surface and hereby give rise to "etching" of the wire. In the second etching blunt tips become sharp while tips which are already sharp apparently stay sharp. Therefore, the second etching scheme with pulses separated by pauses is found to be a very important factor...... for the production of sharp tips. After being etched the tips are ready for use in scanning tunneling microscopes, or they may be bent to form integrated tip/cantilever systems in ordinary commercial atomic force microscopes, being applicable as tapping mode tips and as electrostatic force microscopy tips. ©1999...

  3. Active current-noise cancellation for Scanning Tunneling Microscopy

    Science.gov (United States)

    Pabbi, Lavish; Shoop, Conner; Banerjee, Riju; Dusch, Bill; Hudson, E. W.

    The high sensitivity of the scanning tunneling microscope (STM) poses a barrier to its use in a noisy environment. Vibrational noise, whether structural or acoustic in source, manifests as relative motion between the probe tip and the sample, then appearing in the Z feedback that tries to cancel it. Here we describe an active noise cancellation process that nullifies this motion by adding a drive signal into the existing Z feedback loop. The drive is digitally calculated by actively monitoring vibrations measured by an accelerometer placed in-situ close to the STM head. By transferring the vibration cancellation effort to this drive signal, vibration-created noise in the Z-feedback (during topography) or current (during spectroscopy) is significantly reduced. This inexpensive and easy solution, requiring no major instrumental modifications, is ideal for those looking to place their STM in a noisier environment, for example in the presence of active refrigeration systems (e.g. pulse tube cryocoolers) or coupled to high-vibration instrumentation. This material is based upon work supported by the National Science Foundation under Grant No. 1229138.

  4. Three-Dimensional scanning transmission electron microscopy of biological specimens

    KAUST Repository

    De Jonge, Niels

    2010-01-18

    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. © 2010 Microscopy Society of America.

  5. Scanning Surface Potential Microscopy of Spore Adhesion on Surfaces

    Energy Technology Data Exchange (ETDEWEB)

    Lee, Ida [University of Tennessee, Knoxville (UTK); Chung, Eunhyea [Georgia Institute of Technology; Kweon, Hyojin [Georgia Institute of Technology; Yiacoumi, Sotira [Georgia Institute of Technology; Tsouris, Costas [ORNL

    2012-01-01

    The adhesion of spores of Bacillus anthracis - the cause of anthrax and a likely biological threat - to solid surfaces is an important consideration in cleanup after an accidental or deliberate release. However, because of safety concerns, directly studying B. anthracis spores with advanced instrumentation is problematic. As a first step, we are examining the electrostatic potential of Bacillus thuringiensis (Bt), which is a closely related species that is often used as a simulant to study B. anthracis. Scanning surface potential microscopy (SSPM), also known as Kelvin probe force microscopy (KPFM), was used to investigate the influence of relative humidity (RH) on the surface electrostatic potential of Bt that had adhered to silica, mica, or gold substrates. AFM/SSPM side-by-side images were obtained separately in air, at various values of RH, after an aqueous droplet with spores was applied on each surface and allowed to dry before measurements. In the SSPM images, a negative potential on the surface of the spores was observed compared with that of the substrates. The surface potential decreased as the humidity increased. Spores were unable to adhere to a surface with an extremely negative potential, such as mica.

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

    Energy Technology Data Exchange (ETDEWEB)

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

    2011-01-01

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

  7. Scanning electron microscopy of the neuropathology of murine cerebral malaria

    Directory of Open Access Journals (Sweden)

    Brenneis Christian

    2006-11-01

    Full Text Available Abstract Background The mechanisms leading to death and functional impairments due to cerebral malaria (CM are yet not fully understood. Most of the knowledge about the pathomechanisms of CM originates from studies in animal models. Though extensive histopathological studies of the murine brain during CM are existing, alterations have not been visualized by scanning electron microscopy (SEM so far. The present study investigates the neuropathological features of murine CM by applying SEM. Methods C57BL/6J mice were infected with Plasmodium berghei ANKA blood stages. When typical symptoms of CM developed perfused brains were processed for SEM or light microscopy, respectively. Results Ultrastructural hallmarks were disruption of vessel walls, parenchymal haemorrhage, leukocyte sequestration to the endothelium, and diapedesis of macrophages and lymphocytes into the Virchow-Robin space. Villous appearance of observed lymphocytes were indicative of activated state. Cerebral oedema was evidenced by enlargement of perivascular spaces. Conclusion The results of the present study corroborate the current understanding of CM pathophysiology, further support the prominent role of the local immune system in the neuropathology of CM and might expose new perspectives for further interventional studies.

  8. Scanning electron microscopy physics of image formation and microanalysis

    CERN Document Server

    Reimer, Ludwig

    1985-01-01

    The aim of this book is to outline the physics of image formation, electron­ specimen interactions, imaging modes, the interpretation of micrographs and the use of quantitative modes "in scanning electron microscopy (SEM). lt forms a counterpart to Transmission Electron Microscopy (Vol. 36 of this Springer Series in Optical Sciences) . The book evolved from lectures delivered at the University of Münster and from a German text entitled Raster-Elektronenmikroskopie (Springer-Verlag), published in collaboration with my colleague Gerhard Pfefferkorn. In the introductory chapter, the principles of the SEM and of electron­ specimen interactions are described, the most important imaging modes and their associated contrast are summarized, and general aspects of eiemental analysis by x-ray and Auger electron emission are discussed. The electron gun and electron optics are discussed in Chap. 2 in order to show how an electron probe of small diameter can be formed, how the elec­ tron beam can be blanked at high fre...

  9. Confocal laser scanning microscopy in study of bone calcification

    Energy Technology Data Exchange (ETDEWEB)

    Nishikawa, Tetsunari, E-mail: tetsu-n@cc.osaka-dent.ac.jp [Department of Oral Pathology, Osaka Dental University, Osaka (Japan); Kokubu, Mayu; Kato, Hirohito [Department of Oral Pathology, Osaka Dental University, Osaka (Japan); Imai, Koichi [Department of Biomaterials, Osaka Dental University, Osaka (Japan); Tanaka, Akio [Department of Oral Pathology, Osaka Dental University, Osaka (Japan)

    2012-12-01

    Highlights: Black-Right-Pointing-Pointer High-magnification images with depth selection, and thin sections were observed using CLSM. Black-Right-Pointing-Pointer The direction and velocity of calcification of the bone was observed by administration of 2 fluorescent dyes. Black-Right-Pointing-Pointer In dog femora grafted with coral blocks, newly-formed bone was observed in the coral block space with a rough surface. Black-Right-Pointing-Pointer Twelve weeks after dental implant was grafted in dog femora, the space between screws was filled with newly-formed bones. - Abstract: Bone regeneration in mandible and maxillae after extraction of teeth or tumor resection and the use of rough surface implants in bone induction must be investigated to elucidate the mechanism of calcification. The calcified tissues are subjected to chemical decalcification or physical grinding to observe their microscopic features with light microscopy and transmission electron microscopy where the microscopic tissue morphology is significantly altered. We investigated the usefulness of confocal laser scanning microscopy (CLSM) for this purpose. After staggering the time of administration of calcein and alizarin red to experimental rats and dogs, rat alveolar bone and dog femur grafted with coral as scaffold or dental implants were observed with CLSM. In rat alveolar bone, the calcification of newly-formed bone and net-like canaliculi was observed at the mesial bone from the roots progressed at the rate of 15 {mu}m/day. In dog femur grafted with coral, newly-formed bones along the space of coral were observed in an orderly manner. In dog femur with dental implants, after 8 weeks, newly-formed bone proceeded along the rough surface of the implants. CLSM produced high-magnification images of newly-formed bone and thin sections were not needed.

  10. Computer vision distortion correction of scanning probe microscopy images.

    Science.gov (United States)

    Gaponenko, Iaroslav; Tückmantel, Philippe; Ziegler, Benedikt; Rapin, Guillaume; Chhikara, Manisha; Paruch, Patrycja

    2017-04-06

    Since its inception, scanning probe microscopy (SPM) has established itself as the tool of choice for probing surfaces and functionalities at the nanoscale. Although recent developments in the instrumentation have greatly improved the metrological aspects of SPM, it is still plagued by the drifts and nonlinearities of the piezoelectric actuators underlying the precise nanoscale motion. In this work, we present an innovative computer-vision-based distortion correction algorithm for offline processing of functional SPM measurements, allowing two images to be directly overlaid with minimal error - thus correlating position with time evolution and local functionality. To demonstrate its versatility, the algorithm is applied to two very different systems. First, we show the tracking of polarisation switching in an epitaxial Pb(Zr0.2Ti0.8)O3 thin film during high-speed continuous scanning under applied tip bias. Thanks to the precise time-location-polarisation correlation we can extract the regions of domain nucleation and track the motion of domain walls until the merging of the latter in avalanche-like events. Secondly, the morphology of surface folds and wrinkles in graphene deposited on a PET substrate is probed as a function of applied strain, allowing the relaxation of individual wrinkles to be tracked.

  11. Scanning X-ray microscopy of superconductor/ferromagnet bilayers

    Energy Technology Data Exchange (ETDEWEB)

    Stahl, Claudia; Ruoss, Stephen; Weigand, Markus; Schuetz, Gisela [Max Planck Institute for Intelligent Systems, Stuttgart (Germany); Zahn, Patrick; Bayer, Jonas [Max Planck Institute for Intelligent Systems, Stuttgart (Germany); Research Institute for Innovative Surfaces, FINO, Aalen University (Germany); Albrecht, Joachim [Research Institute for Innovative Surfaces, FINO, Aalen University (Germany)

    2016-07-01

    The magnetic flux distribution arising from a high-T{sub c} superconductor is detected and visualized with high spatial resolution using scanning x-ray microscopy (SXM). Therefore, we introduce a sensor layer, namely, an amorphous, soft-magnetic CoFeB cover layer. The magnetic stray fields of the supercurrents lead to a local reorientation of the magnetic moments in the ferromagnet, which is visualized using the large x-ray magnetic circular dichroism (XMCD) effect of the Co and Fe L3-edge. We show that the XMCD contrast in the sensor layer corresponds to the in-plane magnetic flux distribution of the superconductor and can hence be used to image magnetic structures in superconductors with high spatial resolution. Using the total electron yield (TEY) mode the surface structure and the magnetic domains can be imaged simultaneously and can be correlated. The measurements are carried out at our scanning x-ray microscope MAXYMUS at Bessy II, Berlin with the new low temperature setup.

  12. Post-processing strategies in image scanning microscopy.

    Science.gov (United States)

    McGregor, J E; Mitchell, C A; Hartell, N A

    2015-10-15

    Image scanning microscopy (ISM) coupled with pixel reassignment offers a resolution improvement of √2 over standard widefield imaging. By scanning point-wise across the specimen and capturing an image of the fluorescent signal generated at each scan position, additional information about specimen structure is recorded and the highest accessible spatial frequency is doubled. Pixel reassignment can be achieved optically in real time or computationally a posteriori and is frequently combined with the use of a physical or digital pinhole to reject out of focus light. Here, we simulate an ISM dataset using a test image and apply standard and non-standard processing methods to address problems typically encountered in computational pixel reassignment and pinholing. We demonstrate that the predicted improvement in resolution is achieved by applying standard pixel reassignment to a simulated dataset and explore the effect of realistic displacements between the reference and true excitation positions. By identifying the position of the detected fluorescence maximum using localisation software and centring the digital pinhole on this co-ordinate before scaling around translated excitation positions, we can recover signal that would otherwise be degraded by the use of a pinhole aligned to an inaccurate excitation reference. This strategy is demonstrated using experimental data from a multiphoton ISM instrument. Finally we investigate the effect that imaging through tissue has on the positions of excitation foci at depth and observe a global scaling with respect to the applied reference grid. Using simulated and experimental data we explore the impact of a globally scaled reference on the ISM image and, by pinholing around the detected maxima, recover the signal across the whole field of view. Copyright © 2015 Elsevier Inc. All rights reserved.

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

    Science.gov (United States)

    Konopka, K

    2006-09-01

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

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

    Science.gov (United States)

    Seacor, Taylor; Howell, Carina

    2013-03-01

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

  15. Scanning probe microscopy investigation of complex-oxide heterostructures

    Science.gov (United States)

    Bi, Feng

    Advances in the growth of precisely tailored complex-oxide heterostructures have led to new emergent behavior and associated discoveries. One of the most successful examples consists of an ultrathin layer of LaAlO 3 (LAO) deposited on TiO2-terminated SrTiO3 (STO), where a high mobility quasi-two dimensional electron liquid (2DEL) is formed at the interface. Such 2DEL demonstrates a variety of novel properties, including field tunable metal-insulator transition, superconductivity, strong spin-orbit coupling, magnetic and ferroelectric like behavior. Particularly, for 3-unit-cell (3 u.c.) LAO/STO heterostructures, it was demonstrated that a conductive atomic force microscope (c-AFM) tip can be used to "write" or "erase" nanoscale conducting channels at the interface, making LAO/STO a highly flexible platform to fabricate novel nanoelectronics. This thesis is focused on scanning probe microscopy studies of LAO/STO properties. We investigate the mechanism of c-AFM lithography over 3 u.c. LAO/STO in controlled ambient conditions by using a vacuum AFM, and find that the water molecules dissociated on the LAO surface play a critical role during the c-AFM lithography process. We also perform electro-mechanical response measurements over top-gated LAO/STO devices. Simultaneous piezoresponse force microscopy (PFM) and capacitance measurements reveal a correlation between LAO lattice distortion and interfacial carrier density, which suggests that PFM could not only serve as a powerful tool to map the carrier density at the interface but also provide insight into previously reported frequency dependence of capacitance enhancement of top-gated LAO/STO structures. To study magnetism at the LAO/STO interface, magnetic force microscopy (MFM) and magnetoelectric force microscopy (MeFM) are carried out to search for magnetic signatures that depend on the carrier density at the interface. Results demonstrate an electronicallycontrolled ferromagnetic phase on top-gated LAO

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

    Science.gov (United States)

    Tomitori, Masahiko; Sasahara, Akira

    2014-11-01

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

  17. Preliminary Study of In Vivo Formed Dental Plaque Using Confocal Microscopy and Scanning Electron Microscopy

    Directory of Open Access Journals (Sweden)

    KA. Al-Salihi

    2009-12-01

    Full Text Available Objective: Confocal laser scanning microscopy (CLSM is relatively a new light microscopical imaging technique with a wide range of applications in biological sciences. The primary value of CLSM for the biologist is its ability to provide optical sections from athree-dimensional specimen. The present study was designed to assess the thickness and content of in vivo accumulated dental plaque using CLSM and scanning electron microscopy (SEM.Materials and Methods: Acroflat lower arch splints (acrylic appliance were worn by five participants for three days without any disturbance. The formed plaques were assessed using CLSM combined with vital fluorescence technique and SEM.Results: In this study accumulated dental plaque revealed varied plaque microflora vitality and thickness according to participant’s oral hygiene. The thickness of plaque smears ranged from 40.32 to 140.72 μm and 65.00 to 128.88 μm for live (vital and dead accumulated microorganisms, respectively. Meanwhile, the thickness of plaque on the appliance ranged from 101 μm to 653 μm. CLSM revealed both dead and vital bacteria on the surface of the dental plaque. In addition, SEM revealed layers of various bacterial aggregations in all dental plaques.Conclusion: This study offers a potent non-invasive tool to evaluate and assess the dental plaque biofilm, which is a very important factor in the development of dental caries.

  18. Reflection across plant cell boundaries in confocal laser scanning microscopy.

    Science.gov (United States)

    Liu, D Y T; Kuhlmey, B T; Smith, P M C; Day, D A; Faulkner, C R; Overall, R L

    2008-08-01

    The fluorescence patterns of proteins tagged with the green fluorescent protein (GFP) and its derivatives are routinely used in conjunction with confocal laser scanning microscopy to identify their sub-cellular localization in plant cells. GFP-tagged proteins localized to plasmodesmata, the intercellular junctions of plants, are often identified by single or paired punctate labelling across the cell wall. The observation of paired puncta, or 'doublets', across cell boundaries in tissues that have been transformed through biolistic bombardment is unexpected if there is no intercellular movement of the GFP-tagged protein, since bombardment usually leads to the transformation of single, isolated cells. We expressed a putative plasmodesmal protein tagged with GFP by bombarding Allium porrum epidermal cells and assessed the nature of the doublets observed at the cell boundaries. Doublets were formed when fluorescent spots were abutting a cell boundary and were only observable at certain focal planes. Fluorescence emitted from the half of a doublet lying outside the transformed cells was polarized. Optical simulations performed using finite-difference time-domain computations showed a dramatic distortion of the confocal microscope's point spread function when imaging voxels close to the plant cell wall due to refractive index differences between the wall and the cytosol. Consequently, axially and radially out-of-focus light could be detected. A model of this phenomenon suggests how a doublet may form when imaging only a single real fluorescent body in the vicinity of a plant cell wall using confocal microscopy. We suggest, therefore, that the appearance of doublets across cell boundaries is insufficient evidence for plasmodesmal localization due to the effects of the cell wall on the reflection and scattering of light.

  19. 3D correlative light and electron microscopy of cultured cells using serial blockface scanning electron microscopy

    Science.gov (United States)

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

    2017-01-01

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

  20. Scanning near-field infrared microscopy on semiconductor structures

    Energy Technology Data Exchange (ETDEWEB)

    Jacob, Rainer

    2011-01-15

    literature. While the structures of the first system were in the micrometer regime, the capability to probe buried nanostructures is demonstrated at a sample of indium arsenide quantum dots. Those dots are covered by a thick layer of gallium arsenide. For the first time ever, it is shown experimentally that transitions between electron states in single quantum dots can be investigated by near-field microscopy. By monitoring the near-field response of these quantum dots while scanning the wavelength of the incident light beam, it was possible to obtain characteristic near-field signatures of single dots. Near-field contrasts up to 30 % could be measured for resonant excitation of electrons in the conduction band of the indium arsenide dots. (orig.)

  1. Cryo-planing for cryo-scanning electron microscopy.

    Science.gov (United States)

    Nijsse, J; van Aelst, A C

    1999-01-01

    In the past decade, investigators of cryo-planing for low-temperature scanning electron microscopy (cryo-SEM) have developed techniques that enable observations of flat sample surfaces. This study reviews these sample preparation techniques, compares and contrasts their results, and introduces modifications that improve results from cryo-planing. A prerequisite for all successful cryo-planing required a stable attachment of the specimen to a holder. In most cases, clamping with a screw mechanism and using indium as space-filler sufficed. Once this problem was solved, any of three existing cryo-planing methods could be used to provide successful results: cryo-milling, microtomy in a cold room, and cryo-ultramicrotomy. This study introduces modifications to the cryo-planing technique that produces flat surfaces of any desired plane through a specimen. These flat surfaces of frozen, fully hydrated samples can be used to improve observations from cryo-SEM as well as to enhance results from x-ray microanalysis and (digital) image analysis. Cryo-planing results of chrysanthemum (Dendranthema x grandiflorum Tzvelev) stems, hazel (Corylus avelane L.) stems, and repeseed (Brassica napus L.) pistils are presented to illustrate the use of the planing method on fibrous, hard, and delicate materials, respectively.

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

    Energy Technology Data Exchange (ETDEWEB)

    Lunov, O., E-mail: lunov@fzu.cz; Churpita, O.; Zablotskii, V.; Jäger, A.; Dejneka, A. [Institute of Physics AS CR, Prague 18221 (Czech Republic); Deyneka, I. G.; Meshkovskii, I. K. [St. Petersburg State University of Information Technologies, Mechanics and Optics, St. Petersburg 197101 (Russian Federation); Syková, E. [Institute of Experimental Medicine AS CR, Prague 14220 (Czech Republic); Kubinová, Š. [Institute of Physics AS CR, Prague 18221 (Czech Republic); Institute of Experimental Medicine AS CR, Prague 14220 (Czech Republic)

    2015-02-02

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

  3. Band excitation method applicable to scanning probe microscopy

    Science.gov (United States)

    Jesse, Stephen [Knoxville, TN; Kalinin, Sergei V [Knoxville, TN

    2010-08-17

    Methods and apparatus are described for scanning probe microscopy. A method includes generating a band excitation (BE) signal having finite and predefined amplitude and phase spectrum in at least a first predefined frequency band; exciting a probe using the band excitation signal; obtaining data by measuring a response of the probe in at least a second predefined frequency band; and extracting at least one relevant dynamic parameter of the response of the probe in a predefined range including analyzing the obtained data. The BE signal can be synthesized prior to imaging (static band excitation), or adjusted at each pixel or spectroscopy step to accommodate changes in sample properties (adaptive band excitation). An apparatus includes a band excitation signal generator; a probe coupled to the band excitation signal generator; a detector coupled to the probe; and a relevant dynamic parameter extractor component coupled to the detector, the relevant dynamic parameter extractor including a processor that performs a mathematical transform selected from the group consisting of an integral transform and a discrete transform.

  4. Humidity effects on scanning polarization force microscopy imaging

    Science.gov (United States)

    Shen, Yue; Zhou, Yuan; Sun, Yanxia; Zhang, Lijuan; Wang, Ying; Hu, Jun; Zhang, Yi

    2017-08-01

    Scanning polarization force microscopy (SPFM) is a useful surface characterization technique to visually characterize and distinguish nanomaterial with different local dielectric properties at nanometer scale. In this paper, taking the individual one-atom-thick graphene oxide (GO) and reduced graphene oxide (rGO) sheets on mica as examples, we described the influences of environmental humidity on SPFM imaging. We found that the apparent heights (AHs) or contrast of SPFM imaging was influenced significantly by relative humidity (RH) at a response time of a few seconds. And this influence rooted in the sensitive dielectric constant of mica surface to the RH change. While dielectric properties of GO and rGO sheets were almost immune to the humidity change. In addition, we gave the method to determine the critical humidity at which the contrast conversion happened under different conditions. And this is important to the contrast control and repeatable imaging of SPFM through RH adjusting. These findings suggest a strategy of controllable and repeatable imaging the local dielectric properties of nanomaterials with SPFM, which is critically important for further distinguishment, manipulation, electronic applications, etc.

  5. Scanning reflection ion microscopy in a helium ion microscope

    Directory of Open Access Journals (Sweden)

    Yuri V. Petrov

    2015-05-01

    Full Text Available Reflection ion microscopy (RIM is a technique that uses a low angle of incidence and scattered ions to form an image of the specimen surface. This paper reports on the development of the instrumentation and the analysis of the capabilities and limitations of the scanning RIM in a helium ion microscope (HIM. The reflected ions were detected by their “conversion” to secondary electrons on a platinum surface. An angle of incidence in the range 5–10° was used in the experimental setup. It was shown that the RIM image contrast was determined mostly by surface morphology but not by the atomic composition. A simple geometrical analysis of the reflection process was performed together with a Monte Carlo simulation of the angular dependence of the reflected ion yield. An interpretation of the RIM image formation and a quantification of the height of the surface steps were performed. The minimum detectable step height was found to be approximately 5 nm. RIM imaging of an insulator surface without the need for charge compensation was successfully demonstrated.

  6. An overview on bioaerosols viewed by scanning electron microscopy

    Energy Technology Data Exchange (ETDEWEB)

    Wittmaack, K. [GSF-National Research Centre for Environment and Health, Institute of Radiation Protection, 85758 Neuherberg (Germany)]. E-mail: wittmaack@gsf.de; Wehnes, H. [GSF-National Research Centre for Environment and Health, Institute of Pathology, 85758 Neuherberg (Germany); Heinzmann, U. [GSF-National Research Centre for Environment and Health, Institute of Pathology, 85758 Neuherberg (Germany); Agerer, R. [Ludwig-Maximilians University Munich, Department Biology, Biodiversity Research: Mycology, Menzinger Stasse 67, 80638 Munich (Germany)

    2005-06-15

    Bioaerosols suspended in ambient air were collected with single-stage impactors at a semiurban site in southern Germany during late summer and early autumn. Sampling was mostly carried out at a nozzle velocity of 35 m/s, corresponding to a minimum aerodynamic diameter (cut-off diameter) of aerosol particles of 0.8 {mu}m. The collected particles, sampled for short periods ({approx}15 min) to avoid pile-up, were characterized by scanning electron microscopy (SEM). The observed bioaerosols include brochosomes, fungal spores, hyphae, insect scales, hairs of plants and, less commonly, bacteria and epicuticular wax. Brochosomes, which serve as a highly water repellent body coating of leafhoppers, are hollow spheroids with diameters around 400 nm, resembling C{sub 60} or footballs (soccer balls). They are usually airborne not as individuals but in the form of large clusters containing up to 10,000 individual species or even more. Various types of spores and scales were observed, but assignment turned out be difficult due to the large number of fungi and insects from which they may have originated. Pollens were observed only once. The absence these presumably elastic particles suggests that they are frequently lost, at the comparatively high velocities, due to bounce-off from the nonadhesive impaction surfaces.

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

    Science.gov (United States)

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

    2017-08-01

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

  8. Scanning electron microscopy of eggs of Sabethes cyaneus.

    Science.gov (United States)

    Santos-Mallet, Jacenir; Sarmento, Juliana Soares; Alencar, Jeronimo; Müller, Gerson Azulim; Oliveira, Eliana Medeiros; Foster, Woodbridge A; Marcondes, Carlos Brisola

    2013-03-01

    Mosquitoes of the Neotropical genus Sabethes, some species of which are yellow fever vectors, most often develop through the immature stages in tree holes. Sabethes eggs have not been previously characterized using scanning electron microscopy. Eggs of Sabethes cyaneus (length: 349.6 +/- 2.7 microm; width: 172.6 +/- 1.14 microm; n = 10) are almost biconical when examined from the top. From a lateral perspective 2 surfaces can be seen. One surface is smooth and more convex, whereas the other is less convex and partially covered by a network from which many fungiform tubercles arise. The micropyle is situated on the smooth surface of the pointed anterior tip and is surrounded by an irregular row of tubercles, some of which are leaf shaped. No structures possibly involved in adhesion to surfaces are visible. When hatching, the egg splits dorsoventrally approximately two-thirds of the length from the anterior end. The tubercles appear to be water repellent, and the more convex/smoother surface is downturned, and this position on water was confirmed by direct observation. The eggs float free on the water surface.

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

    Science.gov (United States)

    Alves, Marcelo de Carvalho; Pozza, Edson Ampélio

    2009-07-01

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

  10. [Scanning electron microscopy findings in titanium middle ear prostheses].

    Science.gov (United States)

    Schwager, K

    2000-12-01

    Titanium as a biomaterial in ossicular replacement has widely spread within the last couple of years. 23 prostheses (12 PORPs, partial ossicular replacement prostheses and 11 TORPs total ossicular replacement prostheses) removed during revision surgery were studied using scanning electron microscopy. The average implantation time was 8 (range 3-15) months. The specimens were investigated regarding tissue growth, epithelialization, inflammation and cellular signs of rejection. Only few prostheses were totally covered by connective tissue or epithelium due to technical problems in removing the implant and the covering tissue as one specimen. But this offered the possibility to study the interface at the edges where the tissue was torn off. The connective tissue looked unremarkable. Polygonal squamous epithelium was detected on several implants. Respiratory epithelium with ciliated cells and mucus producing goblet cells was seen in two specimens. In cases of cholesteatoma or protrusion the explanted prostheses showed typical rosette-like formation of hornifying squamous epithelium. According to underlying disease a lymphocytic infiltration could be seen. There were no cellular signs of incompatibility noticed neither macrophages nor foreign body giant cells. From these investigations titanium seems to be a favorable biomaterial for ossicular replacement with good acceptance also in an implantation site showing chronic inflammation.

  11. Improved Visualization of Vertebrate Nuclear Pore Complexes by Field Emission Scanning Electron Microscopy

    National Research Council Canada - National Science Library

    Shaulov, Lihi; Harel, Amnon

    2012-01-01

    Field emission scanning electron microscopy (FESEM) can provide high-resolution three-dimensional surface imaging of many biological structures, including nuclear envelopes and nuclear pore complexes (NPCs...

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

    Science.gov (United States)

    Sindern, Sven; Meyer, F. Michael

    2016-09-01

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

  13. Outwitting the series resistance in scanning spreading resistance microscopy.

    Science.gov (United States)

    Schulze, A; Cao, R; Eyben, P; Hantschel, T; Vandervorst, W

    2016-02-01

    The performance of nanoelectronics devices critically depends on the distribution of active dopants inside these structures. For this reason, dopant profiling has been defined as one of the major metrology challenges by the international technology roadmap of semiconductors. Scanning spreading resistance microscopy (SSRM) has evolved as one of the most viable approaches over the last decade due to its excellent spatial resolution, sensitivity and quantification accuracy. However, in case of advanced device architectures like fins and nanowires a proper measurement of the spreading resistance is often hampered by the increasing impact of parasitic series resistances (e.g. bulk series resistance) arising from the confined nature of the aforementioned structures. In order to overcome this limitation we report in this paper the development and implementation of a novel SSRM mode (fast Fourier transform-SSRM: FFT-SSRM) which essentially decouples the spreading resistance from parasitic series resistance components. We show that this can be achieved by a force modulation (leading to a modulated spreading resistance signal) in combination with a lock-in deconvolution concept. In this paper we first introduce the principle of operation of the technique. We discuss in detail the underlying physical mechanisms as well as the technical implementation on a state-of-the-art atomic force microscope (AFM). We demonstrate the performance of FFT-SSRM and its ability to remove substantial series resistance components in practice. Eventually, the possibility of decoupling the spreading resistance from the intrinsic probe resistance will be demonstrated and discussed. Copyright © 2015 Elsevier B.V. All rights reserved.

  14. Environmental scanning electron microscopy observation of the ultrastructure of Demodex.

    Science.gov (United States)

    Jing, Xu; Shuling, Guo; Ying, Liu

    2005-12-01

    In this study, numbers of Demodex of hair follicles and sebaceous glands were prepared and the ultrastructure (especially the mouthparts) of Demodex was observed firstly with environmental scanning electron microscopy (ESEM). The most suitable treatment methods and optimal environmental condition for observing the genus samples were found. The samples were washed with detergent and rinsed with distilled water, and then were taken to the specimen stage, on which there was carbon adhesive tape, using special tools. When the temperature was at 5 degrees C and chamber pressure at 5 mbar respectively, the surface of the samples could be fully imaged without covering water or dehydration. The sample surfaces were plump and clear without postmortem changes and charging artifacts. Detailed information about each part of Demodex was observed by ESEM, and clear three-dimensional images were recorded. The mouthparts of D. folliculorum were composed of a complex set of structures, which included a round oral opening, a sharp oral needle, and a special hypostome that looked like a longitudinal spindle in the central position. On the end segment of palpus, there were seven strong palpal claws located on each side of the mouthparts. D. folliculorum had special piercing mouthparts, while the mouthparts of D. brevis were a simpler structure. We could not observe the oral needle of D. brevis, and there were only five pairs of palpal claws on the end segment of palpus. The offensive organs of Demodex resulted in its pathogenic effects. After studying hundreds of Demodex, we identified both female and male species of D. folliculorum, but only females of D. brevis in our sample. (c) 2005 Wiley-Liss, Inc.

  15. Spectral analysis of irregular roughness artifacts measured by atomic force microscopy and laser scanning microscopy.

    Science.gov (United States)

    Chen, Yuhang; Luo, Tingting; Ma, Chengfu; Huang, Wenhao; Gao, Sitian

    2014-12-01

    Atomic force microscopy (AFM) and laser scanning microscopy (LSM) measurements on a series of specially designed roughness artifacts were performed and the results characterized by spectral analysis. As demonstrated by comparisons, both AFM and LSM can image the complex structures with high resolution and fidelity. When the surface autocorrelation length increases from 200 to 500 nm, the cumulative power spectral density spectra of the design, AFM and LSM data reach a better agreement with each other. The critical wavelength of AFM characterization is smaller than that of LSM, and the gap between the measured and designed critical wavelengths is reduced with an increase in the surface autocorrelation length. Topography measurements of surfaces with a near zero or negatively skewed height distribution were determined to be accurate. However, obvious discrepancies were found for surfaces with a positive skewness owing to more severe dilations of either the solid tip of the AFM or the laser tip of the LSM. Further surface parameter evaluation and template matching analysis verified that the main distortions in AFM measurements are tip dilations while those in LSM are generally larger and more complex.

  16. Scanning tunneling microscopy III theory of STM and related scanning probe methods

    CERN Document Server

    Güntherodt, Hans-Joachim

    1993-01-01

    While the first two volumes on Scanning Tunneling Microscopy (STM) and its related scanning probe (SXM) methods have mainly concentrated on intro­ ducing the experimental techniques, as well as their various applications in different research fields, this third volume is exclusively devoted to the theory of STM and related SXM methods. As the experimental techniques including the reproducibility of the experimental results have advanced, more and more theorists have become attracted to focus on issues related to STM and SXM. The increasing effort in the development of theoretical concepts for STM/SXM has led to considerable improvements in understanding the contrast mechanism as well as the experimental conditions necessary to obtain reliable data. Therefore, this third volume on STM/SXM is not written by theorists for theorists, but rather for every scientist who is not satisfied by just obtaining real­ space images of surface structures by STM/SXM. After a brief introduction (Chap. 1), N. D. Lang first co...

  17. Plastic-to-Elastic Transition in Aggregated Emulsion Networks, Studied with Atomic Force Microscopy-Confocal Scanning Laser Microscopy Microrheology

    NARCIS (Netherlands)

    Filip, D.; Duits, Michael H.G.; Uricanu, V.I.; Mellema, J.

    2006-01-01

    In this paper, we demonstrate how the simultaneous application of atomic force microscopy (AFM) and confocal scanning laser microscopy (CSLM) can be used to characterize the (local) rheological properties of soft condensed matter at micrometer length scales. Measurement of AFM force curves as a

  18. Observation of the sweating in lipstick by scanning electron microscopy.

    Science.gov (United States)

    Seo, S Y; Lee, I S; Shin, H Y; Choi, K Y; Kang, S H; Ahn, H J

    1999-06-01

    The relationship between the wax matrix in lipstick and sweating has been investigated by observing the change of size and shape of the wax matrix due to sweating by Scanning Electron Microscopy (SEM). For observation by SEM, a lipstick sample was frozen in liquid nitrogen. The oil in the lipstick was then extracted in cold isopropanol (-70 degrees C) for 1-3 days. After the isopropanol was evaporated, the sample was sputtered with gold and examined by SEM. The change of wax matrix underneath the surface from fine, uniform structure to coarse, nonuniform structure resulted from the caking of surrounding wax matrix. The oil underneath the surface migrated to the surface of lipstick with sweating; consequently the wax matrix in that region was rearranged into the coarse matrix. In case of flamed lipstick, sweating was delayed and the wax matrix was much coarser than that of the unflamed one. The larger wax matrix at the surface region was good for including oil. The effect of molding temperature on sweating was also studied. As the molding temperature rose, sweating was greatly reduced and the size of the wax matrix increased. It was found that sweating was influenced by the compatibility of wax and oil. A formula consisting of wax and oil that have good compatibility has a tendency to reduce sweating and increase the size of the wax matrix. When pigments were added to wax and oil, the size of the wax matrix was changed, but in all cases sweating was increased due to the weakening of the binding force between wax and oil. On observing the thick membrane of wax at the surface of lipstick a month after molding it was also found that sweating was influenced by ageing. In conclusion, the structure of the wax matrix at the surface region of lipstick was changed with the process of flaming, molding temperature, compatibility of wax and oil, addition of pigment, and ageing. In most cases, as the size of the wax matrix was increased, sweating was reduced and delayed.

  19. Imaging by in situ Scanning Tunnelling Microscopy and its Nanotechnological Perspectives

    DEFF Research Database (Denmark)

    Andersen, Jens Enevold Thaulov

    2002-01-01

    in the interpretation of the imaging procedure. Other methods of in situ Scanning Probe Microscopy (in situ SPM), such as in situ Scanning Force Microscopy (in situ AFM) are considered for the sake of comparison and they are applied to imaging of non-conducting systems. Major results include demonstration of atomic...

  20. Nanolithography on hydrogen terminateed silicon by scanning probe microscopy

    NARCIS (Netherlands)

    Schönenberger, Christian; Kramer, Niels; Kramer, N.

    1996-01-01

    Scanning-probe microscopes (SPM), i.e. the scanning-tunneling and force microscopes, can be used to locally oxidize hydrogen-terminated silicon and hydrogenated amorphous silicon. Because of its reliability and potential for pattern transfer, this lithography process has found great attention and

  1. Adaptive remodeling of trabecular bone core cultured in 3-D bioreactor providing cyclic loading: an acoustic microscopy study.

    Science.gov (United States)

    Rupin, Fabienne; Bossis, Dorothée; Vico, Laurence; Peyrin, Françoise; Raum, Kay; Laugier, Pascal; Saïed, Amena

    2010-06-01

    Scanning acoustic microscopy (SAM) provides high-resolution mapping of acoustic impedance related to tissue stiffness. This study investigates changes in tissue acoustic impedance resulting from mechanical loading in trabecular bone cores cultured in 3-D bioreactor. Trabecular bone cores were extracted from bovine sternum (n = 15) and ulna metaphysis (n = 15). From each bone, the samples were divided in three groups. The basal control (BC) group was fixed post-extraction, the control (C) and loaded (L) groups were maintained as viable in a controlled culture-loading cell over three weeks. Samples of L group underwent a dynamic compressive strain, whereas C samples were left free from loading. After three weeks, L and C samples were embedded in polymethylmethacrylate and all samples were explored with a 200-MHz SAM. For each specimen, the acoustic impedance distribution was obtained over flat and polished section of bone blocks prepared parallel to the loading axis. Our results showed that in basal controls, the acoustic impedance varied with bone anatomical location and was 15% higher in weight-bearing ulna compared with nonweight-bearing sternum. The comparison between loaded and nonloaded groups showed that sternum-only exhibited significant change in acoustic impedance (L vs. C sternum: +9%). This result suggests that when the applied load is comparable with the stress naturally experienced by a weight-bearing bone (ulna), the tissue material properties (manifested by acoustic impedance) remained unchanged. In conclusion, SAM is a potentially relevant tool for the assessment of subtle changes in intrinsic microelastic properties of bone induced by adaptive remodeling process in response to mechanical loading. Copyright 2010 World Federation for Ultrasound in Medicine & Biology. Published by Elsevier Inc. All rights reserved.

  2. Big Data Analytics for Scanning Transmission Electron Microscopy Ptychography

    OpenAIRE

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

    2016-01-01

    Electron microscopy is undergoing a transition; from the model of producing only a few micrographs, through the current state where many images and spectra can be digitally recorded, to a new mode where very large volumes of data (movies, ptychographic and multi-dimensional series) can be rapidly obtained. Here, we discuss the application of so-called ?big-data? methods to high dimensional microscopy data, using unsupervised multivariate statistical techniques, in order to explore salient ima...

  3. A simplified model to estimate thermal resistance between carbon nanotube and sample in scanning thermal microscopy

    Science.gov (United States)

    Nazarenko, Maxim; Rosamond, Mark C.; Gallant, Andrew J.; Kolosov, Oleg V.; Dubrovskii, Vladimir G.; Zeze, Dagou A.

    2017-12-01

    Scanning thermal microscopy (SThM) is an attractive technique for nanoscale thermal measurements. Multiwalled carbon nanotubes (MWCNT) can be used to enhance a SThM probe in order to drastically increase spatial resolution while keeping required thermal sensitivity. However, an accurate prediction of the thermal resistance at the interface between the MWCNT-enhanced probe tip and a sample under study is essential for the accurate interpretation of experimental measurements. Unfortunately, there is very little literature on Kapitza interfacial resistance involving carbon nanotubes under SThM configuration. We propose a model for heat conductance through an interface between the MWCNT tip and the sample, which estimates the thermal resistance based on phonon and geometrical properties of the MWCNT and the sample, without neglecting the diamond-like carbon layer covering the MWCNT tip. The model considers acoustic phonons as the main heat carriers and account for their scattering at the interface based on a fundamental quantum mechanical approach. The predicted value of the thermal resistance is then compared with experimental data available in the literature. Theoretical predictions and experimental results are found to be of the same order of magnitude, suggesting a simplified, yet realistic model to approximate thermal resistance between carbon nanotube and sample in SThM, albeit low temperature measurements are needed to achieve a better match between theory and experiment. As a result, several possible avenues are outlined to achieve more accurate predictions and to generalize the model.

  4. Second-harmonic scanning optical microscopy of semiconductor quantum dots

    DEFF Research Database (Denmark)

    Vohnsen, B.; Bozhevolnyi, S.I.; Pedersen, K.

    2001-01-01

    Second-harmonic (SH) optical imaging of self-assembled InAlGaAs quantum dots (QD's) grown on a GaAs(0 0 1) substrate has been accomplished at room temperature by use of respectively a scanning far-field optical microscope in reflection mode and a scanning near-field optical microscope...... in transmission mode. In both cases the SH signal peaks at a pump wavelength of similar to 885 nm in correspondence to the maximum in the photoluminescence spectrum of the QD sample. SH near-field optical images exhibit spatial signal variations on a subwavelength scale that depend on the pump wavelength. We...

  5. Focusing and scanning microscopy with propagating surface plasmons

    NARCIS (Netherlands)

    Gjonaj, B.; Aulbach, Jochen; Johnson, P.M.; Mosk, Allard; Kuipers, L.; Lagendijk, Aart

    2013-01-01

    Here we demonstrate a novel surface plasmon polariton (SPP) microscope which is capable of imaging below the optical diffraction limit. A plasmonic lens, generated through phase-structured illumination, focuses SPPs down to their diffraction limit and scans the focus with steps as small as 10 nm.

  6. Scanning Emitter Lifetime Imaging Microscopy for Spontaneous Emission Control

    DEFF Research Database (Denmark)

    Frimmer, Martin; Chen, Yuntian; Koenderink, A. Femius

    2011-01-01

    We report an experimental technique to map and exploit the local density of optical states of arbitrary planar nanophotonic structures. The method relies on positioning a spontaneous emitter attached to a scanning probe deterministically and reversibly with respect to its photonic environment while...

  7. Apparent Barrier Height in Scanning Tunneling Microscopy Revisited

    DEFF Research Database (Denmark)

    Olesen, L.; Brandbyge, Mads; Sørensen, Mads Reinholdt

    1996-01-01

    The apparent barrier height phi(ap), that is, the rate of change of the logarithm of the conductance with tip-sample separation in a scanning tunneling microscope (STM), has been measured for Ni, Pt, and Au single crystal surfaces. The results show that phi(ap) is constant until point contact is ...

  8. Big Data Analytics for Scanning Transmission Electron Microscopy Ptychography

    Science.gov (United States)

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

    2016-05-01

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

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

    Science.gov (United States)

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

    2016-05-23

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

  10. Investigation of acoustic waves generated in an elastic solid by a pulsed ion beam and their application in a FIB based scanning ion acoustic microscope

    Energy Technology Data Exchange (ETDEWEB)

    Akhmadaliev, C.

    2004-12-01

    The aim of this work is to investigate the acoustic wave generation by pulsed and periodically modulated ion beams in different solid materials depending on the beam parameters and to demonstrate the possibility to apply an intensity modulated focused ion beam (FIB) for acoustic emission and for nondestructive investigation of the internal structure of materials on a microscopic scale. The combination of a FIB and an ultrasound microscope in one device can provide the opportunity of nondestructive investigation, production and modification of micro- and nanostructures simultaneously. This work consists of the two main experimental parts. In the first part the process of elastic wave generation during the irradiation of metallic samples by a pulsed beam of energetic ions was investigated in an energy range from 1.5 to 10 MeV and pulse durations of 0.5-5 {mu}s, applying ions with different masses, e.g. oxygen, silicon and gold, in charge states from 1{sup +} to 4{sup +}. The acoustic amplitude dependence on the ion beam parameters like the ion mass and energy, the ion charge state, the beam spot size and the pulse duration were of interest. This work deals with ultrasound transmitted in a solid, i.e. bulk waves, because of their importance for acoustic transmission microscopy and nondestructive inspection of internal structure of a sample. The second part of this work was carried out using the IMSA-100 FIB system operating in an energy range from 30 to 70 keV. The scanning ion acoustic microscope based on this FIB system was developed and tested. (orig.)

  11. Simulation study of secondary electron images in scanning ion microscopy

    CERN Document Server

    Ohya, K

    2003-01-01

    The target atomic number, Z sub 2 , dependence of secondary electron yield is simulated by applying a Monte Carlo code for 17 species of metals bombarded by Ga ions and electrons in order to study the contrast difference between scanning ion microscopes (SIM) and scanning electron microscopes (SEM). In addition to the remarkable reversal of the Z sub 2 dependence between the Ga ion and electron bombardment, a fine structure, which is correlated to the density of the conduction band electrons in the metal, is calculated for both. The brightness changes of the secondary electron images in SIM and SEM are simulated using Au and Al surfaces adjacent to each other. The results indicate that the image contrast in SIM is much more sensitive to the material species and is clearer than that for SEM. The origin of the difference between SIM and SEM comes from the difference in the lateral distribution of secondary electrons excited within the escape depth.

  12. Optical characterication of probes for photon scanning tunnelling microscopy

    DEFF Research Database (Denmark)

    Vohnsen, Brian; Bozhevolnyi, Sergey I.

    1999-01-01

    The photon scanning tunnelling microscope is a well-established member of the family of scanning near-field optical microscopes used for optical imaging at the sub-wavelength scale. The quality of the probes, typically pointed uncoated optical fibres, used is however difficult to evaluate...... in a direct manner and has most often been inferred from the apparent quality of recorded optical images. Complicated near-field optical imaging characteristics, together with the possibility of topographically induced artefacts, however, has increased demands for a more reliable probe characterization...... technique. Here we present experimental results obtained for optical characterization of two different probes by imaging of a well-specified near-field intensity distribution at various spatial frequencies. In particular, we observe that a sharply pointed dielectric probe can be highly suitable for imaging...

  13. Video rate near-field scanning optical microscopy

    Science.gov (United States)

    Bukofsky, S. J.; Grober, R. D.

    1997-11-01

    The enhanced transmission efficiency of chemically etched near-field optical fiber probes makes it possible to greatly increase the scanning speed of near-field optical microscopes. This increase in system bandwidth allows sub-diffraction limit imaging of samples at video rates. We demonstrate image acquisition at 10 frames/s, rate-limited by mechanical resonances in our scanner. It is demonstrated that the optical signal to noise ratio is large enough for megahertz single pixel acquisition rates.

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

    Science.gov (United States)

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

    2014-09-01

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

  15. Contribution of Metal Layer Thickness for Quantitative Backscattered Electron Imaging of Field Emission Scanning Electron Microscopy

    National Research Council Canada - National Science Library

    Kim, Hyonchol; Takei, Hiroyuki; Negishi, Tsutomu; Kudo, Masato; Terazono, Hideyuki; Yasuda, Kenji

    2012-01-01

    ...) imaging in field emission scanning electron microscopy (FE-SEM) were studied to evaluate the potential of using these particles as simultaneously distinguishable labels of target molecules in FE-SEM studies...

  16. Quantitative detection of gold nanoparticles on individual, unstained cancer cells by Scanning Electron Microscopy

    NARCIS (Netherlands)

    Hartsuiker, Liesbeth; van Es, Peter; Petersen, Wilhelmina; van Leeuwen, Ton; Terstappen, Leonardus Wendelinus Mathias Marie; Otto, Cornelis

    2011-01-01

    Gold nanoparticles are rapidly emerging for use in biomedical applications. Characterization of the interaction and delivery of nanoparticles to cells through microscopy is important. Scanning electron microscopes have the intrinsic resolution to visualize gold nanoparticles on cells. A novel sample

  17. Combination of scanning probe microscopy techniques for evaluating the electrical parameters of individual multiwalled carbon nanotubes

    Science.gov (United States)

    Sokolov, D. V.; Davletkildeev, N. A.; Bolotov, V. V.; Lobov, I. A.

    2017-10-01

    Using two techniques of scanning probe microscopy, the electrical properties (work function, Fermi level position, free carriers’ concentration, electrical resistance, conductivity, and carriers’ mobility) of individual multiwalled carbon nanotubes were evaluated.

  18. Mechanical characterization of porous nano-thin films by use of atomic force acoustic microscopy

    Energy Technology Data Exchange (ETDEWEB)

    Kopycinska-Müller, M., E-mail: Malgorzata.Kopycinska-Mueller@ikts.fraunhofer.de [Fraunhofer Institute for Ceramic Technologies and Systems – Material Diagnostics IKTS-MD, Maria Reiche Str. 2, 01109 Dresden (Germany); Faculty of Electrical Engineering and Information Technology, Technical University Dresden, Helmholtz Str. 18, 01069 Dresden (Germany); Clausner, A.; Yeap, K.-B.; Köhler, B.; Kuzeyeva, N. [Fraunhofer Institute for Ceramic Technologies and Systems – Material Diagnostics IKTS-MD, Maria Reiche Str. 2, 01109 Dresden (Germany); Mahajan, S.; Savage, T. [SBA Materials, Inc., 9430-H San Mateo Blvd NE, Albuquerque, NM 87113 (United States); Zschech, E. [Fraunhofer Institute for Ceramic Technologies and Systems – Material Diagnostics IKTS-MD, Maria Reiche Str. 2, 01109 Dresden (Germany); Faculty of Electrical Engineering and Information Technology, Technical University Dresden, Helmholtz Str. 18, 01069 Dresden (Germany); Wolter, K.-J. [Faculty of Electrical Engineering and Information Technology, Technical University Dresden, Helmholtz Str. 18, 01069 Dresden (Germany)

    2016-03-15

    The indentation modulus of thin films of porous organosilicate glass with a nominal porosity content of 30% and thicknesses of 350 nm, 200 nm, and 46 nm is determined with help of atomic force acoustic microscopy (AFAM). This scanning probe microscopy based technique provides the highest possible depth resolution. The values of the indentation modulus obtained for the 350 nm and 200 nm thin films were respectively 6.3 GPa±0.2 GPa and 7.2 GPa±0.2 GPa and free of the substrate influence. The sample with the thickness of 46 nm was tested in four independent measurement sets. Cantilevers with two different tip radii of about 150 nm and less than 50 nm were applied in different force ranges to obtain a result for the indentation modulus that was free of the substrate influence. A detailed data analysis yielded value of 8.3 GPa±0.4 GPa for the thinnest film. The values of the indentation modulus obtained for the thin films of porous organosilicate glasses increased with the decreasing film thickness. The stiffening observed for the porous films could be explained by evolution of the pore topology as a function of the film thickness. To ensure that our results were free of the substrate influence, we analyzed the ratio of the sample deformation as well as the tip radius to the film thickness. The results obtained for the substrate parameter were compared for all the measurement series and showed, which ones could be declared as free of the substrate influence. - Highlights: • We characterized mechanical properties of nano-thin films of porous organosilicate glass. • We observed an increase of the indentation modulus of the films with decreasing film thickness. • Substrate influence was evaluated with help of a dimensionless parameter.

  19. Time-resolved scanning electron microscopy with polarization analysis

    Energy Technology Data Exchange (ETDEWEB)

    Frömter, Robert, E-mail: rfroemte@physik.uni-hamburg.de; Oepen, Hans Peter [Institut für Nanostruktur-und Festkörperphysik, Universität Hamburg, Jungiusstraße 11, 20355 Hamburg (Germany); The Hamburg Centre for Ultrafast Imaging, Luruper Chaussee 149, 22761 Hamburg (Germany); Kloodt, Fabian; Rößler, Stefan; Frauen, Axel; Staeck, Philipp; Cavicchia, Demetrio R. [Institut für Nanostruktur-und Festkörperphysik, Universität Hamburg, Jungiusstraße 11, 20355 Hamburg (Germany); Bocklage, Lars [Deutsches Elektronen-Synchrotron DESY, Notkestraße 85, 22607 Hamburg (Germany); The Hamburg Centre for Ultrafast Imaging, Luruper Chaussee 149, 22761 Hamburg (Germany); Röbisch, Volker; Quandt, Eckhard [Institute for Materials Science, Christian-Albrechts-Universität zu Kiel, 24143 Kiel (Germany)

    2016-04-04

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

  20. Observation of diamond turned OFHC copper using Scanning Tunneling Microscopy

    Energy Technology Data Exchange (ETDEWEB)

    Grigg, D.A.; Russell, P.E.; Dow, T.A.

    1988-12-01

    Diamond turned OFHC copper samples have been observed within the past few months using the Scanning Tunneling Microscope. Initial results have shown evidence of artifacts which may be used to better understand the diamond turning process. The STM`s high resolution capability and three dimensional data representation allows observation and study of surface features unobtainable with conventional profilometry systems. Also, the STM offers a better quantitative means by which to analyze surface structures than the SEM. This paper discusses findings on several diamond turned OFHC copper samples having different cutting conditions. Each sample has been cross referenced using STM and SEM.

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

    Science.gov (United States)

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

    2012-12-01

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

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

    Science.gov (United States)

    Harsanyl, L

    1977-02-01

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

  3. Evaluation of the bleached human enamel by Scanning Electron Microscopy

    DEFF Research Database (Denmark)

    Miranda, Carolina Baptista; Pagani, Clovis; Benetti, Ana Raquel

    2005-01-01

    Since bleaching has become a popular procedure, the effect of peroxides on dental hard tissues is of great interest in research. Purpose: The aim of this in vitro study was to perform a qualitative analysis of the human enamel after the application of in-office bleaching agents, using Scanning...... characteristic of an erosive process that took place on human enamel. Depression areas, including the formation of craters, and exposure of enamel rods could also be detected. Conclusion: Bleaching effects on enamel morphology were randomly distributed throughout enamel surface and various degrees of enamel...

  4. Special raster scanning for reduction of charging effects in scanning electron microscopy.

    Science.gov (United States)

    Suzuki, Kazuhiko; Oho, Eisaku

    2014-01-01

    A special raster scanning (SRS) method for reduction of charging effects is developed for the field of SEM. Both a conventional fast scan (horizontal direction) and an unusual scan (vertical direction) are adopted for acquiring raw data consisting of many sub-images. These data are converted to a proper SEM image using digital image processing techniques. About sharpness of the image and reduction of charging effects, the SRS is compared with the conventional fast scan (with frame-averaging) and the conventional slow scan. Experimental results show the effectiveness of SRS images. By a successful combination of the proposed scanning method and low accelerating voltage (LV)-SEMs, it is expected that higher-quality SEM images can be more easily acquired by the considerable reduction of charging effects, while maintaining the resolution. © 2013 Wiley Periodicals, Inc.

  5. Anticipating, measuring, and minimizing MEMS mirror scan error to improve laser scanning microscopy's speed and accuracy.

    Science.gov (United States)

    Giannini, John P; York, Andrew G; Shroff, Hari

    2017-01-01

    We describe a method to speed up microelectromechanical system (MEMS) mirror scanning by > 20x, while also improving scan accuracy. We use Landweber deconvolution to determine an input voltage which would produce a desired output, based on the measured MEMS impulse response. Since the MEMS is weakly nonlinear, the observed behavior deviates from expectations, and we iteratively improve our input to minimize this deviation. This allows customizable MEMS angle vs. time with <1% deviation from the desired scan pattern. We demonstrate our technique by optimizing a point scanning microscope's raster patterns to image mammal submandibular gland and pollen at ~10 frames/s.

  6. Advantages of environmental scanning electron microscopy in studies of microorganisms.

    Science.gov (United States)

    Collins, S P; Pope, R K; Scheetz, R W; Ray, R I; Wagner, P A; Little, B J

    1993-08-01

    Microorganisms, including bacteria, fungi, protozoa, and microalgae, are composed predominantly of water which prohibits direct observation in a traditional scanning electron microscope (SEM). Preparation for SEM requires that microorganisms be fixed, frozen or dehydrated, and coated with a conductive film before observation in a high vacuum environment. Sample preparation may mechanically disturb delicate samples, compromise morphological information, and introduce other artifacts. The environmental scanning electron microscope (ESEM) provides a technology for imaging hydrated or dehydrated biological samples with minimal manipulation and without the need for conductive coatings. Sporulating cultures of three fungi, Aspergillus sp., Cunninghamella sp., and Mucor sp., were imaged in the ESEM to assess usefulness of the instrument in the direct observation of delicate, uncoated, biological specimens. Asexual sporophores showed no evidence of conidial displacement or disruption of sporangia. Uncoated algal cells of Euglena gracilis and Spirogyra sp. were examined using the backscatter electron detector (BSE) and the environmental secondary electron detector (ESD) of the ESEM. BSE images had more clearly defined intracellular structures, whereas ESD gave a clearer view of the surface E. gracilis cells fixed with potassium permanganate, Spirogyra sp. stained with Lugol's solution, and Saprolegnia sp. fixed with osmium tetroxide were compared using BSE and ESD to demonstrate that cellular details could be enhanced by the introduction of heavy metals. The effect of cellular water on signal quality was evaluated by comparing hydrated to critical point dried specimens.

  7. In-situ Scanning Transmission X-Ray Microscopy of Catalytic Solids and Related Nanomaterials

    NARCIS (Netherlands)

    de Groot, F.M.F.; de Smit, E.; van Schooneveld, M.M.; Aramburo, L.R.; Weckhuysen, B.M.

    2013-01-01

    The present status of in-situ scanning transmission X-ray microscopy (STXM) is reviewed, with an emphasis on the abilities of the STXM technique in comparison with electron microscopy. The experimental aspects and interpretation of X-ray absorption spectroscopy (XAS) are briefly introduced and the

  8. Core/shell nanofiber characterization by Raman scanning microscopy

    Science.gov (United States)

    Sfakis, Lauren; Sharikova, Anna; Tuschel, David; Costa, Felipe Xavier; Larsen, Melinda; Khmaladze, Alexander; Castracane, James

    2017-01-01

    Core/shell nanofibers are becoming increasingly popular for applications in tissue engineering. Nanofibers alone provide surface topography and increased surface area that promote cellular attachment; however, core/shell nanofibers provide the versatility of incorporating two materials with different properties into one. Such synthetic materials can provide the mechanical and degradation properties required to make a construct that mimics in vivo tissue. Many variations of these fibers can be produced. The challenge lies in the ability to characterize and quantify these nanofibers post fabrication. We developed a non-invasive method for the composition characterization and quantification at the nanoscale level of fibers using Confocal Raman microscopy. The biodegradable/biocompatible nanofibers, Poly (glycerol-sebacate)/Poly (lactic-co-glycolic) (PGS/PLGA), were characterized as a part of a fiber scaffold to quickly and efficiently analyze the quality of the substrate used for tissue engineering. PMID:28271000

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

    Science.gov (United States)

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

    2014-01-01

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

  10. PSD microscopy: a new technique for adaptive local scanning of microscale objects.

    Science.gov (United States)

    Rahimi, Mehdi; Shen, Yantao

    2017-01-01

    A position-sensitive detector/device (PSD) is a sensor that is capable of tracking the location of a laser beam on its surface. PSDs are used in many scientific instruments and technical applications including but not limited to atomic force microscopy, human eye movement monitoring, mirrors or machine tool alignment, vibration analysis, beam position control and so on. This work intends to propose a new application using the PSD. That is a new microscopy system called scanning PSD microscopy. The working mechanism is about putting an object on the surface of the PSD and fast scanning its area with a laser beam. To achieve a high degree of accuracy and precision, a reliable framework was designed using the PSD. In this work, we first tried to improve the PSD reading and its measurement performance. This was done by minimizing the effects of noise, distortion and other disturbing parameters. After achieving a high degree of confidence, the microscopy system can be implemented based on the improved PSD measurement performance. Later to improve the scanning efficiency, we developed an adaptive local scanning system to scan the whole area of the PSD in a short matter of time. It was validated that our comprehensive and adaptive local scanning method can shorten the scanning time in order of hundreds of times in comparison with the traditional raster scanning without losing any important information about the scanned 2D objects. Methods are also introduced to scan very complicated objects with bifurcations and crossings. By incorporating all these methods, the new microscopy system is capable of scanning very complicated objects in the matter of a few seconds with a resolution that is in order of a few micrometers.

  11. Three-dimensional computer-controlled acoustic pressure scanning and quantification of focused ultrasound.

    Science.gov (United States)

    Seo, Joonho; Koizumi, Norihiro; Yoshinaka, Kiyoshi; Sugita, Naohikoa; Nomiya, Akira; Homma, Yukio; Matsumoto, Yoichiro; Mitsuishi, Mamoru

    2010-04-01

    We propose an automated needle hydrophone-based scanning system to measure high-resolution 3-D acoustic pressure distributions generated by high-intensity focused ultrasound (HIFU). The hardware consists of a host computer, subsystems for HIFU generation, and an oscilloscope to sample the pressure response from a sensor in a water tank. Software was developed to control the hardware subsystems, to search for the initial scan position, and to design the scanning path and volume. A preoperative scanning plan with three perpendicular planes is used to manipulate the position of the HIFU transducer and to automate the acquisition of the spatial acoustic pressure distribution. The post-processing process displays the scanning results, compensates time delays caused by continuous linear scans, and quantifies the focal region. A method to minimize the displacement error induced by the time delay improves the scanning speed of a conventional needle hydrophone-based scanning system. Moreover, a noise-robust, automatic-focus searching algorithm using Gaussian function fitting reduces the total number of iterations and prevents the initial scanning position search from diverging. Finally, the minimum-volume enclosing ellipsoid approximation is used to quantify the size and orientation of the 3-D focal region thresholded by the minimum pressure of interest for various input conditions and to test the reproducibility of the scanning system.

  12. Micromachining and validation of the scanning acoustic microscope spatial resolution and sensitivity calibration block for 20-230 MHz frequency range.

    Science.gov (United States)

    Tamulevičius, Tomas; Šimatonis, Linas; Ulčinas, Orestas; Tamulevičius, Sigitas; Žukauskas, Egidijus; Rekuvienė, Regina; Mažeika, Liudas

    2016-10-01

    Scanning acoustic microscopy (SAM) is used as a routine non-destructive test tool for different diagnostic examinations: detection of defects such as microcracks, delamination, disbonding, inclusions, subsurface features in materials such as pores and cracks. SAM can be operated in a wide frequency range from Megahertz to Gigahertz. SAM measurement spatial resolution is diffraction limited by the wavelength of the acoustic wave in particular medium and also depends on individual transducers geometry. Actual SAM spatial resolution can be determined by measuring calibrated lithographically formed microstructures in high acoustic impedance materials. Numerical acoustic signal simulation method, based on the diffraction approach, was employed for the selection of the calibration block pattern geometry and linear dimensions of the elements. Universal calibration block for SAM operating in a 20-230 MHz frequency range was micromachined in high acoustic impedance ceramic substrates. Differently spaced (from 18 to 185 µm) lines of the same width and different widths (from 17 to 113 µm) but similar spacing lines were imposed in alumina ceramics employing one step lithography process, i.e. femtosecond laser ablation. Proposed SAM calibration pattern linear dimensions were characterized employing optical and scanning electron microscopy. Finally the samples were measured with SAM employing different frequency transducers and results were compared with the numerical simulations. It was obtained that resolution of SAM operating with 230 MHz transducer is not worse than 40 µm. © The Author (2016). Published by Oxford University Press on behalf of Journal of Molecular Cell Biology, IBCB, SIBS, CAS. All rights reserved.

  13. Identification of sandstone core damage using scanning electron microscopy

    Science.gov (United States)

    Ismail, Abdul Razak; Jaafar, Mohd Zaidi; Sulaiman, Wan Rosli Wan; Ismail, Issham; Shiunn, Ng Yinn

    2017-12-01

    Particles and fluids invasion into the pore spaces causes serious damage to the formation, resulting reduction in petroleum production. In order to prevent permeability damage for a well effectively, the damage mechanisms should be identified. In this study, water-based drilling fluid was compared to oil-based drilling fluids based on microscopic observation. The cores were damaged by several drilling fluid systems. Scanning electron microscope (SEM) was used to observe the damage mechanism caused by the drilling fluids. Results showed that the ester based drilling fluid system caused the most serious damage followed by synthetic oil based system and KCI-polymer system. Fine solids and filtrate migration and emulsion blockage are believed to be the major mechanisms controlling the changes in flow properties for the sandstone samples.

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

    Energy Technology Data Exchange (ETDEWEB)

    Joy, D.C. (Tennessee Univ., Knoxville, TN (USA) Oak Ridge National Lab., TN (USA))

    1990-01-01

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

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

    Energy Technology Data Exchange (ETDEWEB)

    Prabhakaran, Ramprashad [Pacific Northwest National Lab. (PNNL), Richland, WA (United States); Joshi, Vineet V. [Pacific Northwest National Lab. (PNNL), Richland, WA (United States); Rhodes, Mark A. [Pacific Northwest National Lab. (PNNL), Richland, WA (United States); Schemer-Kohrn, Alan L. [Pacific Northwest National Lab. (PNNL), Richland, WA (United States); Guzman, Anthony D. [Pacific Northwest National Lab. (PNNL), Richland, WA (United States); Lavender, Curt A. [Pacific Northwest National Lab. (PNNL), Richland, WA (United States)

    2016-03-30

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

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

    Energy Technology Data Exchange (ETDEWEB)

    Prabhakaran, Ramprashad [Pacific Northwest National Lab. (PNNL), Richland, WA (United States); Joshi, Vineet V. [Pacific Northwest National Lab. (PNNL), Richland, WA (United States); Rhodes, Mark A. [Pacific Northwest National Lab. (PNNL), Richland, WA (United States); Schemer-Kohrn, Alan L. [Pacific Northwest National Lab. (PNNL), Richland, WA (United States); Guzman, Anthony D. [Pacific Northwest National Lab. (PNNL), Richland, WA (United States); Lavender, Curt A. [Pacific Northwest National Lab. (PNNL), Richland, WA (United States)

    2016-10-01

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

  17. Quantitative three-dimensional ice roughness from scanning electron microscopy

    Science.gov (United States)

    Butterfield, Nicholas; Rowe, Penny M.; Stewart, Emily; Roesel, David; Neshyba, Steven

    2017-03-01

    We present a method for inferring surface morphology of ice from scanning electron microscope images. We first develop a novel functional form for the backscattered electron intensity as a function of ice facet orientation; this form is parameterized using smooth ice facets of known orientation. Three-dimensional representations of rough surfaces are retrieved at approximately micrometer resolution using Gauss-Newton inversion within a Bayesian framework. Statistical analysis of the resulting data sets permits characterization of ice surface roughness with a much higher statistical confidence than previously possible. A survey of results in the range -39°C to -29°C shows that characteristics of the roughness (e.g., Weibull parameters) are sensitive not only to the degree of roughening but also to the symmetry of the roughening. These results suggest that roughening characteristics obtained by remote sensing and in situ measurements of atmospheric ice clouds can potentially provide more facet-specific information than has previously been appreciated.

  18. Scanning Electron Microscopy with Samples in an Electric Field

    Science.gov (United States)

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

    2012-01-01

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

  19. Sequencing of adenine in DNA by scanning tunneling microscopy

    Science.gov (United States)

    Tanaka, Hiroyuki; Taniguchi, Masateru

    2017-08-01

    The development of DNA sequencing technology utilizing the detection of a tunnel current is important for next-generation sequencer technologies based on single-molecule analysis technology. Using a scanning tunneling microscope, we previously reported that dI/dV measurements and dI/dV mapping revealed that the guanine base (purine base) of DNA adsorbed onto the Cu(111) surface has a characteristic peak at V s = -1.6 V. If, in addition to guanine, the other purine base of DNA, namely, adenine, can be distinguished, then by reading all the purine bases of each single strand of a DNA double helix, the entire base sequence of the original double helix can be determined due to the complementarity of the DNA base pair. Therefore, the ability to read adenine is important from the viewpoint of sequencing. Here, we report on the identification of adenine by STM topographic and spectroscopic measurements using a synthetic DNA oligomer and viral DNA.

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

    Science.gov (United States)

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

    2016-06-01

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

  1. Scanning electron microscopy investigations regarding Adonis vernalis L. flower morphology

    Directory of Open Access Journals (Sweden)

    Irina Neta GOSTIN

    2009-11-01

    Full Text Available The floral morphology of Adonis vernalis L. was observed with a scanning electron microscope (SEM. The investigations are important to clarify some taxonomical problems and also could provide useful diagnostic elements for the identification of this medicinal plant in powdered materials. All floral organs are initiated spirally and centripetally and develop centripetally. The petals (8-12 are shorter than the sepals (5-6 in early developmental stages. The petals are disposed on spiral (with 3-4 whorls. The stamens (numerous are unbranched and reach maturity centripetally; they are free of the perianth. The anther walls consisting of a single layer epidermis in the anther wall surrounding the sporagenous tissue, one row of endothecium, two to four rows of middle layer and one row of tapetum layer. In the anther walls, the tapetal cells, by glandular type, persist later in ontogenesis. Pollen grains are tricolpate with echinate surface. The gynoecium is multiple, apocarpous with distinct carpels. The carpels are ascidiate from the beginning. At the base of each carpel, numerousness short, unicellular, trichomes are present. The stigma differentiates as two crests along the ventral slit of the ovary. Each carpel contains a single ovule inside the ovary cavity. The mature ovule is anatropous, with two integuments. It is almost parallel to the funicle.

  2. Fabrication and characterization of probes for combined scanning electrochemical/optical microscopy experiments.

    Science.gov (United States)

    Lee, Youngmi; Bard, Allen J

    2002-08-01

    A technique that combines scanning electrochemical microscopy (SECM) and optical microscopy (OM) was implemented with a new probe tip. The tip for scanning electrochemicaVoptical microscopy (SECM/OM) was constructed by insulating a typical gold-coated near-field scanning optical microscopy tip using electrophoretic anodic paint. Once fabricated, the tip was characterized by steady-state cyclic voltammetry, as well as optical and electrochemical approach experiments. This tip generated a stable steady-state current and well-defined SECM approach curves for both conductive and insulating substrates. Durable tips whose geometry was a ring with < 1 microm as outer ring radius could be consistently fabricated. Simultaneous electrochemical and optical images of an interdigitated array electrode were obtained with a resolution on the micrometer scale, demonstrating good performance of the tip as both an optical and an electrochemical probe for imaging microstructures. The SECM feedback current measurements were successfully employed to determine tip-substrate distances for imaging.

  3. Candida albicans morphologies revealed by scanning electron microscopy analysis

    Directory of Open Access Journals (Sweden)

    M. Staniszewska

    2013-09-01

    Full Text Available Scanning electron microscope (SEM observations were used to analyze particular morphologies of Candida albicans clinical isolate (strain 82 and mutants defective in hyphae-promoting genes EFG1 (strain HLC52 and/ or CPH1 (strains HLC54 and Can16. Transcription factors Efg1 and Cph1 play role in regulating filamentation and adhesion of C. albicans' morphologies. Comparative analysis of such mutants and clinical isolate showed that Efg1 is required for human serum-induced cell growth and morphological switching. In the study, distinct differences between ultrastructural patterns of clinical strain's and null mutants' morphologies were observed (spherical vs tube-like blastoconidia, or solid and fragile constricted septa vs only the latter observed in strains with EFG1 deleted. In addition, wild type strain displayed smooth colonies of cells in comparison to mutants which exhibited wrinkled phenotype. It was observed that blastoconidia of clinical strain exhibited either polarly or randomly located budding. Contrariwise, morphotypes of mutants showed either multiple polar budding or a centrally located single bud scar (mother-daughter cell junction distinguishing tube-like yeast/ pseudohyphal growth (the length-to-width ratios larger than 1.5. In their planktonic form of growth, blastoconidia of clinical bloodstream isolate formed constitutively true hyphae under undiluted human serum inducing conditions. It was found that true hyphae are essential elements for developing structural integrity of conglomerate, as mutants displaying defects in their flocculation and conglomerate-forming abilities in serum. While filamentation is an important virulence trait in C. albicans the true hyphae are the morphologies which may be expected to play a role in bloodstream infections.

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

    Directory of Open Access Journals (Sweden)

    Pia C. Lansåker

    2014-10-01

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

  5. Cryo-Scanning Electron Microscopy (SEM) and Scanning Transmission Electron Microscopy (STEM)-in-SEM for Bio- and Organo-Mineral Interface Characterization in the Environment.

    Science.gov (United States)

    Wille, Guillaume; Hellal, Jennifer; Ollivier, Patrick; Richard, Annie; Burel, Agnes; Jolly, Louis; Crampon, Marc; Michel, Caroline

    2017-11-16

    Understanding biofilm interactions with surrounding substratum and pollutants/particles can benefit from the application of existing microscopy tools. Using the example of biofilm interactions with zero-valent iron nanoparticles (nZVI), this study aims to apply various approaches in biofilm preparation and labeling for fluorescent or electron microscopy and energy dispersive X-ray spectrometry (EDS) microanalysis for accurate observations. According to the targeted microscopy method, biofilms were sampled as flocs or attached biofilm, submitted to labeling using 4',6-diamidino-2-phenylindol, lectins PNA and ConA coupled to fluorescent dye or gold nanoparticles, and prepared for observation (fixation, cross-section, freezing, ultramicrotomy). Fluorescent microscopy revealed that nZVI were embedded in the biofilm structure as aggregates but the resolution was insufficient to observe individual nZVI. Cryo-scanning electron microscopy (SEM) observations showed nZVI aggregates close to bacteria, but it was not possible to confirm direct interactions between nZVI and cell membranes. Scanning transmission electron microscopy in the SEM (STEM-in-SEM) showed that nZVI aggregates could enter the biofilm to a depth of 7-11 µm. Bacteria were surrounded by a ring of extracellular polymeric substances (EPS) preventing direct nZVI/membrane interactions. STEM/EDS mapping revealed a co-localization of nZVI aggregates with lectins suggesting a potential role of EPS in nZVI embedding. Thus, the combination of divergent microscopy approaches is a good approach to better understand and characterize biofilm/metal interactions.

  6. Nonperturbing measurements of spatially distributed underwater acoustic fields using a scanning laser Doppler vibrometer.

    Science.gov (United States)

    Harland, Andy R; Petzing, Jon N; Tyrer, John R

    2004-01-01

    Localized changes in the density of water induced by the presence of an acoustic field cause perturbations in the localized refractive index. This relationship has given rise to a number of nonperturbing optical metrology techniques for recording measurement parameters from underwater acoustic fields. A method that has been recently developed involves the use of a Laser Doppler Vibrometer (LDV) targeted at a fixed, nonvibrating, plate through an underwater acoustic field. Measurements of the rate of change of optical pathlength along a line section enable the identification of the temporal and frequency characteristics of the acoustic wave front. This approach has been extended through the use of a scanning LDV, which facilitates the measurement of a range of spatially distributed parameters. A mathematical model is presented that relates the distribution of pressure amplitude and phase in a planar wave front with the rate of change of optical pathlength measured by the LDV along a specifically orientated laser line section. Measurements of a 1 MHz acoustic tone burst generated by a focused transducer are described and the results presented. Graphical depictions of the acoustic power and phase distribution recorded by the LDV are shown, together with images representing time history during the acoustic wave propagation.

  7. Resonant Scanning with Large Field of View Reduces Photobleaching and Enhances Fluorescence Yield in STED Microscopy.

    Science.gov (United States)

    Wu, Yong; Wu, Xundong; Lu, Rong; Zhang, Jin; Toro, Ligia; Stefani, Enrico

    2015-10-01

    Photobleaching is a major limitation of superresolution Stimulated Depletion Emission (STED) microscopy. Fast scanning has long been considered an effective means to reduce photobleaching in fluorescence microscopy, but a careful quantitative study of this issue is missing. In this paper, we show that the photobleaching rate in STED microscopy can be slowed down and the fluorescence yield be enhanced by scanning with high speed, enabled by using large field of view in a custom-built resonant-scanning STED microscope. The effect of scanning speed on photobleaching and fluorescence yield is more remarkable at higher levels of depletion laser irradiance, and virtually disappears in conventional confocal microscopy. With ≥6 GW∙cm(-2) depletion irradiance, we were able to extend the fluorophore survival time of Atto 647N and Abberior STAR 635P by ~80% with 8-fold wider field of view. We confirm that STED Photobleaching is primarily caused by the depletion light acting upon the excited fluorophores. Experimental data agree with a theoretical model. Our results encourage further increasing the linear scanning speed for photobleaching reduction in STED microscopy.

  8. Artifact mitigation of ptychography integrated with on-the-fly scanning probe microscopy

    Science.gov (United States)

    Huang, Xiaojing; Yan, Hanfei; Ge, Mingyuan; Öztürk, Hande; Nazaretski, Evgeny; Robinson, Ian K.; Chu, Yong S.

    2017-07-01

    We report our experiences with conducting ptychography simultaneously with the X-ray fluorescence measurement using the on-the-fly mode for efficient multi-modality imaging. We demonstrate that the periodic artifact inherent to the raster scan pattern can be mitigated using a sufficiently fine scan step size to provide an overlap ratio of >70%. This allows us to obtain transmitted phase contrast images with enhanced spatial resolution from ptychography while maintaining the fluorescence imaging with continuous-motion scans on pixelated grids. This capability will greatly improve the competence and throughput of scanning probe X-ray microscopy.

  9. Scanning electron microscopy and transmission electron microscopy study of hot-deformed gamma-TiAl-based alloy microstructure.

    Science.gov (United States)

    Chrapoński, J; Rodak, K

    2006-09-01

    The aim of this work was to assess the changes in the microstructure of hot-deformed specimens made of alloys containing 46-50 at.% Al, 2 at.% Cr and 2 at.% Nb (and alloying additions such as carbon and boron) with the aid of scanning electron microscopy and transmission electron microscopy techniques. After homogenization and heat treatment performed in order to make diverse lamellae thickness, the specimens were compressed at 1000 degrees C. Transmission electron microscopy examinations of specimens after the compression test revealed the presence of heavily deformed areas with a high density of dislocation. Deformation twins were also observed. Dynamically recrystallized grains were revealed. For alloys no. 2 and no. 3, the recovery and recrystallization processes were more extensive than for alloy no. 1.

  10. Spontaneous Polarization in Bio-organic Materials Studied by Scanning Pyroelectric Microscopy (SPEM) and Second Harmonic Generation Microscopy (SHGM)

    Science.gov (United States)

    Putzeys, T.; Wübbenhorst, M.; van der Veen, M. A.

    2015-06-01

    Bio-organic materials such as bones, teeth, and tendon generally show nonlinear optical (Masters and So in Handbook of Biomedical Nonlinear Optical Microscopy, 2008), pyro- and piezoelectric (Fukada and Yasuda in J Phys Soc Jpn 12:1158, 1957) properties, implying a permanent polarization, the presence of which can be rationalized by describing the growth of the sample and the creation of a polar axis according to Markov's theory of stochastic processes (Hulliger in Biophys J 84:3501, 2003; Batagiannis et al. in Curr Opin Solid State Mater Sci 17:107, 2010). Two proven, versatile techniques for probing spontaneous polarization distributions in solids are scanning pyroelectric microscopy (SPEM) and second harmonic generation microscopy (SHGM). The combination of pyroelectric scanning with SHG-microscopy in a single experimental setup leading to complementary pyroelectric and nonlinear optical data is demonstrated, providing us with a more complete image of the polarization in organic materials. Crystals consisting of a known polar and hyperpolarizable material, CNS (4-chloro-4-nitrostilbene) are used as a reference sample, to verify the functionality of the setup, with both SPEM and SHGM images revealing the same polarization domain information. In contrast, feline and human nails exhibit a pyroelectric response, but a second harmonic response is absent for both keratin containing materials, implying that there may be symmetry-allowed SHG, but with very inefficient second harmonophores. This new approach to polarity detection provides additional information on the polar and hyperpolar nature in a variety of (bio) materials.

  11. Observation of silicon carbide Schottky barrier diode under applied reverse bias using atomic force microscopy/Kelvin probe force microscopy/scanning capacitance force microscopy

    Science.gov (United States)

    Uruma, Takeshi; Satoh, Nobuo; Yamamoto, Hidekazu

    2017-08-01

    We have observed a commercial silicon-carbide Schottky barrier diode (SiC-SBD) using our novel analysis system, in which atomic force microscopy (AFM) is combined with both Kelvin probe force microscopy (KFM; for surface-potential measurement) and scanning capacitance force microscopy (SCFM; for differential-capacitance measurement). The results obtained for the SiC-SBD under an applied reverse bias indicate both the scan area in the sample and a peak value of the SCFM signal in the region where the existence of trapped electrons is deduced from the KFM analysis. Thus, our measurement system can be used to examine commercial power devices; however, novel polishing procedures are required in order to investigate the Schottky contact region.

  12. Scanning ion conductance microscopy for visualizing the three-dimensional surface topography of cells and tissues.

    Science.gov (United States)

    Nakajima, Masato; Mizutani, Yusuke; Iwata, Futoshi; Ushiki, Tatsuo

    2018-01-01

    Scanning ion conductance microscopy (SICM), which belongs to the family of scanning probe microscopy, regulates the tip-sample distance by monitoring the ion current through the use of an electrolyte-filled nanopipette as the probing tip. Thus, SICM enables "contact-free" imaging of cell surface topography in liquid conditions. In this paper, we applied hopping mode SICM for obtaining topographical images of convoluted tissue samples such as trachea and kidney in phosphate buffered saline. Some of the SICM images were compared with the images obtained by scanning electron microscopy (SEM) after drying the same samples. We showed that the imaging quality of hopping mode SICM was excellent enough for investigating the three-dimensional surface structure of the soft tissue samples. Thus, SICM is expected to be used for imaging a wide variety of cells and tissues - either fixed or alive- at high resolution under physiologically relevant liquid conditions. Copyright © 2017 Elsevier Ltd. All rights reserved.

  13. Emulation and design of terahertz reflection-mode confocal scanning microscopy based on virtual pinhole

    Science.gov (United States)

    Yang, Yong-fa; Li, Qi

    2014-12-01

    In the practical application of terahertz reflection-mode confocal scanning microscopy, the size of detector pinhole is an important factor that determines the performance of spatial resolution characteristic of the microscopic system. However, the use of physical pinhole brings some inconvenience to the experiment and the adjustment error has a great influence on the experiment result. Through reasonably selecting the parameter of matrix detector virtual pinhole (VPH), it can efficiently approximate the physical pinhole. By using this approach, the difficulty of experimental calibration is reduced significantly. In this article, an imaging scheme of terahertz reflection-mode confocal scanning microscopy that is based on the matrix detector VPH is put forward. The influence of detector pinhole size on the axial resolution of confocal scanning microscopy is emulated and analyzed. Then, the parameter of VPH is emulated when the best axial imaging performance is reached.

  14. Studying Dynamic Processes of Nano-sized Objects in Liquid using Scanning Transmission Electron Microscopy

    OpenAIRE

    Hermannsd?rfer, Justus; de Jonge, Niels

    2017-01-01

    Samples fully embedded in liquid can be studied at a nanoscale spatial resolution with Scanning Transmission Electron Microscopy (STEM) using a microfluidic chamber assembled in the specimen holder for Transmission Electron Microscopy (TEM) and STEM. The microfluidic system consists of two silicon microchips supporting thin Silicon Nitride (SiN) membrane windows. This article describes the basic steps of sample loading and data acquisition. Most important of all is to ensure that the liquid c...

  15. Data analysis using the Internet: the World Wide Web scanning probe microscopy data analysis system.

    Science.gov (United States)

    Williams, P M; Davies, M C; Roberts, C J; Tendler, S J

    1997-10-01

    The first interactive world-wide web-based image analysis system is presented (http://pharm6.pharm.nottingham.ac.uk/processing/main. html). The system, currently tailored to scanning probe microscopy image data, has been developed to permit the use of software algorithms developed within our laboratory by researchers throughout the world. The implementation and functionality of the scanning probe microscopy server is described. Feedback from users of the facility has demonstrated its value within the research community, and highlighted key operational issues which are to be addressed. A future role of Internet-based data processing software is also discussed.

  16. Scanning tunneling microscopy I general principles and applications to clean and adsorbate-covered surfaces

    CERN Document Server

    Wiesendanger, Roland

    1992-01-01

    Scanning Tunneling Microscopy I provides a unique introduction to a novel and fascinating technique that produces beautiful images of nature on an atomic scale. It is the first of three volumes that together offer a comprehensive treatment of scanning tunneling microscopy, its diverse applications, and its theoretical treatment. In this volume the reader will find a detailed description of the technique itself and of its applications to metals, semiconductors, layered materials, adsorbed molecules and superconductors. In addition to the many representative results reviewed, extensive references to original work will help to make accessible the vast body of knowledge already accumulated in this field.

  17. Local analysis of semiconductor nanoobjects by scanning tunneling atomic force microscopy

    Directory of Open Access Journals (Sweden)

    Natalia A. Lashkova

    2015-03-01

    Full Text Available The features of the current–voltage (I–V measurements in local regions of semiconductor nanostructures by conductive atomic force microscopy (AFM are discussed. The standard procedure of I–V measurements in conductive AFM leads not infrequently to the thermomechanical stresses in the sample and, as a consequence, nonreproducibility and unreliability of measurements. The technique of obtaining reproducible current–voltage characteristics is proposed. According to the technique, a series of measurements of the selected scanning area in the mode of conducting AFM should be taken, each at the certain value of the potential. According to a series of scans I–V curve at a particular point (for any point of the scan was plotted. The program is realized in the LabVIEW software. The proposed method extends the capabilities of scanning probe microscopy in the diagnosis of nanostructured semiconductor materials.

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

    Science.gov (United States)

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

    2015-01-01

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

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

    OpenAIRE

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

    2014-01-01

    Background Limbal ring (also known as ‘circle’) contact lenses are becoming increasingly popular, especially in Asian markets because of their eye-enhancing effects. The pigment particles that give the eye-enhancing effects of these lenses can be found on the front or back surface of the contact lens or ‘enclosed’ within the lens matrix. The purpose of this research was to evaluate the pigment location and surface roughness of seven types of ‘circle’ contact lenses. Methods Scanning electron ...

  20. Superresolution upgrade for confocal spinning disk systems using image scanning microscopy (Conference Presentation)

    Science.gov (United States)

    Isbaner, Sebastian; Hähnel, Dirk; Gregor, Ingo; Enderlein, Jörg

    2017-02-01

    Confocal Spinning Disk Systems are widely used for 3D cell imaging because they offer the advantage of optical sectioning at high framerates and are easy to use. However, as in confocal microscopy, the imaging resolution is diffraction limited, which can be theoretically improved by a factor of 2 using the principle of Image Scanning Microscopy (ISM) [1]. ISM with a Confocal Spinning Disk setup (CSDISM) has been shown to improve contrast as well as lateral resolution (FWHM) from 201 +/- 20 nm to 130 +/- 10 nm at 488 nm excitation. A minimum total acquisition time of one second per ISM image makes this method highly suitable for 3D live cell imaging [2]. Here, we present a multicolor implementation of CSDISM for the popular Micro-Manager Open Source Microscopy platform. Since changes in the optical path are not necessary, this will allow any researcher to easily upgrade their standard Confocal Spinning Disk system at remarkable low cost ( 5000 USD) with an ISM superresolution option. [1]. Müller, C.B. and Enderlein, J. Image Scanning Microscopy. Physical Review Letters 104, (2010). [2]. Schulz, O. et al. Resolution doubling in fluorescence microscopy with confocal spinning-disk image scanning microscopy. Proceedings of the National Academy of Sciences of the United States of America 110, 21000-5 (2013).

  1. Artifact characterization and reduction in scanning X-ray Zernike phase contrast microscopy.

    Science.gov (United States)

    Vartiainen, Ismo; Holzner, Christian; Mohacsi, Istvan; Karvinen, Petri; Diaz, Ana; Pigino, Gaia; David, Christian

    2015-05-18

    Zernike phase contrast microscopy is a well-established method for imaging specimens with low absorption contrast. It has been successfully implemented in full-field microscopy using visible light and X-rays. In microscopy Cowley's reciprocity principle connects scanning and full-field imaging. Even though the reciprocity in Zernike phase contrast has been discussed by several authors over the past thirty years, only recently it was experimentally verified using scanning X-ray microscopy. In this paper, we investigate the image and contrast formation in scanning Zernike phase contrast microscopy with a particular and detailed focus on the origin of imaging artifacts that are typically associated with Zernike phase contrast. We demonstrate experimentally with X-rays the effect of the phase mask design on the contrast and halo artifacts and present an optimized design of the phase mask with respect to photon efficiency and artifact reduction. Similarly, due to the principle of reciprocity the observations and conclusions of this work have direct applicability to Zernike phase contrast in full-field microscopy as well.

  2. Simultaneous Bright-Field and Dark-Field Scanning Transmission Electron Microscopy in Scanning Electron Microscopy: A New Approach for Analyzing Polymer System Morphology

    Science.gov (United States)

    Patel, Binay S.

    Scanning transmission electron microscopy in scanning electron microscopy (STEM-IN-SEM) is a convenient technique for polymer characterization. Utilizing the lower accelerating voltages, larger field of view and, exclusion of post-specimen projection lens in an SEM; STEM-IN-SEM has shown results comparable to transmission electron microscopy (TEM) observation of polymer morphology. Various specimen-holder geometries and detector arrangements have been used for bright field (BF) STEM-IN-SEM imaging. To further the characterization potential of STEM-IN-SEM a new specimen holder has been developed to facilitate simultaneous BF and dark field (DF) STEM-IN-SEM imaging. A new specimen holder and a new microscope configuration were designed for this new imaging technique. BF and DF signals were maximized for optimal STEM-IN-SEM imaging. BF signal intensities were found to be twice as large as DF signal intensities. BF and DF STEM-IN-SEM imaging spatial resolutions are limited to 1.8 nm and approximately 5 nm, respectively. Simultaneous BF & DF STEM-IN-SEM imaging is applicable to both industrial and academic research environments. Examples of commodity and engineering polymer morphology characterization are provided. Results are comparable to TEM observation and may serve as a suitable precursor to STEM characterization of polymer systems. Finally, future developments of various accessories for this technique are discussed.

  3. Confocal laser scanning microscopy. Using new technology to answer old questions in forensic investigations.

    Science.gov (United States)

    Turillazzi, Emanuela; Karch, Steven B; Neri, Margherita; Pomara, Cristoforo; Riezzo, Irene; Fineschi, Vittorio

    2008-03-01

    Confocal laser scanning microscopy (CLSM) is a relatively new technique for microscopic imaging. It has found a wide field of application in the general sphere of biological sciences. It has completely changed the study of cells and tissues by allowing greater resolution, optical sectioning of the sample and three-dimensional sanoke reconstruction. Confocal microscopy represents a valid, precious and useful tool capable of providing data (images) of unrivalled clearness and definition. This review discusses the possible applications of confocal microscopy in specific fields of forensic investigation, with specific regard to ballistics, forensic histopathology and toxicological pathology.

  4. Ultrafast axial scanning for two-photon microscopy via a digital micromirror device and binary holography.

    Science.gov (United States)

    Cheng, Jiyi; Gu, Chenglin; Zhang, Dapeng; Wang, Dien; Chen, Shih-Chi

    2016-04-01

    In this Letter, we present an ultrafast nonmechanical axial scanning method for two-photon excitation (TPE) microscopy based on binary holography using a digital micromirror device (DMD), achieving a scanning rate of 4.2 kHz, scanning range of ∼180  μm, and scanning resolution (minimum step size) of ∼270  nm. Axial scanning is achieved by projecting the femtosecond laser to a DMD programmed with binary holograms of spherical wavefronts of increasing/decreasing radii. To guide the scanner design, we have derived the parametric relationships between the DMD parameters (i.e., aperture and pixel size), and the axial scanning characteristics, including (1) maximum optical power, (2) minimum step size, and (3) scan range. To verify the results, the DMD scanner is integrated with a custom-built TPE microscope that operates at 60 frames per second. In the experiment, we scanned a pollen sample via both the DMD scanner and a precision z-stage. The results show the DMD scanner generates images of equal quality throughout the scanning range. The overall efficiency of the TPE system was measured to be ∼3%. With the high scanning rate, the DMD scanner may find important applications in random-access imaging or high-speed volumetric imaging that enables visualization of highly dynamic biological processes in 3D with submillisecond temporal resolution.

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

    Science.gov (United States)

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

    2011-09-01

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

  6. Alternative configuration scheme for signal amplification with scanning ion conductance microscopy

    Science.gov (United States)

    Kim, Joonhui; Kim, Seong-Oh; Cho, Nam-Joon

    2015-02-01

    Scanning Ion Conductance Microscopy (SICM) is an emerging nanotechnology tool to investigate the morphology and charge transport properties of nanomaterials, including soft matter. SICM uses an electrolyte filled nanopipette as a scanning probe and detects current changes based on the distance between the nanopipette apex and the target sample in an electrolyte solution. In conventional SICM, the pipette sensor is excited by applying voltage as it raster scans near the surface. There have been attempts to improve upon raster scanning because it can induce collisions between the pipette sidewalls and target sample, especially for soft, dynamic materials (e.g., biological cells). Recently, Novak et al. demonstrated that hopping probe ion conductance microscopy (HPICM) with an adaptive scan method can improve the image quality obtained by SICM for such materials. However, HPICM is inherently slower than conventional raster scanning. In order to optimize both image quality and scanning speed, we report the development of an alternative configuration scheme for SICM signal amplification that is based on applying current to the nanopipette. This scheme overcomes traditional challenges associated with low bandwidth requirements of conventional SICM. Using our alternative scheme, we demonstrate successful imaging of L929 fibroblast cells and discuss the capabilities of this instrument configuration for future applications.

  7. Alternative configuration scheme for signal amplification with scanning ion conductance microscopy.

    Science.gov (United States)

    Kim, Joonhui; Kim, Seong-Oh; Cho, Nam-Joon

    2015-02-01

    Scanning Ion Conductance Microscopy (SICM) is an emerging nanotechnology tool to investigate the morphology and charge transport properties of nanomaterials, including soft matter. SICM uses an electrolyte filled nanopipette as a scanning probe and detects current changes based on the distance between the nanopipette apex and the target sample in an electrolyte solution. In conventional SICM, the pipette sensor is excited by applying voltage as it raster scans near the surface. There have been attempts to improve upon raster scanning because it can induce collisions between the pipette sidewalls and target sample, especially for soft, dynamic materials (e.g., biological cells). Recently, Novak et al. demonstrated that hopping probe ion conductance microscopy (HPICM) with an adaptive scan method can improve the image quality obtained by SICM for such materials. However, HPICM is inherently slower than conventional raster scanning. In order to optimize both image quality and scanning speed, we report the development of an alternative configuration scheme for SICM signal amplification that is based on applying current to the nanopipette. This scheme overcomes traditional challenges associated with low bandwidth requirements of conventional SICM. Using our alternative scheme, we demonstrate successful imaging of L929 fibroblast cells and discuss the capabilities of this instrument configuration for future applications.

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

    Science.gov (United States)

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

  9. Scanning electron microscopy and X-ray spectroscopy applied to mycelial phase of sporothrix schenckii

    Directory of Open Access Journals (Sweden)

    M. Thibaut

    1975-04-01

    Full Text Available Scanning electron microscopy applied to the mycelial phase of Sporothrix schenckii shows a matted mycelium with conidia of a regular pattern. X-Ray microanalysis applied in energy dispersive spectroscopy and also in wavelength dispersive spectroscopy reveals the presence of several elements of Mendeleef's classification.

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

    Energy Technology Data Exchange (ETDEWEB)

    Wang, Peng; Behan, Gavin; Kirkland, Angus I. [Department of Materials, University of Oxford, Parks Road, Oxford OX1 3PH (United Kingdom); Nellist, Peter D., E-mail: peter.nellist@materials.ox.ac.uk [Department of Materials, University of Oxford, Parks Road, Oxford OX1 3PH (United Kingdom); Cosgriff, Eireann C.; D' Alfonso, Adrian J.; Morgan, Andrew J.; Allen, Leslie J. [School of Physics, University of Melbourne, Parkville, Victoria 3010 (Australia); Hashimoto, Ayako [Advanced Nano-characterization Center, National Institute for Materials Science (NIMS), 3-13 Sakura, Tsukuba 305-0003 (Japan); Takeguchi, Masaki [Advanced Nano-characterization Center, National Institute for Materials Science (NIMS), 3-13 Sakura, Tsukuba 305-0003 (Japan); High Voltage Electron Microscopy Station, NIMS, 3-13 Sakura, Tsukuba 305-0003 (Japan); Mitsuishi, Kazutaka [Advanced Nano-characterization Center, National Institute for Materials Science (NIMS), 3-13 Sakura, Tsukuba 305-0003 (Japan); Quantum Dot Research Center, NIMS, 3-13 Sakura, Tsukuba 305-0003 (Japan); Shimojo, Masayuki [High Voltage Electron Microscopy Station, NIMS, 3-13 Sakura, Tsukuba 305-0003 (Japan); Advanced Science Research Laboratory, Saitama Institute of Technology, 1690 Fusaiji, Fukaya 369-0293 (Japan)

    2011-06-15

    Scanning confocal electron microscopy (SCEM) offers a mechanism for three-dimensional imaging of materials, which makes use of the reduced depth of field in an aberration-corrected transmission electron microscope. The simplest configuration of SCEM is the bright-field mode. In this paper we present experimental data and simulations showing the form of bright-field SCEM images. We show that the depth dependence of the three-dimensional image can be explained in terms of two-dimensional images formed in the detector plane. For a crystalline sample, this so-called probe image is shown to be similar to a conventional diffraction pattern. Experimental results and simulations show how the diffracted probes in this image are elongated in thicker crystals and the use of this elongation to estimate sample thickness is explored. -- Research Highlights: {yields} The confocal probe image in a scanning confocal electron microscopy image reveals information about the thickness and height of the crystalline layer. {yields} The form of the contrast in a three-dimensional bright-field scanning confocal electron microscopy image can be explained in terms of the confocal probe image. {yields} Despite the complicated form of the contrast in bright-field scanning confocal electron microscopy, we see that depth information is transferred on a 10 nm scale.

  11. Field-emission scanning electron microscopy of the internal cellular organization of fungi

    NARCIS (Netherlands)

    Muller, W.H.; Aelst, van A.C.; Humbel, B.M.; Krift, van der T.P.; Boekhout, T.

    2000-01-01

    Internal viewing of the cellular organization of hyphae by scanning electron microscopy is an alternative to observing sectioned fungal material with a transmission electron microscope. To study cytoplasmic organelles in the hyphal cells of fungi by SEM, colonies were chemically fixed with

  12. Morphologic differences observed by scanning electron microscopy according to the reason for pseudophakic IOL explantation

    DEFF Research Database (Denmark)

    Fernandez-Buenaga, Roberto; Alio, Jorge L.; Ramirez, Jose M.

    2015-01-01

    Purpose To compare variations in surface morphology, as studied by scanning electron microscopy (SEM), of explanted intraocular lenses (IOLs) concerning the cause leading to the explantation surgery. Methods In this prospective multicenter study, explanted IOLs were analyzed by SEM and energy-dis...

  13. DTAF: an efficient probe to study cyanobacterial-plant interaction using confocal laser scanning microscopy (CLSM)

    NARCIS (Netherlands)

    Ahmed, M.; Stal, L.J.; Hasnain, S.

    2011-01-01

    A variety of microscopic techniques have been utilized to study cyanobacterial associations with plant roots, but confocal laser scanning microscopy (CLSM) is the least used due to the unavailability of a suitable fluorescent dye. Commonly used lectins have problems with their binding ability with

  14. DTAF: an efficient probe to study cyanobacterial-plant interaction using confocal laser scanning microscopy (CLSM).

    NARCIS (Netherlands)

    Ahmed, M.; Stal, L.J.; Hasnain, S.

    2011-01-01

    A variety of microscopic techniques have been utilized to study cyanobacterial associations with plant roots, but confocal laser scanning microscopy (CLSM) is the least used due to the unavailability of a suitable fluorescent dye. Commonly used lectins have problems with their binding ability with

  15. Scanning electron microscopy with polarization analysis for multilayered chiral spin textures

    NARCIS (Netherlands)

    Lucassen, Juriaan; Kloodt-Twesten, Fabian; Frömter, Robert; Oepen, Hans Peter; Duine, Rembert A.|info:eu-repo/dai/nl/304830127; Swagten, Henk J. M.; Koopmans, Bert; Lavrijsen, Reinoud

    We show that scanning electron microscopy with polarization analysis (SEMPA) that is sensitive to both in-plane magnetization components can be used to image the out-of-plane magnetized multi-domain state in multilayered chiral spin textures. By depositing a thin layer of Fe on top of the multilayer

  16. Batch fabrication of scanning microscopy probes for thermal and magnetic imaging using standard micromachining

    NARCIS (Netherlands)

    Sarajlic, Edin; Vermeer, Rolf; Delalande, M.Y.; Siekman, Martin Herman; Huijink, R.; Fujita, H.; Abelmann, Leon

    2010-01-01

    We present a process for batch fabrication of a novel scanning microscopy probe for thermal and magnetic imaging using standard micromachining and conventional optical contact lithography. The probe features an AFM-type cantilever with a sharp pyramidal tip composed of four freestanding silicon

  17. Investigation of whispering gallery modes in microlasers by scanning near-field optical microscopy

    Science.gov (United States)

    Polubavkina, Yu S.; Kryzhanovskaya, N. V.; Nadtochiy, A. M.; Mintairov, A. M.; Lipovsky, A. A.; Scherbak, S. A.; Kulagina, M. M.; Maximov, M. V.; Zhukov, A. E.

    2017-11-01

    Near-field scanning optical microscopy (NSOM) with a spatial resolution below the light diffraction limit was used to study intensity distributions of the whispering gallery modes (WGMs) in quantum dot-based microdisk and microring lasers on GaAs with different outer diameters. Room temperature microphotoluminescence study (μPL) reveal lasing in microlasers of both geometries.

  18. Preparation of Chemically Etched Tips for Ambient Instructional Scanning Tunneling Microscopy

    Science.gov (United States)

    Zaccardi, Margot J.; Winkelmann, Kurt; Olson, Joel A.

    2010-01-01

    A first-year laboratory experiment that utilizes concepts of electrochemical tip etching for scanning tunneling microscopy (STM) is described. This experiment can be used in conjunction with any STM experiment. Students electrochemically etch gold STM tips using a time-efficient method, which can then be used in an instructional grade STM that…

  19. RGB color coded images in scanning electron microscopy of biological surfaces

    Czech Academy of Sciences Publication Activity Database

    Kofroňová, Olga; Benada, Oldřich

    2017-01-01

    Roč. 61, č. 3 (2017), s. 349-352 ISSN 0001-723X R&D Projects: GA MŠk(CZ) LO1509; GA ČR(CZ) GA16-20229S Institutional support: RVO:61388971 Keywords : Biological surfaces * Color images * Scanning electron microscopy Subject RIV: EE - Microbiology, Virology Impact factor: 0.673, year: 2016

  20. Imaging inclusion complex formation in starch granules using confocal laser scanning microscopy

    NARCIS (Netherlands)

    Manca, Marianna; Woortman, Albert J. J.; Loos, Katja; Loi, Maria A.

    The tendency of amylose to form inclusion complexes with guest molecules has been an object of wide interest due to its fundamental role in food processing. Here we investigated the features of starch granules from several botanical sources using confocal laser scanning microscopy (CLSM) and

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

    Energy Technology Data Exchange (ETDEWEB)

    Zaka, Fowzia [Lawrence Livermore National Lab. (LLNL), Livermore, CA (United States). Energetic Materials Center

    2016-03-08

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

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

    Energy Technology Data Exchange (ETDEWEB)

    Zaka, Fowzia [Lawrence Livermore National Lab. (LLNL), Livermore, CA (United States). Energetic Materials Center

    2016-03-21

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

  3. Covalently Immobilised Cytochrome C Imaged by In Situ Scanning Tunnelling Microscopy

    DEFF Research Database (Denmark)

    Andersen, Jens Enevold Thaulov; Olesen, Klaus G.; Danilov, Alexey I.

    1997-01-01

    In situ scanning tunnelling microscopy (STM) imaging of cytochrome c (cyt c) on polycrystalline Pt surfaces and on Au(lll) was achieved first by covalent immobilisation of 3-aminopropyltriethoxysilane (3-APTS) brought to react with oxide present on the Pt surfaces. Covalently bound 3-APTS forms a...

  4. Pollen grain surface in Vaccinium myrtillus as seen in scanning electron microscopy

    Directory of Open Access Journals (Sweden)

    Józef Kocoń

    2014-01-01

    Full Text Available Pollen grain surface of Vaccinium myrtillus L. was analyzed by scanning electron microscopy. Pollen grains remain in tetrahedral tetrads. Grain surface is verrucose, consisting of thick, irregularly shaped muri, surrounding small, round or oval lumina. The surface of the muri is fissured, and minute papillae can also be noted.

  5. Carbon induced metal dusting of iron-nickel-chromium alloy surfaces : a scanning auger microscopy study

    NARCIS (Netherlands)

    Palasantzas, G; DeHosson, JTM

    2004-01-01

    In this work, we present an investigation on metal dusting of iron-nickel-chromium (Fe-Ni-Cr) alloy surfaces using scanning auger microscopy. It is shown that the formation of surface Cr-oxide and the surface finish condition can strongly influence and interrupt this catastrophic phenomenon. The

  6. Second-Harmonic Generation Scanning Microscopy on Domains in Al Surfaces

    DEFF Research Database (Denmark)

    Pedersen, Kjeld; Bozhevolnyi, Sergey I.

    1999-01-01

    Scanning optical second-harmonic generation microscopy has been used to investigate domains in the surface of polycrystaline Al. Strong contrast among the crystalline grains is obtained due to variations in their crystallographic orientations and thus also nonlinear response. The origin of the co...

  7. THALLUS SURFACES IN COCCOCARPIACEAE AND PANNARIACEAE (LICHENIZED ASCOMYCETES) VIEWED WITH SCANNING ELECTRON-MICROSCOPY

    NARCIS (Netherlands)

    LUMBSCH, HT; KOTHE, HW

    1992-01-01

    The thallus surfaces of species belonging to the Coccocarpiaceae and Pannariaceae were studied using scanning electron microscopy (SEM). A pored epicortex was shown in Coccocarpia ssp., Degelia gayana and D. plumbea. In the other species studied no definite pores were found. The probable systematic

  8. Cold-induced imbibition damage of lettuce embryos: A study using cryo-scanning electron microscopy

    NARCIS (Netherlands)

    Nijsse, J.; Walther, P.; Hoekstra, F.

    2004-01-01

    The impact of rehydration on a multicellular organism was studied in lettuce (Lactuca sativa L.) embryos, using cryo-scanning electron microscopy (cryo-SEM). Naked embryos were sensitive to imbibitional stress, whereas embryos with an intact, thick-walled endosperm were not. Imbibitional injury to

  9. Scanning Tunneling Microscopy Studies of Topological Insulators Grown by Molecular Beam Epitaxy

    Directory of Open Access Journals (Sweden)

    Xue Qikun

    2012-03-01

    Full Text Available We summarize our recent scanning tunneling microscopy (STM study of topological insulator thin films grown by molecular beam epitaxy (MBE, which includes the observation of electron standing waves on topological insulator surface and the Landau quantization of topological surface states. The work has provided valuable information to the understanding of intriguing properties of topological insulators, as predicted by theory.

  10. Combining optical tweezers and scanning probe microscopy to study DNA-protein interactions

    NARCIS (Netherlands)

    Huisstede, Jurgen H G; Subramaniam, Vinod; Bennink, Martin L

    We present the first results obtained with a new instrument designed and built to study DNA-protein interactions at the single molecule level. This microscope combines optical tweezers with scanning probe microscopy and allows us to locate DNA-binding proteins on a single suspended DNA molecule. A

  11. Near-Field Scanning Optical Microscopy of Single Fluorescent Dendritic Molecules

    NARCIS (Netherlands)

    Veerman, J.A.; Levi, S.; van Veggel, F.C.J.M.; Reinhoudt, David; van Hulst, N.F.

    1999-01-01

    Individual dendritic molecules adsorbed o­n glass containing a single fluorescent rhodamine B core have been observed with near-field scanning optical microscopy (NSOM); height and fluorescence images were obtained simultaneously. The dendritic assemblies can be discriminated from free fluorescent

  12. Adsorption of Cu phthalocyanine on Pt modified Ge(001): A scanning tunneling microscopy study

    NARCIS (Netherlands)

    Saedi, A.; Berkelaar, Robin P.; Kumar, Avijit; Poelsema, Bene; Zandvliet, Henricus J.W.

    2010-01-01

    The adsorption configurations of copper phthalocyanine (CuPc) molecules on platinum-modified Ge(001) have been studied using scanning tunneling microscopy. After deposition at room temperature and cooling down to 77 K the CuPc molecules are still dynamic. However, after annealing at 550±50 K, the

  13. PROBING STRESS EFFECTS IN SINGLE CRYSTAL ORGANIC TRANSISTORS BY SCANNING KELVIN PROBE MICROSCOPY

    Energy Technology Data Exchange (ETDEWEB)

    Teague, L

    2010-06-11

    We report scanning Kelvin probe microscopy (SKPM) of single crystal difluoro bis(triethylsilylethynyl) anthradithiophene (diF-TESADT) organic transistors. SKPM provides a direct measurement of the intrinsic charge transport in the crystals independent of contact effects and reveals that degradation of device performance occurs over a time period of minutes as the diF-TESADT crystal becomes charged.

  14. Acoustic Noise of MRI Scans of the Internal Auditory Canal and Potential for Intracochlear Physiological Changes

    CERN Document Server

    Busada, M A; Ibrahim, G; Huckans, J H

    2012-01-01

    Magnetic resonance imaging (MRI) is a widely used medical imaging technique to assess the health of the auditory (vestibulocochlear) nerve. A well known problem with MRI machines is that the acoustic noise they generate during a scan can cause auditory temporary threshold shifts (TTS) in humans. In addition, studies have shown that excessive noise in general can cause rapid physiological changes of constituents of the auditory within the cochlea. Here, we report in-situ measurements of the acoustic noise from a 1.5 Tesla MRI machine (GE Signa) during scans specific to auditory nerve assessment. The measured average and maximum noise levels corroborate earlier investigations where TTS occurred. We briefly discuss the potential for physiological changes to the intracochlear branches of the auditory nerve as well as iatrogenic misdiagnoses of intralabyrinthine and intracochlear schwannomas due to hypertrophe of the auditory nerve within the cochlea during MRI assessment.

  15. High-contrast en bloc staining of neuronal tissue for field emission scanning electron microscopy.

    Science.gov (United States)

    Tapia, Juan Carlos; Kasthuri, Narayanan; Hayworth, Kenneth J; Schalek, Richard; Lichtman, Jeff W; Smith, Stephen J; Buchanan, JoAnn

    2012-01-12

    Conventional heavy metal poststaining methods on thin sections lend contrast but often cause contamination. To avoid this problem, we tested several en bloc staining techniques to contrast tissue in serial sections mounted on solid substrates for examination by field emission scanning electron microscopy (FESEM). Because FESEM section imaging requires that specimens have higher contrast and greater electrical conductivity than transmission electron microscopy (TEM) samples, our technique uses osmium impregnation (OTO) to make the samples conductive while heavily staining membranes for segmentation studies. Combining this step with other classic heavy metal en bloc stains, including uranyl acetate (UA), lead aspartate, copper sulfate and lead citrate, produced clean, highly contrasted TEM and scanning electron microscopy (SEM) samples of insect, fish and mammalian nervous systems. This protocol takes 7-15 d to prepare resin-embedded tissue, cut sections and produce serial section images.

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

    Science.gov (United States)

    Platek, Michael J.; Gregory, Otto J.

    2015-10-01

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

  17. Growth of Pd-Filled Carbon Nanotubes on the Tip of Scanning Probe Microscopy

    Directory of Open Access Journals (Sweden)

    Tomokazu Sakamoto

    2009-01-01

    Full Text Available We have synthesized Pd-filled carbon nanotubes (CNTs oriented perpendicular to Si substrates using a microwave plasma-enhanced chemical vapor deposition (MPECVD for the application of scanning probe microscopy (SPM tip. Prior to the CVD growth, Al thin film (10 nm was coated on the substrate as a buffer layer followed by depositing a 5∼40 nm-thick Pd film as a catalyst. The diameter and areal density of CNTs grown depend largely on the initial Pd thickness. Scanning electron microscopy (SEM and transmission electron microscopy (TEM images clearly show that Pd is successfully encapsulated into the CNTs, probably leading to higher conductivity. Using optimum growth conditions, Pd-filled CNTs are successfully grown on the apex of the conventional SPM cantilever.

  18. EDITORIAL: Three decades of scanning tunnelling microscopy that changed the course of surface science Three decades of scanning tunnelling microscopy that changed the course of surface science

    Science.gov (United States)

    Ramachandra Rao, M. S.; Margaritondo, Giorgio

    2011-11-01

    Three decades ago, with a tiny tip of platinum, the scientific world saw the real space imaging of single atoms with unprecedented spatial resolution. This signalled the birth of one of the most versatile surface probes, based on the physics of quantum mechanical tunnelling: the scanning tunnelling microscope (STM). Invented in 1981 by Gerd Binnig and Heinrich Rohrer of IBM, Zurich, it led to their award of the 1986 Nobel Prize. Atoms, once speculated to be abstract entities used by theoreticians for mere calculations, can be seen to exist for real with the nano-eye of an STM tip that also gives real-space images of molecules and adsorbed complexes on surfaces. From a very fundamental perspective, the STM changed the course of surface science and engineering. STM also emerged as a powerful tool to study various fundamental phenomena relevant to the properties of surfaces in technological applications such as tribology, medical implants, catalysis, sensors and biology—besides elucidating the importance of local bonding geometries and defects, non-periodic structures and the co-existence of nano-scale phases. Atom-level probing, once considered a dream, has seen the light with the evolution of STM. An important off-shoot of STM was the atomic force microscope (AFM) for surface mapping of insulating samples. Then followed the development of a flurry of techniques under the general name of scanning probe microscopy (SPM). These techniques (STM, AFM, MFM, PFM etc) designed for atomic-scale-resolution imaging and spectroscopy, have led to brand new developments in surface analysis. All of these novel methods enabled researchers in recent years to image and analyse complex surfaces on microscopic and nanoscopic scales. All of them utilize a small probe for sensing the surface. The invention of AFM by Gerd Binnig, Calvin Quate and Christopher Gerber opened up new opportunities for characterization of a variety of materials, and various industrial applications could be

  19. An endolithic microbial community in dolomite rock in central Switzerland: characterization by reflection spectroscopy, pigment analyses, scanning electron microscopy, and laser scanning microscopy.

    Science.gov (United States)

    Horath, T; Neu, T R; Bachofen, R

    2006-04-01

    A community of endolithic microorganisms dominated by phototrophs was found as a distinct band a few millimeters below the surface of bare exposed dolomite rocks in the Piora Valley in the Alps. Using in situ reflectance spectroscopy, we detected chlorophyll a (Chl a), phycobilins, carotenoids, and an unknown type of bacteriochlorophyll-like pigment absorbing in vivo at about 720 nm. In cross sections, the data indicated a defined distribution of different groups of organisms perpendicular to the rock surface. High-performance liquid chromatography analyses of pigments extracted with organic solvents confirmed the presence of two types of bacteriochlorophylls besides chlorophylls and various carotenoids. Spherical organisms of varying sizes and small filaments were observed in situ with scanning electron microscopy and confocal laser scanning microscopy (one- and two-photon technique). The latter allowed visualization of the distribution of phototrophic microorganisms by the autofluorescence of their pigments within the rock. Coccoid cyanobacteria of various sizes predominated over filamentous ones. Application of fluorescence-labeled lectins demonstrated that most cyanobacteria were embedded in an exopolymeric matrix. Nucleic acid stains revealed a wide distribution of small heterotrophs. Some biological structures emitting a green autofluorescence remain to be identified.

  20. Effects of anticancer drugs on glia-glioma brain tumor model characterized by acoustic impedance microscopy

    Science.gov (United States)

    Soon, Thomas Tiong Kwong; Chean, Tan Wei; Yamada, Hikari; Takahashi, Kenta; Hozumi, Naohiro; Kobayashi, Kazuto; Yoshida, Sachiko

    2017-07-01

    An ultrasonic microscope is a useful tool for observing living tissue without chemical fixation or histochemical processing. Two-dimensional (2D) acoustic impedance microscopy developed in our previous study for living cell observation was employed to visualize intracellular changes. We proposed a brain tumor model by cocultivating rat glial cells and C6 gliomas to quantitatively analyze the effects of two types of anticancer drugs, cytochalasin B (CyB) and temozolomide (TMZ), when they were applied. We reported that CyB treatment (25 µg/ml, T = 90 min) significantly reduced the acoustic impedance of gliomas and has little effect on glial cells. Meanwhile, TMZ treatment (2 mg/ml, T = 90 min) impacted both cells equally, in which both cells’ acoustic impedances were decreased. As CyB targets the actin filament polymerization of the cells, we have concluded that the decrease in acoustic impedance was in fact due to actin filament depolymerization and the data can be quantitatively assessed for future studies in novel drug development.

  1. Spatially resolved quantitative mapping of thermomechanical properties and phase transition temperatures using scanning probe microscopy

    Science.gov (United States)

    Jesse, Stephen; Kalinin, Sergei V; Nikiforov, Maxim P

    2013-07-09

    An approach for the thermomechanical characterization of phase transitions in polymeric materials (polyethyleneterephthalate) by band excitation acoustic force microscopy is developed. This methodology allows the independent measurement of resonance frequency, Q factor, and oscillation amplitude of a tip-surface contact area as a function of tip temperature, from which the thermal evolution of tip-surface spring constant and mechanical dissipation can be extracted. A heating protocol maintained a constant tip-surface contact area and constant contact force, thereby allowing for reproducible measurements and quantitative extraction of material properties including temperature dependence of indentation-based elastic and loss moduli.

  2. Plasma-deposited fluorocarbon films: insulation material for microelectrodes and combined atomic force microscopy-scanning electrochemical microscopy probes.

    Science.gov (United States)

    Wiedemair, Justyna; Balu, Balamurali; Moon, Jong-Seok; Hess, Dennis W; Mizaikoff, Boris; Kranz, Christine

    2008-07-01

    Pinhole-free insulation of micro- and nanoelectrodes is the key to successful microelectrochemical experiments performed in vivo or in combination with scanning probe experiments. A novel insulation technique based on fluorocarbon insulation layers deposited from pentafluoroethane (PFE, CF3CHF2) plasmas is presented as a promising electrical insulation approach for microelectrodes and combined atomic force microscopy-scanning electrochemical microscopy (AFM-SECM) probes. The deposition allows reproducible and uniform coating, which is essential for many analytical applications of micro- and nanoelectrodes such as, e.g., in vivo experiments and SECM experiments. Disk-shaped microelectrodes and frame-shaped AFM tip-integrated electrodes have been fabricated by postinsulation focused ion beam (FIB) milling. The thin insulation layer for combined AFM-SECM probes renders this fabrication technique particularly useful for submicro insulation providing radius ratios of the outer insulation versus the disk electrode (RG values) suitable for SECM experiments. Characterization of PFE-insulated AFM-SECM probes will be presented along with combined AFM-SECM approach curves and imaging.

  3. Scanning Acoustic Microscopy—A Novel Noninvasive Method to Determine Tumor Interstitial Fluid Pressure in a Xenograft Tumor Model

    Directory of Open Access Journals (Sweden)

    Matthias Hofmann

    2016-06-01

    Full Text Available Elevated tumor interstitial fluid pressure (TIFP is a prominent feature of solid tumors and hampers the transmigration of therapeutic macromolecules, for example, large monoclonal antibodies, from tumor-supplying vessels into the tumor interstitium. TIFP values of up to 40 mm Hg have been measured in experimental solid tumors using two conventional invasive techniques: the wick-in-needle and the micropuncture technique. We propose a novel noninvasive method of determining TIFP via ultrasonic investigation with scanning acoustic microscopy at 30-MHz frequency. In our experimental setup, we observed for the impedance fluctuations in the outer tumor hull of A431-vulva carcinoma–derived tumor xenograft mice. The gain dependence of signal strength was quantified, and the relaxation of tissue was calibrated with simultaneous hydrostatic pressure measurements. Signal patterns from the acoustical images were translated into TIFP curves, and a putative saturation effect was found for tumor pressures larger than 3 mm Hg. This is the first noninvasive approach to determine TIFP values in tumors. This technique can provide a potentially promising noninvasive assessment of TIFP and, therefore, can be used to determine the TIFP before treatment approach as well to measure therapeutic efficacy highlighted by lowered TFP values.

  4. Lateral spatial resolution of thermal lens microscopy during continuous scanning for nonstaining biofilm imaging

    Science.gov (United States)

    Rossteuscher, T. T. J.; Hibara, A.; Mawatari, K.; Kitamori, T.

    2009-05-01

    The possible application of continuous scanning thermal lens microscopy (TLM) as alternative online biofilm observation method is studied. As biofilm is a heterogeneous sample, the influence of spatially limited thermal flow at the sample heterogeneities and the biofilm-environment border has to be considered. The influence of the edges on the lateral resolution with respect to scanning velocity during continuous scanning TLM was therefore evaluated. Lateral scanning experiments on 100 nm thin gold stripes showed that the maximum scan speed can be predicted from a time constant of a lock-in amplifier and the beamwidth. Since three-dimensional mapping is needed to fully characterize the biofilm structure, depth scanning experiments with stained 4 μm thick polystyrene samples with the coaxial TLM setup were evaluated for signal width at full width at half maximum. Thus, a minimum step width for depth scanning of 10 μm for observation has been acquired. A three-dimensional image of unstained biofilm grown in a flow chamber was acquired using continuous scanning TLM.

  5. The Use Of Scanning Probe Microscopy To Investigate Crystal-Fluid Interfaces

    Energy Technology Data Exchange (ETDEWEB)

    Orme, C A; Giocondi, J L

    2007-04-16

    Over the past decade there has been a natural drive to extend the investigation of dynamic surfaces in fluid environments to higher resolution characterization tools. Various aspects of solution crystal growth have been directly visualized for the first time. These include island nucleation and growth using transmission electron microscopy and scanning tunneling microscopy; elemental step motion using scanning probe microscopy; and the time evolution of interfacial atomic structure using various diffraction techniques. In this lecture we will discuss the use of one such in situ method, scanning probe microscopy, as a means of measuring surface dynamics during crystal growth and dissolution. We will cover both practical aspects of imaging such as environmental control, fluid flow, and electrochemical manipulation, as well as the types of physical measurements that can be made. Measurements such as step motion, critical lengths, nucleation density, and step fluctuations, will be put in context of the information they provide about mechanistic processes at surfaces using examples from metal and mineral crystal growth.

  6. Characterization of tip size and geometry of the pipettes used in scanning ion conductance microscopy.

    Science.gov (United States)

    Tognoni, Elisabetta; Baschieri, Paolo; Ascoli, Cesare; Pellegrini, Monica; Pellegrino, Mario

    2016-04-01

    Scanning ion-conductance microscopy (SICM) belongs to the family of scanning-probe microscopies. The spatial resolution of these techniques is limited by the size of the probe. In SICM the probe is a pipette, obtained by heating and pulling a glass capillary tubing. The size of the pipette tip is therefore an important parameter in SICM experiments. However, the characterization of the tip is not a consolidated routine in SICM experimental practice. In addition, potential and limitations of the different methods available for this characterization may not be known to all users. We present an overview of different methods for characterizing size and geometry of the pipette tip, with the aim of collecting and facilitating the use of several pieces of information appeared in the literature in a wide interval of time under different disciplines. In fact, several methods that have been developed for pipettes used in cell physiology can be also fruitfully employed in the characterization of the SICM probes. The overview includes imaging techniques, such as scanning electron microscopy and atomic Force microscopy, and indirect methods, which measure some physical parameter related to the size of the pipette. Examples of these parameters are the electrical resistance of the pipette filled with a saline solution and the surface tension at the pipette tip. We discuss advantages and drawbacks of the methods, which may be helpful in answering a wide range of experimental questions. Copyright © 2016 Elsevier Ltd. All rights reserved.

  7. Scanning probe microscopy: instrumentation and applications on thin films and magnetic multilayers.

    Science.gov (United States)

    Karoutsos, Vagelis

    2009-12-01

    In this article we present a review on instrumentation and the modes of operation of a scanning probe microscope. In detail, we review the main techniques of Scanning Probe Microscopy (SPM), which are Scanning Tunneling Microscopy (STM) and Atomic Force Microscopy (AFM), focusing our attention on the latter one. The AFM instrument provides information on the roughness and grain size of thin films. As an example we review recent results on two metallic thin film systems: thin Ag films deposited on glass, and Ni/Pt compositionally modulated multilayers deposited on glass, Si, and polyimide substrates. To show the validity of the grain size measurements, we compare the data with the ones resulting from X-ray diffraction (XRD) measurements. We show that the AFM results are reliable for grain diameters as small as 14 nm, which is approximately comparable to the tip radius. Finally, we deal with Magnetic Force Microscopy (MFM) results on Co/Pt and Co/Au multilayers. We observe perpendicularly magnetized domains. The domain configurations are correlated to the magnetization hysteresis curves.

  8. Combined nanoprobes for scanning probe microscopy: laser technology for processing and testing

    Science.gov (United States)

    Veiko, V. P.; Golubok, A. O.; Zuong, Z.; Varkentina, N. V.; Yakovlev, E. B.

    2008-02-01

    Scanning probe microscopy (SPM) is a high spatial resolution method of surface topography visualization and measurement of its local properties. The detecting of interaction arising between the sharp solid-state probe and the sample surface is the foundation of SPM. In dependence from nature of this interaction the scanning tunnel microscopy (STM), scanning force microscopy (SFM), scanning near field optical microscopy (SNOM), etc. are distinguished. The spatial resolution of all types of probe microscopy determins both sharpness of increasing of interaction between a probe and a sample at their approach, and shape and size of a top of a solid-state probe. So, the progress in SPM information capabilities is highly depends from probe properties and first of all from properly fabricated aperture size. Fabrication procedures are rather complicated because of nanometric scale size of aperture and hard requirements to reproducibility and need to be improved. The way how to do it is involving of feed-back in a processing procedure-results in two types of feedback for the process of drawing-out has been suggested, tested and installed into the technological set-up. Different probes have been fabricated by laser-assisted drawing-out during this work: SNOM types from optical fibers, micropipettes from quartz glass capillaries, micropipettes with microwires inside and with metallic covers outside. Some examples of application of above mentioned combined probes for cell membrane technology are described. Most important from them are topographical studying of cells and bacteria in living condition (in liquid) and studying of the mechanical properties of cell (rigidity of cell membrane) using the nanopipette as a tip of a force sensor. Also measurement of ion current that runs through cell membrane during its metabolic process using the nanopipette as well as in the well-known patch-clamp method have been done.

  9. The use of scanning ion conductance microscopy to image A6 cells.

    Science.gov (United States)

    Gorelik, Julia; Zhang, Yanjun; Shevchuk, Andrew I; Frolenkov, Gregory I; Sánchez, Daniel; Lab, Max J; Vodyanoy, Igor; Edwards, Christopher R W; Klenerman, David; Korchev, Yuri E

    2004-03-31

    Continuous high spatial resolution observations of living A6 cells would greatly aid the elucidation of the relationship between structure and function and facilitate the study of major physiological processes such as the mechanism of action of aldosterone. Unfortunately, observing the micro-structural and functional changes in the membrane of living cells is still a formidable challenge for a microscopist. Scanning ion conductance microscopy (SICM), which uses a glass nanopipette as a sensitive probe, has been shown to be suitable for imaging non-conducting surfaces bathed in electrolytes. A specialized version of this microscopy has been developed by our group and has been applied to image live cells at high-resolution for the first time. This method can also be used in conjunction with patch clamping to study both anatomy and function and identify ion channels in single cells. This new microscopy provides high-resolution images of living renal cells which are comparable with those obtained by scanning electron microscopy (SEM) and atomic force microscopy (AFM). Continuous 24h observations under normal physiological conditions showed how A6 kidney epithelial cells changed their height, volume, and reshaped their borders. The changes in cell area correlated with the density of microvilli on the surface. Surface microvilli density ranged from 0.5 microm(-2) for extended cells to 2.5 microm(2) for shrunk cells. Patch clamping of individual cells enabled anatomy and function to be correlated. Scanning ion conductance microscopy provides unique information about living cells that helps to understand cellular function. It has the potential to become a powerful tool for research on living renal cells.

  10. Image scanning fluorescence emission difference microscopy based on a detector array.

    Science.gov (United States)

    Li, Y; Liu, S; Liu, D; Sun, S; Kuang, C; Ding, Z; Liu, X

    2017-06-01

    We propose a novel imaging method that enables the enhancement of three-dimensional resolution of confocal microscopy significantly and achieve experimentally a new fluorescence emission difference method for the first time, based on the parallel detection with a detector array. Following the principles of photon reassignment in image scanning microscopy, images captured by the detector array were arranged. And by selecting appropriate reassign patterns, the imaging result with enhanced resolution can be achieved with the method of fluorescence emission difference. Two specific methods are proposed in this paper, showing that the difference between an image scanning microscopy image and a confocal image will achieve an improvement of transverse resolution by approximately 43% compared with that in confocal microscopy, and the axial resolution can also be enhanced by at least 22% experimentally and 35% theoretically. Moreover, the methods presented in this paper can improve the lateral resolution by around 10% than fluorescence emission difference and 15% than Airyscan. The mechanism of our methods is verified by numerical simulations and experimental results, and it has significant potential in biomedical applications. © 2017 The Authors Journal of Microscopy © 2017 Royal Microscopical Society.

  11. Imaging metazoan nuclear pore complexes by field emission scanning electron microscopy.

    Science.gov (United States)

    Fichtman, Boris; Shaulov, Lihi; Harel, Amnon

    2014-01-01

    High resolution three-dimensional surface images of nuclear pore complexes (NPCs) can be obtained by field emission scanning electron microscopy. We present a short retrospective view starting from the early roots of microscopy, through the discovery of the cell nucleus and the development of some modern techniques for sample preparation and imaging. Detailed protocols are presented for assembling anchored nuclei in a Xenopus cell-free reconstitution system and for the exposure of the nuclear surface in mammalian cell nuclei. Immunogold labeling of metazoan NPCs and a promising new technique for delicate coating with iridium are also discussed. Copyright © 2014 Elsevier Inc. All rights reserved.

  12. Line scan--structured illumination microscopy super-resolution imaging in thick fluorescent samples.

    Science.gov (United States)

    Mandula, Ondrej; Kielhorn, Martin; Wicker, Kai; Krampert, Gerhard; Kleppe, Ingo; Heintzmann, Rainer

    2012-10-22

    Structured illumination microscopy in thick fluorescent samples is a challenging task. The out-of-focus fluorescence background deteriorates the illumination pattern and the reconstructed images suffer from influence of noise. We present a combination of structured illumination microscopy with line scanning. This technique reduces the out-of-focus fluorescence background, which improves the modulation and the quality of the illumination pattern and therefore facilitates the reconstruction. We present super-resolution, optically sectioned images of a thick fluorescent sample, revealing details of the specimen's inner structure.

  13. Optical microscope illumination analysis using through-focus scanning optical microscopy.

    Science.gov (United States)

    Attota, Ravi Kiran; Park, Haesung

    2017-06-15

    Misalignment of the aperture diaphragm present in optical microscopes results in angular illumination asymmetry (ANILAS) at the sample plane. Here we show that through-focus propagation of ANILAS results in a lateral image shift with a focus position. This could lead to substantial errors in quantitative results for optical methods that use through-focus images such as three-dimensional nanoparticle tracking, confocal microscopy, and through-focus scanning optical microscopy (TSOM). A correlation exists between ANILAS and the slant in TSOM images. Hence, the slant in the TSOM image can be used to detect, analyze, and rectify the presence of ANILAS.

  14. Application of scanning force and near field microscopies to the characterization of minimally adhesive polymer surfaces.

    Science.gov (United States)

    Akhremitchev, Boris B; Bemis, Jason E; al-Maawali, Sabah; Sun, Yujie; Stebounova, Larissa; Walker, Gilbert C

    2003-04-01

    This mini-review reports efforts to develop new scanning probe microscopies to characterize the function and aging of textured, minimally adhesive polymer surfaces used for antifouling applications in the marine environment. Novel atomic force and infrared near field microscopy techniques have been used to characterize the polymer surface adhesion and structural properties. These techniques may find promise for characterizing the deposition of the extracellular matrix of organisms as well as aging of the polymer coating itself. The reported work is part of a larger effort to reduce biofouling on ships' hulls through the development and use of improved coating materials.

  15. In Situ Scanning Probe Microscopy and New Perspectives in Analytical Chemistry

    DEFF Research Database (Denmark)

    Andersen, Jens Enevold Thaulov; Zhang, Jingdong; Chi, Qijin

    1999-01-01

    for molecular- and mesoscopic-scale analytical chemistry, are then reviewed. They are illustrated by metallic electro-crystallisation and -dissolution, and in situ STM spectroscopy of large redox molecules. The biophysically oriented analytical options of in situ atomic force microscopy, and analytical chemical......The resolution of scanning probe microscopies is unpresedented but the techniques are fraught with limitations as analytical tools. These limitations and their relationship to the physical mechanisms of image contrast are first discussed. Some new options based on in situ STM, which hold prospects...

  16. Development of an add-on kit for scanning confocal microscopy (Conference Presentation)

    Science.gov (United States)

    Guo, Kaikai; Zheng, Guoan

    2017-03-01

    Scanning confocal microscopy is a standard choice for many fluorescence imaging applications in basic biomedical research. It is able to produce optically sectioned images and provide acquisition versatility to address many samples and application demands. However, scanning a focused point across the specimen limits the speed of image acquisition. As a result, scanning confocal microscope only works well with stationary samples. Researchers have performed parallel confocal scanning using digital-micromirror-device (DMD), which was used to project a scanning multi-point pattern across the sample. The DMD based parallel confocal systems increase the imaging speed while maintaining the optical sectioning ability. In this paper, we report the development of an add-on kit for high-speed and low-cost confocal microscopy. By adapting this add-on kit to an existing regular microscope, one can convert it into a confocal microscope without significant hardware modifications. Compared with current DMD-based implementations, the reported approach is able to recover multiple layers along the z axis simultaneously. It may find applications in wafer inspection and 3D metrology of semiconductor circuit. The dissemination of the proposed add-on kit under $1000 budget could also lead to new types of experimental designs for biological research labs, e.g., cytology analysis in cell culture experiments, genetic studies on multicellular organisms, pharmaceutical drug profiling, RNA interference studies, investigation of microbial communities in environmental systems, and etc.

  17. Observations of liver cancer cells in scanning probe acoustic microscope: a preliminary study

    Science.gov (United States)

    Chen, Xiaohui; Fang, Xiaoyue; Xi, Qing; Guo, Hua; Zhang, Ning; Ding, Mingyue

    2016-04-01

    Scanning probe acoustic microscope (SPAM) can be used to acquire the morphology image as well as the non-destructive internal structures acoustic image. However, the observations of the morphology image as well as the internal structures acoustic image of liver cancer cells in SPAM are few. In this paper, we cultured 4 different types of liver cancer cells on the silicon wafer and coverslip to observe their morphology images as well as acoustic images in SPAM, and made a preliminary study of the 8 types of cells specimens (hereinafter referred to as the silicon specimens and coverslips specimens). The experimental measurement results showed that some cellular pseudopodium were observed in the morphology images of the coverslip specimens while no such cellular pseupodium were appeared in the morphology images of the silicon specimens, which concluded that the living liver cancer cells were less likely to grow on the silicon wafer. SPAM provides a rapid and sensitive visual method for studying the morphology and internal structures of the cancer cells. The proposed method can be also used to obtain the morphology and internal information in both solid and soft material wafers, such as silicon and cells, with the resolution of nanometer scale.

  18. Observation of mesenteric microcirculatory disturbance in rat by laser oblique scanning optical microscopy

    Science.gov (United States)

    Ding, Yichen; Zhang, Yu; Peng, Tong; Lu, Yiqing; Jin, Dayong; Ren, Qiushi; Liu, Yuying; Han, Jingyan; Xi, Peng

    2013-05-01

    Ischemia-reperfusion (I/R) injury model has been widely applied to the study of microcirculation disturbance. In this work, we used laser oblique scanning optical microscopy (LOSOM) to observe the microcirculation system in the mesentery of rat model. Utilizing a localized point-scanning detection scheme, high-contrast images of leukocytes were obtained. The extended detection capability facilitated both the automatic in vivo cell counting and the accurate measurement of the rolling velocity of leukocytes. Statistical analysis of the different treatment groups suggested that the distinction between I/R and sham groups with time lapse is significant.

  19. Practical aspects of single-pass scan Kelvin probe force microscopy

    Science.gov (United States)

    Li, Guangyong; Mao, Bin; Lan, Fei; Liu, Liming

    2012-11-01

    The single-pass scan Kelvin probe force microscopy (KPFM) in ambient condition has a few advantages over the dual-pass lift-up scan KPFM. For example, its spatial resolution is expected to be higher; and its topographical errors caused by electrostatic forces are minimized because electrostatic forces are actively suppressed during the simultaneous topographical and KPFM measurement. Because single-pass scan KPFM in ambient condition is relatively new, it received little attention in the literature so far. In this article, we discuss several major practical aspects of single-pass scan KPFM especially in ambient condition. First, we define the resolution using a point spread function. With this definition, we analyze the relation between the resolution and the scanning parameters such as tip apex radius and tip-surface distance. We further study the accuracy of KPFM based on the point spread function. Then, we analyze the sensitivity of KPFM under different operation modes. Finally, we investigate the crosstalk between the topographical image and the surface potential image and demonstrate the practical ways to minimize the crosstalk. These discussions not only help us to understand the single-pass scan KPFM but also provide practical guidance in using single-pass scan KPFM.

  20. Electron beam confinement and image contrast enhancement in near field emission scanning electron microscopy.

    Science.gov (United States)

    Kirk, T L; De Pietro, L G; Pescia, D; Ramsperger, U

    2009-04-01

    In conventional scanning electron microscopy (SEM), the lateral resolution is limited by the electron beam diameter impinging on the specimen surface. Near field emission scanning electron microscopy (NFESEM) provides a simple means of overcoming this limit; however, the most suitable field emitter remains to be determined. NFESEM has been used in this work to investigate the W (110) surface with single-crystal tungsten tips of (310), (111), and (100)-orientations. The topographic images generated from both the electron intensity variations and the field emission current indicate higher resolution capabilities with decreasing tip work function than with polycrystalline tungsten tips. The confinement of the electron beam transcends the resolution limitations of the geometrical models, which are determined by the minimum beam width.

  1. Field emission scanning electron microscopy of biofilm-growing bacteria involved in nosocomial infections.

    Science.gov (United States)

    Vuotto, Claudia; Donelli, Gianfranco

    2014-01-01

    Scanning electron microscopy (SEM) provides useful information on the shape, size, and localization within the biofilm of single bacteria as well as on the steps of biofilm formation process, on bacterial interactions, and on production of extracellular polymeric substances.When biofilms are constituted by microbial species involved in health care-associated infections, information provided by SEM can be fruitfully used not only for basic researches but also for diagnostic purposes.The protocols currently used in our laboratory for biofilm investigation by SEM are reported here. Particularly, the procedures to fix, dehydrate, and metalize in vitro-developed biofilms or ex vivo clinical specimens colonized by biofilm-growing microorganisms are described as well as the advantages of the observation of these samples by field emission scanning electron microscopy.

  2. Improved depth resolution in video-rate line-scanning multiphoton microscopy using temporal focusing

    Science.gov (United States)

    Tal, Eran; Oron, Dan; Silberberg, Yaron

    2005-07-01

    By introducing spatiotemporal pulse shaping techniques to multiphoton microscopy it is possible to obtain video-rate images with depth resolution similar to point-by-point scanning multiphoton microscopy while mechanically scanning in only one dimension. This is achieved by temporal focusing of the illumination pulse: The pulsed excitation field is compressed as it propagates through the sample, reaching its shortest duration (and highest peak intensity) at the focal plane before stretching again beyond it. This method is applied to produce, in a simple and scalable setup, video-rate two-photon excitation fluorescence images of Drosophila egg chambers with nearly 100,000 effective pixels and 1.5 μm depth resolution.

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

    Science.gov (United States)

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

    2010-12-01

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

  4. Spatiotemporal Rank Filtering Improves Image Quality Compared to Frame Averaging in 2-Photon Laser Scanning Microscopy.

    Directory of Open Access Journals (Sweden)

    Henry Pinkard

    Full Text Available Live imaging of biological specimens using optical microscopy is limited by tradeoffs between spatial and temporal resolution, depth into intact samples, and phototoxicity. Two-photon laser scanning microscopy (2P-LSM, the gold standard for imaging turbid samples in vivo, has conventionally constructed images with sufficient signal-to-noise ratio (SNR generated by sequential raster scans of the focal plane and temporal integration of the collected signals. Here, we describe spatiotemporal rank filtering, a nonlinear alternative to temporal integration, which makes more efficient use of collected photons by selectively reducing noise in 2P-LSM images during acquisition. This results in much higher SNR while preserving image edges and fine details. Practically, this allows for at least a four fold decrease in collection times, a substantial improvement for time-course imaging in biological systems.

  5. Scanning near-field optical microscopy on rough surfaces: Applications in chemistry, biology, and medicine

    OpenAIRE

    Kaupp, Gerd

    2006-01-01

    Shear-force apertureless scanning near-field optical microscopy (SNOM) with very sharp uncoated tapered waveguides relies on the unexpected enhancement of reflection in the shear-force gap. It is the technique for obtaining chemical (materials) contrast in the optical image of “real world” surfaces that are rough and very rough without topographical artifacts, and it is by far less complicated than other SNOM techniques that can only be used for very flat surfaces. The ex...

  6. Evaluation of Yogurt Microstructure Using Confocal Laser Scanning Microscopy and Image Analysis

    DEFF Research Database (Denmark)

    Skytte, Jacob Lercke; Ghita, Ovidiu; Whelan, Paul F.

    2015-01-01

    The microstructure of protein networks in yogurts defines important physical properties of the yogurt and hereby partly its quality. Imaging this protein network using confocal scanning laser microscopy (CSLM) has shown good results, and CSLM has become a standard measuring technique for fermented...... to image texture description. Here, CSLM images from a yogurt fermentation study are investigated, where production factors including fat content, protein content, heat treatment, and incubation temperature are varied. The descriptors are evaluated through nearest neighbor classification, variance analysis...

  7. Scanning Auger microscopy analysis of 90 K Y--Ba--Cu--O superconductors

    Energy Technology Data Exchange (ETDEWEB)

    Cota, L.; Morales de la Garza, L.; Hirata, G.; Martinez, L.; Orozco, E.; Carrillo, E.; Mendoza, A.; Albarran, J.L.; Fuentes-Maya, J.; Boldu, J.L.; and others

    1988-05-01

    The oxide superconductor Y--Ba--Cu--O is studied using Auger scanning microscopy. The chemical depth profiles of the samples were obtained. It is concluded that two phases are present in the sample, one corresponding to the standard composition and another that is Ba enriched. The first shows a platelet shape and the second a granular appearence that covers the surface of the sample.

  8. Effect of Autoclave Cycles on Surface Characteristics of S-File Evaluated by Scanning Electron Microscopy

    OpenAIRE

    Razavian, Hamid; Iranmanesh, Pedram; Mojtahedi, Hamid; Nazeri, Rahman

    2015-01-01

    Introduction: Presence of surface defects in endodontic instruments can lead to unwanted complications such as instrument fracture and incomplete preparation of the canal. The current study was conducted to evaluate the effect of autoclave cycles on surface characteristics of S-File by scanning electron microscopy (SEM). Methods and Materials: In this experimental study, 17 brand new S-Files (#30) were used. The surface characteristics of the files were examined in four steps (without autocla...

  9. Pulse Plating on Gold Surfaces Studied by In Situ Scanning Tunneling Microscopy

    DEFF Research Database (Denmark)

    Andersen, Jens Enevold Thaulov; Bech-Nielsen, Gregers; Møller, Per

    1994-01-01

    Deposition of bulk copper on thin film gold surfaces is carried out by computer-aided pulse plating. It is demonstrated that the morphology of the copper deposit can be studied by in situ scanning tunnelling microscopy both in potentiostatic experiments and in galvanostatic experiments. Optimized...... procedures for obtaining smooth deposits by pulse plating are explained in terms of a levelling effect. Possible non-faradaic processes observed in measurements with high frequency pulse plating are discussed....

  10. Scanning electron microscopy and calcification in amelogenesis imperfecta in anterior and posterior human teeth

    OpenAIRE

    Sánchez-Quevedo, M.C.; Ceballos, G.; García, J. M.; Rodriguez, I. A.; Gómez de Ferraris, M.E.; Campos, Antonio

    2001-01-01

    Teeth fragments from members of a famil? clinically and genetically diagnosed as having amelogenesis imperfecta were studied by scanning electron microscopy and X-ray microprobe analysis to establish the morphological patterns and the quantitative concentration of calcium in the enamel of anterior (canine, incisor) and posterior (premolar and molar) teeth. The prism patterns in the enamel of teeth from both regions were parallel or irregularly decussate, with ...

  11. Scanning electron microscopy of the egg-shell of the fly synthesiomyia Nudiseta (wulp)

    OpenAIRE

    El Alfy, Nagla Z. [نجلاء زكي الالفي

    1994-01-01

    The ultrastructure of the egg-shell of the fly Synthesiomyia nudiseta has been studied by scanning electron microscopy. The surface of the outer chorion is highly ridged, grooved and has a reticulate appearance except at the collar surrounding the micropyle. There are few aeropyles at the antimicropylar pole. The structure of the egg-shell outside the hatching lines is composed of three layers. The outer layer consists of vertical columns that arise from the middle layer, these columns bra...

  12. Scanning electron microscopy of the teguments of males from five populations of Schistosoma mattheei.

    Science.gov (United States)

    Kruger, F J; Hamilton-Attwell, V L; Schutte, C H

    1986-06-01

    The teguments of males from 5 populations of S. mattheei, of which 3 were sympatric and 2 allopatric with S. haematobium, were studied by means of scanning electron microscopy (SEM). A certain percentage of the males of each sympatric population bore tubercle spines while the allopatric populations were spineless. It is postulated that the presence of tubercle spines is a characteristic inherited from S. haematobium.

  13. Scanning transmission x-ray microscopy: A new ``looking glass`` into coal chemical structure

    Energy Technology Data Exchange (ETDEWEB)

    Botto, R.E.; Cody, G.D.

    1994-02-01

    This paper reports the use of scanning transmission x-ray microscopy to spatially map the chemistry of aromatic and aliphatic carbon functionalities in coal to a resolution of less than 0.1 {mu}m. Localized x-ray absorption spectroscopy recorded at the carbon K absorption edge was also used to facilitate analysis of variations in fundamental chemistry at maceral interfaces and within maceral boundaries.

  14. Scanning Electron Microscopy study of Macbat regeneration effect on lead-acid battery electrodes

    OpenAIRE

    Emanuelsson, Christian

    2013-01-01

    Electrodes from lead-acid batteries were studied using scanning electron microscopy and energy dispersive spectroscopy. This to observe the effects of cycling on the batteries and how a capacity recovery process, known as Macbat regeneration, affected the active material with focus on hard sulphation. First, two new batteries were cycled for two months and electrodes from them were studied when the batteries were new, cycled, fully charged after cycling and regenerated after cycling. Then ele...

  15. Corneal endothelium of the Magellanic penguin (Spheniscus magellanicus) by scanning electron microscopy.

    Science.gov (United States)

    Pigatto, João A T; Laus, José L; Santos, Jaime M; Cerva, Cristine; Cunha, Luciana S; Ruoppolo, Valéria; Barros, Paulo S M

    2005-12-01

    The corneal endothelium is essential for the maintenance of the corneal transparency. The aim of this study was to examine the morphology of the endothelial surface and perform morphometric analysis of the normal corneal endothelial cells of the Magellanic penguin (Spheniscus magellanicus) using scanning electron microscopy. The present work demonstrates that the corneal endothelium of the Magellanic penguin is similar to those described in other vertebrates.

  16. Virtual microscopy, data management and image analysis in Aperio ScanScope system.

    Directory of Open Access Journals (Sweden)

    Wojciech Staniszewski

    2010-05-01

    Full Text Available The histology and the pathology clinical practice undergo a digital revolution. Essential change in laboratory practice - from classical light microscopes, thousands of glass specimens waiting on plates to a virtual microscope and onscreen diagnosis is right now. Currently there are more than 30 different systems for the Virtual Microscopy available on the market. However none of them is so oriented for the practical matters as Aperio ScanScope system.

  17. Histology and scanning electron microscopy observations of cryopreserved protocorm-like bodies of Dendrobium sonia-28

    OpenAIRE

    POOBATHY, Ranjetta; Sinniah, Uma Rani; RATHINAM, Xavier; Subramaniam, Sreeramanan

    2013-01-01

    The genus Dendrobium possesses horticultural importance. Dendrobium sonia-28 is an important ornamental orchid in the flower industry. Cryopreservation is a favoured long-term storage method for orchids with propagation problems. Protocorm-like bodies (PLBs) of Dendrobium sonia-28 were cryopreserved using the vitrification technique. Histology and scanning electron microscopy (SEM) observations were conducted on stock, non-cryopreserved (control), and cryopreserved PLBs of Dendrobium sonia-28...

  18. Tuning Localized Surface Plasmon Resonance in Scanning Near-Field Optical Microscopy Probes.

    Science.gov (United States)

    Vasconcelos, Thiago L; Archanjo, Bráulio S; Fragneaud, Benjamin; Oliveira, Bruno S; Riikonen, Juha; Li, Changfeng; Ribeiro, Douglas S; Rabelo, Cassiano; Rodrigues, Wagner N; Jorio, Ado; Achete, Carlos A; Cançado, Luiz Gustavo

    2015-06-23

    A reproducible route for tuning localized surface plasmon resonance in scattering type near-field optical microscopy probes is presented. The method is based on the production of a focused-ion-beam milled single groove near the apex of electrochemically etched gold tips. Electron energy-loss spectroscopy and scanning transmission electron microscopy are employed to obtain highly spatially and spectroscopically resolved maps of the milled probes, revealing localized surface plasmon resonance at visible and near-infrared wavelengths. By changing the distance L between the groove and the probe apex, the localized surface plasmon resonance energy can be fine-tuned at a desired absorption channel. Tip-enhanced Raman spectroscopy is applied as a test platform, and the results prove the reliability of the method to produce efficient scattering type near-field optical microscopy probes.

  19. The detection and influence of food soils on microorganisms on stainless steel using scanning electron microscopy and epifluorescence microscopy.

    Science.gov (United States)

    Whitehead, Kathryn A; Smith, Lindsay A; Verran, Joanna

    2010-07-31

    A range of food soils and components (complex [meat extract, fish extract, and cottage cheese extract]; oils [cholesterol, fish oil, and mixed fatty acids]; proteins [bovine serum albumin (BSA), fish peptones, and casein]; and carbohydrates [glycogen, starch, and lactose]) were deposited onto 304 2B finish stainless steel surfaces at different concentrations (10-0.001%). Scanning electron microscopy (SEM) and epifluorescence microscopy were used to visualise the cell and food soil distribution across the surface. Epifluorescence microscopy was also used to quantify the percentage of a field covered by cells or soil. At 10% concentration, most soils, with the exception of BSA and fish peptone were easily visualised using SEM, presenting differences in gross soil morphology and distribution. When soil was stained with acridine orange and visualised by epifluorescence microscopy, the limit of detection of the method varied between soils, but some (meat, cottage cheese and glycogen) were detected at the lowest concentrations used (0.001%). The decrease in soil concentration did not always relate to the surface coverage measurement. When 10% food soil was applied to a surface with Escherichia coli and compared, cell attachment differed depending on the nature of the soil. The highest percentage coverage of cells was observed on surfaces with fish extract and related products (fish peptone and fish oil), followed by carbohydrates, meat extract/meat protein, cottage cheese/casein and the least to the oils (cholesterol and mixed fatty acids). Cells could not be clearly observed in the presence of some food soils using SEM. Findings demonstrate that food soils heterogeneously covered stainless steel surfaces in differing patterns. The pattern and amount of cell attachment was related to food soil type rather than to the amount of food soil detected. This work demonstrates that in the study of conditioning film and cell retention on the hygienic properties of surfaces, SEM

  20. Observation of photodynamically-induced cell destruction probed by video microscopy, laser-scanning microscopy, and fluorescence spectroscopy

    Science.gov (United States)

    Rueck, Angelika C.; Strauss, Wolfgang S. L.; Gschwend, Michael H.; Koenig, Karsten; Brunner, B.; Schneckenburger, Herbert; Walt, Heinrich; Steiner, Rudolf W.

    1993-07-01

    In order to study light-induced reactions during PDT, the fluorescence response of the photosensitizer meso-tetra(4-sulfonatophenyl)porphyrin (TPPS4) was observed in different cell systems and correlated with the sensitivity to photodynamic induced destructions. RR 1022 epithelial cells from the rat were grown on microscopic slides at a high and low cell density. Using video microscopy in combination with microspectrofluorometry we observed a different fluorescence behavior for high and low cell conditions during light exposure. A fluorescence relocalization from the cytoplasm to the nucleus and an intensity increase-- correlated with the formation of a new molecular species--could be detected only for low cell density. Moreover, cell cultures at a high density showed to be less sensitive to photodynamic destructions. In addition to cell culture-experiments, we observed the light-induced reactions of TPPS4 accumulated in multicellular tumor spheroids. For these measurements laser scanning microscopy was used. Fluorescence relocalization and intensity increase could be detected only for the peripheric parts of the spheroids. The different fluorescence response seems to reflect different metabolic and physiologic states of the cells.

  1. Adaptive and robust statistical methods for processing near-field scanning microwave microscopy images.

    Science.gov (United States)

    Coakley, K J; Imtiaz, A; Wallis, T M; Weber, J C; Berweger, S; Kabos, P

    2015-03-01

    Near-field scanning microwave microscopy offers great potential to facilitate characterization, development and modeling of materials. By acquiring microwave images at multiple frequencies and amplitudes (along with the other modalities) one can study material and device physics at different lateral and depth scales. Images are typically noisy and contaminated by artifacts that can vary from scan line to scan line and planar-like trends due to sample tilt errors. Here, we level images based on an estimate of a smooth 2-d trend determined with a robust implementation of a local regression method. In this robust approach, features and outliers which are not due to the trend are automatically downweighted. We denoise images with the Adaptive Weights Smoothing method. This method smooths out additive noise while preserving edge-like features in images. We demonstrate the feasibility of our methods on topography images and microwave |S11| images. For one challenging test case, we demonstrate that our method outperforms alternative methods from the scanning probe microscopy data analysis software package Gwyddion. Our methods should be useful for massive image data sets where manual selection of landmarks or image subsets by a user is impractical. Published by Elsevier B.V.

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

    Science.gov (United States)

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

    2009-04-01

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

  3. SLAM examination of solar cells and solar cell welds. [Scanning Laser Acoustic Microscope

    Science.gov (United States)

    Stella, P. M.; Vorres, C. L.; Yuhas, D. E.

    1981-01-01

    The scanning laser acoustic microscope (SLAM) has been evaluated for non-destructive examination of solar cells and interconnector bonds. Using this technique, it is possible to view through materials in order to reveal regions of discontinuity such as microcracks and voids. Of particular interest is the ability to evaluate, in a unique manner, the bonds produced by parallel gap welding. It is possible to not only determine the area and geometry of the bond between the tab and cell, but also to reveal any microcracks incurred during the welding. By correlating the SLAM results with conventional techniques of weld evaluation a more confident weld parameter optimization can be obtained.

  4. Scanning electron microscopy of individual nanoparticle bio-markers in liquid

    Energy Technology Data Exchange (ETDEWEB)

    Liv, Nalan, E-mail: n.liv@tudelft.nl; Lazić, Ivan; Kruit, Pieter; Hoogenboom, Jacob P.

    2014-08-01

    We investigated SEM imaging of nanoparticle biomarkers suspended below a thin membrane, with the ultimate goal of integrating functional fluorescence and structural SEM measurements of samples kept at ambient or hydrated conditions. In particular, we investigated how resolving power in liquid SEM is affected by the interaction of the electron beam with the membrane. Simulations with the Geant4-based Monte Carlo scheme developed by Kieft and Bosch (2008) [1] are compared to experimental results with suspended nanoparticles. For 20 nm and 50 nm thin membranes, we found a beam broadening of 1.5 nm and 3 nm, respectively, with an excellent agreement between simulations and experiments. 15 nm Au nanoparticles and bio-functionalized core-shell quantum dots can be individually resolved in denser clusters. We demonstrated the imaging of single EGF-conjugated quantum dots docked at filopodia during cellular uptake with both fluorescence microscopy and SEM simultaneously. These results open novel opportunities for correlating live fluorescence microscopy with structural electron microscopy. - Highlights: • We investigate the achievable resolution in liquid scanning electron microscopy (SEM). • We demonstrate liquid SEM imaging of individual fluorescent nanoparticle bio-markers • We show imaging of cellular QDot uptake with simultaneous fluorescence microscopy and SEM. • The positions of individual QDots can be resolved with details on cellular structure.

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

    Science.gov (United States)

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

    2014-01-01

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

  6. Potential Challenges in Near-Field Scanning Optical Microscopy for Space Applications

    Science.gov (United States)

    Vikram, Chandra S.; Witherow, William K.; Rose, M. Franklin (Technical Monitor)

    2000-01-01

    Near-field scanning optical microscopy (NSOM) also called scanning near-field optical microscopy (SNOM) is now well accepted as a powerful tool for sub-wavelength (nanoscale in the optical region) spatial resolution microscopy and a large number of related tasks. The importance lies in the fact of strategic advantages of standard microscopy but with significantly enhanced resolution. Since many modern optical diagnostic techniques have found useful applications in space, it is logical to consider the future role of NSOM in such situations. For example, protein crystal growth study under microgravity conditions is a valid candidate. If applied successfully, processes at molecular level can be studied during the growth. NSOM has already been demonstrated to be useful for the study of such crystals here on earth. The basic principle of NSOM can be illustrated. The illumination-collection mode is shown although several other possible approaches exist. In this, the sample is illuminated and the light from the sample is collected through the same tiny aperture opening. A tapered optical fiber is scanned near the sample surface. The tip is coated generally with a metal with a sub-wavelength aperture opening. The tip-sample distance is maintained constant while scanning. Thus, the optical signal available for collection is generally a function of the optical properties of the sample surface. Since the aperture is sub-wavelength in diameter and the tip is held very close (again in the sub-wavelength domain) to the surface, the lateral resolution in the sub-wavelength domain is obtained. Thus, the typical wavelength- order resolution of ordinary microscopy can be significantly enhanced while maintaining the strategic advantages (no need of sample in vacuum chamber, electron beams, etc). Commercial NSOM systems play a key role in the success and widespread acceptance of the tool. These commercial systems work fairly well in laboratory conditions on earth. However, they may

  7. Fast axial scanning for 2-photon microscopy using liquid lens technology

    Science.gov (United States)

    Tehrani, Kayvan Forouhesh; Sun, Min Kyoung; Karumbaiah, Lohitash; Mortensen, Luke J.

    2017-02-01

    Scanning microscopy methods require movement of the focus in Z coordinates to produce an image of a 3-dimensional volume. In a typical imaging system, the optical setup is kept fixed and either the sample or the objective is translated with a mechanical stage driven by a stepper motor or a piezoelectric element. Mechanical Z scanning is precise, but its slow response and vulnerability to mechanical vibrations and stress make it disadvantageous to image dynamic, time-varying samples such as live cell structures. An alternative method less susceptible to these problems is to change the focal plane using conjugate optics. Deformable mirrors, acousto-optics, and electrically tunable lenses have been experimented with to achieve this goal and have attained very fast and precise Z-scanning without physically moving the sample. Here, we present the use of a liquid lens for fast axial scanning. Liquid lenses have a long functional life, high degree of phase shift, and low sensitivity to mechanical stress. They work on the principle of refraction at a liquid-liquid interface. At the boundary of a polar and an apolar liquid a spherical surface is formed whose curvature can be controlled by adjusting its relative wettability using electro-wetting. We characterize the effects of the lens on attainable Z displacement, beam spectral characteristics, and pulse duration as compared with mechanical scanning.

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

    Science.gov (United States)

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

    2016-04-01

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

  9. Three-dimensional imaging of cerebellar mossy fiber rosettes by ion-abrasion scanning electron microscopy.

    Science.gov (United States)

    Kim, Hyun-Wook; Kim, Namkug; Kim, Ki Woo; Rhyu, Im Joo

    2013-08-01

    The detailed knowledge of the three-dimensional (3D) organization of the nervous tissue provides essential information on its functional elucidation. We used serial block-face scanning electron microscopy with focused ion beam (FIB) milling to reveal 3D morphologies of the mossy fiber rosettes in the mice cerebellum. Three-week-old C57 black mice were perfused with a fixative of 1% paraformaldehyde/1% glutaraldehyde in phosphate buffer; the cerebellum was osmicated and embedded in the Araldite. The block containing granule cell layer was sliced with FIB and observed by field-emission scanning electron microscopy. The contrast of backscattered electron image of the block-face was similar to that of transmission electron microscopy and processed using 3D visualization software for further analysis. The mossy fiber rosettes on each image were segmented and rendered to visualize the 3D model. The complete 3D characters of the mossy fiber rosette could be browsed on the A-Works, in-house software, and some preliminary quantitative data on synapse of the rosette could be extracted from these models. Thanks to the development of two-beam imaging and optimized software, we could get 3D information on cerebellar mossy fiber rosettes with ease and speedily, which would be an additive choice to explore 3D structures of the nervous systems and their networks.

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

    Science.gov (United States)

    Crowther, Susan D.

    1988-01-01

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

  11. Determining the resolution of scanning microwave impedance microscopy using atomic-precision buried donor structures

    Science.gov (United States)

    Scrymgeour, D. A.; Baca, A.; Fishgrab, K.; Simonson, R. J.; Marshall, M.; Bussmann, E.; Nakakura, C. Y.; Anderson, M.; Misra, S.

    2017-11-01

    To quantify the resolution limits of scanning microwave impedance microscopy (sMIM), we created scanning tunneling microscope (STM)-patterned donor nanostructures in silicon composed of 10 nm lines of highly conductive silicon buried under a protective top cap of silicon, and imaged them with sMIM. This dopant pattern is an ideal test of the resolution and sensitivity of the sMIM technique, as it is made with nm-resolution and offers minimal complications from topography convolution. It has been determined that typical sMIM tips can resolve lines down to ∼80 nm spacing, while resolution is independent of tip geometry as extreme tip wear does not change the resolving power, contrary to traditional scanning capacitance microscopy (SCM). Going forward, sMIM is an ideal technique for qualifying buried patterned devices, potentially allowing for quantitative post-fabrication characterization of donor structures, which may be an important tool for the study of atomic-scale transistors and state of the art quantum computation schemes.

  12. Analysis of enamel microbiopsies in shed primary teeth by Scanning Electron Microscopy (SEM) and Polarizing Microscopy (PM)

    Energy Technology Data Exchange (ETDEWEB)

    Costa de Almeida, Glauce Regina; Molina, Gabriela Ferian; Meschiari, Cesar Arruda [Department of Morphology, Stomatology and Physiology, Dental School of Ribeirao Preto, University of Sao Paulo - FORP/USP, Av. do Cafe, S/N, Monte Alegre, CEP 14040-904, Ribeirao Preto, SP (Brazil); Barbosa de Sousa, Frederico [Department of Morphology, Dental School of Joao Pessoa, Federal University of Paraiba - UFPB, Av Castelo Branco - Campus I, CEP 58.059-900, Joao Pessoa, PB (Brazil); Gerlach, Raquel Fernanda, E-mail: rfgerlach@forp.usp.br [Department of Morphology, Stomatology and Physiology, Dental School of Ribeirao Preto, University of Sao Paulo - FORP/USP, Av. do Cafe, S/N, Monte Alegre, CEP 14040-904, Ribeirao Preto, SP (Brazil)

    2009-09-01

    The aims of this study were 1) to verify how close to the theoretically presumed areas are the areas of enamel microbiopsies carried out in vivo or in exfoliated teeth; 2) to test whether the etching solution penetrates beyond the tape borders; 3) to test whether the etching solution demineralizes the enamel in depth. 24 shed upper primary central incisors were randomly divided into two groups: the Rehydrated Teeth Group and the Dry Teeth Group. An enamel microbiopsy was performed, and the enamel microbiopsies were then analyzed by Scanning Electron Microscopy (SEM) and Polarizing Microscopy (PM). Quantitative birefringence measurements were performed. The 'true' etched area was determined by measuring the etched enamel using the NIH Image analysis program. Enamel birefringence was compared using the paired t test. There was a statistically significant difference when the etched areas in the Rehydrated teeth were compared with those of the Dry teeth (p = 0.04). The etched areas varied from - 11.6% to 73.5% of the presumed area in the Rehydrated teeth, and from 6.6% to 61.3% in the Dry teeth. The mean percentage of variation in each group could be used as a correction factor for the etched area. Analysis of PM pictures shows no evidence of in-depth enamel demineralization by the etching solution. No statistically significant differences in enamel birefringence were observed between values underneath and outside the microbiopsy area in the same tooth, showing that no mineral loss occurred below the enamel superficial layer. Our data showed no evidence of in-depth enamel demineralization by the etching solution used in the enamel microbiopsy proposed for primary enamel. This study also showed a variation in the measured diameter of the enamel microbiopsy in nineteen teeth out of twenty four, indicating that in most cases the etching solution penetrated beyond the tape borders.

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

    Science.gov (United States)

    Lu, Kangbo; Sourty, Erwan; Loos, Joachim

    2010-08-01

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

  14. Scanning Transmission X-ray Microscopy: Applications in Atmospheric Aerosol Research

    Energy Technology Data Exchange (ETDEWEB)

    Moffet, Ryan C.; Tivanski, Alexei V.; Gilles, Mary K.

    2011-01-20

    Scanning transmission x-ray microscopy (STXM) combines x-ray microscopy and near edge x-ray absorption fine structure spectroscopy (NEXAFS). This combination provides spatially resolved bonding and oxidation state information. While there are reviews relevant to STXM/NEXAFS applications in other environmental fields (and magnetic materials) this chapter focuses on atmospheric aerosols. It provides an introduction to this technique in a manner approachable to non-experts. It begins with relevant background information on synchrotron radiation sources and a description of NEXAFS spectroscopy. The bulk of the chapter provides a survey of STXM/NEXAFS aerosol studies and is organized according to the type of aerosol investigated. The purpose is to illustrate the current range and recent growth of scientific investigations employing STXM-NEXAFS to probe atmospheric aerosol morphology, surface coatings, mixing states, and atmospheric processing.

  15. Morphology of the dentin structure of sloths Bradypus tridactylus: a light and scanning electron microscopy investigation.

    Science.gov (United States)

    Santana, L N S; Barbosa, L V M; Teixeira, F B; Costa, A M P; Fernandes, L M P; Lima, R R

    2013-12-01

    The aim of this study was to describe the dentine morphology of sloths (Bradypus tridactylus). The sloth teeth were removed and prepared for light microscopy (LM) and scanning electron microscopy analyses (SEM). LM revealed two patterns of tubular dentins: an outer with dentinary tubules over the all tooth length and one in the inner part with larger diameter and more spaced tubules, when compared to those present in the outer dentine. These findings were confirmed by SEM, which revealed a tubular pattern in the outer dentine like in humans. The inner dentine displayed pared grouped tubules that were characterized as vascular channels. It can be concluded that this sloth species present two types of dentins: an inner dentin (ortodentin) and an outer dentin characterized as a vascular dentin. This suggests a partial evolutive/adaptive process of this dental tissue, as compared to other mammalian species. © 2013 Blackwell Verlag GmbH.

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

    Science.gov (United States)

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

    2017-05-01

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

  17. Atomic species identification at the (101) anatase surface by simultaneous scanning tunnelling and atomic force microscopy

    Science.gov (United States)

    Stetsovych, Oleksandr; Todorović, Milica; Shimizu, Tomoko K.; Moreno, César; Ryan, James William; León, Carmen Pérez; Sagisaka, Keisuke; Palomares, Emilio; Matolín, Vladimír; Fujita, Daisuke; Perez, Ruben; Custance, Oscar

    2015-01-01

    Anatase is a pivotal material in devices for energy-harvesting applications and catalysis. Methods for the accurate characterization of this reducible oxide at the atomic scale are critical in the exploration of outstanding properties for technological developments. Here we combine atomic force microscopy (AFM) and scanning tunnelling microscopy (STM), supported by first-principles calculations, for the simultaneous imaging and unambiguous identification of atomic species at the (101) anatase surface. We demonstrate that dynamic AFM-STM operation allows atomic resolution imaging within the material's band gap. Based on key distinguishing features extracted from calculations and experiments, we identify candidates for the most common surface defects. Our results pave the way for the understanding of surface processes, like adsorption of metal dopants and photoactive molecules, that are fundamental for the catalytic and photovoltaic applications of anatase, and demonstrate the potential of dynamic AFM-STM for the characterization of wide band gap materials. PMID:26118408

  18. Two-dimensional dopant profiling of gallium nitride p–n junctions by scanning capacitance microscopy

    Energy Technology Data Exchange (ETDEWEB)

    Lamhamdi, M. [GREMAN UMR 7347-Université de Tours, 10 Rue Thales de Milet, BP 7155, 37071 Tours (France); Ecole national des sciences appliquées khouribga, Université Hassan 1er, 26000 Settat (Morocco); Cayrel, F. [GREMAN UMR 7347-Université de Tours, 10 Rue Thales de Milet, BP 7155, 37071 Tours (France); Frayssinet, E. [CRHEA-CNRS, Rue Bernard Grégory, Sophia Antipolis, 06560 Valbonne (France); Bazin, A.E.; Yvon, A.; Collard, E. [STMicroelectronics, 16 Rue Pierre et Marie Curie, BP 7155, 37071 Tours (France); Cordier, Y. [CRHEA-CNRS, Rue Bernard Grégory, Sophia Antipolis, 06560 Valbonne (France); Alquier, D. [GREMAN UMR 7347-Université de Tours, 10 Rue Thales de Milet, BP 7155, 37071 Tours (France)

    2016-04-01

    Two-dimensional imaging of dopant profiles for n and p-type regions are relevant for the development of new power semiconductors, especially for gallium nitride (GaN) for which classical profiling techniques are not adapted. This is a challenging task since it needs a technique with simultaneously good sensitivity, high spatial resolution and high dopant gradient resolution. To face these challenges, scanning capacitance microscopy combined with Atomic Force Microscopy is a good candidate, presenting reproducible results, as demonstrated in literature. In this work, we attempt to distinguish reliably and qualitatively the various doping concentrations and type at p–n and unipolar junctions. For both p–n and unipolar junctions three kinds of samples were prepared and measured separately. The space-charge region of the p–n metallurgical junction, giving rise to different contrasts under SCM imaging, is clearly observed, enlightening the interest of the SCM technique.

  19. Submonolayer growth of Pd on Cu(111) studied by scanning tunneling microscopy

    DEFF Research Database (Denmark)

    Lægsgaard, E.; Ruban, Andrei; Stensgaard, I.

    1998-01-01

    The growth mode of sub-monolayer amounts of Pd on Cu(111) in the temperature range - 80-300 degrees C has been investigated by scanning tunneling microscopy (STM), Rutherford backscattering spectroscopy (RBS) and Auger electron spectroscopy (AES). Below approximate to 100 degrees C, the Pd induced...... is dug out from the surface in extended, monolayer deep pits, and concurrently, the brims and islands increase in height by one layer. High-resolution STM images of brims and islands in this phase are interpreted as evidence for Cu capping. For Pd evaporation at temperatures of 220-300 degrees C...

  20. Scanning electron microscopy and X-ray spectroscopy applied to mycelial phase of sporothrix schenckii

    Directory of Open Access Journals (Sweden)

    M. Thibaut

    1975-04-01

    Full Text Available Scanning electron microscopy applied to the mycelial phase of Sporothrix schenckii shows a matted mycelium with conidia of a regular pattern. X-Ray microanalysis applied in energy dispersive spectroscopy and also in wavelength dispersive spectroscopy reveals the presence of several elements of Mendeleef's classification.Sporothrix schenckii foi estudado em microscopia eletrônica. Foram observados caracteres das hífas e dos esporos, vários elementos da classificação periódica foram postos em evidência graças à micro-análise a raios X.

  1. Atomic bonding effects in annular dark field scanning transmission electron microscopy. II. Experiments

    Energy Technology Data Exchange (ETDEWEB)

    Odlyzko, Michael L.; Held, Jacob T.; Mkhoyan, K. Andre, E-mail: mkhoyan@umn.edu [Department of Chemical Engineering and Materials Science, University of Minnesota, Minneapolis, Minnesota 55455 (United States)

    2016-07-15

    Quantitatively calibrated annular dark field scanning transmission electron microscopy (ADF-STEM) imaging experiments were compared to frozen phonon multislice simulations adapted to include chemical bonding effects. Having carefully matched simulation parameters to experimental conditions, a depth-dependent bonding effect was observed for high-angle ADF-STEM imaging of aluminum nitride. This result is explained by computational predictions, systematically examined in the preceding portion of this study, showing the propagation of the converged STEM beam to be highly sensitive to net interatomic charge transfer. Thus, although uncertainties in experimental conditions and simulation accuracy remain, the computationally predicted experimental bonding effect withstands the experimental testing reported here.

  2. Phase stabilized homodyne of infrared scattering type scanning near-field optical microscopy

    Energy Technology Data Exchange (ETDEWEB)

    Xu, Xiaoji G., E-mail: xgx214@lehigh.edu [Department of Chemistry, Lehigh University, Bethlehem, Pennsylvania 18015 (United States); Gilburd, Leonid; Walker, Gilbert C., E-mail: gwalker@chem.utoronto.ca [Department of Chemistry, University of Toronto, Toronto, Ontario M5S 3H6 (Canada)

    2014-12-29

    Scattering type scanning near-field optical microscopy (s-SNOM) allows sub diffraction limited spatial resolution. Interferometric homodyne detection in s-SNOM can amplify the signal and extract vibrational responses based on sample absorption. A stable reference phase is required for a high quality homodyne-detected near-field signal. This work presents the development of a phase stabilization mechanism for s-SNOM to provide stable homodyne conditions. The phase stability is found to be better than 0.05 rad for the mid infrared light source. Phase stabilization results in improved near field images and vibrational spectroscopies. Spatial inhomogeneities of the boron nitride nanotubes are measured and compared.

  3. Vortex imaging and local magnetization studies in HTS by scanning Hall probe microscopy

    Energy Technology Data Exchange (ETDEWEB)

    Crisan, A.; Pross, A.; Cole, D.; Bending, S

    2004-08-01

    We have used scanning Hall probe microscopy to correlate vortex images and local magnetisation measurements in two different high temperature superconducting samples. Near the edge of a continuous YBCO thin film we have observed local hysteresis inversion and negative remanent fields, which can be semi-quantitatively explained in terms of a theoretical model of flux penetration in an infinitely long superconducting strip. In a YBCO film containing a regular square array of antidots we have further find that vortices trapped at antidots exhibit an unusual behaviour upon field sweep reversal.

  4. Low-Level Detection of Poly(amidoamine PAMAM Dendrimers Using Immunoimaging Scanning Probe Microscopy

    Directory of Open Access Journals (Sweden)

    Chevelle A. Cason

    2012-01-01

    Full Text Available Immunoimaging scanning probe microscopy was utilized for the low-level detection and quantification of biotinylated G4 poly(amidoamine PAMAM dendrimers. Results were compared to those of high-performance liquid chromatography (HPLC and found to provide a vastly improved analytical method for the low-level detection of dendrimers, improving the limit of detection by a factor of 1000 (LOD=2.5×10−13 moles. The biorecognition method is reproducible and shows high specificity and good accuracy. In addition, the capture assay platform shows a promising approach to patterning dendrimers for nanotechnology applications.

  5. In vivo measurements of skin barrier: comparison of different methods and advantages of laser scanning microscopy

    Science.gov (United States)

    Patzelt, A.; Sterry, W.; Lademann, J.

    2010-12-01

    A major function of the skin is to provide a protective barrier at the interface between external environment and the organism. For skin barrier measurement, a multiplicity of methods is available. As standard methods, the determination of the transepidermal water loss (TEWL) as well as the measurement of the stratum corneum hydration, are widely accepted, although they offer some obvious disadvantages such as increased interference liability. Recently, new optical and spectroscopic methods have been introduced to investigate skin barrier properties in vivo. Especially, laser scanning microscopy has been shown to represent an excellent tool to study skin barrier integrity in many areas of relevance such as cosmetology, occupation, diseased skin, and wound healing.

  6. Scanning probe microscopy estimation of the wear resistance of the surface of a modified PVC film

    Science.gov (United States)

    Kochetkova, A. S.; Gorbushin, P. N.; Sosnov, E. A.; Kolert, K.; Malygin, A. A.

    2017-04-01

    An atomic force microscopy technique is proposed to determine the wear resistance of a protective coating deposited by the sol-gel method on the surface of a polyvinylchloride film. The force of action of a probe on a sample is estimated under various scanning conditions. It is shown that the obtained data on the resistance of a coating to the action of a probe in the contact mode can be used to qualitatively estimate the adhesion of the coating to the surface of a polymer matrix.

  7. Structure and Reactions of Carbon and Hydrogen on Ru(0001): A Scanning Tunneling Microscopy Study

    Energy Technology Data Exchange (ETDEWEB)

    Shimizu, Tomoko K.; Mugarza, Aitor; Cerda, Jorge; Salmeron, Miquel

    2008-09-09

    The interaction between carbon and hydrogen atoms on a Ru(0001) surface was studied using scanning tunneling microscopy (STM), Density Functional Theory (DFT) and STM image calculations. Formation of CH species by reaction between adsorbed H and C was observed to occur readily at 100 K. When the coverage of H increased new complexes of the form CH+nH (n = 1, 2 and 3) were observed. These complexes, never observed before, might be precursors for further hydrogenation reactions. DFT analysis reveals that a considerable energy barrier exists for the CH+H {yields} CH{sub 2} reaction.

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

    Energy Technology Data Exchange (ETDEWEB)

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

    2013-12-02

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

  9. Metadislocations in complex metallic alloys: A high-resolution scanning transmission electron microscopy study

    Energy Technology Data Exchange (ETDEWEB)

    Heggen, Marc; Houben, Lothar; Feuerbacher, Michael [Ernst Ruska Centre for Microscopy and Spectroscopy with Electrons, Forschungszentrum Juelich GmbH, D-52425 Juelich (Germany)

    2011-07-01

    Metadislocations are highly complex defects which involve several hundreds of atoms in their core. We present a microstructural investigation on Metadislocations using aberration-corrected high-resolution scanning transmission electron microscopy. A novel and highly complex deformation mechanism is found which is based on the movement of a metadislocation core mediating strain and separate escort defects. Upon deformation, the escort defects move along with the metadislocation core and locally transform the material structure. This mechanism implies the coordinated movement of hundreds of atoms per elementary step. Although the mechanism is very complex, it can be described by a simple jigsaw-puzzle-like rearrangement of basic structural subunits.

  10. Cytochrome C Dynamics at Gold and Glassy Carbon Surfaces Monitored by in Situ Scanning Tunnel Microscopy

    DEFF Research Database (Denmark)

    Andersen, Jens Enevold Thaulov; Møller, Per; Pedersen, Marianne Vind

    1995-01-01

    We have investigated the absorption of cytochrome c on gold and glassy carbon substrates by in situ scanning tunnel microscopy under potentiostatic control of both substrate and tip. Low ionic strength and potential ranges where no Faradaic current flows were used. Cyt c aggregates into flat...... composite structures of about 50 nm lateral extension at gold surfaces. The aggregates evolve in time, and structures resembling individual cyt c molecules can be distinguished in the space between the 50 nm structures. Cyt c aggregates also form at glassy carbon but have a different, unbroken character...

  11. Monolithically Integrated, Mechanically Resilient Carbon-Based Probes for Scanning Probe Microscopy

    Science.gov (United States)

    Kaul, Anupama B.; Megerian, Krikor G.; Jennings, Andrew T.; Greer, Julia R.

    2010-01-01

    Scanning probe microscopy (SPM) is an important tool for performing measurements at the nanoscale in imaging bacteria or proteins in biology, as well as in the electronics industry. An essential element of SPM is a sharp, stable tip that possesses a small radius of curvature to enhance spatial resolution. Existing techniques for forming such tips are not ideal. High-aspect-ratio, monolithically integrated, as-grown carbon nanofibers (CNFs) have been formed that show promise for SPM applications by overcoming the limitations present in wet chemical and separate substrate etching processes.

  12. Calibration of Electrochemical Capacitance-voltage Method on Pyramid Texture Surface Using Scanning Electron Microscopy

    Energy Technology Data Exchange (ETDEWEB)

    Komatsu, Y.; Cesar, I. [ECN Solar Energy, P.O.Box 1, 1755ZG Petten (Netherlands); Harata, D. [Department of Electronic Science and Engineering, Kyoto University, Nishikyo-ku, Kyoto 615-8530 (Japan); Schuring, E.W. [ECN Environment and Energy Engineering, P.O.Box 1, 1755ZG Petten (Netherlands); Vlooswijk, A.H.G.; Venema, P.R. [Tempress Systems BV, Radeweg 31, 8171MD Vaassen (Netherlands); Katori, S.; Fujita, S. [Photonics and Electronics Science and Engineering Center, Kyoto University, Nishikyo-ku, Kyoto 615-8530 (Japan)

    2013-07-01

    The electrochemical capacitance-voltage (ECV) technique can practically profile carrier concentrations on textured surfaces, but reliable calibration of the surface area is strongly demanded since it plays a decisive role in calculating both the carrier concentration and the profiling depth. In this work, we calibrate the area factor of pyramidally textured surfaces by comparing ECV profiles with cross-sectional scanning electron microscopy image, and found out it is 1.66, and not 1.73 which was formerly assumed. Furthermore, the calibrated area factor was applied to POCl3 and BBr3 diffusions which resulted in comparable diffusion profiles for both textured and polished surfaces.

  13. Observations on mouthparts of Dermatobia hominis (Linneaus Jr., 1781) (Diptera: Cuterebridae) by scanning electron microscopy.

    Science.gov (United States)

    Fernandes, Fernando de Freitas; Linardi, Pedro Marcos

    2002-02-01

    The ultrastructure of the mouthparts of Dermatobia hominis was studied using scanning electron microscopy. The morphological characteristics of the segments, articulations, sensory organs, and pilose covering are described. Mechanoreceptors of the long trichoid sensillum and smaller trichoid sensillum types were observed, as well as labellar gustatory receptors of the basiconic sensillum type, which differed between the sexes. These observations are discussed with reference to the current literature on the morphology and sense organs of dipteran mouthparts, and the prevailing view that the adult mouthparts of this species are non-functional is challenged.

  14. Atomic-scale structure of dislocations revealed by scanning tunneling microscopy and molecular dynamics

    DEFF Research Database (Denmark)

    Christiansen, Jesper; Morgenstern, K.; Schiøtz, Jakob

    2002-01-01

    The intersection between dislocations and a Ag(111) surface has been studied using an interplay of scanning tunneling microscopy (STM) and molecular dynamics. Whereas the STM provides atomically resolved information about the surface structure and Burgers vectors of the dislocations......, the simulations can be used to determine dislocation structure and orientation in the near-surface region. In a similar way, the subsurface structure of other extended defects can be studied. The simulations show dislocations to reorient the partials in the surface region leading to an increased splitting width...

  15. Low-Level Detection of Poly(amidoamine) PAMAM Dendrimers Using Immunoimaging Scanning Probe Microscopy.

    Science.gov (United States)

    Cason, Chevelle A; Fabré, Thomas A; Buhrlage, Andrew; Haik, Kristi L; Bullen, Heather A

    2012-01-01

    Immunoimaging scanning probe microscopy was utilized for the low-level detection and quantification of biotinylated G4 poly(amidoamine) PAMAM dendrimers. Results were compared to those of high-performance liquid chromatography (HPLC) and found to provide a vastly improved analytical method for the low-level detection of dendrimers, improving the limit of detection by a factor of 1000 (LOD = 2.5 × 10(-13) moles). The biorecognition method is reproducible and shows high specificity and good accuracy. In addition, the capture assay platform shows a promising approach to patterning dendrimers for nanotechnology applications.

  16. Teaching Plasmonics, Scanning Probe Microscopy and Other Useful Experiments at the Upper Level

    Science.gov (United States)

    Sanchez, Erik

    2012-10-01

    It is important to teach students concepts and experimental skills relating to modern research being performed today. Experiments that help educate students about the latest research helps them get jobs and into the doors at many great academic institutions. PSU's Advanced Experimental Class for physics undergraduates offers many novel experiments to help the students accomplish this task. Labs involving Plasmonics, thin film deposition, scanning probe microscopy (SPM) and more will be discussed. In addition, a new NSF funded project involving the building of a Do-It-Yourself (DIY) SPM will be discussed.

  17. Creating Nanoscale Pits on Solid Surfaces in Aqueous Environment with Scanning Tunnelling Microscopy

    DEFF Research Database (Denmark)

    Chi, Qijin; Zhang, Jingdong; Friis, Esben P.

    2000-01-01

    A novel method has been developed to fabricate nanoscale pits on Au(111) in aqueous environments by in situ scanning tunnelling microscopy (STM), based on critical interactions between tip and substrate. The most striking advantages of the present method are that the dimension and position...... of the pits can be controlled well in aqueous environments, and the operations are simple. Parameters affecting the pit formation and size have been systematically characterized to show that pit formation is dominated by bias voltage. A mechanism is proposed based on local surface reconstruction induced...

  18. Scanning electron microscopy of the collodion membrane from a self-healing collodion baby*

    Science.gov (United States)

    de Almeida Jr., Hiram Larangeira; Isaacsson, Henrique; Guarenti, Isabelle Maffei; Silva, Ricardo Marques e; de Castro, Luis Antônio Suita

    2015-01-01

    Abstract Self-healing collodion baby is a well-established subtype of this condition. We examined a male newborn, who was covered by a collodion membrane. The shed membrane was examined with scanning electron microscopy. The outer surface showed a very compact keratin without the normal elimination of corneocytes. The lateral view of the specimen revealed a very thick, horny layer. The inner surface showed the structure of lower corneocytes with polygonal contour. With higher magnifications villous projections were seen in the cell membrane. PMID:26375232

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

    Directory of Open Access Journals (Sweden)

    Suhyun Kim

    2013-09-01

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

  20. Identification of nitrogen dopants in single-walled carbon nanotubes by scanning tunneling microscopy.

    Science.gov (United States)

    Tison, Yann; Lin, Hong; Lagoute, Jérôme; Repain, Vincent; Chacon, Cyril; Girard, Yann; Rousset, Sylvie; Henrard, Luc; Zheng, Bing; Susi, Toma; Kauppinen, Esko I; Ducastelle, François; Loiseau, Annick

    2013-08-27

    Using scanning tunnelling microscopy and spectroscopy, we investigated the atomic and electronic structure of nitrogen-doped single walled carbon nanotubes synthesized by chemical vapor deposition. The insertion of nitrogen in the carbon lattice induces several types of point defects involving different atomic configurations. Spectroscopic measurements on semiconducting nanotubes reveal that these local structures can induce either extended shallow levels or more localized deep levels. In a metallic tube, a single doping site associated with a donor state was observed in the gap at an energy close to that of the first van Hove singularity. Density functional theory calculations reveal that this feature corresponds to a substitutional nitrogen atom in the carbon network.

  1. Dopant migration in silicon during implantation/annealing measured by scanning tunneling microscopy

    Energy Technology Data Exchange (ETDEWEB)

    Hessel, H.E.; Memmert, U.; Behm, R.J. (Univ. Muenchen (West Germany)); Cerva, H. (Siemens Research Lab., Muenchen (West Germany))

    In this paper spatial correlation between the lateral distribution of the doping type and the former implantation mask edge was investigated by scanning tunneling microscopy (STM) measurements. The position of the former mask edge was determined from surface steps resolved by STM topography measurements. Current imaging tunneling spectroscopy (CITS) data recorded simultaneously allowed to detect the transition from a high doping level with an ohmic I-V curve to a lower doping level displaying a Schottky barrier behavior. The influence of different annealing treatments on the position of this transition was investigated.

  2. Direct Measurement of Built-in Electrical Potential in Photovoltaic Devices by Scanning Kelvin Probe Microscopy

    Energy Technology Data Exchange (ETDEWEB)

    Jiang, C. S.; Mutinho, H. R.; Hasoon, F. S.; Al-Thani, H. A.; Friedman, D. J.; Geisz, J. F.; Wang, Q.; Romero, M. J.; Al-Jassim, M. M.

    2003-05-01

    We report on direct measurements of the built-in electrical potential in Cu(In,Ga)Se2, GaInP2 single-junction, and GaInP2/GaAs tandem-junction solar cells, by using scanning Kelvin probe microscopy. Potential profiles on cross sections of the devices were measured quantitatively and spatially resolved in open and short circuit, under and without illuminations, with selective photon energies matching the band gaps of the junctions. The measurements provide valuable information about the electrical properties of the devices and are useful for understanding the performance and improving the design of solar cells.

  3. Calibration and examination of piezoresistive Wheatstone bridge cantilevers for scanning probe microscopy.

    Science.gov (United States)

    Gotszalk, Teodor; Grabiec, Piotr; Rangelow, Ivo W

    2003-01-01

    This paper describes the method of determining the force constant and displacement sensitivity of piezoresistive Wheatstone bridge cantilevers applied in scanning probe microscopy (SPM). In the procedure presented here, the force constant for beams with various geometry is determined based on resonance frequency measurement. The displacement sensitivity is measured by the deflection of the cantilever with the calibrated piezoactuator stage. Preliminary results show that our method is capable of measuring the force constant of Wheatstone bridge cantilevers with an accuracy of better than 5% and this is used as feedback for improvement of sensor micromachining process.

  4. Quantitative study of mammalian cells by scanning transmission soft X-ray microscopy

    Science.gov (United States)

    Shinohara, K.; Ohigashi, T.; Toné, S.; Kado, M.; Ito, A.

    2017-06-01

    Molecular distribution in mammalian cells was studied by soft X-ray scanning transmission microscopy with respect to the quantitative aspect of analysis. NEXAFS profiles at the C, N and O K-absorption edges were combined and used for the analysis. For the estimation of quantity for nucleic acids and proteins, NEXAFS profiles of DNA and bovine serum albumin (BSA) at the N K-absorption edge were applied assuming that those were their representatives. The method has a potential to explore the other molecular components than nucleic acids and proteins.

  5. Circular photocurrent response of a topological insulator thin film probed by scanning photocurrent microscopy

    Science.gov (United States)

    Qu, Dong-Xia; Kou, Xufeng; Lang, Murong; Crowhurst, Jonathan; Armstrong, Michael; Zaug, Joseph; Wang, Kang L.; Chapline, George

    2015-03-01

    The remarkable nature of surface states in topological insulators is expected to have a unique photocurrent response to electromagnetic radiation. However, the surface and bulk photo-excited charge transport mechanisms, in relation to the band bending at the electrode-topological insulator interface, have not been well understood. Here, we present scanning photocurrent microscopy measurements on a gated topological insulator microdevice and show that the spin-polarized photocurrent displays direction reversal near the electrical contact interfaces. We discuss two possible mechanisms, which alternatively play dominant roles in the helicity-dependent photocurrent map. Our analysis determines the magnitude of each contribution, and reveals the governing process under different gate conditions.

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

    Science.gov (United States)

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

    1990-10-01

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

  7. First-principles modelling of scanning tunneling microscopy using non-equilibrium Green's functions

    DEFF Research Database (Denmark)

    Lin, H.P.; Rauba, J.M.C.; Thygesen, Kristian Sommer

    2010-01-01

    The investigation of electron transport processes in nano-scale architectures plays a crucial role in the development of surface chemistry and nano-technology. Experimentally, an important driving force within this research area has been the concurrent refinements of scanning tunneling microscopy...... into account. As an illustrating example we apply the NEGF-STM method to the Si(001)(2x1):H surface with sub-surface P doping and discuss the results in comparison to the Bardeen and Tersoff-Hamann methods....

  8. Doppler-scanning tunneling microscopy current imaging in superconductor-ferromagnet hybrids

    Energy Technology Data Exchange (ETDEWEB)

    Moore, S. A.; Plummer, G.; Fedor, J.; Pearson, J. E.; Novosad, V.; Karapetrov, G.; Iavarone, M.

    2016-01-25

    Mapping the distribution of currents inside a superconductor is usually performed indirectly through imaging of the stray magnetic fields above the surface. Here, we show that by direct imaging of the Doppler shift contribution to the quasiparticle excitation spectrum in the superconductor using low temperature scanning tunneling microscopy, we obtain directly the distribution of supercurrents inside the superconductor. We demonstrate the technique at the example of superconductor/ferromagnet hybrid structure that produces intricate current pattern consisting of combination Meissner shielding currents and Abrikosov vortex currents.

  9. Imaging plant nuclei and membrane-associated cytoskeleton by field emission scanning electron microscopy.

    Science.gov (United States)

    Fišerová, Jindřiška; Goldberg, Martin W

    2014-01-01

    Scanning electron microscopy (SEM) is a powerful technique that can image exposed surfaces in 3D. Modern scanning electron microscopes, with field emission electron sources and in-lens specimen chambers, achieve resolutions of better than 0.5 nm and thus offer views of ultrastructural details of subcellular structures or even macromolecular complexes. Obtaining a reliable image is, however, dependent on sample preparation methods that robustly but accurately preserve biological structures. In plants, exposing the object of interest may be difficult due to the existence of a cell wall. This protocol shows how to isolate plant nuclei for SEM imaging of the nuclear envelope and associated structures from both sides of the nuclear envelope in cultured cells as well as in leaf or root cells. Further, it provides a method for uncovering membrane-associated cytoskeletal structures.

  10. High resolution characterizations of fine structure of semiconductor device and material using scanning nonlinear dielectric microscopy

    Science.gov (United States)

    Cho, Yasuo

    2017-10-01

    Scanning nonlinear dielectric microscopy (SNDM) can easily distinguish the dopant type (PN) and has a wide dynamic range of sensitivity from low to high concentrations of dopants, because it has a high sensitivity to capacitance variation on the order of 10-22 F/\\sqrt{\\text{Hz}} . It is also applicable to the analysis of compound semiconductors with much lower signal levels than Si. We can avoid misjudgments from the two-valued function (contrast reversal) problem of dC/dV signals. Under an ultrahigh-vacuum condition, SNDM has atomic resolution. As the extended versions of SNDM, super-higher-order SNDM, local-deep-level transient spectroscopy, noncontact SNDM, and scanning nonlinear dielectric potentiometory have been developed and introduced. The favorable features of SNDM originate from its significantly high sensitivity.

  11. Revisit laser scanning fluorescence microscopy performance under fluorescence-lifetime-limited regime

    Science.gov (United States)

    Chan, Antony C.; Wong, Terence T. W.; Wong, Kenneth K. Y.; Lam, Edmund Y.; Tsia, Kevin K.

    2014-03-01

    Continuing desire for higher-speed laser scanning fluorescence microscopy (LSFM) and progressive advancement in ultrafast and sensitive photodetectors might imply that our conventional understanding of LSFM is not adequate when approaching to the intrinsic speed limit — fluorescence lifetime. In this regard, we here revisit the theoretical framework of LSFM and evaluate its general performance in lifetime-limited and noise-limited regimes. Our model suggests that there still exists an order-of-magnitude gap between the current LSFM speed and the intrinsic limit. An imaging frame rate of > 100 kHz could be viable with the emerging laser-scanning techniques using ultrafast wavelength-swept sources, or optical time-stretch.

  12. High Speed Scanning Ion Conductance Microscopy for Quantitative Analysis of Nanoscale Dynamics of Microvilli.

    Science.gov (United States)

    Ida, Hiroki; Takahashi, Yasufumi; Kumatani, Akichika; Shiku, Hitoshi; Matsue, Tomokazu

    2017-06-06

    Observation of nanoscale structure dynamics on cell surfaces is essential to understanding cell functions. Hopping-mode scanning ion conductance microscopy (SICM) was used to visualize the topography of fragile convoluted nanoscale structures on cell surfaces under noninvasive conditions. However, conventional hopping mode SICM does not have sufficient temporal resolution to observe cell-surface dynamics in situ because of the additional time required for performing vertical probe movements of the nanopipette. Here, we introduce a new scanning algorithm for high speed SICM measurements using low capacitance and high-resonance-frequency piezo stages. As a result, a topographic image is taken within 18 s with a 64 × 64 pixel resolution at 10 × 10 μm. The high speed SICM is applied to the characterization of microvilli dynamics on surfaces, which shows clear structural changes after the epidermal growth factor stimulation.

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

    Science.gov (United States)

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

    2016-12-01

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

  14. MRT letter: An extended scanning probe microscopy system for macroscopic topography imaging.

    Science.gov (United States)

    Fu, Ji; Li, Faxin

    2014-10-01

    Enlightened by the principle of scanning probe microscopy or atomic force microscope (AFM), we proposed a novel surface topography imaging system based on the scanning of a piezoelectric unimorph cantilever. The height of sample surface can be obtained by recording the cantilever's strain using an ultra-sensitive strain gauge and the Z-axis movement is realized by electric bending of the cantilever. This system can be operated in the way similar to the contact mode in AFM, with the practical height detection resolution better than 100 nm. Imaging of the inner surface of a steel tube and on a transparent wing of a honey bee were conducted and the obtained results showed that this proposed system is a very promising solution for in situ topography mapping. © 2014 Wiley Periodicals, Inc.

  15. Scanning tunneling microscopy investigation of different porphynoids on a Ni-prestructured Cu(111) surface

    Energy Technology Data Exchange (ETDEWEB)

    Roeckert, Michael; Buchner, Florian; Zillner, Elisabeth; Glaessel, Stefanie; Steinrueck, Hans-Peter; Marbach, Hubertus [Lehrstuhl fuer Physikalische Chemie II and Interdisciplinary Center for Molecular Materials (ICMM), Universitaet Erlangen-Nuernberg, Egerlandstrasse 3, D-91058 Erlangen (Germany)

    2010-07-01

    The assembly of organic molecules on single-crystal surfaces is an approach towards the creation of novel materials with outstanding properties. Porphyrins appear to be ideal candidates to generate functional molecular devices, due to their self-assembly properties and their versatile functionality. In the present work we study the possibility to locally anchor or functionalize porphyrins on a prestructured surface, namely a composite Ni/Cu(111) surface, by scanning tunneling microscopy (STM) in ultra-high vacuum at room temperature. Based on scanning tunneling micrographs and movies the dynamics, assembly and intramolecular conformation of the corresponding porphyrins (2HTPP,CoTPP,OEP) as well as the role of molecule-molecule and molecule-substrate interactions are discussed. The obtained findings indeed indicate the possibility to locally anchor and/or functionalize (e.g. metalate) the porphyrins on a Cu(111) surface prestructured either with atomically flat Ni- or oxygen-islands.

  16. Frequency-Modulated Magneto-Acoustic Detection and Imaging: Challenges, Experimental Procedures, and B-Scan Images

    CERN Document Server

    Aliroteh, Miaad S; Arbabian, Amin

    2016-01-01

    Magneto-acoustic tomography combines near-field radio-frequency (RF) and ultrasound with the aim of creating a safe, high resolution, high contrast hybrid imaging technique. We present continuous-wave magneto-acoustic imaging techniques, which improve SNR and/or reduce the required peak-to-average excitation power ratio, to make further integration and larger fields of view feasible. This method relies on the coherency between RF excitation and the resulting ultrasound generated through Lorentz force interactions, which was confirmed by our previous work. We provide detailed methodology, clarify the details of experiments, and explain how the presence of magneto-acoustic phenomenon was verified. An example magneto-acoustic B-scan image is acquired in order to illustrate the capability of magneto-acoustic tomography in highlighting boundaries where electrical conductivity alters, such as between different tissues.

  17. Ultra-High Resolution Thin Film Thickness Delineation Using Reflection Phase-Sensitive Acoustic Microscopy

    Science.gov (United States)

    Mohamed, E. A.; Kamanyi, A.; von Buttlar, M.; Wannemacher, R.; Hillmann, K.; Ngwa, W.; Grill, W.

    The acoustic phase and magnitude data of a planar homogenous sample of smoothly varying thickness deposited on a glass substrate can best be represented by a polar plot. In this work, the method is extended to achieve topographical mapping of thin films with a height resolution beyond the diffraction limit of optical confocal microscopy. The radial dependence of the polar graph describes the regression of the magnitude of the reflected signal due to the attenuation. The later increases with the gradual increase of the thickness and is additionally influenced by interference effects. The angular dependence of the polar plot reveals the rotation of the phase angle of the signal due to reflection from different thicknesses of the sample. Model calculations are employed, and input values are varied until an optimum agreement with the measurement data points is achieved and the primary acoustic properties (speed of longitudinally polarized ultrasound, mechanical density of the sample and the attenuation within the material) are obtained. The model manifests the variation of the magnitude and phase of the reflected signal due to variation in thickness. After optimum adjustment of the model parameters, the thickness corresponding to each measured value of the reflectivity is obtained.

  18. Comparative analysis of Trichuris muris surface using conventional, low vacuum, environmental and field emission scanning electron microscopy

    National Research Council Canada - National Science Library

    Lopes Torres, Eduardo José; de Souza, Wanderley; Miranda, Kildare

    2013-01-01

    .... The morphology of Trichuris spp. and other helminths has been mostly studied using conventional scanning electron microscopy of chemically fixed, dried and metal-coated specimens, although this kind of preparation has been shown...

  19. THE INTEGRATED USE OF COMPUTATIONAL CHEMISTRY, SCANNING PROBE MICROSCOPY, AND VIRTUAL REALITY TO PREDICT THE CHEMICAL REACTIVITY OF ENVIRONMENTAL SURFACES

    Science.gov (United States)

    In the last decade three new techniques scanning probe microscopy (SPM), virtual reality (YR) and computational chemistry ave emerged with the combined capability of a priori predicting the chemically reactivity of environmental surfaces. Computational chemistry provides the cap...

  20. Critical-point drying and gold sputtering as applied to scanning electron microscopy of human reproductive tissues.

    Science.gov (United States)

    Ludwig, H; Metzger, H; Hafez, E S

    1976-01-01

    Improved techniques have been developed for washing, pinning, fixation, dehydration and critical-point drying of tissues of female and male reproductive tracts, gametes, for viewing by scanning electron microscopy. The sputtering method, performed by an ion gun, is applied to shadow and produce a thin film of gold. Technical details and metal coating are noted which are suitable to improve images of surface ultrastructure of cilia and microvilli. Studies using scanning electron microscopy, histological and histochemical techniques, and cinematography are described.

  1. Simultaneous X-ray fluorescence and scanning X-ray diffraction microscopy at the Australian Synchrotron XFM beamline

    Energy Technology Data Exchange (ETDEWEB)

    Jones, Michael W. M.; Phillips, Nicholas W.; van Riessen, Grant A.; Abbey, Brian; Vine, David J.; Nashed, Youssef S. G.; Mudie, Stephen T.; Afshar, Nader; Kirkham, Robin; Chen, Bo; Balaur, Eugeniu; de Jonge, Martin D.

    2016-08-11

    Owing to its extreme sensitivity, quantitative mapping of elemental distributionsviaX-ray fluorescence microscopy (XFM) has become a key microanalytical technique. The recent realisation of scanning X-ray diffraction microscopy (SXDM) meanwhile provides an avenue for quantitative super-resolved ultra-structural visualization. The similarity of their experimental geometries indicates excellent prospects for simultaneous acquisition. Here, in both step- and fly-scanning modes, robust, simultaneous XFM-SXDM is demonstrated.

  2. Scanning tunneling microscopy I general principles and applications to clean and absorbate-covered surfaces

    CERN Document Server

    Wiesendanger, Roland

    1994-01-01

    Since the first edition of "Scanning 'funneling Microscopy I" has been pub­ lished, considerable progress has been made in the application of STM to the various classes of materials treated in this volume, most notably in the field of adsorbates and molecular systems. An update of the most recent develop­ ments will be given in an additional Chapter 9. The editors would like to thank all the contributors who have supplied up­ dating material, and those who have provided us with suggestions for further improvements. We also thank Springer-Verlag for the decision to publish this second edition in paperback, thereby making this book affordable for an even wider circle of readers. Hamburg, July 1994 R. Wiesendanger Preface to the First Edition Since its invention in 1981 by G. Binnig, H. Rohrer and coworkers at the IBM Zurich Research Laboratory, scanning tunneling microscopy (STM) has devel­ oped into an invaluable surface analytical technique allowing the investigation of real-space surface structures at th...

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

    Science.gov (United States)

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

    2016-11-01

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

  4. Scanning two-photon microscopy with upconverting lanthanide nanoparticles via Richardson-Lucy deconvolution

    Science.gov (United States)

    Gainer, Christian F.; Utzinger, Urs

    2012-01-01

    Abstract. The use of upconverting lanthanide nanoparticles in fast-scanning microscopy is hindered by a long luminescence decay time, which greatly blurs images acquired in a nondescanned mode. We demonstrate herein an image processing method based on Richardson-Lucy deconvolution that mitigates the detrimental effects of their luminescence lifetime. This technique generates images with lateral resolution on par with the system’s performance, ∼1.2  μm, while maintaining an axial resolution of 5 μm or better at a scan rate comparable with traditional two-photon microscopy. Remarkably, this can be accomplished with near infrared excitation power densities of 850  W/cm2, several orders of magnitude below those used in two-photon imaging with molecular fluorophores. By way of illustration, we introduce the use of lipids to coat and functionalize these nanoparticles, rendering them water dispersible and readily conjugated to biologically relevant ligands, in this case epidermal growth factor receptor antibody. This deconvolution technique combined with the functionalized nanoparticles will enable three-dimensional functional tissue imaging at exceptionally low excitation power densities. PMID:22894486

  5. Magnetic scanning gate microscopy of CoFeB lateral spin valve

    Science.gov (United States)

    Corte-León, Héctor; Scarioni, Alexander Fernandez; Mansell, Rhodri; Krzysteczko, Patryk; Cox, David; McGrouther, Damien; McVitie, Stephen; Cowburn, Russell; Schumacher, Hans W.; Antonov, Vladimir; Kazakova, Olga

    2017-05-01

    Devices comprised of CoFeB nanostructures with perpendicular magnetic anisotropy and non-magnetic Ta channel were operated in thermal lateral spin valve (LSV) mode and studied by magnetotransport measurements and magnetic scanning gate microscopy (SGM). Due to the short spin diffusion length of Ta, the spin diffusion signal was suppressed, allowing the study of the contribution from the anomalous Nernst (ANE) and anomalous Hall effects (AHE). The magnetotransport measurements identified the switching fields of the CoFeB nanostructures and demonstrated a combination of AHE and ANE when the devices were operated in thermally-driven spin-injection mode. Modified scanning probe microscopy probes were fabricated by placing a NdFeB magnetic bead (MB) on the apex of a commercial Si probe. The dipole magnetic field distribution around the MB was characterized by using differential phase contrast technique and direct measurement of the switching field induced by the bead in the CoFeB nanodevices. Using SGM we demonstrate the influence of localized magnetic field on the CoFeB nanostructures near the non-magnetic channel. This approach provides a promising route towards the study of thermal and spin diffusion effects using local magnetic fields.

  6. Magnetic scanning gate microscopy of CoFeB lateral spin valve

    Directory of Open Access Journals (Sweden)

    Héctor Corte-León

    2017-05-01

    Full Text Available Devices comprised of CoFeB nanostructures with perpendicular magnetic anisotropy and non-magnetic Ta channel were operated in thermal lateral spin valve (LSV mode and studied by magnetotransport measurements and magnetic scanning gate microscopy (SGM. Due to the short spin diffusion length of Ta, the spin diffusion signal was suppressed, allowing the study of the contribution from the anomalous Nernst (ANE and anomalous Hall effects (AHE. The magnetotransport measurements identified the switching fields of the CoFeB nanostructures and demonstrated a combination of AHE and ANE when the devices were operated in thermally-driven spin-injection mode. Modified scanning probe microscopy probes were fabricated by placing a NdFeB magnetic bead (MB on the apex of a commercial Si probe. The dipole magnetic field distribution around the MB was characterized by using differential phase contrast technique and direct measurement of the switching field induced by the bead in the CoFeB nanodevices. Using SGM we demonstrate the influence of localized magnetic field on the CoFeB nanostructures near the non-magnetic channel. This approach provides a promising route towards the study of thermal and spin diffusion effects using local magnetic fields.

  7. Chemical-state imaging of Li using scanning Auger electron microscopy

    Energy Technology Data Exchange (ETDEWEB)

    Ishida, Nobuyuki, E-mail: ISHIDA.Nobuyuki@nims.go.jp [Global Research Center for Environment and Energy based on Nanomaterials Science (GREEN), National Institute for Materials Science, 1-2-1 Sengen, Tsukuba, Ibaraki 305-0047 (Japan); Fujita, Daisuke [Global Research Center for Environment and Energy based on Nanomaterials Science (GREEN), National Institute for Materials Science, 1-2-1 Sengen, Tsukuba, Ibaraki 305-0047 (Japan); Advanced Nanocharacterization Unit, National Institute for Materials Science, 1-2-1 Sengen, Tsukuba, Ibaraki 305-0047 (Japan)

    2013-02-15

    Highlights: •Scanning Auger electron microscopy is used to image chemical states of Li. •The combined use of AES and EELS signals for the elemental mapping is powerful. •Distribution corresponding to metallic and oxidized states of Li can be imaged. -- Abstract: The demand for measurement tools to detect Li with high spatial resolution and precise chemical sensitivity is increasing with the spread of lithium-ion batteries (LIBs) for use in a wide range of applications. In this work, scanning Auger electron microscopy (SAM) is used to image chemical states of a partially oxidized Li surface on the basis of the Auger electron spectroscopy (AES) and electron energy loss spectroscopy (EELS) data obtained during an oxidation process of a metal Li. We show that distribution of metallic and oxidized states of Li is clearly imaged by mapping the intensity of the corresponding AES and EELS peaks. Furthermore, a tiny difference in the extent of oxidation can be distinguished by comparing the elemental map of an AES peak with that of an EELS peak owing to the different behaviors of those signals to the chemical states of Li.

  8. A streaming multi-GPU implementation of image simulation algorithms for scanning transmission electron microscopy.

    Science.gov (United States)

    Pryor, Alan; Ophus, Colin; Miao, Jianwei

    2017-01-01

    Simulation of atomic-resolution image formation in scanning transmission electron microscopy can require significant computation times using traditional methods. A recently developed method, termed plane-wave reciprocal-space interpolated scattering matrix (PRISM), demonstrates potential for significant acceleration of such simulations with negligible loss of accuracy. Here, we present a software package called Prismatic for parallelized simulation of image formation in scanning transmission electron microscopy (STEM) using both the PRISM and multislice methods. By distributing the workload between multiple CUDA-enabled GPUs and multicore processors, accelerations as high as 1000 × for PRISM and 15 × for multislice are achieved relative to traditional multislice implementations using a single 4-GPU machine. We demonstrate a potentially important application of Prismatic, using it to compute images for atomic electron tomography at sufficient speeds to include in the reconstruction pipeline. Prismatic is freely available both as an open-source CUDA/C++ package with a graphical user interface and as a Python package, PyPrismatic.

  9. Bulk crystalline copper electrodeposition on polycrystalline gold surfaces observed by in-situ scanning tunneling microscopy

    DEFF Research Database (Denmark)

    Andersen, Jens Enevold Thaulov; Bech-Nielsen, Gregers; Møller, Per

    1994-01-01

    Bulk copper electrodeposition onto technical gold surfaces in electrolytes of 0.05 M H2SO4 and 1 mM CuSO4 was investigated by in-situ scanning tunnelling microscopy at fixed overpotentials. At potentials between -60 and -30 mV the growth of bulk copper proceeds in cycles of nucleation, agglomerat......Bulk copper electrodeposition onto technical gold surfaces in electrolytes of 0.05 M H2SO4 and 1 mM CuSO4 was investigated by in-situ scanning tunnelling microscopy at fixed overpotentials. At potentials between -60 and -30 mV the growth of bulk copper proceeds in cycles of nucleation......, agglomeration and crystallization. Crystalline copper is seen as involving an intermediate stage in the progress of growth. The final stage in the growth involves an equilibrium of copper electrochemically dissolving and precipitating. The drift velocity was measured for a gold surface subjected to flame...... annealing and subsequently installed in the cell compartment. It was found that the drift velocity decays with time in an exponential-like manner, and a 70 min waiting time before experiments with atomic resolution is recommended. Atomic resolution on Au(111) has been obtained, and an apparent surface...

  10. EPS composition and calcification potential of tufa-dominating cyanobacteria investigated by Scanning Transmission X-ray Microscopy (STXM) and Laser Scanning Microscopy (LSM)

    Science.gov (United States)

    Zippel, Barbara; Dynes, James J.; Obst, Martin; Lawrence, John R.; Neu, Thomas R.

    2010-05-01

    Tufa deposits in freshwater habitats are the result of calcium carbonate precipitation within interfacial microbial ecosystems. Calcite precipitation is influenced by the saturation index and the occurrence of extracellular polymeric substances (EPS) which are produced by a variety of microorganisms. In theory, the first important step of biologically induced calcification processes is the adsorption of calcium ions by extracellular polymeric substances (EPS) produced by cyanobacteria. In the present study we take advantage of Laser Scanning Microscopy (LSM) and combine it with Synchrotron imaging using Scanning Transmission X-ray Microscopy (STXM). STXM represents a technique that allows simultaneous analysis of inorganic and organic constituents as a scale of 50 nm. By means of STXM it is possible to differentiate between calcium carbonate phases at the Ca L-edge. Furthermore, STXM has also been used at the C K-edge to map the major biomolecules (proteins, lipids, and polysaccharides). The purpose of this study is to find out if there are differences in calcium adsorption depending on specific composition of the EPS produced by filamentous cyanobacteria isolated from a German hard water creek (Westerhöfer Bach, Harz Mountains). The goal was to elucidate the potential of biofilms constituents, including microbial cell surfaces as well as extracellular polymeric substances, in triggering the formation of calcium carbonate in tufa systems. For this purpose three filamentous cyanobacteria (Pseudanabaena sp., Leptolyngbya sp. and Nostoc sp.) were cultivated in creek-adapted as well as standard media (BG11) on polycarbonate slides. In situ EPS composition was detected by means of fluorescence lectin-binding approach (FLBA) using 23 commercially available lectins with different specificities for mono- and disaccharides and amino sugars. For CaCO3 nucleation experiments cyanobacterial biofilms grown on polycarbonate slides were deposited in NaHCO3/CaCl2 solutions

  11. Field emission scanning electron microscopy (FE-SEM) as an approach for nanoparticle detection inside cells.

    Science.gov (United States)

    Havrdova, M; Polakova, K; Skopalik, J; Vujtek, M; Mokdad, A; Homolkova, M; Tucek, J; Nebesarova, J; Zboril, R

    2014-12-01

    When developing new nanoparticles for bio-applications, it is important to fully characterize the nanoparticle's behavior in biological systems. The most common techniques employed for mapping nanoparticles inside cells include transmission electron microscopy (TEM) and scanning transmission electron microscopy (STEM). These techniques entail passing an electron beam through a thin specimen. STEM or TEM imaging is often used for the detection of nanoparticles inside cellular organelles. However, lengthy sample preparation is required (i.e., fixation, dehydration, drying, resin embedding, and cutting). In the present work, a new matrix (FTO glass) for biological samples was used and characterized by field emission scanning electron microscopy (FE-SEM) to generate images comparable to those obtained by TEM. Using FE-SEM, nanoparticle images were acquired inside endo/lysosomes without disruption of the cellular shape. Furthermore, the initial steps of nanoparticle incorporation into the cells were captured. In addition, the conductive FTO glass endowed the sample with high stability under the required accelerating voltage. Owing to these features of the sample, further analyses could be performed (material contrast and energy-dispersive X-ray spectroscopy (EDS)), which confirmed the presence of nanoparticles inside the cells. The results showed that FE-SEM can enable detailed characterization of nanoparticles in endosomes without the need for contrast staining or metal coating of the sample. Images showing the intracellular distribution of nanoparticles together with cellular morphology can give important information on the biocompatibility and demonstrate the potential of nanoparticle utilization in medicine. Copyright © 2014 Elsevier Ltd. All rights reserved.

  12. Investigating the use of in situ liquid cell scanning transmission electron microscopy

    Energy Technology Data Exchange (ETDEWEB)

    Nguy, Amanda [Iowa State Univ., Ames, IA (United States)

    2016-02-19

    Engineering nanoparticles with desired shape-dependent properties is the key to many applications in nanotechnology. Although many synthetic procedures exist to produce anisotropic gold nanoparticles, the dynamics of growth are typically unknown or hypothetical. In the case of seed-mediated growth in the presence of DNA into anisotropic nanoparticles, it is not known exactly how DNA directs growth into specific morphologies. A series of preliminary experiments were carried out to contribute to the investigation of the possible mechanism of DNA-mediated growth of gold nanoprisms into gold nanostars using liquid cell scanning transmission electron microscopy (STEM). Imaging in the liquid phase was achieved through the use of a liquid cell platform and liquid cell holder that allow the sample to be contained within a “chip sandwich” between two electron transparent windows. Ex situ growth experiments were performed using Au-T30 NPrisms (30-base thymine oligonucleotide-coated gold nanoprisms) that are expected to grow into gold nanostars. Growth to form these nanostars were imaged using TEM (transmission electron microscopy) and liquid cell STEM (scanning transmission electron microscopy). An attempt to perform in situ growth experiments with the same Au-T30 nanoprisms revealed challenges in obtaining desired morphology results due to the environmental differences within the liquid cell compared to the ex situ environment. Different parameters in the experimental method were explored including fluid line set up, simultaneous and alternating reagent addition, and the effect of different liquid cell volumes to ensure adequate flow of reagents into the liquid cell. Lastly, the binding affinities were compared for T30 and A30 DNA incubated with gold nanoparticles using zeta potential measurements, absorption spectroscopy, and isothermal titration calorimetry (ITC). It was previously reported thymine bases have a lower binding affinity to gold surfaces than adenine

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

    Science.gov (United States)

    Chemes, Hector E

    2013-01-01

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

  14. Novel scanning electron microscopy methods for analyzing the 3D structure of the Golgi apparatus.

    Science.gov (United States)

    Koga, Daisuke; Ushiki, Tatsuo; Watanabe, Tsuyoshi

    2017-01-01

    The structure of the Golgi apparatus has been extensively examined by light and electron microscopy, but details of its three-dimensional (3D) structure have remained unclear because of the technical limitations of conventional microscopy techniques. To overcome this problem, we have developed several novel scanning electron microscopy (SEM) methods for observing the 3D structure of subcellular organelles including the Golgi apparatus: (1) an osmium maceration method that facilitates SEM observation of membranous organelles, including the Golgi apparatus, by selectively removing soluble cytoplasmic proteins, (2) an osmium impregnation/maceration method that combines an osmium impregnation method with the osmium maceration method to determine the polarity of the Golgi apparatus by SEM, (3) a correlative light and SEM method that combines a cryosectioning technique with the osmium maceration method to enable correlation of the immunocytochemical distribution of molecules with the 3D ultrastructure of the Golgi apparatus, and (4) array tomography based on the systematic collection and integration of SEM images of serial ultrathin sections on glass slides for revealing the 3D ultrastructure of the entire Golgi apparatus. Together, the novel SEM techniques listed above can reveal the complete 3D structure of the Golgi apparatus in different cell types.

  15. Immunogold scanning electron microscopy can reveal the polysaccharide architecture of xylem cell walls.

    Science.gov (United States)

    Sun, Qiang; Sun, Yuliang; Juzenas, Kevin

    2017-04-01

    Immunofluorescence microscopy (IFM) and immunogold transmission electron microscopy (TEM) are the two main techniques commonly used to detect polysaccharides in plant cell walls. Both are important in localizing cell wall polysaccharides, but both have major limitations, such as low resolution in IFM and restricted sample size for immunogold TEM. In this study, we have developed a robust technique that combines immunocytochemistry with scanning electron microscopy (SEM) to study cell wall polysaccharide architecture in xylem cells at high resolution over large areas of sample. Using multiple cell wall monoclonal antibodies (mAbs), this immunogold SEM technique reliably localized groups of hemicellulosic and pectic polysaccharides in the cell walls of five different xylem structures (vessel elements, fibers, axial and ray parenchyma cells, and tyloses). This demonstrates its important advantages over the other two methods for studying cell wall polysaccharide composition and distribution in these structures. In addition, it can show the three-dimensional distribution of a polysaccharide group in the vessel lateral wall and the polysaccharide components in the cell wall of developing tyloses. This technique, therefore, should be valuable for understanding the cell wall polysaccharide composition, architecture and functions of diverse cell types. © The Author 2017. Published by Oxford University Press on behalf of the Society for Experimental Biology.

  16. Scanning a microhabitat: plant-microbe interactions revealed by confocal laser microscopy.

    Science.gov (United States)

    Cardinale, Massimiliano

    2014-01-01

    No plant or cryptogam exists in nature without microorganisms associated with its tissues. Plants as microbial hosts are puzzles of different microhabitats, each of them colonized by specifically adapted microbiomes. The interactions with such microorganisms have drastic effects on the host fitness. Since the last 20 years, the combination of microscopic tools and molecular approaches contributed to new insights into microbe-host interactions. Particularly, confocal laser scanning microscopy (CLSM) facilitated the exploration of microbial habitats and allowed the observation of host-associated microorganisms in situ with an unprecedented accuracy. Here I present an overview of the progresses made in the study of the interactions between microorganisms and plants or plant-like organisms, focusing on the role of CLSM for the understanding of their significance. I critically discuss risks of misinterpretation when procedures of CLSM are not properly optimized. I also review approaches for quantitative and statistical analyses of CLSM images, the combination with other molecular and microscopic methods, and suggest the re-evaluation of natural autofluorescence. In this review, technical aspects were coupled with scientific outcomes, to facilitate the readers in identifying possible CLSM applications in their research or to expand their existing potential. The scope of this review is to highlight the importance of confocal microscopy in the study of plant-microbe interactions and also to be an inspiration for integrating microscopy with molecular techniques in future researches of microbial ecology.

  17. Scanning a microhabitat: plant-microbe interactions revealed by confocal laser microscopy

    Directory of Open Access Journals (Sweden)

    Massimiliano eCardinale

    2014-03-01

    Full Text Available No plant or cryptogam exists in nature without microorganisms associated with its tissues. Plants as microbial hosts are puzzles of different microhabitats, each of them colonized by specifically adapted microbiomes. The interactions with such microorganisms have drastic effects on the host fitness. Since the last 20 years, the combination of microscopic tools and molecular approaches contributed to new insights into microbe-host interactions. Particularly, confocal laser scanning microscopy (CLSM facilitated the exploration of microbial habitats and allowed the observation of host-associated microorganisms in situ with an unprecedented accuracy. Here I present an overview of the progresses made in the study of the interactions between microorganisms and plants or plant-like organisms, focusing on the role of CLSM for the understanding of their significance. I critically discuss risks of misinterpretation when procedures of CLSM are not properly optimized. I also review approaches for quantitative and statistical analyses of CLSM images, the combination with other molecular and microscopic methods, and suggest the re-evaluation of natural autofluorescence. In this review, technical aspects were coupled with scientific outcomes, to facilitate the readers in identifying possible CLSM applications in their research or to expand their existing potential. The scope of this review is to highlight the importance of confocal microscopy in the study of plant-microbe interactions and also to be an inspiration for integrating microscopy with molecular techniques in future researches of microbial ecology.

  18. Redescription of Cichlidogyrus philander (Monogenea, Ancyrocephalidae using scanning electron microscopy (SEM and molecular analysis

    Directory of Open Access Journals (Sweden)

    Igeh Patience C

    2017-01-01

    Full Text Available The sclerotized structures of monogeneans have traditionally been studied by light microscopy and different staining techniques. Recently, enzymatic digestion followed by scanning electron microscopy (SEM has enabled the examination of structural details not visible with light microscopy. In order to obtain better, and more accurate, morphological information on sclerotized structures not affected by mounting medium or cover slip pressure, the sclerites of Cichlidogyrus philander Douëllou, 1993 (Monogenea, Ancyrocephalidae, collected from Pseudocrenilabrus philander (Weber, 1897, were redescribed using SEM. Parasites were collected from Padda Dam, Gauteng, South Africa and soft tissue was digested to release the sclerotized structures. The digested tissue also provided sufficient genetic material for molecular characterization of this species. Cichlidogyrus philander is characterised by a penis with a sharp, curved, lateral termination, an accessory piece with a hook-like extremity that may appear forked terminally, and lack of a visible vagina. The transverse bars have concave and convex surfaces with ribs on the concave surface. The dorsal bar bears fenestrations at the base of the auricles and the ventral and dorsal gripi are dissimilar. Furthermore, the large first pair of uncinuli shows lateral wings on the left side of the base. On top of this wing, a ball-like structure with a small fenestration is visible. Genetic characters derived from the 28S rDNA, the COI mitochondrial DNA and ITS1 rDNA regions distinguish C. philander from all other Cichlidogyrus sequenced species.

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

    Science.gov (United States)

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

    2017-04-01

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

  20. Advantages of indium-tin oxide-coated glass slides in correlative scanning electron microscopy applications of uncoated cultured cells.

    NARCIS (Netherlands)

    Pluk, H.; Stokes, D.J.; Lich, B.; Wieringa, B.; Fransen, J.A.M.

    2009-01-01

    A method of direct visualization by correlative scanning electron microscopy (SEM) and fluorescence light microscopy of cell structures of tissue cultured cells grown on conductive glass slides is described. We show that by growing cells on indium-tin oxide (ITO)-coated glass slides, secondary

  1. Imaging hyphal growth of Physisporinus vitreus in Norway spruce wood by means of confocal laser scanning microscopy (CLSM)

    OpenAIRE

    Schubert, Mark; Stührk, Chris; Fuhr, Matthias J.; Schwarze, Francis W.M.R.

    2017-01-01

    Light microscopy and electron microscopy are the most common methods for analyzing wood-decay fungi. However, the 3D visualization and quantification of the filamentous structure of fungi in wood is difficult to realize by means of these traditional techniques. In the present work, confocal laser scanning microscopy (CLSM) was further developed for the quantitative imaging of the 3D microscopic hyphal growth of Physisporinus vitreus, a versatile fungus for engineering value-added wood product...

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

    Science.gov (United States)

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

    2016-02-01

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

  3. High frame-rate multichannel beam-scanning microscopy based on Lissajous trajectories.

    Science.gov (United States)

    Sullivan, Shane Z; Muir, Ryan D; Newman, Justin A; Carlsen, Mark S; Sreehari, Suhas; Doerge, Chris; Begue, Nathan J; Everly, R Michael; Bouman, Charles A; Simpson, Garth J

    2014-10-06

    A simple beam-scanning optical design based on Lissajous trajectory imaging is described for achieving up to kHz frame-rate optical imaging on multiple simultaneous data acquisition channels. In brief, two fast-scan resonant mirrors direct the optical beam on a circuitous trajectory through the field of view, with the trajectory repeat-time given by the least common multiplier of the mirror periods. Dicing the raw time-domain data into sub-trajectories combined with model-based image reconstruction (MBIR) 3D in-painting algorithms allows for effective frame-rates much higher than the repeat time of the Lissajous trajectory. Since sub-trajectory and full-trajectory imaging are simply different methods of analyzing the same data, both high-frame rate images with relatively low resolution and low frame rate images with high resolution are simultaneously acquired. The optical hardware required to perform Lissajous imaging represents only a minor modification to established beam-scanning hardware, combined with additional control and data acquisition electronics. Preliminary studies based on laser transmittance imaging and polarization-dependent second harmonic generation microscopy support the viability of the approach both for detection of subtle changes in large signals and for trace-light detection of transient fluctuations.

  4. Helium ion microscopy and ultra-high-resolution scanning electron microscopy analysis of membrane-extracted cells reveals novel characteristics of the cytoskeleton of Giardia intestinalis.

    Science.gov (United States)

    Gadelha, Ana Paula Rocha; Benchimol, Marlene; de Souza, Wanderley

    2015-06-01

    Giardia intestinalis presents a complex microtubular cytoskeleton formed by specialized structures, such as the adhesive disk, four pairs of flagella, the funis and the median body. The ultrastructural organization of the Giardia cytoskeleton has been analyzed using different microscopic techniques, including high-resolution scanning electron microscopy. Recent advances in scanning microscopy technology have opened a new venue for the characterization of cellular structures and include scanning probe microscopy techniques such as ultra-high-resolution scanning electron microscopy (UHRSEM) and helium ion microscopy (HIM). Here, we studied the organization of the cytoskeleton of G. intestinalis trophozoites using UHRSEM and HIM in membrane-extracted cells. The results revealed a number of new cytoskeletal elements associated with the lateral crest and the dorsal surface of the parasite. The fine structure of the banded collar was also observed. The marginal plates were seen linked to a network of filaments, which were continuous with filaments parallel to the main cell axis. Cytoplasmic filaments that supported the internal structures were seen by the first time. Using anti-actin antibody, we observed a labeling in these filamentous structures. Taken together, these data revealed new surface characteristics of the cytoskeleton of G. intestinalis and may contribute to an improved understanding of the structural organization of trophozoites. Copyright © 2015 Elsevier Inc. All rights reserved.

  5. Imaging of metal ion dissolution and electrodeposition by anodic stripping voltammetry-scanning electrochemical microscopy.

    Science.gov (United States)

    Alpuche-Aviles, Mario A; Baur, John E; Wipf, David O

    2008-05-15

    We have developed a new imaging method for scanning electrochemical microscopy (SECM) employing fast-scan anodic stripping voltammetry (ASV) to provide sensitive and selective imaging of multiple chemical species at interfaces immersed in solution. A rapid cyclic voltammetry scan (100 V/s) is used along with a short preconcentration time (300-750 ms) to allow images to be acquired in a normal SECM time frame. A Hg-Pt film electrode is developed having an equivalent Hg thickness of 40 nm that has good sensitivity at short preconcentration times and also retains thin-film behavior with high-speed voltammetric stripping. Fast-scan anodic stripping currents are shown to be linear for 1-100 microM of Pb (2+) and Cd (2+) solutions using a preconcentration time of 300 ms. SECM images showing the presence of Pb (2+) and Cd (2+) at concentrations as low as 1 microM are presented. In addition, a single ASV-SECM image is shown to produce unique concentration maps indicating Cd (2+) and Pb (2+), generated in situ from a corroding sample, while simultaneously detecting the depletion of O 2 at this sample. The transient voltammetric response at the film electrode is simulated and shows good agreement with the experimental behavior. We discuss the behavior of images and concentration profiles obtained with different imaging conditions and show that mass-transport limitations in the tip-substrate gap can induce dissolution. ASV-SECM can thus be used to detect and study induced dissolution not only at bulk metal surfaces but also on underpotential deposition layers, in this case Cd and Pb on Pt. In addition, we discuss how surface diffusion phenomena may relate to the observed ASV-SECM behavior.

  6. Rapid imaging of mycoplasma in solution using Atmospheric Scanning Electron Microscopy (ASEM)

    Energy Technology Data Exchange (ETDEWEB)

    Sato, Chikara, E-mail: ti-sato@aist.go.jp [Biomedical Research Institute, National Institute of Advanced Industrial Science and Technology (AIST), 1-1-1 Higashi, Tsukuba, Ibaraki 305-8566 (Japan); Manaka, Sachie [Biomedical Research Institute, National Institute of Advanced Industrial Science and Technology (AIST), 1-1-1 Higashi, Tsukuba, Ibaraki 305-8566 (Japan); Nakane, Daisuke [Department of Biology, Graduate School of Science, Osaka City University, Sumiyoshi-ku, Osaka 558-8585 (Japan); Nishiyama, Hidetoshi; Suga, Mitsuo [Advanced Technology Division, JEOL Ltd., Akishima, Tokyo 196-8558 (Japan); Nishizaka, Takayuki [Department of Physics, Faculty of Science, Gakushuin University, 1-5-1 Mejiro, Toshima-ku, Tokyo 171-8588 (Japan); Miyata, Makoto [Department of Biology, Graduate School of Science, Osaka City University, Sumiyoshi-ku, Osaka 558-8585 (Japan); Maruyama, Yuusuke [Biomedical Research Institute, National Institute of Advanced Industrial Science and Technology (AIST), 1-1-1 Higashi, Tsukuba, Ibaraki 305-8566 (Japan)

    2012-01-27

    Highlights: Black-Right-Pointing-Pointer Mycoplasma mobile was observed in buffer with the Atmospheric Scanning Electron Microscope. Black-Right-Pointing-Pointer Characteristic protein localizations were visualized using immuno-labeling. Black-Right-Pointing-Pointer M. mobile attached to sialic acid on the SiN film surface within minutes. Black-Right-Pointing-Pointer Cells were observed at low concentrations. Black-Right-Pointing-Pointer ASEM should promote study and early-stage diagnosis of mycoplasma. -- Abstract: Mycoplasma is a genus of bacterial pathogen that causes disease in vertebrates. In humans, the species Mycoplasma pneumoniae causes 15% or more of community-acquired pneumonia. Because this bacterium is tiny, corresponding in size to a large virus, diagnosis using optical microscopy is not easy. In current methods, chest X-rays are usually the first action, followed by serology, PCR amplification, and/or culture, but all of these are particularly difficult at an early stage of the disease. Using Mycoplasma mobile as a model species, we directly observed mycoplasma in buffer with the newly developed Atmospheric Scanning Electron Microscope (ASEM). This microscope features an open sample dish with a pressure-resistant thin film window in its base, through which the SEM beam scans samples in solution, from below. Because of its 2-3 {mu}m-deep scanning capability, it can observe the whole internal structure of mycoplasma cells stained with metal solutions. Characteristic protein localizations were visualized using immuno-labeling. Cells were observed at low concentrations, because suspended cells concentrate in the observable zone by attaching to sialic acid on the silicon nitride (SiN) film surface within minutes. These results suggest the applicability of the ASEM for the study of mycoplasmas as well as for early-stage mycoplasma infection diagnosis.

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

    Energy Technology Data Exchange (ETDEWEB)

    Forslind, B.

    1984-01-01

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

  8. Scanning electron microscopy of Leucochloridiomorpha constantiae during development from metacercaria to adult.

    Science.gov (United States)

    Font, W F; Wittrock, D D

    1980-12-01

    Scanning electron microscopy revealed that the tegument of Leucochloridiomorpha constantiae metacercariae was marked with interconnected longitudinal and transverse ridges. Alteration of the ridges began within 1 hr after implantation into the chick coelom. Ridges were completely lost by the 4th hr, leaving a smooth tegument. Body shape altered from that of a tapered metacercaria to a plump, rounded adult within 3 days, and was accompanied by doubling of the acetabular diameter. Papillae, present in both the metacercaria and adult, were located on the suckers, genital orifice, and body surface. The microtopography of the tegument consisted of knoblike protuberances that gave the surface a cobblestonelike appearance. Tegumental knobs in te vicinity of the genital pore contained minute, rounded inclusions.

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

    Science.gov (United States)

    Jiang, Nan

    2017-08-01

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

  10. Improved visualization of vertebrate nuclear pore complexes by field emission scanning electron microscopy.

    Science.gov (United States)

    Shaulov, Lihi; Harel, Amnon

    2012-03-07

    Field emission scanning electron microscopy (FESEM) can provide high-resolution three-dimensional surface imaging of many biological structures, including nuclear envelopes and nuclear pore complexes (NPCs). For this purpose, it is important to preserve NPCs as close as possible to their native morphology, embedded in undamaged nuclear membranes. We present optimized methodologies for FESEM imaging in a cell-free reconstitution system and for the direct visualization of mammalian cell nuclei. The use of anchored chromatin templates in the cell-free system is particularly advantageous for imaging fragile intermediates inhibited at early stages of assembly. Our new method for exposing the surface of mammalian nuclei results in an unprecedented quality of NPC images, avoiding detergent-induced and physical damage. These new methodologies pave the way for the combined use of FESEM imaging with biochemical and genetic manipulation, in cell-free systems and in mammalian cells. Copyright © 2012 Elsevier Ltd. All rights reserved.

  11. Atomic bonding effects in annular dark field scanning transmission electron microscopy. I. Computational predictions

    Energy Technology Data Exchange (ETDEWEB)

    Odlyzko, Michael L.; Mkhoyan, K. Andre, E-mail: mkhoyan@umn.edu [Department of Chemical Engineering and Materials Science, University of Minnesota, Minneapolis, Minnesota 55455 (United States); Himmetoglu, Burak [Department of Chemical Engineering and Materials Science, University of Minnesota, Minneapolis, Minnesota 55455 and Materials Department, University of California, Santa Barbara, California 93106 (United States); Cococcioni, Matteo [Department of Chemical Engineering and Materials Science, University of Minnesota, Minneapolis, Minnesota 55455 and Theory and Simulations of Materials, National Centre for Computational Design and Discovery of Novel Materials, École polytechnique fédérale de Lausanne (EPFL), CH-1015 Lausanne (Switzerland)

    2016-07-15

    Annular dark field scanning transmission electron microscopy (ADF-STEM) image simulations were performed for zone-axis-oriented light-element single crystals, using a multislice method adapted to include charge redistribution due to chemical bonding. Examination of these image simulations alongside calculations of the propagation of the focused electron probe reveal that the evolution of the probe intensity with thickness exhibits significant sensitivity to interatomic charge transfer, accounting for observed thickness-dependent bonding sensitivity of contrast in all ADF-STEM imaging conditions. Because changes in image contrast relative to conventional neutral atom simulations scale directly with the net interatomic charge transfer, the strongest effects are seen in crystals with highly polar bonding, while no effects are seen for nonpolar bonding. Although the bonding dependence of ADF-STEM image contrast varies with detector geometry, imaging parameters, and material temperature, these simulations predict the bonding effects to be experimentally measureable.

  12. Scanning tunneling microscopy studies of organic monolayers adsorbed on the rhodium(111) crystal surface

    Energy Technology Data Exchange (ETDEWEB)

    Cernota, Paul Davis [Univ. of California, Berkeley, CA (United States)

    1999-08-01

    Scanning Tunneling Microscopy studies were carried out on ordered overlayers on the (111) surface of rhodium. These adsorbates include carbon monoxide (CO), cyclohexane, cyclohexene, 1,4-cyclohexadiene, para-xylene, and meta-xylene. Coadsorbate systems included: CO with ethylidyne, CO with para- and meta-xylene, and para-xylene with meta-xylene. In the case of CO, the structure of the low coverage (2x2) overlayer has been observed. The symmetry of the unit cell in this layer suggests that the CO is adsorbed in the 3-fold hollow sites. There were also two higher coverage surface structures with (√7x√7) unit cells. One of these is composed of trimers of CO and has three CO molecules in each unit cell. The other structure has an additional CO molecule, making a total of four. This extra CO sits on a top site.

  13. Electrochemical Size Measurement and Characterization of Electrodeposited Platinum Nanoparticles at Nanometer Resolution with Scanning Electrochemical Microscopy.

    Science.gov (United States)

    Ma, Wei; Hu, Keke; Chen, QianJin; Zhou, Min; Mirkin, Michael V; Bard, Allen J

    2017-07-12

    The properties of nanoparticles (NPs) are determined by their size and geometric structures. A reliable determination of NP dimension is critical for understanding their physical and chemical properties, but sizing ultrasmall particles on the order of nanometer (nm) scale in the solution is still challenging. Here, we report the size measurement of PtNP at nanometer resolution by in situ scanning electrochemical microscopy (SECM), performed with the electrochemical generation and removal of H2 bubble at a reasonably small distance between tip and substrate electrodes in 200 or 500 mM HClO4 solution. A series of different PtNPs or nanoclusters were electrodeposited and in situ measured in the solution, proving the concept of sizing ultrasmall particles using tip generation/substrate collection mode of SECM. This technique could be also used for investigations of other supported ultrasmall metal nanocluster systems.

  14. Scanning electron microscopy of the tegumental surface of adult Schistosoma spindale.

    Science.gov (United States)

    Kruatrachue, M; Riengrojpitak, S; Upatham, E S; Sahaphong, S

    1983-09-01

    The tegumental surfaces of adult male and female of Schistosoma spindale were studied by scanning electron microscopy. In general, the body surface of the male appears to be fairly uniform from anterior end to posterior end. It is characterized by the presence of transverse ridges and papillae of various types. These papillae are distributed fairly regularly over the whole body surface of the worm. The tegument lining the gynecophoral canal of the male worm is covered with numerous spines interspersed with papillae, some without cilia and some with crater-like holes in the centres and apical cilia. The tegument of the female worm is covered with smooth and perforated ridges and sensory bulbs with apical nodules.

  15. A leveling method based on current feedback mode of scanning electrochemical microscopy.

    Science.gov (United States)

    Han, Lianhuan; Yuan, Ye; Zhang, Jie; Zhao, Xuesen; Cao, Yongzhi; Hu, Zhenjiang; Yan, Yongda; Dong, Shen; Tian, Zhong-Qun; Tian, Zhao-Wu; Zhan, Dongping

    2013-02-05

    Substrate leveling is an essential but neglected instrumental technique of scanning electrochemical microscopy (SECM). In this technical note, we provide an effective substrate leveling method based on the current feedback mode of SECM. By using an air-bearing rotary stage as the supporter of an electrolytic cell, the current feedback presents a periodic waveform signal, which can be used to characterize the levelness of the substrate. Tuning the adjusting screws of the tilt stage, substrate leveling can be completed in minutes by observing the decreased current amplitude. The obtained high-quality SECM feedback curves and images prove that this leveling technique is valuable in not only SECM studies but also electrochemical machining.

  16. Hydrogen adsorption on Ru(001) studied by Scanning TunnelingMicroscopy

    Energy Technology Data Exchange (ETDEWEB)

    Tatarkhanov, Mous; Rose, Franck; Fomin, Evgeny; Ogletree, D.Frank; Salmeron, Miquel

    2008-01-18

    The adsorption of hydrogen on Ru(001) was studied by scanning tunneling microscopy at temperatures around 50 K. Hydrogen was found to adsorb dissociatively forming different ordered structures as a function of coverage. In order of increasing coverage {theta} in monolayers (ML) these were ({radical}3 x {radical}3)r30{sup o} at {theta} = 0.3 ML; (2 x 1) at {theta} = 0.50 ML, (2 x 2)-3H at {theta} = 0.75, and (1 x 1) at {theta} = 1.00. Some of these structures were observed to coexist at intermediate coverage values. Close to saturation of 1 ML, H-vacancies (unoccupied three fold fcc hollow Ru sites) were observed either as single entities or forming transient aggregations. These vacancies diffuse and aggregate to form active sites for the dissociative adsorption of hydrogen.

  17. Interfacial self-assembly of amino acids and peptides: Scanning tunneling microscopy investigation

    Science.gov (United States)

    Xu, Li-Ping; Liu, Yibiao; Zhang, Xueji

    2011-12-01

    Proteins play important roles in human daily life. To take advantage of the lessons learned from nature, it is essential to investigate the self-assembly of subunits of proteins, i.e., amino acids and polypeptides. Due to its high resolution and versatility of working environment, scanning tunneling microscopy (STM) has become a powerful tool for studying interfacial molecular assembly structures. This review is intended to reflect the progress in studying interfacial self-assembly of amino acids and peptides by STM. In particular, we focus on environment-induced polymorphism, chiral recognition, and coadsorption behavior with molecular templates. These studies would be highly beneficial to research endeavors exploring the mechanism and nanoscale-controlling molecular assemblies of amino acids and polypeptides on surfaces, understanding the origin of life, unravelling the essence of disease at the molecular level and deeming what is necessary for the ``bottom-up'' nanofabrication of molecular devices and biosensors being constructed with useful properties and desired performance.

  18. Anterior lens epithelium in cataract patients with retinitis pigmentosa - scanning and transmission electron microscopy study.

    Science.gov (United States)

    Andjelic, Sofija; Drašlar, Kazimir; Hvala, Anastazija; Hawlina, Marko

    2017-05-01

    In retinitis pigmentosa (RP) patients, relatively minor lens opacity in central part of posterior pole of the lens may cause disproportionate functional symptoms requiring cataract operation. To investigate the possible structural reasons for this opacity development, we studied the structure of the lens epithelium of patients with RP. The anterior lens capsule (aLC: basement membrane and associated lens epithelial cells, LECs) was obtained from cataract surgery and prepared for scanning and transmission electron microscopy (SEM and TEM). Both SEM and TEM show a number of abnormal features in the anterior lens epithelium of cataract patients with RP. The abnormalities appear mainly as holes, thinning and degradation of the epithelium, with the dimensions from cataractous lens. We suggest that the lens epithelium has a role in the development of the cataract in patients with RP. © 2016 Acta Ophthalmologica Scandinavica Foundation. Published by John Wiley & Sons Ltd.

  19. Ultra-High Vacuum Compatible Optical Chopper System for Synchrotron X-ray Scanning Tunneling Microscopy

    Energy Technology Data Exchange (ETDEWEB)

    Chang, Hao; Cummings, Marvin L.; Shirato, Nozomi; Stripe, Benjamin D.; Rosenmann, Daniel; Preissner, Curt A.; Freeland, John W.; Kersell, Heath R.; Hla, Saw Wai; Rose, Volker

    2015-01-01

    High-speed beam choppers are a crucial part of time-resolved x-ray studies as well as a necessary component to enable elemental contrast in synchrotron x-ray scanning tunneling microscopy (SX-STM). However, many chopper systems are not capable of operation in vacuum, which restricts their application to x-ray studies with high photon energies, where air absorption does not present a significant problem. To overcome this limitation, we present a fully ultra-high vacuum (UHV) compatible chopper system capable of operating at variable chopping frequencies up to 4 kHz. The lightweight aluminum chopper disk is coated with Ti and Au films to provide the required beam attenuation for soft and hard x-rays with photon energies up to about 12 keV. The chopper is used for lock-in detection of x-ray enhanced signals in SX-STM.

  20. Ultra-high vacuum compatible optical chopper system for synchrotron x-ray scanning tunneling microscopy

    Energy Technology Data Exchange (ETDEWEB)

    Chang, Hao, E-mail: hc000211@ohio.edu [Advanced Photon Source, Argonne National Laboratory, 9700 South Cass Avenue, Argonne, Illinois 60439 (United States); Nanoscale and Quantum Phenomena Institute, Physics & Astronomy Department, Ohio University, Athens, Ohio 45701 (United States); Cummings, Marvin; Shirato, Nozomi; Stripe, Benjamin; Preissner, Curt; Freeland, John W. [Advanced Photon Source, Argonne National Laboratory, 9700 South Cass Avenue, Argonne, Illinois 60439 (United States); Rosenmann, Daniel [Center for Nanoscale Materials, Argonne National Laboratory, 9700 South Cass Avenue, Argonne, Illinois 60439 (United States); Kersell, Heath; Hla, Saw-Wai [Nanoscale and Quantum Phenomena Institute, Physics & Astronomy Department, Ohio University, Athens, Ohio 45701 (United States); Center for Nanoscale Materials, Argonne National Laboratory, 9700 South Cass Avenue, Argonne, Illinois 60439 (United States); Rose, Volker, E-mail: vrose@anl.gov [Advanced Photon Source, Argonne National Laboratory, 9700 South Cass Avenue, Argonne, Illinois 60439 (United States); Center for Nanoscale Materials, Argonne National Laboratory, 9700 South Cass Avenue, Argonne, Illinois 60439 (United States)

    2016-01-28

    High-speed beam choppers are a crucial part of time-resolved x-ray studies as well as a necessary component to enable elemental contrast in synchrotron x-ray scanning tunneling microscopy (SX-STM). However, many chopper systems are not capable of operation in vacuum, which restricts their application to x-ray studies with high photon energies, where air absorption does not present a significant problem. To overcome this limitation, we present a fully ultra-high vacuum (UHV) compatible chopper system capable of operating at variable chopping frequencies up to 4 kHz. The lightweight aluminum chopper disk is coated with Ti and Au films to provide the required beam attenuation for soft and hard x-rays with photon energies up to about 12 keV. The chopper is used for lock-in detection of x-ray enhanced signals in SX-STM.

  1. Large area strain analysis using scanning transmission electron microscopy across multiple images

    Energy Technology Data Exchange (ETDEWEB)

    Oni, A. A.; Sang, X.; LeBeau, J. M., E-mail: jmlebeau@ncsu.edu [Department of Materials Science and Engineering, North Carolina State University, Raleigh, North Carolina 27695-7907 (United States); Raju, S. V.; Saxena, S. [Center for the Study of Matter under Extreme Conditions, Florida International University, Miami, Florida 33199 (United States); Dumpala, S.; Broderick, S.; Rajan, K. [Department of Materials Science and Engineering, Iowa State University, Ames, Iowa 50011 (United States); Kumar, A.; Sinnott, S. [Department of Materials Science and Engineering, University of Florida, Gainesville, Florida 32611 (United States)

    2015-01-05

    Here, we apply revolving scanning transmission electron microscopy to measure lattice strain across a sample using a single reference area. To do so, we remove image distortion introduced by sample drift, which usually restricts strain analysis to a single image. Overcoming this challenge, we show that it is possible to use strain reference areas elsewhere in the sample, thereby enabling reliable strain mapping across large areas. As a prototypical example, we determine the strain present within the microstructure of a Ni-based superalloy directly from atom column positions as well as geometric phase analysis. While maintaining atomic resolution, we quantify strain within nanoscale regions and demonstrate that large, unit-cell level strain fluctuations are present within the intermetallic phase.

  2. The memory effect of nanoscale memristors investigated by conducting scanning probe microscopy methods

    Directory of Open Access Journals (Sweden)

    César Moreno

    2012-11-01

    Full Text Available We report on the use of scanning force microscopy as a versatile tool for the electrical characterization of nanoscale memristors fabricated on ultrathin La0.7Sr0.3MnO3 (LSMO films. Combining conventional conductive imaging and nanoscale lithography, reversible switching between low-resistive (ON and high-resistive (OFF states was locally achieved by applying voltages within the range of a few volts. Retention times of several months were tested for both ON and OFF states. Spectroscopy modes were used to investigate the I–V characteristics of the different resistive states. This permitted the correlation of device rectification (reset with the voltage employed to induce each particular state. Analytical simulations by using a nonlinear dopant drift within a memristor device explain the experimental I–V bipolar cycles.

  3. Modeling and de-embedding the interferometric scanning microwave microscopy by means of dopant profile calibration

    Energy Technology Data Exchange (ETDEWEB)

    Michalas, L., E-mail: loukas.michalas@artov.imm.cnr.it; Marcelli, R. [National Research Council, Institute for Microelectronics and Microsystems (CNR-IMM), Via del Fosso del Cavaliere 100, 00133 Rome (Italy); Wang, F.; Brillard, C.; Theron, D. [Institut d' Electronique, de Microélectronique et de Nanotechnologie, CNRS UMR 8520/University of Lille 1, Avenue Poincaré, CS 60069, 59652 Villeneuve d' Ascq (France); Chevalier, N.; Hartmann, J. M. [Univ. Grenoble Alpes, F-38000 Grenoble, France and CEA, LETI, MINATEC Campus, F-38054 Grenoble (France)

    2015-11-30

    This paper presents the full modeling and a methodology for de-embedding the interferometric scanning microwave microscopy measurements by means of dopant profile calibration. A Si calibration sample with different boron-doping level areas is used to that end. The analysis of the experimentally obtained S{sub 11} amplitudes based on the proposed model confirms the validity of the methodology. As a specific finding, changes in the tip radius between new and used tips have been clearly identified, leading to values for the effective tip radius in the range of 45 nm to 85 nm, respectively. Experimental results are also discussed in terms of the effective area concept, taking into consideration details related to the nature of tip-to-sample interaction.

  4. Nanoscale Electric Permittivity of Single Bacterial Cells at Gigahertz Frequencies by Scanning Microwave Microscopy.

    Science.gov (United States)

    Biagi, Maria Chiara; Fabregas, Rene; Gramse, Georg; Van Der Hofstadt, Marc; Juárez, Antonio; Kienberger, Ferry; Fumagalli, Laura; Gomila, Gabriel

    2016-01-26

    We quantified the electric permittivity of single bacterial cells at microwave frequencies and nanoscale spatial resolution by means of near-field scanning microwave microscopy. To this end, calibrated complex admittance images have been obtained at ∼19 GHz and analyzed with a methodology that removes the nonlocal topographic cross-talk contributions and thus provides quantifiable intrinsic dielectric images of the bacterial cells. Results for single Escherichia coli cells provide a relative electric permittivity of ∼4 in dry conditions and ∼20 in humid conditions, with no significant loss contributions. Present findings, together with the ability of microwaves to penetrate the cell membrane, open an important avenue in the microwave label-free imaging of single cells with nanoscale spatial resolution.

  5. Real-time phase error compensation in phase sensitive scanning near-field optical microscopy.

    Science.gov (United States)

    Wu, Xiaoyu; Sun, Lin; Wang, Jia; Tan, Qiaofeng

    2015-07-01

    Phase measurements are critical for investigations on the optical properties of surface plasmon polariton (SPP) nanostructures. In this paper, a real-time phase error compensation method based on a phase sensitive scanning near-field optical microscopy (SNOM) measurement system is proposed. The method adopts the common optical path configuration and CMR (common-mode rejection) principle. It can be seen that the phase error compensation is real-time and mainly relies on optical devices, therefore neither post processing nor previous knowledge of environmental effects is required. The causes of the phase drift errors are discussed. We demonstrate experimentally the effectiveness of this method by measuring a SPP focusing device. Regardless of the drift velocity, degree of linearity, or phase accuracy, the compensation method shows great improvement compared to the previous phase sensitive SNOMs. All the measured distributions are in good agreement with theoretical simulations obtained by the finite-different time-domain (FDTD) method.

  6. Atomic resolution on the (111 )B surface of mercury cadmium telluride by scanning tunneling microscopy

    Science.gov (United States)

    Zha, Fang-Xing; Hong, Feng; Pan, Bi-Cai; Wang, Yin; Shao, Jun; Shen, Xue-Chu

    2018-01-01

    The real-space atomic surface structure of mercury cadmium telluride was successfully achieved on the (111 )B surface of H g0.78C d0.22Te by ultrahigh-vacuum scanning tunneling microscopy (STM). The work casts light on the reconstructions of the (111 )B surface unraveling a (2 ×2 ) surface reconstruction induced by adatom adsorption of Cd. The other (2 ×2 ) surface reconstruction is clarified to be induced by the single Te vacancy, which is more stable than the reconstruction of multivacancies in contrast to the prevailing view. The simulated STM images are in good agreement with the experiments. We also observed an in situ morphology transition from the (1 ×1 ) structure to those (2 ×2 ) reconstructions, implying the stability of the reconstructions.

  7. NATO Advanced Study Institute on Scanning Probe Microscopy : Characterization, Nanofabrication and Device Application of Functional Materials

    CERN Document Server

    Vilarinho, Paula Maria; Kingon, Angus; Scanning Probe Microscopy : Characterization, Nanofabrication and Device Application of Functional Materials

    2005-01-01

    As the characteristic dimensions of electronic devices continue to shrink, the ability to characterize their electronic properties at the nanometer scale has come to be of outstanding importance. In this sense, Scanning Probe Microscopy (SPM) is becoming an indispensable tool, playing a key role in nanoscience and nanotechnology. SPM is opening new opportunities to measure semiconductor electronic properties with unprecedented spatial resolution. SPM is being successfully applied for nanoscale characterization of ferroelectric thin films. In the area of functional molecular materials it is being used as a probe to contact molecular structures in order to characterize their electrical properties, as a manipulator to assemble nanoparticles and nanotubes into simple devices, and as a tool to pattern molecular nanostructures. This book provides in-depth information on new and emerging applications of SPM to the field of materials science, namely in the areas of characterisation, device application and nanofabrica...

  8. An environmental sample chamber for reliable scanning transmission x-ray microscopy measurements under water vapor

    Energy Technology Data Exchange (ETDEWEB)

    Kelly, Stephen T.; Nigge, P.; Prakash, Shruti; Laskin, Alexander; Wang, Bingbing; Tyliszczak, Tolek; Leone, Stephen R.; Gilles, Mary K.

    2013-08-01

    We have designed, fabricated, and tested a compact gas-phase reactor for performing in situ soft x-ray scanning transmission x-ray microscopy (STXM) measurements. The reactor accommodates many gas atmospheres, including reactive or corrosive gasses, but was designed specically to address the needs of measurements under water vapor. An on-board sensor measures the relative humidity and temperature inside the reactor, minimizing uncertainties associated with measuring these quantities outside the instrument. The reactor mounts directly to the existing sample holder used in the majority of STXM instruments around the world and installs with minimal instrument reconguration. Using the reactor contributes over 85% less additional absorption compared to lling the STXM chamber with process gas, and results in much more stable imaging conditions. The reactor is in use at the STXM instruments at beamlines 11.0.2 and 5.3.2.2 at the Advanced Light Source.

  9. Evaluation of vermicompost maturity using scanning electron microscopy and paper chromatography analysis.

    Science.gov (United States)

    Senthil Kumar, D; Satheesh Kumar, P; Rajendran, N M; Uthaya Kumar, V; Anbuganapathi, G

    2014-04-02

    Vermicompost was produced from flower waste inoculated with biofertilizers using the earthworm Eisenia fetida. Principal component analysis (PCA) and cluster analysis (CA) were carried out on the basis of physicochemical parameters of vermicomposted samples. From the results of the PCA and CA, it was possible to classify two different groups of vermicompost samples in the following categories: E2 and E5; and E1, E3, E4, and control. Scanning electron microscopy and biodynamic circular paper chromatography analysis were used to investigate the changes in surface morphology and functional groups in the control and vermicompost products. SEM analysis of E1-E5 shows more fragment and pores than the control. Chromatographic analysis of vermicompost indicated the mature condition of the compost materials.

  10. Spectroscopic infrared scanning near-field optical microscopy (IR-SNOM)

    Energy Technology Data Exchange (ETDEWEB)

    Vobornik, D. [Institut de Physique de la Matiere Complexe, Ecole Polytechnique Federale de Lausanne (EPFL), Station 3, CH-1015 Lausanne (Switzerland)]. E-mail: dusan.vobornik@epfl.ch; Margaritondo, G. [Institut de Physique de la Matiere Complexe, Ecole Polytechnique Federale de Lausanne (EPFL), Station 3, CH-1015 Lausanne (Switzerland); Sanghera, J.S. [Optical Sciences Division, U.S. Naval Research Laboratory, 4555 Overlook Avenue SE, Washington, DC 20375 (United States); Thielen, P. [Optical Sciences Division, U.S. Naval Research Laboratory, 4555 Overlook Avenue SE, Washington, DC 20375 (United States); Aggarwal, I.D. [Optical Sciences Division, U.S. Naval Research Laboratory, 4555 Overlook Avenue SE, Washington, DC 20375 (United States); Ivanov, B. [Department of Physics and Astronomy, Vanderbilt University, Nashville, TN 31235 (United States); Tolk, N.H. [Department of Physics and Astronomy, Vanderbilt University, Nashville, TN 31235 (United States); Manni, V. [Institute of Neurobiology and Molecular Medicine, 00133 Rome (Italy); Grimaldi, S. [Institute of Neurobiology and Molecular Medicine, 00133 Rome (Italy); Lisi, A. [Institute of Neurobiology and Molecular Medicine, 00133 Rome (Italy); Rieti, S. [Institute of Neurobiology and Molecular Medicine, 00133 Rome (Italy); Piston, D.W. [Department of Molecular Physiology and Biophysics, Vanderbilt University, Nashville, TN 37232 (United States); Generosi, R. [Istituto di Stuttura della Materia, via Fosso del Cavaliere 100, 00133 Rome (Italy); Luce, M. [Istituto di Stuttura della Materia, via Fosso del Cavaliere 100, 00133 Rome (Italy); Perfetti, P. [Istituto di Stuttura della Materia, via Fosso del Cavaliere 100, 00133 Rome (Italy); Cricenti, A. [Istituto di Stuttura della Materia, via Fosso del Cavaliere 100, 00133 Rome (Italy)

    2005-09-29

    Scanning near-field optical microscopy (SNOM or NSOM) is the technique with the highest lateral optical resolution available today, while infrared (IR) spectroscopy has a high chemical specificity. Combining SNOM with a tunable IR source produces a unique tool, IR-SNOM, capable of imaging distributions of chemical species with a 100 nm spatial resolution. We present in this paper boron nitride (BN) thin film images, where IR-SNOM shows the distribution of hexagonal and cubic phases within the sample. Exciting potential applications in biophysics and medical sciences are illustrated with SNOM images of the distribution of different chemical species within cells. We present in this article images with resolutions of the order of {lambda}/60 with SNOM working with infrared light. With our SNOM setup, we routinely get optical resolutions between 50 and 150 nm, regardless of the wavelength of the light used to illuminate the sample.

  11. Investigation of thermal effects in through-silicon vias using scanning thermal microscopy.

    Science.gov (United States)

    Wielgoszewski, Grzegorz; Jóźwiak, Grzegorz; Babij, Michał; Baraniecki, Tomasz; Geer, Robert; Gotszalk, Teodor

    2014-11-01

    Results of quantitative investigations of copper through-silicon vias (TSVs) are presented. The experiments were performed using scanning thermal microscopy (SThM), enabling highly localized imaging of thermal contrast between the copper TSVs and the surrounding material. Both dc and ac active-mode SThM was used and differences between these variants are shown. SThM investigations of TSVs may provide information on copper quality in TSV, as well as may lead to quantitative investigation of thermal boundaries in micro- and nanoelectronic structures. A proposal for heat flow analysis in a TSV, which includes the influence of the boundary region between the TSV and the silicon substrate, is presented; estimation of contact resistance and boundary thermal conductance is also given. Copyright © 2014 Elsevier Ltd. All rights reserved.

  12. Fossilization in Geopark Araripe studied through X-ray diffraction, scanning microscopy and thermogravimetric analysis

    CERN Document Server

    Lima, Ricardo J C; Macedo, Zélia S; Sasaki, José M; Saraiva, Antônio A F

    2008-01-01

    The Geopark Araripe, located in Northeastern Brazil, is the first UNESCO Natural Park in the South hemisphere and a world-famous fossil deposit of the Early Cretaceous period (approximately 120 million years). Fossilized fish fauna in Geopark Araripe is found inside of sedimentary rocks in three-dimensional forms. In the present study sedimentary rocks and fossil fish Rhacolepis bucalis have been carefully analysed by means of X-ray powder diffraction, scanning electron microscopy and termogravimetric analysis. Mineralogical composition of the fossil fish was explained in terms of facts occurred at the initial stages of the opening of the South Atlantic and the oceanic hydrothermal phenomena (``black smoker'', ``white smoker'' and warm-water events). The occurrence of organic substance was, for the first time, evaluated in collapsed internal elements (intestinal and muscles) by termogravimetric analysis.

  13. High-resolution characterization of multiferroic heterojunction using aberration-corrected scanning transmission electron microscopy

    Science.gov (United States)

    Yuan, Zhoushen; Ruan, Jieji; Xie, Lin; Pan, Xiaoqing; Wu, Di; Wang, Peng

    2017-04-01

    Multiferroic tunnel junctions have been considered as potential candidates for nonvolatile memory devices. Understanding the atomic structure at the interface is crucial for optimizing the performances in such oxide electronics. Spatially resolved electron energy loss spectroscopy (EELS) combined with aberration-corrected scanning transmission electron microscopy is employed to measure the compositional profiles across the interfaces of different layers with atomic resolution. Two-dimensional elemental imaging with atomic resolution is demonstrated, and the influences of the interface sharpness, the terminal layer, and cation intermixing are investigated. An asymmetric sublattice intermixing at the Pr0.8Ca0.2MnO3/BaTiO3/La0.7Sr0.3MnO3 interface is observed, which can affect the local Mn valence and coupling. The reduction in the Mn valence at the interface is further studied using EELS near-edge fine structures.

  14. Simultaneous scanning tunneling microscopy and synchrotron X-ray measurements in a gas environment.

    Science.gov (United States)

    Mom, Rik V; Onderwaater, Willem G; Rost, Marcel J; Jankowski, Maciej; Wenzel, Sabine; Jacobse, Leon; Alkemade, Paul F A; Vandalon, Vincent; van Spronsen, Matthijs A; van Weeren, Matthijs; Crama, Bert; van der Tuijn, Peter; Felici, Roberto; Kessels, Wilhelmus M M; Carlà, Francesco; Frenken, Joost W M; Groot, Irene M N

    2017-11-01

    A combined X-ray and scanning tunneling microscopy (STM) instrument is presented that enables the local detection of X-ray absorption on surfaces in a gas environment. To suppress the collection of ion currents generated in the gas phase, coaxially shielded STM tips were used. The conductive outer shield of the coaxial tips can be biased to deflect ions away from the tip core. When tunneling, the X-ray-induced current is separated from the regular, 'topographic' tunneling current using a novel high-speed separation scheme. We demonstrate the capabilities of the instrument by measuring the local X-ray-induced current on Au(1 1 1) in 800 mbar Ar. Copyright © 2017 Elsevier B.V. All rights reserved.

  15. Analysis of the melanin distribution in different ethnic groups by in vivo laser scanning microscopy

    Science.gov (United States)

    Antoniou, C.; Lademann, J.; Richter, H.; Astner, S.; Patzelt, A.; Zastrow, L.; Sterry, W.; Koch, S.

    2009-05-01

    The aim of this study was to determine whether Laser scanning confocal microscopy (LSM) is able to visualize differences in melanin content and distribution in different Skin Phototypes. The investigations were carried out on six healthy volunteers with Skin Phototypes II, IV, and VI. Representative skin samples of Skin Phototypes II, V, and VI were obtained for histological analysis from remaining tissue of skin grafts and were used for LSM-pathologic correlation. LSM evaluation showed significant differences in melanin distribution in Skin Phototypes II, IV, and VI, respectively. Based on the differences in overall reflectivity and image brightness, a visual evaluation scheme showed increasing brightness of the basal and suprabasal layers with increasing Skin Phototypes. The findings correlated well with histological analysis. The results demonstrate that LSM may serve as a promising adjunctive tool for real time assessment of melanin content and distribution in human skin, with numerous clinical applications and therapeutic and preventive implications.

  16. Colorimeter and scanning electron microscopy analysis of teeth submitted to internal bleaching.

    Science.gov (United States)

    Martin-Biedma, Benjamin; Gonzalez-Gonzalez, Teresa; Lopes, Manuela; Lopes, Luis; Vilar, Rui; Bahillo, José; Varela-Patiño, Purificación

    2010-02-01

    This in vitro study compared the tooth color and the ultrastructure of internal dental tissues before and after internal bleaching. Sodium perborate was placed in the pulp chamber of endodontically treated molars and sealed with intermediate restorative material. The test samples were stored in a physiologic solution, and the bleaching agent was replaced every 7 days. A control group was used. After 1 month, the colors of the test and control samples were measured with a colorimeter, and the internal surfaces were observed under field emission scanning electron microscopy (FESEM). Statistically significant differences were found between the test and control sample colors. The FESEM ultrastructure analysis of the internal enamel and dentin surfaces did not show any changes after the internal bleaching. The results of the present study show that sodium perborate is effective in bleaching nonvital teeth and does not produce ultrastructural changes in the dental tissues. Copyright 2010 American Association of Endodontists. Published by Elsevier Inc. All rights reserved.

  17. Use of scanning electron microscopy to monitor nanofibre/cell interaction in digestive epithelial cells.

    Science.gov (United States)

    Millaku, Agron; Drobne, Damjana; Torkar, Matjaz; Novak, Sara; Remškar, Maja; Pipan-Tkalec, Živa

    2013-09-15

    We provide data obtained by scanning electron microscopy (SEM) and energy dispersive X-ray spectroscopy (EDS) on the interaction of ingested tungsten nanofibers with epithelial cells of the digestive tubes of a test organism Porcellio scaber. Conventional toxicity endpoints including feeding behaviour, weight loss and mortality were also measured in each investigated animal. No toxicity was detected in any of exposed animals after 14 days of feeding on tungsten nanofiber dosed food, but when nanofibers enter the digestive system they can react with epithelial cells of the digestive tubes, becoming physically inserted into the cells. In this way, nanofibers can injure the epithelial cells of digestive gland tubes when they are ingested with food. Our SEM data suggest that peristaltic forces may have an important role, not predicted by in vitro experiments, in the interactions of nanomaterials with digestive intestinal cells. Copyright © 2013 Elsevier B.V. All rights reserved.

  18. Fluorescence Readout of a Patch Clamped Membrane by Laser Scanning Microscopy.

    Science.gov (United States)

    Gerhardt, Matthias; Walz, Michael; Beta, Carsten

    2016-01-01

    In this chapter, we describe how to shield a patch of a cell membrane against extracellularly applied chemoattractant stimuli. Classical patch clamp methodology is applied to allow for controlled shielding of a membrane patch by measuring the seal resistivity. In Dictyostelium cells, a seal resistivity of 50 MΩ proved to be tight enough to exclude molecules from diffusing into the shielded membrane region. This allowed for separating a shielded and a non-shielded region of a cell membrane to study the spatiotemporal dynamics of intracellular chemotactic signaling events at the interface between shielded and non-shielded areas. The spatiotemporal dynamics of signaling events in the membrane was read out by means of appropriate fluorescent markers using laser scanning confocal microscopy.

  19. Fluorescence confocal laser scanning microscopy for in vivo imaging of epidermal reactions to two experimental irritants

    DEFF Research Database (Denmark)

    Suihko, C.; Serup, J.

    2008-01-01

    Background: Fibre-optic fluorescence confocal laser scanning microscopy (CLSM) is a novel non-invasive technique for in vivo imaging of skin. The cellular structure of the epidermis can be studied. A fluorophore, e.g. fluorescein sodium, is introduced by an intradermal injection or applied...... dermatitis reactions caused by established model irritants, e.g. sodium lauryl sulphate (SLS) and pelargonic acid (PA). Methods: Twelve healthy individuals volunteered. The flexor aspect of the right and the left forearm was exposed to SLS in water and PA in isopropanol and occluded under Finn Chambers...... for 24 h. The reactions were rated clinically and, following epicutaneous and intra-dermal application of fluorescein sodium, studied by fluorescence CLSM, magnification x 1000. Results: Both irritants disturbed the epidermal intercellular borders, which became blurred, thickened and variably altered...

  20. High-Resolution Scanning Tunneling Microscopy Studies of Molecular Nanostructures on Surfaces

    DEFF Research Database (Denmark)

    Song, Xin

    . First, to study the role of hydrogen bonding in self-assembly, we investigate the monomolecular self-assembled system of pyrene-4,5,9,10-tetrone and phenanthrene- 9,10-dione molecules on Au(111) and HOPG surface respectively and the binary molecular self-assembled system of stearic acid and guanine...... molecules co-adsorbed on HOPG surface. Hydrogen bonding plays a key role in these self-assembled nanostructures and the substrate could also give an effect in the self-assembly. Second, to study the self-assembly across the terrace steps as a defect on surface, the chiral self-assembled supramolecular...... of nanostructures requires deeper insight into the adsorption sites, adsorption configurations, diffusion behaviour and driving forces for self-assembly of different molecules or atoms on different substrates. To study these fundamental issues, scanning tunneling microscopy (STM) has proven to be an ideal choice...

  1. Scanning tunneling microscopy on the formation of lipoamide-cyclodextrin monolayer on Au(111)

    Science.gov (United States)

    Yasuda, Satoshi; Shigekawa, Hidemi; Suzuki, Iwao; Nakamura, Tohru; Matsumoto, Mutsuyoshi; Komiyama, Makoto

    2000-01-01

    β-cyclodextrin molecules modified with lipoamide residue (LP-β-CyD) were self-assembled on an Au(111) surface in ethanol solution, and the growth process was studied by scanning tunneling microscopy. At the initial stage, adsorption sites were not only random, but also partially linear ordering, which suggests the existence of some influence by the herringbone structure of the Au(111) surface. According to the macroscopic analysis, the subsequent growth process was explained by the Elovich model, which is based on the repulsive interaction between adsorbed molecules. However, when the immersion time increased, island structures began forming. This result suggests the interaction between LP-β-CyD molecules as attractive, which in fact is more probable in consideration of the possibility of the hydrophobic and the hydrogen bonding interactions between CyD molecules. Finally, formation of a single LP-β-CyD layer was clearly confirmed.

  2. Development of scanning electrochemical microscopy for the investigation of photocatalysis at semiconductor surfaces

    CERN Document Server

    Fonseca, S M M C D

    2002-01-01

    This thesis is concerned with the development and application of scanning electrochemical microscopy (SECM) to investigate interfacial photoelectrochemical processes occurring at supported TiO sub 2 surfaces. The new SECM approach, involving both amperometric and potentiometric electrodes, was used to monitor interfacial photoprocesses with high spatial resolution. A new in situ photoelectrochemical approach to chemical actinometry has been developed and used to determine the light flux through a quartz fibre employed in the SECM system. In this system an ultramicroelectrode (UME) probe is positioned with high precision at a known distance close to a TiO sub 2 -coated fibre and used to detect reactants or products of the ongoing photodegradation process. The microelectrochemical actinometry approach was developed using the well-known liquid phase potassium ferrioxalate actinometer. The approach involved recording the steady-state current for Fe(lll) reduction at an SECM tip positioned close to the fibre. A st...

  3. Trypan blue as a fluorochrome for confocal laser scanning microscopy of arbuscular mycorrhizae in three mangroves.

    Science.gov (United States)

    Kumar, T; Majumdar, A; Das, P; Sarafis, V; Ghose, M

    2008-06-01

    Roots of three mangroves, Acanthus ilicifolius, Ceriops tagal and Excoecaria agallocha, collected from forests of the Sundarbans of India were stained with trypan blue to observe arbuscular mycorrhizal colonization. Spores of arbuscular mycorrhizal fungi isolated from rhizospheric soil, collected together with the root samples, also were stained for testing the suitability of the dye as a fluorochrome. Confocal laser scanning microscopy images were constructed. A. ilicifolius and E. agallocha exhibited "Arum" type colonization with highly branched arbuscules, whereas C. tagal showed "Paris" type association with clumped and collapsed arbuscules. We demonstrated that trypan blue is a suitable fluorochrome for staining arbuscular mycorrhizal fungal spores, fungal hyphae, arbuscules and vesicles, which presumably have a considerable amount of surface chitin. It appears that as the integration of chitin into the fungal cell wall changes, its accessibility to trypan blue dye also changes.

  4. Uncertainty assessment for measurements performed in the determination of thermal conductivity by scanning thermal microscopy

    Science.gov (United States)

    Ramiandrisoa, Liana; Allard, Alexandre; Hay, Bruno; Gomés, Séverine

    2017-11-01

    Although its use has been restricted to relative studies, scanning thermal microscopy (SThM) is presented today as a candidate technique for performing quantitative measurement of thermal properties at the nanoscale, thanks to the development of relevant calibration protocols. Based on the principle behind near-field microscopes, SThM uses a miniaturized probe to quantify heat transfers versus samples of various thermal conductivities: since the thermal conductivity of a sample cannot be directly estimated, a direct measurand related to the heat transfer must be defined and measured for each sample. That is the reason why the SThM technique applied to thermal conductivity determination belongs to the family of inverse methods. In this work we aim to qualify the technique from a metrological point of view. For the first time, assessment of uncertainty associated with the direct measurand Δ R is performed, yielding a result of less than 2%.

  5. Effects of industrial noise on circumpulpar dentin - a field emission scanning electron microscopy and energy dispersive spectroscopy analysis

    Science.gov (United States)

    Cavacas, Maria Alzira; Tavares, Vitor; Oliveira, Maria João; Oliveira, Pedro; Sezinando, Ana; Martins dos Santos, José

    2013-01-01

    Chronic exposure to Industrial Noise (IN), rich in Low Frequency Noise (LFN), causes systemic fibrotic transformation and sustained stress. Dental wear, significantly increased with exposure to LFN, affects the teeth particularly through the circumpulpar dentin. Our goal is to understand the consequences of IN exposure on the circumpulpar dentin of Wistar rats. 10 Wistar rats were exposed to IN for 4 months, according to an occupationally simulated time schedule and 10 animals were used as age-matched controls. The first and the second upper and lower molars of each animal were processed for observation by Field Emission Scanning Electron Microscopy (FESEM) and Energy Dispersive Spectroscopy (EDS) analysis was performed. In exposed animals FESEM showed a 2.0 to 6.0 μm-dense mineral band between dentin and the pulp with no regular continuity with the tubules. This structure had a few tubules where the odontoblasts processes could be observed embedded within the band and collagen fibers were trapped inside. EDS analysis revealed that it was hydroxyapatite similar to dentin, with a higher carbon content. FESEM results show that the band may be tertiary reparative dentin formed by odontoblast-like cells, but the increased amount of carbon (EDS) could mean that it is sclerotic dentin. IN should be acknowledge as a strong stimulus, able to cause an injury to odontoblasts and to the formation of reparative tertiary dentin, in a process that may accelerate the aging of the teeth, either by direct impact of acoustic pressure pulsations or by increased stress and dental wear. PMID:24294356

  6. Effects of industrial noise on circumpulpar dentin--a field emission scanning electron microscopy and energy dispersive spectroscopy analysis.

    Science.gov (United States)

    Cavacas, Maria Alzira; Tavares, Vitor; Oliveira, Maria João; Oliveira, Pedro; Sezinando, Ana; Martins dos Santos, José

    2013-01-01

    Chronic exposure to Industrial Noise (IN), rich in Low Frequency Noise (LFN), causes systemic fibrotic transformation and sustained stress. Dental wear, significantly increased with exposure to LFN, affects the teeth particularly through the circumpulpar dentin. Our goal is to understand the consequences of IN exposure on the circumpulpar dentin of Wistar rats. 10 Wistar rats were exposed to IN for 4 months, according to an occupationally simulated time schedule and 10 animals were used as age-matched controls. The first and the second upper and lower molars of each animal were processed for observation by Field Emission Scanning Electron Microscopy (FESEM) and Energy Dispersive Spectroscopy (EDS) analysis was performed. In exposed animals FESEM showed a 2.0 to 6.0 μm-dense mineral band between dentin and the pulp with no regular continuity with the tubules. This structure had a few tubules where the odontoblasts processes could be observed embedded within the band and collagen fibers were trapped inside. EDS analysis revealed that it was hydroxyapatite similar to dentin, with a higher carbon content. FESEM results show that the band may be tertiary reparative dentin formed by odontoblast-like cells, but the increased amount of carbon (EDS) could mean that it is sclerotic dentin. IN should be acknowledge as a strong stimulus, able to cause an injury to odontoblasts and to the formation of reparative tertiary dentin, in a process that may accelerate the aging of the teeth, either by direct impact of acoustic pressure pulsations or by increased stress and dental wear.

  7. Visualising impregnated chitosan in Pinus radiata early wood cells using light and scanning electron microscopy.

    Science.gov (United States)

    Singh, Adya P; Singh, Tripti; Rickard, Catherine L

    2010-04-01

    Chitosan, a deacetylated product of an abundant naturally occurring biopolymer chitin, has been used in a range of applications, particularly in food and health areas, as an antimicrobial agent. In the work reported here Pinus radiata wood was impregnated with chitosan as an environmentally compatible organic biocide (Eikenes et al., 2005a,b) to protect wood against wood deteriorating microorganisms and to thus prolong the service life of wooden products. We developed sample preparation techniques targeted to visualise impregnated chitosan within wood tissues using light microscope and field-emission scanning electron microscope (FE-SEM). Sections were viewed with the light microscope without staining with a dye as well as after staining with the dye toluidine blue. Light microscopy was also undertaken on sections that had been stained with 1% aqueous osmium tetroxide (OsO(4)). For SEM observations, the sections were treated with OsO(4) and then examined with the FE-SEM, first in the secondary electron imaging mode (SEI) and then in the backscattered electron imaging (BEI) mode, imaging the same areas of a section in both SEI and BEI modes. The preparation techniques employed and the combined use of light and scanning electron microscopy provided valuable complementary information, revealing that chitosan had penetrated into the cavities (cell lumens, intercellular spaces) of all sizes present within wood tissues and had also impregnated early wood cell walls. The information obtained is discussed in relation to its importance in further development of chitosan formulations and refinement of impregnation technologies to optimise chitosan impregnation into and distribution within wood tissues as well as in assessing chitosan efficacy. Copyright 2009 Elsevier Ltd. All rights reserved.

  8. Enumeration of leukocyte infiltration in solid tumors by confocal laser scanning microscopy

    Directory of Open Access Journals (Sweden)

    Amirkhosravi A

    2006-07-01

    Full Text Available Abstract Background Leukocytes commonly infiltrate solid tumors, and have been implicated in the mechanism of spontaneous regression in some cancers. Conventional techniques for the quantitative estimation of leukocyte infiltrates in tumors rely on light microscopy of immunostained thin tissue sections, in which an arbitrary assessment (based on low, medium or high levels of infiltration of antigen density is made by the pathologist. These estimates are relatively subjective and often require the opinion of a second pathologist. In addition, since thin tissue sections are cut, no data regarding the three-dimensional distribution of antigen can be obtained. Results To overcome these problems, we have designed a method to enumerate leukocyte infiltration into tumors, using confocal laser scanning microscopy of fluorescently immunostained leukocytes in thick tissue sections. Using image analysis software, a threshold was applied to eliminate unstained tissue and residual noise. The total antigen volume in the scanned tissue was calculated and divided by the mean cell volume (calculated by "seeding" ten individual cells to obtain the cell count. Using this method, we compared the calculated leukocyte counts with those obtained manually by ten laboratory personnel. There was no significant difference (P > 0.05 between the cell counts obtained by either method. We then compared leukocyte infiltration into seven tumors and matched non-malignant tissue obtained from the periphery of the resected tissue. There was a significant increase in the infiltration of all leukocyte subsets into the tumors compared to minimal numbers in the non-malignant tissue. Conclusion From these results we conclude that this method may be of considerable use for the enumeration of cells in tissues. Furthermore, since it can be performed by laboratory technical staff, less time input is required by the pathologist in assessing the degree of leukocyte infiltration into tumors.

  9. All-optical histology using two photon laser scanning microscopy and ablation with ultrashort pulses

    Science.gov (United States)

    Tsai, Philbert S.

    This dissertation discusses the use of ultrashort laser pulses to image and manipulate tissue for the purpose of three-dimensional histological reconstruction of extended brain structures. Two photon laser scanning microscopy (TPLSM) and ultrashort pulsed laser ablation are used to provide in situ three-dimensional imaging through thick preparations of fixed tissue. Surface regions of fixed tissue are first imaged using TPLSM. The imaged regions are then removed by ablation with amplified, ultrashort laser pulses, thereby exposing a previously underlying tissue region for imaging. This process of imaging and ablation proceeds iteratively until the desired tissue volume has been processed. First, the principles, design, and construction of a two photon laser scanning microscope are discussed, followed by a discussion of the physical mechanisms of tissue ablation with ultrashort laser pulses. The compatibility of tissue ablation using ultrashort pulses with subsequent histological analysis, particularly with fluorescent microscopy, is evaluated. Tissue ablation with ultrashort laser pulses is found to produce ablated tissue surfaces that are smooth to within a micrometer. Intrinsic fluorescence as well as immunoreactivity are found to be resilient to the ablation process. The all-optical histological technique is demonstrated on brain tissue from rats and mice, including tissue from embryonic mouse as early at E15. The ablation process is shown to preserve both macroscopic and microscopic structures within tissue. To facilitate the all-optical histological analysis of neuronal vasculature and its relative distribution to surrounding neuronal tissue, a fluorescent gel perfusion technique is developed that provides a temperature-stabilized fluorescent label of the neuronal vasculature. The use of immunohistochemistry to label specific cell populations throughout an 800 micrometer-thick tissue section is demonstrated. Additionally, the immersion of fixed tissue in high

  10. Charge-imaging field-effect transistors for scanned probe microscopy

    Science.gov (United States)

    Chen, Lester Hao-Lin

    This thesis presents experiments on integrating a charge-imaging field-effect transistor onto a scanned probe microscopy cantilever to make a moveable charge-imager. Such an imager would be used for imaging the spatial distribution of electric charge in semiconductor heterostructures and devices. Learning about the spatial distribution of charge yields knowledge about electrical transport at the microscopic level. The information gained from measuring the spatial distribution of charge increases with improvements in the spatial resolution and charge sensitivity of the charge-imaging probes. So, the goal is to devise a charge-imager with sub-micron spatial resolution and single-electron charge sensitivity. To achieve high spatial resolution and excellent charge sensitivity, the charge-imaging field-effect transistors are made with a quantum point contact geometry. The charge response is confined to a disc with full width half-maximum comparable to its channel width, and the charge noise spectrum reaches values "1 e/Hz½ at 30 kHz. Their low power dissipation (deflections of the cantilever to map the sample topography. The strain-sensing field-effect transistors have a white noise value for the deflection noise of 0.5 nm/Hz½ at 10 kHz. This thesis describes the fabrication and characterization of charge-imaging field-effect transistors and scanned microscopy cantilevers with integrated strain-sensing transistors. The transistors and cantilevers were fabricated in a GaAs/AlGaAs heterostructure using electron-beam lithography and were characterized at liquid Helium temperatures. Possible future experiments include demonstrating the charge-imaging FET's sensitivity to single electrons, creating a charge- and topography-imaging cantilever, and directly measuring the electron distributions in nanostructures.

  11. Use of scanning electron microscopy to monitor nanofibre/cell interaction in digestive epithelial cells

    Energy Technology Data Exchange (ETDEWEB)

    Millaku, Agron, E-mail: agron.mi@hotmail.com [Limnos-Company for Applied Ecology Ltd, Podlimbarskega 31, 1000 Ljubljana (Slovenia); Drobne, Damjana [University of Ljubljana, Biotechnical Faculty, Department of Biology, Večna pot 111, 1000 Ljubljana (Slovenia); Centre of Excellence, Advanced Materials and Technologies for the Future (CO NAMASTE), Jamova cesta 39, 1000 Ljubljana (Slovenia); Centre of Excellence, Nanoscience and Nanotechnology (Nanocentre), Jamova cesta 39, 1000 Ljubljana (Slovenia); Torkar, Matjaz [Institute of Metals and Technology IMT, Lepi pot 11, 1000 Ljubljana (Slovenia); Jožef Stefan Institute, Condensed Matter Physics Department, Jamova cesta 39, 1000 Ljubljana (Slovenia); Novak, Sara [University of Ljubljana, Biotechnical Faculty, Department of Biology, Večna pot 111, 1000 Ljubljana (Slovenia); Remškar, Maja [Jožef Stefan Institute, Condensed Matter Physics Department, Jamova cesta 39, 1000 Ljubljana (Slovenia); Pipan-Tkalec, Živa [University of Ljubljana, Biotechnical Faculty, Department of Biology, Večna pot 111, 1000 Ljubljana (Slovenia)

    2013-09-15

    Graphical abstract: Scanning electron microscopy is particularly well suited to the observation of nanofibre/cell interaction in the endothelial cells lining the hepatopancreas. (a) Tungsten oxide nanofibres, (b) test organism Porcellio scaber and schematic appearance of digestive tubes, (c) digestive tube (hepatopancreas) prepared for SEM investigation, (d) digestive gland cells (C) with nanofibres (NF) embedded in the cell membrane and (e) nanofibres inserted deeply in the cells and damaged nanofibres due to peristalsis. -- Highlights: • Tungsten oxide nanofibres react physically with digestive gland epithelial cells in Porcellio scaber. • Physical peristaltic forces of lead to insertion of nanofibres into the cells. • No toxic responses as measured by conventional toxicity biomarkers were detected. • Physical interactions were observed in a majority of the investigated animals. -- Abstract: We provide data obtained by scanning electron microscopy (SEM) and energy dispersive X-ray spectroscopy (EDS) on the interaction of ingested tungsten nanofibers with epithelial cells of the digestive tubes of a test organism Porcellio scaber. Conventional toxicity endpoints including feeding behaviour, weight loss and mortality were also measured in each investigated animal. No toxicity was detected in any of exposed animals after 14 days of feeding on tungsten nanofiber dosed food, but when nanofibers enter the digestive system they can react with epithelial cells of the digestive tubes, becoming physically inserted into the cells. In this way, nanofibers can injure the epithelial cells of digestive gland tubes when they are ingested with food. Our SEM data suggest that peristaltic forces may have an important role, not predicted by in vitro experiments, in the interactions of nanomaterials with digestive intestinal cells.

  12. Studying Dynamic Processes of Nano-sized Objects in Liquid using Scanning Transmission Electron Microscopy.

    Science.gov (United States)

    Hermannsdörfer, Justus; de Jonge, Niels

    2017-02-05

    Samples fully embedded in liquid can be studied at a nanoscale spatial resolution with Scanning Transmission Electron Microscopy (STEM) using a microfluidic chamber assembled in the specimen holder for Transmission Electron Microscopy (TEM) and STEM. The microfluidic system consists of two silicon microchips supporting thin Silicon Nitride (SiN) membrane windows. This article describes the basic steps of sample loading and data acquisition. Most important of all is to ensure that the liquid compartment is correctly assembled, thus providing a thin liquid layer and a vacuum seal. This protocol also includes a number of tests necessary to perform during sample loading in order to ensure correct assembly. Once the sample is loaded in the electron microscope, the liquid thickness needs to be measured. Incorrect assembly may result in a too-thick liquid, while a too-thin liquid may indicate the absence of liquid, such as when a bubble is formed. Finally, the protocol explains how images are taken and how dynamic processes can be studied. A sample containing AuNPs is imaged both in pure water and in saline.

  13. The use of field emission scanning electron microscopy to assess recombinant adenovirus stability.

    Science.gov (United States)

    Obenauer-Kutner, Linda J; Ihnat, Peter M; Yang, Tong-Yuan; Dovey-Hartman, Barbara J; Balu, Arthi; Cullen, Constance; Bordens, Ronald W; Grace, Michael J

    2002-09-20

    A field emission scanning electron microscopy (FESEM) method was developed to assess the stability of a recombinant adenovirus (rAd). This method was designed to simultaneously sort, count, and size the total number of rAd viral species observed within an image field. To test the method, a preparation of p53 transgene-expressing recombinant adenovirus (rAd/p53) was incubated at 37 degrees C and the viral particles were evaluated by number, structure, and degree of aggregation as a function of time. Transmission electron microscopy (TEM) was also used to obtain ultrastructural detail. In addition, the infectious activity of the incubated rAd/p53 samples was determined using flow cytometry. FESEM image-analysis revealed that incubation at 37 degrees C resulted in a time-dependent decrease in the total number of detectable single rAd/p53 virus particles and an increase in apparent aggregates composed of more than three adenovirus particles. There was also an observed decrease in both the diameter and perimeter of the single rAd/p53 viral particles. TEM further revealed the accumulation of damaged single particles with time at 37 degrees C. The results of this study demonstrate that FESEM, coupled with sophisticated image analysis, may be an important tool in quantifying the distribution of aggregated species and assessing the overall stability of rAd samples.

  14. Cryo-field emission scanning electron microscopy imaging of a rigid surfactant mesophase.

    Science.gov (United States)

    Tan, Grace; Xu, Peng; John, Vijay T; He, Jibao; McPherson, Gary L; Agarwal, Vivek; Bose, Arijit

    2008-10-07

    The aerosol OT/ L-alpha-phosphatidylcholine/isooctane/water system forms a rigid mesophase that transitions from reverse hexagonal to multilamellar in structure at specific water contents. This study shows that characteristics of ordered liquid-crystalline mesophases can be distinguished and imaged in high clarity using cryo-field emission scanning electron microscopy (cryo-FESEM). The reverse hexagonal phase consists of bundles of long cylinders, some with length scales of over 2 microm, that are randomly oriented as part of a larger domain. Cryo-imaging allows the visualization of the intercylinder spacings and the details of transitions from one domain to another. The multilamellar structured mesophase consists of spherical vesicles of 100 nm to 10 microm in diameter, with intervening noncrystalline isotropic regions. Coexistence regions containing both the reverse hexagonal and lamellar structures are also observed in the transition from the reverse hexagonal to the lamellar phase. These results complement and qualitatively verify our earlier studies with small-angle neutron scattering, high-field nuclear magnetic resonance spectroscopy, and freeze-fracture direct imaging transmission electron microscopy. The information is useful in understanding materials templating in these rigid systems.

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

    Science.gov (United States)

    Cazaux, Jacques

    2012-01-01

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

  16. Correlative microscopy of Purkinje dendritic spines: a field emission scanning and transmission electron microscopic study.

    Science.gov (United States)

    Castejón, O J; Castellano, A; Arismendi, G; Apkarian, R

    2004-01-01

    Purkinje dendritic spines (Pds) of mouse cerebellar cortex were examined by field emission scanning electron microscopy (FESEM) and by transmission electron microscopy (TEM) using ultrathin sections and freeze-etching replicas, to study their three-dimensional features and intramembrane morphology. FESEM showed unattached mushroom-type, elongated and lanceolate Pds separated by 100-500 nm on the dendritic shaft surface. High resolution FESEM showed 25-50 nm globular subunits at the spine postsynaptic density corresponding to the localization of postsynaptic proteins and/or postsynaptic receptors. TEM images of ultrathin sections showed gem-like, mushroom-shaped, lanceolate and neckless or stubby spines. Freeze etching replicas exposed postsynaptic intramembrane particles that can be correlated with the globular subunits observed at high resolution FESEM. Parallel and climbing fiber endings were observed making asymmetric synaptic contacts with the Pds heads. Simultaneous contacts with the necks and heads were also found. The variety of Pds shapes were interpreted as spine conformational changes related with spine dynamic, and spine plasticity.

  17. Three-dimensional imaging of plant cuticle architecture using confocal scanning laser microscopy.

    Science.gov (United States)

    Buda, Gregory J; Isaacson, Tal; Matas, Antonio J; Paolillo, Dominick J; Rose, Jocelyn K C

    2009-10-01

    Full appreciation of the roles of the plant cuticle in numerous aspects of physiology and development requires a comprehensive understanding of its biosynthesis and deposition; however, much is still not known about cuticle structure, trafficking and assembly. To date, assessment of cuticle organization has been dominated by 2D imaging, using histochemical stains in conjunction with light and fluorescence microscopy. This strategy, while providing valuable information, has limitations because it attempts to describe a complex 3D structure in 2D. An imaging technique that could accurately resolve 3D architecture would provide valuable additions to the growing body of information on cuticle molecular biology and biochemistry. We present a novel application of 3D confocal scanning laser microscopy for visualizing the architecture, deposition patterns and micro-structure of plant cuticles, using the fluorescent stain auramine O. We demonstrate the utility of this technique by contrasting the fruit cuticle of wild-type tomato (Solanum lycopersicum cv. M82) with those of cutin-deficient mutants. We also introduce 3D cuticle modeling based on reconstruction of serial optical sections, and describe its use in identification of several previously unreported features of the tomato fruit cuticle.

  18. Scanning electron microscopy of the attachment of Treponema pallidum to nerve cells in vitro.

    Science.gov (United States)

    Repesh, L A; Fitzgerald, T J; Oakes, S G; Pozos, R S

    1982-01-01

    Treponema pallidum (Nichols strain) was incubated with cultured nerve cells derived from rat embryos. Primary cultures were established from dorsal root ganglia, superior cervical ganglia, and spinal cord. Using phase contrast microscopy treponemes were seen to interact with the nerve cells in a similar manner to other cultured mammalian cells. Organisms began to attach within minutes after inoculation, actively motile organisms attached at the tip of one end, higher numbers of organisms attached with continued incubation, and attached organisms survived longer than unattached organisms. T pallidum attached both to nerve cell bodies and to neuronal processes of each of the three nerve cell cultures. As shown by scanning electron microscopy the mechanism of attachment was identical to that of cultured cells derived from rabbits testis, rat skeletal muscle, and human cervical carcinoma. There was no indentation or swelling of the cultured cell surface at the point of attachment, just a close physical proximity of organisms and cells. These techniques provide a biological means of studying the in-vitro detrimental influences of micro-organisms on nerve tissue. Images PMID:7049315

  19. Evaluation of the infection process by Lecanicillium fungicola in Agaricus bisporus by scanning electron microscopy.

    Science.gov (United States)

    Santana Nunes, Janaira; Rocha de Brito, Manuela; Cunha Zied, Diego; Aparecida das Graças Leite, Eloisa; Souza Dias, Eustáquio; Alves, Eduardo

    Lecanicillium fungicola causes dry bubble disease in Agaricus bisporus mushrooms leading to significant economic losses in commercial production. To monitor the infection process of L. fungicola in Brazilian strains of A. bisporus. The interaction between the mycelium of L. fungicola (LF.1) and three strains of A. bisporus (ABI 7, ABI 11/14 and ABI 11/21) was studied. Electron microscopy and X-ray microanalyses of vegetative growth and basidiocarp infection were evaluated. Micrographs show that the vegetative mycelium of the Brazilian strains of A. bisporus is not infected by the parasite. The images show that the pathogen can interlace the hyphae of A. bisporus without causing damage, which contributes to the presence of L. fungicola during the substrate colonization, allowing their presence during primordial formation of A. bisporus. In the basidiocarp, germ tubes form within 16h of infection with L. fungicola and the beginning of penetration takes place within 18h, both without the formation of specialized structures. Scanning electron microscopy enabled the process of colonization and reproduction to be observed within the formation of phialides, conidiophores and verticils of L. fungicola. The formation of calcium oxalate crystals by the pathogen was also visible using the X-ray microanalysis, both at the hyphae in the Petri plate and at basidiocarp infection site. Copyright © 2016 Asociación Española de Micología. Publicado por Elsevier España, S.L.U. All rights reserved.

  20. Scanning probe acceleration microscopy (SPAM) in fluids: mapping mechanical properties of surfaces at the nanoscale.

    Science.gov (United States)

    Legleiter, Justin; Park, Matthew; Cusick, Brian; Kowalewski, Tomasz

    2006-03-28

    One of the major thrusts in proximal probe techniques is combination of imaging capabilities with simultaneous measurements of physical properties. In tapping mode atomic force microscopy (TMAFM), the most straightforward way to accomplish this goal is to reconstruct the time-resolved force interaction between the tip and surface. These tip-sample forces can be used to detect interactions (e.g., binding sites) and map material properties with nanoscale spatial resolution. Here, we describe a previously unreported approach, which we refer to as scanning probe acceleration microscopy (SPAM), in which the TMAFM cantilever acts as an accelerometer to extract tip-sample forces during imaging. This method utilizes the second derivative of the deflection signal to recover the tip acceleration trajectory. The challenge in such an approach is that with real, noisy data, the second derivative of the signal is strongly dominated by the noise. This problem is solved by taking advantage of the fact that most of the information about the deflection trajectory is contained in the higher harmonics, making it possible to filter the signal by "comb" filtering, i.e., by taking its Fourier transform and inverting it while selectively retaining only the intensities at integer harmonic frequencies. Such a comb filtering method works particularly well in fluid TMAFM because of the highly distorted character of the deflection signal. Numerical simulations and in situ TMAFM experiments on supported lipid bilayer patches on mica are reported to demonstrate the validity of this approach.

  1. Characterisation of biosynthesised silver nanoparticles by scanning electrochemical microscopy (SECM) and voltammetry.

    Science.gov (United States)

    Battistel, Dario; Baldi, Franco; Gallo, Michele; Faleri, Claudia; Daniele, Salvatore

    2015-01-01

    Silver nanoparticles (AgNPs) were biosynthesised by a Klebsiella oxytoca strain BAS-10, which, during its growth, is known to produce a branched exopolysaccharide (EPS). Klebsiella oxytoca cultures, treated with AgNO3 and grown under either aerobic or anaerobic conditions, produced silver nanoparticles embedded in EPS (AgNPs-EPS) containing different amounts of Ag(0) and Ag(I) forms. The average size of the AgNPs-EPS was determined by transmission electron microscopy, while the relative abundance of Ag(0)- or Ag(I)-containing AgNPs-EPS was established by scanning electrochemical microscopy (SECM). Moreover, the release of silver(I) species from the various types of AgNPs-EPS was investigated by combining SECM with anodic stripping voltammetry. These measurements allowed obtaining information on the kinetic of silver ions release from AgNPs-EPS and their concentration profiles at the substrate/water interface. Copyright © 2014 Elsevier B.V. All rights reserved.

  2. Synchronous-digitization for Video Rate Polarization Modulated Beam Scanning Second Harmonic Generation Microscopy.

    Science.gov (United States)

    Sullivan, Shane Z; DeWalt, Emma L; Schmitt, Paul D; Muir, Ryan M; Simpson, Garth J

    2015-03-09

    Fast beam-scanning non-linear optical microscopy, coupled with fast (8 MHz) polarization modulation and analytical modeling have enabled simultaneous nonlinear optical Stokes ellipsometry (NOSE) and linear Stokes ellipsometry imaging at video rate (15 Hz). NOSE enables recovery of the complex-valued Jones tensor that describes the polarization-dependent observables, in contrast to polarimetry, in which the polarization stated of the exciting beam is recorded. Each data acquisition consists of 30 images (10 for each detector, with three detectors operating in parallel), each of which corresponds to polarization-dependent results. Processing of this image set by linear fitting contracts down each set of 10 images to a set of 5 parameters for each detector in second harmonic generation (SHG) and three parameters for the transmittance of the fundamental laser beam. Using these parameters, it is possible to recover the Jones tensor elements of the sample at video rate. Video rate imaging is enabled by performing synchronous digitization (SD), in which a PCIe digital oscilloscope card is synchronized to the laser (the laser is the master clock.) Fast polarization modulation was achieved by modulating an electro-optic modulator synchronously with the laser and digitizer, with a simple sine-wave at 1/10th the period of the laser, producing a repeating pattern of 10 polarization states. This approach was validated using Z-cut quartz, and NOSE microscopy was performed for micro-crystals of naproxen.

  3. Regenerating titanium ventricular assist device surfaces after gold/palladium coating for scanning electron microscopy.

    Science.gov (United States)

    Achneck, Hardean E; Serpe, Michael J; Jamiolkowski, Ryan M; Eibest, Leslie M; Craig, Stephen L; Lawson, Jeffrey H

    2010-01-01

    Titanium is one of the most commonly used materials for implantable devices in humans. Scanning electron microscopy (SEM) serves as an important tool for imaging titanium surfaces and analyzing cells and other organic matter adhering to titanium implants. However, high-vacuum SEM imaging of a nonconductive sample requires a conductive coating on the surface. A gold/palladium coating is commonly used and to date no method has been described to "clean" such gold/palladium covered surfaces for repeated experiments without etching the titanium itself. This constitutes a major problem with titanium-based implantable devices which are very expensive and thus in short supply. Our objective was to devise a protocol to regenerate titaniumsurfaces after SEM analysis. In a series of experiments, titanium samples from implantable cardiac assist devices were coated with fibronectin, seeded with cells and then coated with gold/palladium for SEM analysis. X-ray photoelectron spectroscopy spectra were obtained before and after five different cleaning protocols. Treatment with aqua regia (a 1:3 solution of concentrated nitric and hydrochloric acid), with or without ozonolysis, followed by sonication in soap solution and sonication in deionized water, allowed regenerating titanium surfaces to their original state. Atomic force microscopy confirmed that the established protocol did not alter the titanium microstructure. The protocol described herein is applicable to almost all titanium surfaces used in biomedical sciences and because of its short exposure time to aqua regia, will likely work for many titanium alloys as well. (c) 2009 Wiley-Liss, Inc.

  4. Smoking and fluidity of erythrocyte membranes: a high resolution scanning electron and atomic force microscopy investigation.

    Science.gov (United States)

    Pretorius, Etheresia; du Plooy, Jeanette N; Soma, Prashilla; Keyser, Ina; Buys, Antoinette V

    2013-11-30

    Smoking affects the general health of an individual, however, the red blood cells (RBCs) and their architecture are particularly vulnerable to inhaled toxins related to smoking. Smoking is one of the lifestyle diseases that are responsible for the most deaths worldwide and an individual who smokes is exposed to excessive amounts of oxidants and toxins which generate up to 10(18) free radicals in the human body. Recently, it was reported that smoking decreases RBC membrane fluidity. Here we confirm this and we show changes visible in the topography of RBC membranes, using scanning electron microscopy (SEM). RBC membranes show bubble formation of the phospholipid layer, as well as balloon-like smooth areas; while their general discoid shapes are changed to form pointed extensions. We also investigate membrane roughness using atomic force microscopy (AFM) and these results confirm SEM results. Due to the vast capability of RBCs to be adaptable, their state of well-being is a major indication for the general health status of an individual. We conclude that these changes, using an old technique in a novel application, may provide new insights and new avenues for future improvements in clinical medicine pertaining to conditions like COPD. Copyright © 2013 The Authors. Published by Elsevier Inc. All rights reserved.

  5. In situ observation of water in a fuel cell catalyst using scanning electron microscopy.

    Science.gov (United States)

    Ueda, Satoru; Kobayashi, Yoshio; Koizumi, Satoshi; Tsutsumi, Yasuyuki

    2015-04-01

    To visualize water in the catalyst of polymer electrolyte fuel cells (PEFCs), backscattered electron (BSE) imaging by means of scanning electron microscopy was employed. To confine a wet specimen of catalyst, an environmental wet cell was manufactured with a silicon nitride thin film (∼100 nm) as the beam window. By supplying humidified gas into the cell, a change in BSE brightness was detected in the catalyst attached to the silicon nitride window. As humidification proceeded, the BSE image became darker and returned brighter by switching to a dry gas. Monte Carlo simulations were performed to evaluate the energy and number of BSE obtained after passing through water with thickness d. Combining the results of the Monte Carlo simulation successfully converted the change in brightness to the change in thickness from d = 100 nm to d = 3 μm. This established method of evaluating water with a thickness resolution of the order of Δd = 100 nm can be applied to in situ observations of the catalyst in a PEFC during operation. © 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.

  6. Direct Observation of Protein Microcrystals in Crystallization Buffer by Atmospheric Scanning Electron Microscopy

    Directory of Open Access Journals (Sweden)

    Chikara Sato

    2012-08-01

    Full Text Available X-ray crystallography requires high quality crystals above a given size. This requirement not only limits the proteins to be analyzed, but also reduces the speed of the structure determination. Indeed, the tertiary structures of many physiologically important proteins remain elusive because of the so-called “crystallization bottleneck”. Once microcrystals have been obtained, crystallization conditions can be optimized to produce bigger and better crystals. However, the identification of microcrystals can be difficult due to the resolution limit of optical microscopy. Electron microscopy has sometimes been utilized instead, with the disadvantage that the microcrystals usually must be observed in vacuum, which precludes the usage for crystal screening. The atmospheric scanning electron microscope (ASEM allows samples to be observed in solution. Here, we report the use of this instrument in combination with a special thin-membrane dish with a crystallization well. It was possible to observe protein crystals of lysozyme, lipase B and a histone chaperone TAF-Iβ in crystallization buffers, without the use of staining procedures. The smallest crystals observed with ASEM were a few µm in width, and ASEM can be used with non-transparent solutions. Furthermore, the growth of salt crystals could be monitored in the ASEM, and the difference in contrast between salt and protein crystals made it easy to distinguish between these two types of microcrystals. These results indicate that the ASEM could be an important new tool for the screening of protein microcrystals.

  7. Synchronous-digitization for video rate polarization modulated beam scanning second harmonic generation microscopy

    Science.gov (United States)

    Sullivan, Shane Z.; DeWalt, Emma L.; Schmitt, Paul D.; Muir, Ryan D.; Simpson, Garth J.

    2015-03-01

    Fast beam-scanning non-linear optical microscopy, coupled with fast (8 MHz) polarization modulation and analytical modeling have enabled simultaneous nonlinear optical Stokes ellipsometry (NOSE) and linear Stokes ellipsometry imaging at video rate (15 Hz). NOSE enables recovery of the complex-valued Jones tensor that describes the polarization-dependent observables, in contrast to polarimetry, in which the polarization stated of the exciting beam is recorded. Each data acquisition consists of 30 images (10 for each detector, with three detectors operating in parallel), each of which corresponds to polarization-dependent results. Processing of this image set by linear fitting contracts down each set of 10 images to a set of 5 parameters for each detector in second harmonic generation (SHG) and three parameters for the transmittance of the fundamental laser beam. Using these parameters, it is possible to recover the Jones tensor elements of the sample at video rate. Video rate imaging is enabled by performing synchronous digitization (SD), in which a PCIe digital oscilloscope card is synchronized to the laser (the laser is the master clock.) Fast polarization modulation was achieved by modulating an electro-optic modulator synchronously with the laser and digitizer, with a simple sine-wave at 1/10th the period of the laser, producing a repeating pattern of 10 polarization states. This approach was validated using Z-cut quartz, and NOSE microscopy was performed for micro-crystals of naproxen.

  8. Scanning Capacitance Microscopy Imaging of State-of-the-Art MOSFETs

    Science.gov (United States)

    Kleiman, R. N.; Garno, J. P.; Hergenrother, J. M.; O'Malley, M. L.; Timp, G. L.

    2000-03-01

    We have used a scanning capacitance microscope (SCM) to study cross-sectioned n- and p-MOSFETs with gate lengths as short as 50 nm. The SCM is a contact AFM, coupled to a sensitive capacitance detector. The metallized AFM tip, native oxide and semiconductor form an MOS junction, upon which we perform c-v measurements. We have studied aggressively scaled CMOS transistors as part of an effort to understand and push the limits to CMOS. We have also studied the newly invented Vertical Replacement-Gate MOSFET, in which the gate dimension is defined by a non-lithographic process. In both cases SCM microscopy is beneficial in advancing and optimizing the design of novel device structures. The transistors provide the opportunity to study well-characterized dopant structures with very small dimensions. We describe some recent improvements we have made in SCM microscopy, in the area of modeling1 probe tips2 sample preparation, and instrumentation. 1 M. L. O'Malley, et al, Appl. Phys. Lett., 74, 272 (1999). 2 M. L. O'Malley, et al, Appl. Phys. Lett., 74, 3672 (1999).

  9. Thermal maturity of Tasmanites microfossils from confocal laser scanning fluorescence microscopy

    Science.gov (United States)

    Hackley, Paul C.; Kus, Jolanta

    2015-01-01

    We report here, for the first time, spectral properties of Tasmanites microfossils determined by confocal laser scanning fluorescence microscopy (CLSM, using Ar 458 nm excitation). The Tasmanites occur in a well-characterized natural maturation sequence (Ro 0.48–0.74%) of Devonian shale (n = 3 samples) from the Appalachian Basin. Spectral property λmax shows excellent agreement (r2 = 0.99) with extant spectra from interlaboratory studies which used conventional fluorescence microscopy techniques. This result suggests spectral measurements from CLSM can be used to infer thermal maturity of fluorescent organic materials in geologic samples. Spectra of regions with high fluorescence intensity at fold apices and flanks in individual Tasmanites are blue-shifted relative to less-deformed areas in the same body that have lower fluorescence intensity. This is interpreted to result from decreased quenching moiety concentration at these locations, and indicates caution is needed in the selection of measurement regions in conventional fluorescence microscopy, where it is common practice to select high intensity regions for improved signal intensity and better signal to noise ratios. This study also documents application of CLSM to microstructural characterization of Tasmanites microfossils. Finally, based on an extant empirical relation between conventional λmax values and bitumen reflectance, λmax values from CLSM of Tasmanites microfossils can be used to calculate a bitumen reflectance equivalent value. The results presented herein can be used as a basis to broaden the future application of CLSM in the geological sciences into hydrocarbon prospecting and basin analysis.

  10. Correlative scanning electron and confocal microscopy imaging of labeled cells coated by indium-tin-oxide

    KAUST Repository

    Rodighiero, Simona

    2015-03-22

    Confocal microscopy imaging of cells allows to visualize the presence of specific antigens by using fluorescent tags or fluorescent proteins, with resolution of few hundreds of nanometers, providing their localization in a large field-of-view and the understanding of their cellular function. Conversely, in scanning electron microscopy (SEM), the surface morphology of cells is imaged down to nanometer scale using secondary electrons. Combining both imaging techniques have brought to the correlative light and electron microscopy, contributing to investigate the existing relationships between biological surface structures and functions. Furthermore, in SEM, backscattered electrons (BSE) can image local compositional differences, like those due to nanosized gold particles labeling cellular surface antigens. To perform SEM imaging of cells, they could be grown on conducting substrates, but obtaining images of limited quality. Alternatively, they could be rendered electrically conductive, coating them with a thin metal layer. However, when BSE are collected to detect gold-labeled surface antigens, heavy metals cannot be used as coating material, as they would mask the BSE signal produced by the markers. Cell surface could be then coated with a thin layer of chromium, but this results in a loss of conductivity due to the fast chromium oxidation, if the samples come in contact with air. In order to overcome these major limitations, a thin layer of indium-tin-oxide was deposited by ion-sputtering on gold-decorated HeLa cells and neurons. Indium-tin-oxide was able to provide stable electrical conductivity and preservation of the BSE signal coming from the gold-conjugated markers. © 2015 Wiley Periodicals, Inc.

  11. Correlative scanning electron and confocal microscopy imaging of labeled cells coated by indium-tin-oxide.

    Science.gov (United States)

    Rodighiero, Simona; Torre, Bruno; Sogne, Elisa; Ruffilli, Roberta; Cagnoli, Cinzia; Francolini, Maura; Di Fabrizio, Enzo; Falqui, Andrea

    2015-06-01

    Confocal microscopy imaging of cells allows to visualize the presence of specific antigens by using fluorescent tags or fluorescent proteins, with resolution of few hundreds of nanometers, providing their localization in a large field-of-view and the understanding of their cellular function. Conversely, in scanning electron microscopy (SEM), the surface morphology of cells is imaged down to nanometer scale using secondary electrons. Combining both imaging techniques have brought to the correlative light and electron microscopy, contributing to investigate the existing relationships between biological surface structures and functions. Furthermore, in SEM, backscattered electrons (BSE) can image local compositional differences, like those due to nanosized gold particles labeling cellular surface antigens. To perform SEM imaging of cells, they could be grown on conducting substrates, but obtaining images of limited quality. Alternatively, they could be rendered electrically conductive, coating them with a thin metal layer. However, when BSE are collected to detect gold-labeled surface antigens, heavy metals cannot be used as coating material, as they would mask the BSE signal produced by the markers. Cell surface could be then coated with a thin layer of chromium, but this results in a loss of conductivity due to the fast chromium oxidation, if the samples come in contact with air. In order to overcome these major limitations, a thin layer of indium-tin-oxide was deposited by ion-sputtering on gold-decorated HeLa cells and neurons. Indium-tin-oxide was able to provide stable electrical conductivity and preservation of the BSE signal coming from the gold-conjugated markers. © 2015 Wiley Periodicals, Inc.

  12. Focused ion beam (FIB)/scanning electron microscopy (SEM) in tissue structural research.

    Science.gov (United States)

    Leser, Vladka; Milani, Marziale; Tatti, Francesco; Tkalec, Ziva Pipan; Strus, Jasna; Drobne, Damjana

    2010-10-01

    The focused ion beam (FIB) and scanning electron microscope (SEM) are commonly used in material sciences for imaging and analysis of materials. Over the last decade, the combined FIB/SEM system has proven to be also applicable in the life sciences. We have examined the potential of the focused ion beam/scanning electron microscope system for the investigation of biological tissues of the model organism Porcellio scaber (Crustacea: Isopoda). Tissue from digestive glands was prepared as for conventional SEM or as for transmission electron microscopy (TEM). The samples were transferred into FIB/SEM for FIB milling and an imaging operation. FIB-milled regions were secondary electron imaged, back-scattered electron imaged, or energy dispersive X-ray (EDX) analyzed. Our results demonstrated that FIB/SEM enables simultaneous investigation of sample gross morphology, cell surface characteristics, and subsurface structures. The same FIB-exposed regions were analyzed by EDX to provide basic compositional data. When samples were prepared as for TEM, the information obtained with FIB/SEM is comparable, though at limited magnification, to that obtained from TEM. A combination of imaging, micro-manipulation, and compositional analysis appears of particular interest in the investigation of epithelial tissues, which are subjected to various endogenous and exogenous conditions affecting their structure and function. The FIB/SEM is a promising tool for an overall examination of epithelial tissue under normal, stressed, or pathological conditions.

  13. Facile synthesis and electron transport properties of NiO nanostructures investigated by scanning tunneling microscopy

    Directory of Open Access Journals (Sweden)

    Govind Mallick

    2017-08-01

    Full Text Available Due to their unique chemical, thermal, electronic and photonic properties, low -dimensional transition metal oxides, especially NiO, have attracted great deal of attention for potential applications in a wide range of technologies, such as, sensors, electrochromic coatings and self-healing materials. However, their synthesis involves multi-step complex procedures that in addition to being expensive, further introduce impurities. Here we present a low cost facile approach to synthesize uniform size NiO nanoparticles (NPs from hydrothermally grown Ni(OH2. Detailed transmission electron microscopic analysis reveal the average size of NiO NPs to be around 29 nm. The dimension of NiO NP is also corroborated by the small area scanning tunneling microscope (STM measurements. Further, we investigate electron transport characteristics of newly synthesized Ni(OH2 and NiO nanoparticles on p-type Si substrate using scanning tunneling microscopy. The conductivity of Ni(OH2 and NiO are determined to be 1.46x10-3 S/cm and 2.37x10-5 S/cm, respectively. The NiO NPs exhibit a lower voltage window (∼0.7 V electron tunneling than the parent Ni(OH2.

  14. Invited review article: A 10 mK scanning probe microscopy facility.

    Science.gov (United States)

    Song, Young Jae; Otte, Alexander F; Shvarts, Vladimir; Zhao, Zuyu; Kuk, Young; Blankenship, Steven R; Band, Alan; Hess, Frank M; Stroscio, Joseph A

    2010-12-01

    We describe the design, development and performance of a scanning probe microscopy (SPM) facility operating at a base temperature of 10 mK in magnetic fields up to 15 T. The microscope is cooled by a custom designed, fully ultra-high vacuum (UHV) compatible dilution refrigerator (DR) and is capable of in situ tip and sample exchange. Subpicometer stability at the tip-sample junction is achieved through three independent vibration isolation stages and careful design of the dilution refrigerator. The system can be connected to, or disconnected from, a network of interconnected auxiliary UHV chambers, which include growth chambers for metal and semiconductor samples, a field-ion microscope for tip characterization, and a fully independent additional quick access low temperature scanning tunneling microscope (STM) and atomic force microscope (AFM) system. To characterize the system, we present the cooling performance of the DR, vibrational, tunneling current, and tip-sample displacement noise measurements. In addition, we show the spectral resolution capabilities with tunneling spectroscopy results obtained on an epitaxial graphene sample resolving the quantum Landau levels in a magnetic field, including the sublevels corresponding to the lifting of the electron spin and valley degeneracies.

  15. Touching is believing: interrogating halide perovskite solar cells at the nanoscale via scanning probe microscopy

    Science.gov (United States)

    Li, Jiangyu; Huang, Boyuan; Nasr Esfahani, Ehsan; Wei, Linlin; Yao, Jianjun; Zhao, Jinjin; Chen, Wei

    2017-10-01

    Halide perovskite solar cells based on CH3NH3PbI3 and related materials have emerged as the most exciting development in the next generation photovoltaic technologies, yet the microscopic phenomena involving photo-carriers, ionic defects, spontaneous polarization, and molecular vibration and rotation interacting with numerous grains, grain boundaries, and interfaces are still inadequately understood. In fact, there is still need for an effective method to interrogate the local photovoltaic properties of halide perovskite solar cells that can be directly traced to their microstructures on one hand and linked to their device performance on the other hand. In this perspective, we propose that scanning probe microscopy (SPM) techniques have great potential to realize such promises at the nanoscale, and highlight some of the recent progresses and challenges along this line of investigation toward local probing of photocurrent, work function, ionic activities, polarization switching, and chemical degradation. We also emphasize the importance of multi-modality imaging, in-operando scanning, big data analysis, and multidisciplinary collaboration for further studies toward fully understanding of these complex systems.

  16. Facile synthesis and electron transport properties of NiO nanostructures investigated by scanning tunneling microscopy

    Science.gov (United States)

    Mallick, Govind; Labh, Jyotsna; Giri, Lily; Pandey, Avinash C.; Karna, Shashi P.

    2017-08-01

    Due to their unique chemical, thermal, electronic and photonic properties, low -dimensional transition metal oxides, especially NiO, have attracted great deal of attention for potential applications in a wide range of technologies, such as, sensors, electrochromic coatings and self-healing materials. However, their synthesis involves multi-step complex procedures that in addition to being expensive, further introduce impurities. Here we present a low cost facile approach to synthesize uniform size NiO nanoparticles (NPs) from hydrothermally grown Ni(OH)2. Detailed transmission electron microscopic analysis reveal the average size of NiO NPs to be around 29 nm. The dimension of NiO NP is also corroborated by the small area scanning tunneling microscope (STM) measurements. Further, we investigate electron transport characteristics of newly synthesized Ni(OH)2 and NiO nanoparticles on p-type Si substrate using scanning tunneling microscopy. The conductivity of Ni(OH)2 and NiO are determined to be 1.46x10-3 S/cm and 2.37x10-5 S/cm, respectively. The NiO NPs exhibit a lower voltage window (˜0.7 V) electron tunneling than the parent Ni(OH)2.

  17. Scanning MWCNT-Nanopipette and Probe Microscopy: Li Patterning and Transport Studies.

    Science.gov (United States)

    Larson, Jonathan M; Bharath, Satyaveda C; Cullen, William G; Reutt-Robey, Janice E

    2015-10-07

    A carbon-nanotube-enabling scanning probe technique/nanotechnology for manipulating and measuring lithium at the nano/mesoscale is introduced. Scanning Li-nanopipette and probe microscopy (SLi-NPM) is based on a conductive atomic force microscope (AFM) cantilever with an open-ended multi-walled carbon nanotube (MWCNT) affixed to its apex. SLi-NPM operation is demonstrated with a model system consisting of a Li thin film on a Si(111) substrate. By control of bias, separation distance, and contact time, attograms of Li can be controllably pipetted to or from the MWCNT tip. Patterned surface Li features are then directly probed via noncontact AFM measurements with the MWCNT tip. The subsequent decay of Li features is simulated with a mesoscale continuum model, developed here. The Li surface diffusion coefficient for a four (two) Li layer thick film is measured as D=8(±1.2)×10(-15) cm(2) s(-1) (D=1.75(±0.15)×10(-15) cm(2) s(-1)). Dual-Li pipetting/measuring with SLi-NPM enables a broad range of time-dependent Li and nanoelectrode characterization studies of fundamental importance to energy-storage research. © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  18. Image formation, resolution, and height measurement in scanning ion conductance microscopy

    Science.gov (United States)

    Rheinlaender, Johannes; Schäffer, Tilman E.

    2009-05-01

    Scanning ion conductance microscopy (SICM) is an emerging tool for the noncontact investigation of biological samples such as live cells. It uses an ion current through the opening of a tapered nanopipette filled with an electrolyte for topography measurements. Despite its successful application to numerous systems no systematic investigation of the image formation process has yet been performed. Here, we use finite element modeling to investigate how the scanning ion conductance microscope images small particles on a planar surface, providing a fundamental characterization of the imaging process. We find that a small particle appears with a height that is only a fraction of its actual height. This has significant consequences for the quantitative interpretation of SICM images. Furthermore, small and low particles are imaged as rings in certain cases. This can cause small, closely spaced particles to appear with a lateral orientation that is rotated by 90°. Considering both real space and spatial frequency space we find that a reasonable and useful definition of lateral resolution of SICM is the smallest distance at which two small particles can clearly be resolved from each other in an image. We find that this resolution is approximately equal to three times the inner radius of the pipette tip opening.

  19. High-speed XYZ-nanopositioner for scanning ion conductance microscopy

    Science.gov (United States)

    Watanabe, Shinji; Ando, Toshio

    2017-09-01

    We describe a tip-scan-type high-speed XYZ-nanopositioner designed for scanning ion conductance microscopy (SICM), with a special care being devoted to the way of nanopipette holding. The nanopipette probe is mounted in the center of a hollow piezoactuator, both ends of which are attached to identical diaphragm flexures, for Z-positioning. This design minimizes the generation of undesirable mechanical vibrations. Mechanical amplification is used to increase the XY-travel range of the nanopositioner. The first resonance frequencies of the nanopositioner are measured as ˜100 kHz and ˜2.3 kHz for the Z- and XY-displacements, respectively. The travel ranges are ˜6 μm and ˜34 μm for Z and XY, respectively. When this nanopositioner is used for hopping mode imaging of SICM with a ˜10-nm radius tip, the vertical tip velocity can be increased to 400 nm/ms; hence, the one-pixel acquisition time can be minimized to ˜1 ms.

  20. Scanning tunneling microscopy and spectroscopy of iron suicide epitaxially grown on Si(111)

    Science.gov (United States)

    Raunau, Werner; Niehus, Horst; Schilling, Thomas; Comsa, George

    1993-05-01

    Epitaxial iron suicide films have been grown on Si(111) by solid phase epitaxy (SPE) in UHV. Structural and electronic properties have been investigated with scanning tunneling microscopy (STM) and scanning tunneling spectroscopy (STS). For initial Fe deposition up to 3 Å and annealing at 850 K, metallic γ-FeSi 2 is formed. These films exhibit a perfect (2 × 2) superstructure, which is attributed to γ-FeSi 2(111) with Si termination. SPE at higher initial iron deposition (15 Å) and annealing at 800 K results in ɛ-FeSi showing a (√3 × √3) R30° superstructure. Subsequent annealing above 900 K leads to β-FeSi 2 formation. As by STS, β-FeSi 2 films are semiconducting with Eg = 0.85 eV. STM topographs show that SPE produces rough silicide surfaces wit β-FeSi 2(101) [and not β-FeSi 2(110)] epitaxy. The atomic structure on β-FeSi 2 terraces is complex, consisting domain boundaries and defects.

  1. Noninvasive in vivo detection and quantification of Demodex mites by confocal laser scanning microscopy.

    Science.gov (United States)

    Sattler, E C; Maier, T; Hoffmann, V S; Hegyi, J; Ruzicka, T; Berking, C

    2012-11-01

    In many Demodex-associated skin diseases Demodex mites are present in abundance and seem to be at least partially pathogenic. So far all diagnostic approaches such as scraping or standardized superficial skin biopsy are (semi-)invasive and may cause discomfort to the patient. To see whether confocal laser scanning microscopy (CLSM) - a noninvasive method for the visualization of superficial skin layers - is able to detect and quantify D. folliculorum in facial skin of patients with rosacea. Twenty-five patients (34-72 years of age) with facial rosacea and 25 age- and sex-matched normal controls were examined by CLSM. Mosaics of 8 × 8 mm and 5 × 5 mm were created by scanning horizontal layers of lesional skin and quantification of mites per follicle and per area as well as follicles per area was performed. In all patients D. folliculorum could be detected by CLSM and presented as roundish or lengthy cone-shaped structures. CLSM allowed the quantification of Demodex mites and revealed significant differences (P Demodex mites noninvasively in facial skin of patients with rosacea. © 2012 The Authors. BJD © 2012 British Association of Dermatologists.

  2. Scanning probe microscopy studies on the adsorption of selected molecular dyes on titania

    Directory of Open Access Journals (Sweden)

    Jakub S. Prauzner-Bechcicki

    2016-11-01

    Full Text Available Titanium dioxide, or titania, sensitized with organic dyes is a very attractive platform for photovoltaic applications. In this context, the knowledge of properties of the titania–sensitizer junction is essential for designing efficient devices. Consequently, studies on the adsorption of organic dyes on titania surfaces and on the influence of the adsorption geometry on the energy level alignment between the substrate and an organic adsorbate are necessary. The method of choice for investigating the local environment of a single dye molecule is high-resolution scanning probe microscopy. Microscopic results combined with the outcome of common spectroscopic methods provide a better understanding of the mechanism taking place at the titania–sensitizer interface. In the following paper, we review the recent scanning probe microscopic research of a certain group of molecular assemblies on rutile titania surfaces as it pertains to dye-sensitized solar cell applications. We focus on experiments on adsorption of three types of prototypical dye molecules, i.e., perylene-3,4,9,10-tetracarboxylic dianhydride (PTCDA, phtalocyanines and porphyrins. Two interesting heteromolecular systems comprising molecules that are aligned with the given review are discussed as well.

  3. Diagonally Scanned Light-Sheet Microscopy for Fast Volumetric Imaging of Adherent Cells.

    Science.gov (United States)

    Dean, Kevin M; Roudot, Philippe; Reis, Carlos R; Welf, Erik S; Mettlen, Marcel; Fiolka, Reto

    2016-03-29

    In subcellular light-sheet fluorescence microscopy (LSFM) of adherent cells, glass substrates are advantageously rotated relative to the excitation and emission light paths to avoid glass-induced optical aberrations. Because cells are spread across the sample volume, three-dimensional imaging requires a light-sheet with a long propagation length, or rapid sample scanning. However, the former degrades axial resolution and/or optical sectioning, while the latter mechanically perturbs sensitive biological specimens on pliant biomimetic substrates (e.g., collagen and basement membrane). Here, we use aberration-free remote focusing to diagonally sweep a narrow light-sheet along the sample surface, enabling multicolor imaging with high spatiotemporal resolution. Further, we implement a dithered Gaussian lattice to minimize sample-induced illumination heterogeneities, significantly improving signal uniformity. Compared with mechanical sample scanning, we drastically reduce sample oscillations, allowing us to achieve volumetric imaging at speeds of up to 3.5 Hz for thousands of Z-stacks. We demonstrate the optical performance with live-cell imaging of microtubule and actin cytoskeletal dynamics, phosphoinositide signaling, clathrin-mediated endocytosis, polarized blebbing, and endocytic vesicle sorting. We achieve three-dimensional particle tracking of clathrin-associated structures with velocities up to 4.5 μm/s in a dense intracellular environment, and show that such dynamics cannot be recovered reliably at lower volumetric image acquisition rates using experimental data, numerical simulations, and theoretical modeling. Copyright © 2016 The Authors. Published by Elsevier Inc. All rights reserved.

  4. Analysis of acute impact of oleoresin capsicum on rat nasal mucosa using scanning electron microscopy.

    Science.gov (United States)

    Catli, Tolgahan; Acar, Mustafa; Olgun, Yüksel; Dağ, İlknur; Cengiz, Betül Peker; Cingi, Cemal

    2015-01-01

    Analysis of acute cellular changes seen in nasal mucosa of Wistar-Albino rats exposed to different doses of oleoresin capsicum for various time periods by means of scanning electron microscopy. Thirty-five Wistar-Albino rats were divided into five groups of seven rats each. 6-gram oleoresin capsicum per second was sprayed into cages of the groups except group 1. Spray times and duration of exposure to pepper gasses were different for each group. Thirty minutes after the exposure, the animals were killed and specimens from their nasal mucosas were harvested and examined under scanning electron microscope. Mucosal damage was scored from 0-4 points. Mean values of nasal mucosa damage scores of the groups were calculated and compared statistically. Average damage scores of the groups exposed to identical doses of oleoresin capsicum for various exposure times were compared and a statistically significant difference was seen between Groups 2 and 3 (p 0.05). Average damage scores of the groups exposed to various doses for identical exposure times were compared, and statistically significant differences were observed between Groups 2 and 4 and also Groups 3 and 5 (p < 0.05). Outcomes of our study have demonstrated that pepper gas exerts destructive changes on rat nasal mucosa. The extent of these destructive changes increases with the prolonged exposure to higher doses. Besides, exposure time also stands out as an influential factor on the extent of the destructive changes.

  5. Preparation of scanning tunneling microscopy tips using pulsed alternating current etching

    Energy Technology Data Exchange (ETDEWEB)

    Valencia, Victor A.; Thaker, Avesh A.; Derouin, Jonathan; Valencia, Damian N.; Farber, Rachael G.; Gebel, Dana A.; Killelea, Daniel R., E-mail: dkillelea@luc.edu [Department of Chemistry and Biochemistry, Loyola University Chicago, 1068 W. Sheridan Rd., Chicago, Illinois 60660 (United States)

    2015-03-15

    An electrochemical method using pulsed alternating current etching (PACE) to produce atomically sharp scanning tunneling microscopy (STM) tips is presented. An Arduino Uno microcontroller was used to control the number and duration of the alternating current (AC) pulses, allowing for ready optimization of the procedures for both Pt:Ir and W tips using a single apparatus. W tips prepared using constant and pulsed AC power were compared. Tips fashioned using PACE were sharper than those etched with continuous AC power alone. Pt:Ir tips were prepared with an initial coarse etching stage using continuous AC power followed by fine etching using PACE. The number and potential of the finishing AC pulses was varied and scanning electron microscope imaging was used to compare the results. Finally, tip quality using the optimized procedures was verified by UHV-STM imaging. With PACE, at least 70% of the W tips and 80% of the Pt:Ir tips were of sufficiently high quality to obtain atomically resolved images of HOPG or Ni(111)

  6. Plasmons and Electrons as Nanosecond-Fast Sensors for Scanning Tunneling Microscopy

    Science.gov (United States)

    Loth, Sebastian

    2014-03-01

    The ability to measure the fast dynamical evolution of atomic-scale systems often holds the key to their understanding. We combine fast pump-probe spectroscopy tools with low-temperature scanning tunneling microscopy to study atomically assembled arrays of magnetic atoms. The dynamical information quantifies spin lifetimes, magnetic stability and even allows identifying the cross-over between quantum spins and classical magnetism. The spin relaxation times of transition metal atoms can be measured by all-electronic pump probe spectroscopy in which nanosecond-fast voltage pulses excite the spins and probe the average time-dependent response by variations in the spin-polarized tunnel current. In addition, the fast evolution of the local electrostatic potential can be mapped by detecting plasmonic light emission from the STM tunnel junction with time correlating single photon counting. The combination of electrical stimulus and optical detection provides precise control of the excitation process of individual atoms enabling new experiments to probe charge and spin dynamics in the scanning tunneling microscope.

  7. Laser scanning microscopy of broad freezing interfaces with applications to biological cells

    Science.gov (United States)

    Neils, Christopher Martin

    2000-09-01

    A new, vertical cryostage was used for microscopic observation of broad-front freezing in aqueous solutions. This cryostage complements traditional studies of cell behavior and interface morphology in cryobiology. Traditional systems directionally solidify thin samples perpendicular to the optical axis. Thin samples confer thermal and optical advantages for video brightfield microscopy. However, sample thickness can affect the interface morphology. In the new cryostage, ice propagates parallel to the microscope optical axis. The sample cup is 1 cm tall and 1.5 cm in diameter, with insulated sides and a nitrogen-cooled base to freeze the solution upward. The top of the solution is warmed passively through a cover glass or immersion objective. The freezing solutions contain dilute fluorescein dye, which is visible where it is concentrated by exclusion from the ice. The stage is mounted on a confocal laser-scanning microscope, and thermal control and image capture routines are centralized in a LabView user interface. Filtered water, physiological saline, 9.5% glycerol, and 10% glycerol with PBS were frozen at rates between -2°C/min and -10°C/min and sequential images at one plane were captured. Images distinctly revealed a lamellar interface but could not resolve 3-D morphology. The average lamellar spacing was quantified using image analysis. Physiological saline was frozen in flat glass capillary tubes with 0.05 to 0.4 mm path length, mounted vertically to observe internal ice in cross-section. Lamellae were randomly oriented with respect to the glass, suggesting caution when measuring dendrite spacing in a horizontal cryostage. No correlation between capillary size and lamellar spacing was noted. Cell monolayers and synthetic membranes were mounted horizontally to let a well-developed ice front approach the layer broadly. In transparent membranes, ice-membrane interaction was visible until ice grew over and obscured the membrane. The vertical cryostage improved

  8. Subsurface Examination of a Foliar Biofilm Using Scanning Electron- and Focused-Ion-Beam Microscopy

    Energy Technology Data Exchange (ETDEWEB)

    Wallace, Patricia K.; Arey, Bruce W.; Mahaffee, Walt F.

    2011-08-01

    The dual beam scanning electron microscope, equipped with both a focused ion- and scanning electron- beam (FIB SEM) is a novel tool for the exploration of the subsurface structure of biological tissues. The FIB can remove a predetermined amount of material from a selected site to allow for subsurface exploration and when coupled with SEM or scanning ion- beam microscopy (SIM) could be suitable to examine the subsurface structure of bacterial biofilms on the leaf surface. The suitability of chemical and cryofixation was examined for use with the FIB SEM to examine bacterial biofilms on leaf surfaces. The biological control agent, Burkholderia pyroccinia FP62, that rapidly colonizes the leaf surface and forms biofilms, was inoculated onto geranium leaves and incubated in a greenhouse for 7 or 14 days. Cryofixation was not suitable for examination of leaf biofilms because it created a frozen layer over the leaf surface that cracked when exposed to the electron beam and the protective cap required for FIB milling could not be accurately deposited. With chemically fixed samples, it was possible to precisely FIB mill a single cross section (5 µm) or sequential cross sections from a single site without any damage to the surrounding surface. Biofilms, 7 days post-inoculation (DPI), were composed of 2 to 5 bacterial cell layers while biofilms 14 DPI ranged from 5 to greater than 30 cell layers. Empty spaces between bacteria cells in the subsurface structure were observed in biofilms 7- and 14-DPI. Sequential cross sections inferred that the empty spaces were often continuous between FP62 cells and could possibly make up a network of channels throughout the biofilm. FIB SEM was a useful tool to observe the subsurface composition of a foliar biofilm.

  9. Atomic force acoustic microscopy: Influence of the lateral contact stiffness on the elastic measurements.

    Science.gov (United States)

    Flores-Ruiz, F J; Espinoza-Beltrán, F J; Diliegros-Godines, C J; Siqueiros, J M; Herrera-Gómez, A

    2016-09-01

    Atomic force acoustic microscopy is a dynamic technique where the resonances of a cantilever, that has its tip in contact with the sample, are used to quantify local elastic properties of surfaces. Since the contact resonance frequencies (CRFs) monotonically increase with the tip-sample contact stiffness, they are used to evaluate the local elastic properties of the surfaces through a suitable contact mechanical model. The CRFs depends on both, normal and lateral contact stiffness, kN and kS respectively, where the last one is taken either as constant (kS<1), or as zero, leading to uncertainty in the estimation of the elastic properties of composite materials. In this work, resonance spectra for free and contact vibration were used in a finite element analysis of cantilevers to show the influence of kS in the resonance curves due to changes in the kS/kN ratio. These curves have regions for the different vibrational modes that are both, strongly and weakly dependent on kS, and they can be used in a selective manner to obtain a precise mapping of elastic properties. Copyright © 2016. Published by Elsevier B.V.

  10. Dislocation imaging for orthopyroxene using an atom-resolved scanning transmission electron microscopy.

    Science.gov (United States)

    Kumamoto, Akihito; Kogure, Toshihiro; Raimbourg, Hugues; Ikuhara, Yuichi

    2014-11-01

    Dislocations, one-dimensional lattice defects, appear as a microscopic phenomenon while they are formed in silicate minerals by macroscopic dynamics of the earth crust such as shear stress. To understand ductile deformation mechanisms of silicates, atomic structures of the dislocations have been examined using transmission electron microscopy (TEM). Among them, it has been proposed that {100} primary slip system of orthopyroxene (Opx) is dissociated into partial dislocations, and a stacking fault with the clinopyroxene (Cpx) structure is formed between the dislocations. This model, however, has not been determined completely due to the complex structures of silicates. Scanning transmission electron microscopy (STEM) has a potential to determine the structure of dislocations with single-atomic column sensitivity, particularly by using high-angle annular dark field (HAADF) and annular bright field (ABF) imaging with a probing aberration corrector.[1] Furthermore, successive analyses from light microscopy to atom-resolved STEM have been achieved by focused ion beam (FIB) sampling techniques.[2] In this study, we examined dislocation arrays at a low-angle grain boundary of ∼1° rotation about the b-axis in natural deformed Opx using a simultaneous acquisition of HAADF/ABF (JEM-ARM200F, JEOL) equipped with 100 mm2 silicon drift detector (SDD) for energy dispersive X-ray spectroscopy (EDS). Figure 1 shows averaged STEM images viewed along the b- axis of Opx extracted from repeating units. HAADF provides the cation-site arrangement, and ABF distinguishes the difference of slightly rotated SiO4 tetrahedron around the a- axis. This is useful to distinguish the change of stacking sequence between the partial dislocations. Two types of stacking faults with Cpx and protopyroxene (Ppx) structures were identified between three partial dislocations. Furthermore, Ca accumulation in M2 (Fe) site around the stacking faults was detected by STEM-EDS. Interestingly, Ca is

  11. Scanning Hall Probe Microscopy of Magnetic Vortices inVery Underdoped yttrium-barium-copper-oxide

    Energy Technology Data Exchange (ETDEWEB)

    Guikema, Janice Wynn; /SLAC, SSRL

    2005-12-02

    Since their discovery by Bednorz and Mueller (1986), high-temperature cuprate superconductors have been the subject of intense experimental research and theoretical work. Despite this large-scale effort, agreement on the mechanism of high-T{sub c} has not been reached. Many theories make their strongest predictions for underdoped superconductors with very low superfluid density n{sub s}/m*. For this dissertation I implemented a scanning Hall probe microscope and used it to study magnetic vortices in newly available single crystals of very underdoped YBa{sub 2}Cu{sub 3}O{sub 6+x} (Liang et al. 1998, 2002). These studies have disproved a promising theory of spin-charge separation, measured the apparent vortex size (an upper bound on the penetration depth {lambda}{sub ab}), and revealed an intriguing phenomenon of ''split'' vortices. Scanning Hall probe microscopy is a non-invasive and direct method for magnetic field imaging. It is one of the few techniques capable of submicron spatial resolution coupled with sub-{Phi}{sub 0} (flux quantum) sensitivity, and it operates over a wide temperature range. Chapter 2 introduces the variable temperature scanning microscope and discusses the scanning Hall probe set-up and scanner characterizations. Chapter 3 details my fabrication of submicron GaAs/AlGaAs Hall probes and discusses noise studies for a range of probe sizes, which suggest that sub-100 nm probes could be made without compromising flux sensitivity. The subsequent chapters detail scanning Hall probe (and SQUID) microscopy studies of very underdoped YBa{sub 2}Cu{sub 3}O{sub 6+x} crystals with T{sub c} {le} 15 K. Chapter 4 describes two experimental tests for visons, essential excitations of a spin-charge separation theory proposed by Senthil and Fisher (2000, 2001b). We searched for predicted hc/e vortices (Wynn et al. 2001) and a vortex memory effect (Bonn et al. 2001) with null results, placing upper bounds on the vison energy inconsistent with

  12. Comparison of macroscopic and microscopic (stereomicroscopy and scanning electron microscopy) features of bone lesions due to hatchet hacking trauma.

    Science.gov (United States)

    Nogueira, Luísa; Quatrehomme, Gérald; Bertrand, Marie-France; Rallon, Christophe; Ceinos, Romain; du Jardin, Philippe; Adalian, Pascal; Alunni, Véronique

    2017-03-01

    This experimental study examined the lesions produced by a hatchet on human bones (tibiae). A total of 30 lesions were produced and examined macroscopically (naked eye) and by stereomicroscopy. 13 of them were also analyzed using scanning electron microscopy. The general shape of the lesion, both edges, both walls, the kerf floor and the extremities were described. The length and maximum width of the lesions were also recorded. The microscopic analysis of the lesions led to the description of a sharp-blunt mechanism. Specific criteria were identified (lateral pushing back, fragmentation of the upraising, fossa dug laterally to the edge and vertical striae) enabling the forensic expert to conclude that a hacking instrument was used. These criteria are easily identifiable using scanning electron microscopy, but can also be observed with stereomicroscopy. Overall, lateral pushing back and vertical striae visible using stereomicroscopy and scanning electron microscopy signal the use of a hacking tool.

  13. Analytical Model of the Nonlinear Dynamics of Cantilever Tip-Sample Surface Interactions for Various Acoustic-Atomic Force Microscopies

    Science.gov (United States)

    Cantrell, John H., Jr.; Cantrell, Sean A.

    2008-01-01

    A comprehensive analytical model of the interaction of the cantilever tip of the atomic force microscope (AFM) with the sample surface is developed that accounts for the nonlinearity of the tip-surface interaction force. The interaction is modeled as a nonlinear spring coupled at opposite ends to linear springs representing cantilever and sample surface oscillators. The model leads to a pair of coupled nonlinear differential equations that are solved analytically using a standard iteration procedure. Solutions are obtained for the phase and amplitude signals generated by various acoustic-atomic force microscope (A-AFM) techniques including force modulation microscopy, atomic force acoustic microscopy, ultrasonic force microscopy, heterodyne force microscopy, resonant difference-frequency atomic force ultrasonic microscopy (RDF-AFUM), and the commonly used intermittent contact mode (TappingMode) generally available on AFMs. The solutions are used to obtain a quantitative measure of image contrast resulting from variations in the Young modulus of the sample for the amplitude and phase images generated by the A-AFM techniques. Application of the model to RDF-AFUM and intermittent soft contact phase images of LaRC-cp2 polyimide polymer is discussed. The model predicts variations in the Young modulus of the material of 24 percent from the RDF-AFUM image and 18 percent from the intermittent soft contact image. Both predictions are in good agreement with the literature value of 21 percent obtained from independent, macroscopic measurements of sheet polymer material.

  14. Experimental setup for energy-filtered scanning confocal electron microscopy (EFSCEM) in a double aberration-corrected transmission electron microscope

    Energy Technology Data Exchange (ETDEWEB)

    Wang, P; Behan, G; Kirkland, A I; Nellist, P D, E-mail: peng.wang@materials.ox.ac.u [Department of Materials, University of Oxford, Parks Road, Oxford OX1 3PH (United Kingdom)

    2010-07-01

    Scanning confocal electron microscopy (SCEM) is a new imaging mode in electron microscopy. Spherical aberration corrected electron microscope instruments fitted with two aberration correctors can be used in this mode which provides improved depth resolution and selectivity compared to optical sectioning in a conventional scanning transmission geometry. In this article, we consider a confocal optical configuration for SCEM using inelastically scattered electrons. We lay out the necessary steps for achieving this new operational mode in a double aberration-corrected instrument with uncorrected chromatic aberration and present preliminary experimental results in such mode.

  15. Electropolymerized nanoporous polymeric SPME coatings: preparation and characterization by small angle X-ray scattering and scanning electron microscopy.

    Science.gov (United States)

    Buszewski, Boguslaw; Olszowy, Pawel; Pikus, Stanislaw; Kozak, Maciej

    Polymeric polypyrrole and polythiophene solid phase microextraction (SPME) coatings were prepared using electropolymerization with a linear sweep voltammetry technique. Physicochemical properties were measured using different methods, in particular small angle X-ray scattering and scanning electron microscopy. By using innovative approaches for pore size measurement, we were able to calculate a maximum of the pore size range from 80 to 90 nm. Additionally, film thicknesses measured from 90 to 150 μm. Using scanning electron microscopy, we describe the characteristics of polymer growth on the support surface.

  16. Scanning near-field optical microscopy on rough surfaces: applications in chemistry, biology, and medicine

    Directory of Open Access Journals (Sweden)

    2006-01-01

    Full Text Available Shear-force apertureless scanning near-field optical microscopy (SNOM with very sharp uncoated tapered waveguides relies on the unexpected enhancement of reflection in the shear-force gap. It is the technique for obtaining chemical (materials contrast in the optical image of “real world” surfaces that are rough and very rough without topographical artifacts, and it is by far less complicated than other SNOM techniques that can only be used for very flat surfaces. The experimental use of the new photophysical effect is described. The applications of the new technique are manifold. Important mechanistic questions in solid-state chemistry (oxidation, diazotization, photodimerization, surface hydration, hydrolysis are answered with respect to simultaneous AFM (atomic force microscopy and detailed crystal packing. Prehistoric petrified bacteria and concomitant pyrite inclusions are also investigated with local RAMAN SNOM. Polymer beads and unstained biological objects (rabbit heart, shrimp eye allow for nanoscopic analysis of cell organelles. Similarly, human teeth and a cancerous tissue are analyzed. Bladder cancer tissue is clearly differentiated from healthy tissue without staining and this opens a new highly promising diagnostic tool for precancer diagnosis. Industrial applications are demonstrated at the corrosion behavior of dental alloys (withdrawal of a widely used alloy, harmless substitutes, improvement of paper glazing, behavior of blood bags upon storage, quality assessment of metal particle preparations for surface enhanced RAMAN spectroscopy, and determination of diffusion coefficient and light fastness in textile fiber dyeing. The latter applications include fluorescence SNOM. Local fluorescence SNOM is also used in the study of partly aggregating dye nanoparticles within resin/varnish preparations. Unexpected new insights are obtained in all of the various fields that cannot be obtained by other techniques.

  17. The importance of scanning electron microscopy (sem in taxonomy and morphology of Chironomidae (Diptera

    Directory of Open Access Journals (Sweden)

    Andrzej Kownacki

    2015-07-01

    Full Text Available The paper reports on the value of scanning electron microscopy (SEM in the taxonomy and morphology of Chironomidae. This method has been relatively rarely used in Chironomidae studies. Our studies suggest that the SEM method provides a lot of new information. For example, the plastron plate of the thoracic horn of Macropelopia nebulosa (Meigen under light microscopy is visible as points, while under SEM we have found that it consists of a reticular structure with holes. By using SEM a more precise picture of the body structure of Chironomidae can be revealed. It allows researchers to explain inconsistencies in the existing descriptions of species. Another advantage of the SEM method is obtaining spatial images of the body and organs of Chironomidae. However, the SEM method also has some limitations. The main problem is dirt or debris (e.g. algae, mud, secretions, mucus, bacteria, etc., which often settles on the external surface of structures, especially those which are uneven or covered with hair. The dirt should be removed after collection of chironomid material because if left in place it can become chemically fixed to various surfaces. It unnecessarily remains at the surface and final microscopic images may contain artifacts that obscure chironomid structures being investigated. In this way many details of the surface are thus unreadable. The results reported here indicate that SEM examination helps us to identify new morphological features and details that will facilitate the identification of species of Chironomidae and may help to clarify the function of various parts of the body. Fast development of electron microscope technique allows us to learn more about structure of different organisms.

  18. Neutral Red as a Probe for Confocal Laser Scanning Microscopy Studies of Plant Roots

    Science.gov (United States)

    DUBROVSKY, JOSEPH G.; GUTTENBERGER, MARTIN; SARALEGUI, ANDRES; NAPSUCIALY-MENDIVIL, SELENE; VOIGT, BORIS; BALUŠKA, FRANTIŠEK; MENZEL, DIEDRIK

    2006-01-01

    • Background and Aims Neutral red (NR), a lipophilic phenazine dye, has been widely used in various biological systems as a vital stain for bright-field microscopy. In its unprotonated form it penetrates the plasma membrane and tonoplast of viable plant cells, then due to protonation it becomes trapped in acidic compartments. The possible applications of NR for confocal laser scanning microscopy (CLSM) studies were examined in various aspects of plant root biology. • Methods NR was used as a fluorochrome for living roots of Phaseolus vulgaris, Allium cepa, A. porrum and Arabidopsis thaliana (wild-type and transgenic GFP-carrying lines). The tissues were visualized using CLSM. The effect of NR on the integrity of the cytoskeleton and the growth rate of arabidopsis primary roots was analysed to judge potential toxic effects of the dye. • Key Results The main advantages of the use of NR are related to the fact that NR rapidly penetrates root tissues, has affinity to suberin and lignin, and accumulates in the vacuoles. It is shown that NR is a suitable probe for visualization of proto- and metaxylem elements, Casparian bands in the endodermis, and vacuoles in cells of living roots. The actin cytoskeleton and the microtubule system of the cells, as well as the dynamics of root growth, remain unchanged after short-term application of NR, indicating a relatively low toxicity of this chemical. It was also found that NR is a useful probe for the observation of the internal structures of root nodules and of fungal hyphae in vesicular–arbuscular mycorrhizas. • Conclusions Ease, low cost and absence of tissue processing make NR a useful probe for structural, developmental and vacuole-biogenetic studies of plant roots with CLSM. PMID:16520341

  19. Topographical and electrochemical nanoscale imaging of living cells using voltage-switching mode scanning electrochemical microscopy

    Science.gov (United States)

    Takahashi, Yasufumi; Shevchuk, Andrew I.; Novak, Pavel; Babakinejad, Babak; Macpherson, Julie; Unwin, Patrick R.; Shiku, Hitoshi; Gorelik, Julia; Klenerman, David; Korchev, Yuri E.; Matsue, Tomokazu

    2012-01-01

    We describe voltage-switching mode scanning electrochemical microscopy (VSM-SECM), in which a single SECM tip electrode was used to acquire high-quality topographical and electrochemical images of living cells simultaneously. This was achieved by switching the applied voltage so as to change the faradaic current from a hindered diffusion feedback signal (for distance control and topographical imaging) to the electrochemical flux measurement of interest. This imaging method is robust, and a single nanoscale SECM electrode, which is simple to produce, is used for both topography and activity measurements. In order to minimize the delay at voltage switching, we used pyrolytic carbon nanoelectrodes with 6.5–100 nm radii that rapidly reached a steady-state current, typically in less than 20 ms for the largest electrodes and faster for smaller electrodes. In addition, these carbon nanoelectrodes are suitable for convoluted cell topography imaging because the RG value (ratio of overall probe diameter to active electrode diameter) is typically in the range of 1.5–3.0. We first evaluated the resolution of constant-current mode topography imaging using carbon nanoelectrodes. Next, we performed VSM-SECM measurements to visualize membrane proteins on A431 cells and to detect neurotransmitters from a PC12 cells. We also combined VSM-SECM with surface confocal microscopy to allow simultaneous fluorescence and topographical imaging. VSM-SECM opens up new opportunities in nanoscale chemical mapping at interfaces, and should find wide application in the physical and biological sciences. PMID:22611191

  20. Plasma membrane characterization, by scanning electron microscopy, of multipotent myoblasts-derived populations sorted using dielectrophoresis

    Energy Technology Data Exchange (ETDEWEB)

    Muratore, Massimo, E-mail: M.Muratore@ed.ac.uk [Institute of Integrated Micro and Nano System, School of Engineering, The University of Edinburgh, Edinburgh EH9 3JF (United Kingdom); Mitchell, Steve [Institute of Molecular Plant Science, School of Biological Science, The University of Edinburgh, Edinburgh EH9 3JF (United Kingdom); Waterfall, Martin [Institute of Immunology and Infection Research, School of Biological Science, The University of Edinburgh, Edinburgh EH9 3JT (United Kingdom)

    2013-09-06

    Highlights: •Dielectrophoretic separation/sorting of multipotent cells. •Plasma membrane microvilli structure of C2C12 and fibroblasts by SEM microscopy. •Cell cycle determination by Ki-67 in DEP-sorted cells. •Plasma membrane differences responsible for changes in membrane capacitance. -- Abstract: Multipotent progenitor cells have shown promise for use in biomedical applications and regenerative medicine. The implementation of such cells for clinical application requires a synchronized, phenotypically and/or genotypically, homogenous cell population. Here we have demonstrated the implementation of a biological tag-free dielectrophoretic device used for discrimination of multipotent myoblastic C2C12 model. The multipotent capabilities in differentiation, for these cells, diminishes with higher passage number, so for cultures above 70 passages only a small percentage of cells is able to differentiate into terminal myotubes. In this work we demonstrated that we could recover, above 96% purity, specific cell types from a mixed population of cells at high passage number without any biological tag using dielectrophoresis. The purity of the samples was confirmed by cytometric analysis using the cell specific marker embryonic myosin. To further investigate the dielectric properties of the cell plasma membrane we co-culture C2C12 with similar size, when in suspension, GFP-positive fibroblast as feeder layer. The level of separation between the cell types was above 98% purity which was confirmed by flow cytometry. These levels of separation are assumed to account for cell size and for the plasma membrane morphological differences between C2C12 and fibroblast unrelated to the stages of the cell cycle which was assessed by immunofluorescence staining. Plasma membrane conformational differences were further confirmed by scanning electron microscopy.