Sample records for emission microscopy peem

  1. Ferroelectric domain structures on BaTiO{sub 3}(100) observed by laser-excited PEEM

    Energy Technology Data Exchange (ETDEWEB)

    Hoefer, Anke; Duncker, Klaus; Foerster, Stefan; Widdra, Wolf [Martin-Luther-Universitaet Halle-Wittenberg, Halle (Germany)


    The ferroelectric domain structure at a single crystal BaTiO{sub 3}(100) surface is imaged by photoelectron emission microscopy (PEEM) using a fully tunable femtosecond laser source. For a BaTiO{sub 3}(100) surface which is prepared under UHV conditions by sputtering and annealing in an oxygen atmosphere, at room temperature a stripe like domain pattern is observed with high contrast aligned in the high symmetry [100] direction of the substrate. The PEEM pattern is explained by sequences of 90 -a-c domains. Wavelength-dependent images with UV excitation in the range of 290 - 330 nm show a varying domain contrast and allow the determination of the local photoemission threshold for one ferroelectric domain. For higher laser pulse energies and for wavelengths below the onset of one-photon photoemission, two-photon PEEM images show the ferroelectric domain structure as well.

  2. Time-resolved PEEM measurements on single-crystalline Fe-structures

    Energy Technology Data Exchange (ETDEWEB)

    Kaiser, Alexander; Wiemann, Carsten; Cramm, Stefan; Schneider, Claus M. [Forschungszentrum Juelich, Institut fuer Festkoerperforschung IFF-9, 52425 Juelich (Germany)


    Time-resolved photo-electron emission microscopy (TR-PEEM) provides a method for investigating the spatial and temporal magnetodynamics of micron-sized magnetic elements. By the use of e-beam evaporation thin films of (100)-oriented Iron in bcc-structure can be epitaxially grown on GaAs substrates with a Silver buffer layer. Due to their single-crystallinity the films exhibit a four-fold in-plane magnetocrystalline anisotropy. The films have been microstructured by lithographic techniques and the micromagnetic response on a short magnetic field pulse was investigated by the TR-PEEM technique. Compared to well-studied polycrystalline Permalloy samples the magnetocrystalline anisotropy gives rise to additional terms in the effective magnetic field, leading to different magnetodynamic behaviour. Results of the time-resolved measurements are presented and compared to those of anisotropy-free Permalloy structures.

  3. Operando x-ray photoelectron emission microscopy for studying forward and reverse biased silicon p-n junctions

    Energy Technology Data Exchange (ETDEWEB)

    Barrett, N., E-mail:; Gottlob, D. M.; Mathieu, C.; Lubin, C. [SPEC, CEA, CNRS, Université Paris-Saclay, CEA Saclay, 91191 Gif-sur-Yvette Cedex (France); Passicousset, J. [SPEC, CEA, CNRS, Université Paris-Saclay, CEA Saclay, 91191 Gif-sur-Yvette Cedex (France); IFP Energies nouvelles, Rond-point de l’échangeur de Solaize, BP 3, 69360 Solaize (France); Renault, O.; Martinez, E. [University Grenoble-Alpes, 38000 Grenoble, France and CEA, LETI, MINATEC Campus, 38054 Grenoble (France)


    Significant progress in the understanding of surfaces and interfaces of materials for new technologies requires operando studies, i.e., measurement of chemical, electronic, and magnetic properties under external stimulus (such as mechanical strain, optical illumination, or electric fields) applied in situ in order to approach real operating conditions. Electron microscopy attracts much interest, thanks to its ability to determine semiconductor doping at various scales in devices. Spectroscopic photoelectron emission microscopy (PEEM) is particularly powerful since it combines high spatial and energy resolution, allowing a comprehensive analysis of local work function, chemistry, and electronic structure using secondary, core level, and valence band electrons, respectively. Here we present the first operando spectroscopic PEEM study of a planar Si p-n junction under forward and reverse bias. The method can be used to characterize a vast range of materials at near device scales such as resistive oxides, conducting bridge memories and domain wall arrays in ferroelectrics photovoltaic devices.

  4. Nanoscale Laser Terahertz Emission Microscopy

    DEFF Research Database (Denmark)

    Klarskov, Pernille; Kim, Hyewon; Colvin, Vicki L.


    Laser terahertz emission microscopy (LTEM) has become a powerful tool for studying ultrafast dynamics and local fields in many different types of materials. This technique, which relies on acceleration of charge carriers in a material upon femtosecond excitation, can provide insight into the phys......Laser terahertz emission microscopy (LTEM) has become a powerful tool for studying ultrafast dynamics and local fields in many different types of materials. This technique, which relies on acceleration of charge carriers in a material upon femtosecond excitation, can provide insight...... into the physics of charge transport, built-in fields, grain boundaries or surface states. We describe a new implementation of LTEM with a spatial resolution in the nanoscale regime based on a scattering-type near-field tip-based approach. We observe a spectral reshaping of the signal compared to conventional LTEM......-size-limited spatial resolution of ∼20 nm by imaging a gold nanorod using terahertz emission from the underlying substrate. This work enables for the first time the possibility of performing LTEM measurements on individual nanostructures....

  5. Stimulated emission depletion microscopy on lithographic nanostructures

    Energy Technology Data Exchange (ETDEWEB)

    Westphal, Volker [Department of NanoBiophotonics, Max Planck Institute for Biophysical Chemistry, Goettingen (Germany); Seeger, Jens [Institut fuer Roentgenphysik, Universitaet Goettingen, Goettingen (Germany); Salditt, Tim [Institut fuer Roentgenphysik, Universitaet Goettingen, Goettingen (Germany); Hell, Stefan W [Department of NanoBiophotonics, Max Planck Institute for Biophysical Chemistry, Goettingen (Germany)


    Stimulated emission, predicted by Albert Einstein in 1917, not only prepared the grounds for the invention of the laser, but also for a far-field fluorescence microscopy with diffraction-unlimited resolution. In stimulated emission depletion (STED) microscopy, stimulated emission is not used for light amplification but for a saturated quenching of the fluorescence emission. After demonstrating a five-fold improvement of the lateral (x and y) resolution over the diffraction barrier, we apply STED microscopy to nanostructures of stained PMMA. For the first time, periodic line structures of 80 nm width and 40 nm gaps are resolved with focused visible light.

  6. Preparation of silica films on Ru(0001): A LEEM/PEEM study (United States)

    Klemm, H. W.; Peschel, G.; Madej, E.; Fuhrich, A.; Timm, M.; Menzel, D.; Schmidt, Th.; Freund, H.-J.


    We use an aberration corrected spectro-microscope, the low energy electron microscope/photoelectron emission microscope (LEEM/PEEM) SMART, to follow the preparation and structure of a bilayer silica film on Ru(0001) as a function of temperature and oxidation conditions. This allows us to analyze the growth process at different length scales in order to judge on the overall quality and the morphology of the film. It is found that the film growth occurs in a crystalline and a vitreous phase as previously discovered using scanning tunneling microscopy. However, the present experiment allows an analysis on the sub-micron level to gain insight into the growth process at a mesoscopic scale. We find that the fully oxidized film can be prepared but that this film contains holes. These are unavoidable and are important to consider, if one wants to use the films for ensemble averaging experiments to investigate migration and reaction of molecules between the silica film and the Ru(0001) substrate.

  7. Normal-Incidence PEEM Imaging of Propagating Modes in a Plasmonic Nanocircuit. (United States)

    Razinskas, Gary; Kilbane, Deirdre; Melchior, Pascal; Geisler, Peter; Krauss, Enno; Mathias, Stefan; Hecht, Bert; Aeschlimann, Martin


    The design of noble-metal plasmonic devices and nanocircuitry requires a fundamental understanding and control of the interference of plasmonic modes. Here we report the first visualization of the propagation and interference of guided modes in a showcase plasmonic nanocircuit using normal-incidence nonlinear two-photon photoemission electron microscopy (PEEM). We demonstrate that in contrast to the commonly used grazing-incidence illumination scheme, normal-incidence PEEM provides a direct image of the structure's near-field intensity distribution due to the absence of beating patterns and despite the transverse character of the plasmonic modes. Based on a simple heuristic numerical model for the photoemission yield, we are able to model all experimental findings if global plane wave illumination and coupling to multiple input/output ports, and the resulting interference effects are accounted for.

  8. Standing-wave excited soft x-ray photoemission microscopy: application to Co microdot magnetic arrays

    Energy Technology Data Exchange (ETDEWEB)

    Gray, Alexander; Kronast, Florian; Papp, Christian; Yang, See-Hun; Cramm, Stefan; Krug, Ingo P.; Salmassi, Farhad; Gullikson, Eric M.; Hilken, Dawn L.; Anderson, Erik H.; Fischer, Peter; Durr, Hermann A.; Schneider, Claus M.; Fadley, Charles S.


    We demonstrate the addition of depth resolution to the usual two-dimensional images in photoelectron emission microscopy (PEEM), with application to a square array of circular magnetic Co microdots. The method is based on excitation with soft x-ray standing-waves generated by Bragg reflection from a multilayer mirror substrate. Standing wave is moved vertically through sample simply by varying the photon energy around the Bragg condition. Depth-resolved PEEM images were obtained for all of the observed elements. Photoemission intensities as functions of photon energy were compared to x-ray optical calculations in order to quantitatively derive the depth-resolved film structure of the sample.

  9. Plasmonic nanoprobes for stimulated emission depletion microscopy

    CERN Document Server

    Cortes, Emiliano; Sinclair, Hugo G; Guldbrand, Stina; Peveler, William J; Davies, Timothy; Parrinello, Simona; Görlitz, Frederik; Dunsby, Chris; Neil, Mark A A; Sivan, Yonatan; Parkin, Ivan P; French, Paul M; Maier, Stefan A


    Plasmonic nanoparticles influence the absorption and emission processes of nearby emitters due to local enhancements of the illuminating radiation and the photonic density of states. Here, we use the plasmon resonance of metal nanoparticles in order to enhance the stimulated depletion of excited molecules for super-resolved microscopy. We demonstrate stimulated emission depletion (STED) microscopy with gold nanorods with a long axis of only 26 nm and a width of 8 nm that provide an enhancement of the resolution compared to fluorescent-only probes without plasmonic components irradiated with the same depletion power. These novel nanoparticle-assisted STED probes represent a ~2x10^3 reduction in probe volume compared to previously used nanoparticles and we demonstrate their application to the first plasmon-assisted STED cellular imaging. We also discuss their current limitations.

  10. Design Study on a New Separator for PEEM3

    CERN Document Server

    Wan, Weishi; Padmore, Howard A


    A new aberration-corrected Photoemission Electron Microscope, called PEEM3, is under development at the Advanced Light Source. The resolution and transmission improvement is realized by correcting the lowest order spherical and chromatic aberrations using an electron mirror. A separator is required to separate the incoming uncorrected electron beam to the mirror from the corrected outgoing electron beam to the projector column. In this paper, we present a design study of a new separator for PEEM3. The layout, the Gaussian optics, the analysis of aberrations and the tolerance on power supply stability and alignment errors are reported.

  11. Eliminating deformations in fluorescence emission difference microscopy. (United States)

    You, Shangting; Kuang, Cuifang; Rong, Zihao; Liu, Xu


    We propose a method for eliminating the deformations in fluorescence emission difference microscopy (FED). Due to excessive subtraction, negative values are inevitable in the original FED method, giving rise to deformations. We propose modulating the beam to generate an extended solid focal spot and a hollow focal spot. Negative image values can be avoided by using these two types of excitation spots in FED imaging. Hence, deformations are eliminated, and the signal-to-noise ratio is improved. In deformation-free imaging, the resolution is higher than that of confocal imaging by 32%. Compared to standard FED imaging with the same level of deformations, our method provides superior resolution.

  12. Evanescent excitation and emission in fluorescence microscopy. (United States)

    Axelrod, Daniel


    Evanescent light-light that does not propagate but instead decays in intensity over a subwavelength distance-appears in both excitation (as in total internal reflection) and emission (as in near-field imaging) forms in fluorescence microscopy. This review describes the physical connection between these two forms as a consequence of geometrical squeezing of wavefronts, and describes newly established or speculative applications and combinations of the two. In particular, each can be used in analogous ways to produce surface-selective images, to examine the thickness and refractive index of films (such as lipid multilayers or protein layers) on solid supports, and to measure the absolute distance of a fluorophore to a surface. In combination, the two forms can further increase selectivity and reduce background scattering in surface images. The polarization properties of each lead to more sensitive and accurate measures of fluorophore orientation and membrane micromorphology. The phase properties of the evanescent excitation lead to a method of creating a submicroscopic area of total internal reflection illumination or enhanced-resolution structured illumination. Analogously, the phase properties of evanescent emission lead to a method of producing a smaller point spread function, in a technique called virtual supercritical angle fluorescence. Copyright © 2013 Biophysical Society. Published by Elsevier Inc. All rights reserved.

  13. Using X-PEEM to study biomaterials: Protein and peptide adsorption to a polystyrene-poly(methyl methacrylate)-b-polyacrylic acid blend

    Energy Technology Data Exchange (ETDEWEB)

    Leung, Bonnie O. [Chemistry and Chemical Biology, BIMR, McMaster University, Hamilton, ON, Canada L8S 4M1 (Canada); Hitchcock, Adam P., E-mail: [Chemistry and Chemical Biology, BIMR, McMaster University, Hamilton, ON, Canada L8S 4M1 (Canada); Cornelius, Rena M.; Brash, John L. [School of Biomedical Engineering, McMaster University, Hamilton, ON, Canada L8S 4M1 (Canada); Scholl, Andreas; Doran, Andrew [Advanced Light Source, Berkeley Lab, Berkeley, CA 94720 (United States)


    Highlights: Black-Right-Pointing-Pointer We review applications of synchrotron X-PEEM to biomaterials. Black-Right-Pointing-Pointer We report characterization of a PS/PMMA-b-PAA blend surface by AFM and X-PEEM. Black-Right-Pointing-Pointer We report quantitative mapping of protein (HSA) and peptide adsorption on PS/PMMA-b-PAA. Black-Right-Pointing-Pointer We report how this adsorption changes with pH. -- Abstract: Recent synchrotron-based soft X-ray photoemission electron microscopy (X-PEEM) studies of protein and peptide interaction with phase segregated and patterned polymer surfaces in the context of optimization of candidate biomaterials are reviewed and a study of a new system is reported. X-PEEM and atomic force microscopy (AFM) were used to investigate the morphology of a phase-segregated thin film of a polystyrene/poly(methyl methacrylate)-b-polyacrylic acid (PS/PMMA-PAA) blend, and its interactions with negatively charged human serum albumin (HSA) and positively charged SUB-6 (a cationic antimicrobial peptide, RWWKIWVIRWWR-NH{sub 2}) at several pHs. At neutral pH, where the polymer surface is partially negatively charged, HSA and SUB-6 peptide showed contrasting adsorption behavior which is interpreted in terms of differences in their electrostatic interactions with the polymer surface.

  14. Time-Resolved 2PPE and Time-Resolved PEEM as a Probe of LSP's in Silver Nanoparticles

    Directory of Open Access Journals (Sweden)

    D. Bayer


    Full Text Available The time-resolved two-photon photoemission technique (TR-2PPE has been applied to study static and dynamic properties of localized surface plasmons (LSP in silver nanoparticles. Laterally, integrated measurements show the difference between LSP excitation and nonresonant single electron-hole pair creation. Studies below the optical diffraction limit were performed with the detection method of time-resolved photoemission electron microscopy (TR-PEEM. This microscopy technique with a resolution down to 40 nm enables a systematic study of retardation effects across single nanoparticles. In addition, as will be shown in this paper, it is a highly sensitive sensor for coupling effects between nanoparticles.

  15. Determination of the Goos-Hänchen shift in dielectric waveguides via photo emission electron microscopy in the visible spectrum. (United States)

    Stenmark, Theodore; Word, R C; Könenkamp, R


    Photoemission Electron Microscopy (PEEM) is a versatile tool that relies on the photoelectric effect to produce high-resolution images. Pulse lasers allow for multi-photon PEEM where multiple photons are required excite a single electron. This non-linear process can directly image the near field region of electromagnetic fields in materials. We use this ability here to analyze wave propagation in a linear dielectric waveguide with wavelengths of 410 nm and 780 nm. The propagation constant of the waveguide can be extracted from the interference pattern created by the coupled and incident light and shows distinct polarization dependence. The electromagnetic field interaction at the boundaries can then be deduced which is essential to understand power flow in wave guiding structures. These results match well with simulations using finite element techniques.

  16. In situ, real-time characterization of silicide nanostructure coarsening dynamics by photo-electron emission microscopy (United States)

    Zeman, Matthew Casimir

    Photo-electron emission microscopy (PEEM) was used to observe the growth and coarsening dynamics of transition metal (TM) silicide and rare earth (RE) silicide nanostructures on silicon surfaces in real-time with high lateral resolution during in situ annealing. Evidence was obtained indicating that the coarsening of the silicide islands is strongly influenced by local variations in the size, shape, and number of nanostructures on the surface. The titanium, hafnium, and zirconium silicide nanostructures were observed to grow via Ostwald ripening and attractive migration and coalescence (AMC) at temperatures as high as ~1200°C. Ostwald ripening is a classic coarsening process in which larger nanostructures grow at the expense of smaller surrounding structures as per the Gibbs-Thompson relation. Attractive migration and coalescence is a newly discovered coarsening pathway where nearby islands are observed to migrate attractively towards each other and subsequently coalesce in response to local adatom concentration variations on the surface. A shape distortion of the normally compact and rounded TM silicide islands has been observed during these coarsening processes. The shape distortion suggests that the nanostructures are exchanging mass with each other via diffusion limited processes and these observations support the AMC model. The dysprosium and erbium silicide nanostructures exhibit a distinct faceted morphology and primarily coarsen via Ostwald ripening. The RE silicides form highly elongated nanowires and compact rectangular nanostructures on Si(001) and triangular or hexagonal structures on Si(111). We show that the wires are metastable structures which decay in favor of the larger rectangular islands at high temperatures. Additionally, the rectangular shape and faceted morphology of the RE silicide nanostructures strongly influences their coarsening dynamics. A separate PEEM study explored the thermal stability of thin films of TM oxides (TiO2, ZrO2, HfO2

  17. Saturated virtual fluorescence emission difference microscopy based on detector array (United States)

    Liu, Shaocong; Sun, Shiyi; Kuang, Cuifang; Ge, Baoliang; Wang, Wensheng; Liu, Xu


    Virtual fluorescence emission difference microscopy (vFED) has been proposed recently to enhance the lateral resolution of confocal microscopy with a detector array, implemented by scanning a doughnut-shaped pattern. Theoretically, the resolution can be enhanced by around 1.3-fold compared with that in confocal microscopy. For further improvement of the resolving ability of vFED, a novel method is presented utilizing fluorescence saturation for super-resolution imaging, which we called saturated virtual fluorescence emission difference microscopy (svFED). With a point detector array, matched solid and hollow point spread functions (PSF) can be obtained by photon reassignment, and the difference results between them can be used to boost the transverse resolution. Results show that the diffraction barrier can be surpassed by at least 34% compared with that in vFED and the resolution is around 2-fold higher than that in confocal microscopy.

  18. Ion-induced electron emission microscopy (United States)

    Doyle, Barney L.; Vizkelethy, Gyorgy; Weller, Robert A.


    An ion beam analysis system that creates multidimensional maps of the effects of high energy ions from an unfocussed source upon a sample by correlating the exact entry point of an ion into a sample by projection imaging of the secondary electrons emitted at that point with a signal from a detector that measures the interaction of that ion within the sample. The emitted secondary electrons are collected in a strong electric field perpendicular to the sample surface and (optionally) projected and refocused by the electron lenses found in a photon emission electron microscope, amplified by microchannel plates and then their exact position is sensed by a very sensitive X Y position detector. Position signals from this secondary electron detector are then correlated in time with nuclear, atomic or electrical effects, including the malfunction of digital circuits, detected within the sample that were caused by the individual ion that created these secondary electrons in the fit place.

  19. Experimental assessment of fluorescence microscopy signal enhancement by stimulated emission (United States)

    Dake, Fumihiro; Yazawa, Hiroki


    The quantity of photons generated during fluorescence microscopy is principally determined by the quantum yield of the fluorescence dyes and the optical power of the excitation beam. However, even though low quantum yields can produce poor images, it is challenging to tune this parameter, while increasing the power of the excitation beam often results in photodamage. Here, we propose the use of stimulated emission (SE) as a means of enhancing both the signal intensity and signal-to-noise ratio during confocal fluorescence microscopy. This work experimentally confirmed that both these factors can be enhanced by SE radiation, through generating a greater number of photons than are associated with the standard fluorescence signal. We also propose the concept of stimulated emission enhancing fluorescence (SEEF) microscopy, which employs both the SE and fluorescence signals, and demonstrate that the intensity of an SEEF signal is greater than those of the individual SE and fluorescence signals.

  20. Scanning probe microscopy and field emission schemes for studying electron emission from polycrystalline diamond


    Chubenko, Oksana; Baturin, Stanislav S.; Baryshev, Sergey V.


    The letter introduces a diagram that rationalizes tunneling atomic force microscopy (TUNA) observations of electron emission from polycrystalline diamonds as described in recent publications. The direct observations of electron emission from grain boundary sites by TUNA could indeed be evidence of electrons originating from grain boundaries under external electric fields. At the same time, from the diagram it follows that TUNA and field emission schemes are complimentary rather than equivalen...

  1. Fluorescence microscopy with diffraction resolution barrier broken by stimulated emission


    Klar, Thomas A.; Jakobs, Stefan; Dyba, Marcus; Egner, Alexander; Hell, Stefan W.


    The diffraction barrier responsible for a finite focal spot size and limited resolution in far-field fluorescence microscopy has been fundamentally broken. This is accomplished by quenching excited organic molecules at the rim of the focal spot through stimulated emission. Along the optic axis, the spot size was reduced by up to 6 times beyond the diffraction barrier. The simultaneous 2-fold improvement in the radial direction rendered a nearly spherical fluorescence spot with a diameter of 9...

  2. Stimulated emission depletion microscopy to study amyloid fibril formation (United States)

    Mahou, Pierre; Curry, Nathan; Pinotsi, Dorothea; Kaminski Schierle, Gabriele; Kaminski, Clemens


    Aggregation of misfolded proteins is a characteristic hallmark of many neurodegenerative disorders, such as Parkinson's, Alzheimer's and Huntington's diseases. The ability to observe these aggregation processes and the corresponding structures formed in vitro or in situ is therefore a key requirement to understand the molecular mechanisms of these diseases. We report here on the implementation and application of Stimulated Emission Depletion (STED) microscopy to visualize the formation of amyloid fibrils in vitro.

  3. Structural characterization of colored human iridal melanosomes by photo emission electron microscopy (United States)

    Peles, Dana N.; Hong, Lian; Simon, John D.; Hu, Dan-Ning


    Ocular uveal melanosomes contain both eumelanin and pheomelanin. The ratio of these two melanins has been discussed in relation to the epidemiological data for skin cancer rates, with increased incidence observed for increased relative concentrations of pheomelanin. Recent studies suggest that a similar trend exists underlying the epidemiology of uveal melanomas. In the present study, the biomolecular organization of human iridal melanosomes from different colored irises were examined to determine if the photoreactivity changes with the altered eumelanin:pheomelanin ratio, and whether such changes can account for epidemiological results. Specifically, photoemission electron microscopy (PEEM), a unique surface-sensitive, direct-imaging technique capable of providing chemical information not obtained by other electron microscopies, was used in combination with Duke University's tunable UV free electron laser (FEL) to determine the surface electrochemical properties of melanosomes from blue and dark brown irides. The results demonstrate that the melanins are organized such that pheomelanin is encased by eumelanin. This "casing model" is consistent with kinetic information available on the early steps of melanogenesis and provides new insights into molecular mechanisms underlying the epidemiology of uveal melanoma.

  4. Imaging chromophores with undetectable fluorescence by stimulated emission microscopy. (United States)

    Min, Wei; Lu, Sijia; Chong, Shasha; Roy, Rahul; Holtom, Gary R; Xie, X Sunney


    Fluorescence, that is, spontaneous emission, is generally more sensitive than absorption measurement, and is widely used in optical imaging. However, many chromophores, such as haemoglobin and cytochromes, absorb but have undetectable fluorescence because the spontaneous emission is dominated by their fast non-radiative decay. Yet the detection of their absorption is difficult under a microscope. Here we use stimulated emission, which competes effectively with the nonradiative decay, to make the chromophores detectable, and report a new contrast mechanism for optical microscopy. In a pump-probe experiment, on photoexcitation by a pump pulse, the sample is stimulated down to the ground state by a time-delayed probe pulse, the intensity of which is concurrently increased. We extract the miniscule intensity increase with shot-noise-limited sensitivity by using a lock-in amplifier and intensity modulation of the pump beam at a high megahertz frequency. The signal is generated only at the laser foci owing to the nonlinear dependence on the input intensities, providing intrinsic three-dimensional optical sectioning capability. In contrast, conventional one-beam absorption measurement exhibits low sensitivity, lack of three-dimensional sectioning capability, and complication by linear scattering of heterogeneous samples. We demonstrate a variety of applications of stimulated emission microscopy, such as visualizing chromoproteins, non-fluorescent variants of the green fluorescent protein, monitoring lacZ gene expression with a chromogenic reporter, mapping transdermal drug distributions without histological sectioning, and label-free microvascular imaging based on endogenous contrast of haemoglobin. For all these applications, sensitivity is orders of magnitude higher than for spontaneous emission or absorption contrast, permitting nonfluorescent reporters for molecular imaging.

  5. Fluorescence microscopy with diffraction resolution barrier broken by stimulated emission. (United States)

    Klar, T A; Jakobs, S; Dyba, M; Egner, A; Hell, S W


    The diffraction barrier responsible for a finite focal spot size and limited resolution in far-field fluorescence microscopy has been fundamentally broken. This is accomplished by quenching excited organic molecules at the rim of the focal spot through stimulated emission. Along the optic axis, the spot size was reduced by up to 6 times beyond the diffraction barrier. The simultaneous 2-fold improvement in the radial direction rendered a nearly spherical fluorescence spot with a diameter of 90-110 nm. The spot volume of down to 0.67 attoliters is 18 times smaller than that of confocal microscopy, thus making our results also relevant to three-dimensional photochemistry and single molecule spectroscopy. Images of live cells reveal greater details.

  6. Time-, spin- and energyresolved photoemission microscopy of 3d transition metals

    Energy Technology Data Exchange (ETDEWEB)

    Heitkamp, Bernd; Duerr, H.A.; Eberhardt, W.


    Understanding ultrafast de- and remagnetization processes are of considerable interest, since it allows to shorten the read/write-cycles in magnetism based memories. Our approach is to combine the nm-spatial resolution of a photoelectron emission microscopy (PEEM) with a fs time-resolution using the pump-probe technique. Magnetic sensitivity is obtained by detecting the spin of the emitted photoelectrons. Experiments on nickel and cobalt show a demagnetization below one picosecond. Electron- and spindynamics strongly depend on the dielectric response of the nanostructures.

  7. A Technique for In Situ Ballistic Electron Emission Microscopy (United States)

    Balsano, Robert; Garramone, John; Labella, Vincent


    Ballistic electron emission microscopy (BEEM) is a scanning tunneling microscopy (STM) technique that can measure transport of hot electrons through materials and interfaces with high spatial and energetic resolution. BEEM requires an additional contact to ground the metal base layer of a metal semiconductor junction. Performing BEEM in situ with the sample fabrication requires a custom built STM or modifying a commercial one to facilitate the extra contact, which leaves the technique to highly trained experts. This poster will describe our work to develop a special silicon substrate that has the extra contact built in to enable in situ BEEM without modifications to the STM. Electrically isolated contact traces are lithographically patterned ex situ onto the silicon substrate and connected to the BEEM sample plate which is then inserted into the ultra-high vacuum chamber. The metal is then deposited through a shadow mask and then mounted in situ onto the STM for BEEM measurements. BEEM measurements comparing both in situ and ex situ deposited films will be presented.

  8. Axial ion-electron emission microscopy of IC radiation hardness (United States)

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


    A new system for performing radiation effects microscopy (REM) has been developed at Sandia National Laboratory in Albuquerque. This system combines two entirely new concepts in accelerator physics and nuclear microscopy. A radio frequency quadrupole (RFQ) linac is used to boost the energy of ions accelerated by a conventional Tandem Van de Graaff-Pelletron to velocities of 1.9 MeV/amu. The electronic stopping power for heavy ions is near a maximum at this velocity, and their range is ˜20 μm in Si. These ions therefore represent the most ionizing form of radiation in nature, and are nearly ideal for performing single event effects testing of integrated circuits. Unfortunately, the energy definition of the RFQ-boosted ions is rather poor (˜ a few %), which makes problematic the focussing of such ions to the submicron spots required for REM. To circumvent this problem, we have invented ion electron emission microscopy (IEEM). One can perform REM with the IEEM system without focussing or scanning the ion beam. This is because the position on the sample where each ion strikes is determined by projecting ion-induced secondary electrons at high magnification onto a single electron position sensitive detector. This position signal is then correlated with each REM event. The IEEM system is now mounted along the beam line in an axial geometry so that the ions pass right through the electron detector (which is annular), and all of the electrostatic lenses used for projection. The beam then strikes the sample at normal incidence which results in maximum ion penetration and removes a parallax problem experienced in an earlier system. Details of both the RFQ-booster and the new axial IEEM system are given together with some of the initial results of performing REM on Sandia-manufactured radiation hardened integrated circuits.

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

    National Research Council Canada - National Science Library

    Shaulov, Lihi; Harel, Amnon


    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...

  10. A novel field emission microscopy method to study field emission characteristics of freestanding carbon nanotube arrays (United States)

    Li, Yunhan; Sun, Yonghai; Jaffray, David A.; Yeow, John T. W.


    Field emission (FE) uniformity and the mechanism of emitter failure of freestanding carbon nanotube (CNT) arrays have not been well studied due to the difficulty of observing and quantifying FE performance of each emitter in CNT arrays. Herein a field emission microscopy (FEM) method based on poly(methyl methacrylate) (PMMA) thin film is proposed to study the FE uniformity and CNT emitter failure of freestanding CNT arrays. FE uniformity of freestanding CNT arrays and different levels of FE current contributions from each emitter in the arrays are recorded and visualized. FEM patterns on the PMMA thin film contain the details of the CNT emitter tip shape and whether multiple CNT emitters occur at an emission site. Observation of real-time FE performance and the CNT emitter failure process in freestanding CNT arrays are successfully achieved using a microscopic camera. High emission currents through CNT emitters causes Joule heating and light emission followed by an explosion of the CNTs. The proposed approach is capable of resolving the major challenge of building the relationship between FE performance and CNT morphologies, which can significantly facilitate the study of FE non-uniformity, the emitter failure mechanism and the development of stable and reliable FE devices in practical applications.

  11. Application of a grating coupler for surface plasmon polariton excitation in a photoemission electron microscopy experiment

    DEFF Research Database (Denmark)

    Leißner, Till; Jauernik, Stephan; Lemke, Christoph

    Surface plasmon polariton (SPP) excitation at a gold-vacuum interface via 800 nm light pulses mediated by a periodic array of gold ridges is probed at high lateral resolution by means of photoemission electron microscopy (PEEM). We directly monitor and quantify the coupling properties as a function...... to the grazing incidence excitation geometry intrinsic to a conventional PEEM scheme and the limited propagation distance of the SPP modes at the gold-vacuum interface at the used wavelength....

  12. Laser terahertz emission microscopy with near-field probes

    DEFF Research Database (Denmark)

    Pedersen, Pernille Klarskov; Mittleman, Daniel M.


    Using an AFM, an optical near-field image at 800 nm of a dipole antenna for THz emission is measured, and by simultaneously collecting the emitted THz radiation, the laser light confined under the AFM probe gives a THz emission resolution of less than 50 nm....

  13. 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


    ...) 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...

  14. Immunolabeling for scanning electron microscopy (SEM) and field emission SEM. (United States)

    Goldberg, Martin W


    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.

  15. Towards simultaneous single emission microscopy and magnetic resonance imaging (United States)

    Cai, Liang

    In recent years, the combined nuclear imaging and magnetic resonance imaging (MRI) has drawn extensive research effort. They can provide simultaneously acquired anatomical and functional information inside the human/small animal body in vivo. In this dissertation, the development of an ultrahigh resolution MR-compatible SPECT (Single Photon Emission Computed Tomography) system that can be operated inside a pre-existing clinical MR scanner for simultaneous dual-modality imaging of small animals will be discussed. This system is constructed with 40 small pixel CdTe detector modules assembled in a fully stationary ring SPECT geometry. Series of experiments have demonstrated that this system is capable of providing an imaging resolution of CdTe detector module that we recently developed. Each module consists of CdTe detectors having an overall size of 2.2 cm x 1.1 cm, divided into 64 x 32 pixels of 350 mum in size. A novel hybrid pixel-waveform (HPWF) readout system is also designed to alleviate several challenges for using small-pixel CdTe detectors in ultrahigh-resolution SPECT imaging applications. The HPWF system utilizes a modified version of a 2048-channel 2-D CMOS ASIC to readout the anode pixel, and a digitizing circuitry to sample the signal waveform induced on the cathode. The cathode waveform acquired with the HPWF circuitry offers excellent spatial resolution, energy resolution and depth of interaction (DOI) information, even with the presence of excessive charge-sharing/charge-loss between the small anode pixels. The HPWF CdTe detector is designed and constructed with a minimum amount of ferromagnetic materials, to ensure the MR-compatibility. To achieve sub-500?m imaging resolution, two special designed SPECT apertures have been constructed with different pinhole sizes of 300?m and 500?m respectively. It has 40 pinhole inserts that are made of cast platinum (90%)-iridium (10%) alloy, which provides the maximum stopping power and are compatible with MR

  16. Image scanning fluorescence emission difference microscopy based on a detector array. (United States)

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


    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.

  17. Removal of anti-Stokes emission background in STED microscopy by FPGA-based synchronous detection. (United States)

    Castello, M; Tortarolo, G; Coto Hernández, I; Deguchi, T; Diaspro, A; Vicidomini, G


    In stimulated emission depletion (STED) microscopy, the role of the STED beam is to de-excite, via stimulated emission, the fluorophores that have been previously excited by the excitation beam. This condition, together with specific beam intensity distributions, allows obtaining true sub-diffraction spatial resolution images. However, if the STED beam has a non-negligible probability to excite the fluorophores, a strong fluorescent background signal (anti-Stokes emission) reduces the effective resolution. For STED scanning microscopy, different synchronous detection methods have been proposed to remove this anti-Stokes emission background and recover the resolution. However, every method works only for a specific STED microscopy implementation. Here we present a user-friendly synchronous detection method compatible with any STED scanning microscope. It exploits a data acquisition (DAQ) card based on a field-programmable gate array (FPGA), which is progressively used in STED microscopy. In essence, the FPGA-based DAQ card synchronizes the fluorescent signal registration, the beam deflection, and the excitation beam interruption, providing a fully automatic pixel-by-pixel synchronous detection method. We validate the proposed method in both continuous wave and pulsed STED microscope systems.

  18. Creating infinite contrast in fluorescence microscopy by using lanthanide centered emission

    DEFF Research Database (Denmark)

    R. Carro-Temboury, Miguel; Arppe, Riikka Matleena; Hempel, Casper


    The popularity of fluorescence microscopy arises from the inherent mode of action, where the fluorescence emission from probes is used to visualize selected features on a presumed dark background. However, the background is rarely truly dark, and image processing and analysis is needed to enhance...... for completely removing the background signal in spectrally resolved fluorescence microscopy. The methodology is applicable for all probes with narrow and well-defined emission bands (Full width half-maximum emission lines of europium......(III) and terbium(III) ions. We used a model system with zeolites doped with lanthanides immobilized in a polymer stained with several fluorescent dyes regularly used in bioimaging. After smoothing the spectral data recorded in each pixel, they are differentiated. Method I is based on the direct sum of the gradient...

  19. Rapid subsurface detection of nanoscale defects in live microprocessors by functional infrared emission spectral microscopy. (United States)

    Saloma, Caesar; Tarun, Alvarado; Bailon, Michelle; Soriano, Maricor


    We demonstrate the rapid and nondestructive detection of subsurface nanometer-size defects in 90 nm technology live microprocessors with a new technique called functional infrared emission spectral microscopy. Broken, leaky, and good transistors with similar photoemission images are identified from each other by their different emission spectra that are calculated as linear combinations of weighted basis spectra. The basis spectra are derived from a spectral library by principal component analysis. Leaky transistors do not exhibit apparent morphological damage and are undetectable by optical or scanning probe microscopy alone. The emission signals from two or more transistors combined incoherently, and defect detection is primarily limited by the signal-to-noise ratio of the detected spectrum and not by the separation distance of neighboring transistors.

  20. Photonic and plasmonic surface field distributions characterized with normal- and oblique-incidence multi-photon PEEM. (United States)

    Word, Robert C; Könenkamp, Rolf


    Photonic and plasmonic fields at surfaces can have complicated spatial distributions, which are reflected in the corresponding photoelectron yields imaged in PEEM. These can include the intricate moiré patterns on the surfaces of photonic and plasmonic structures and bright fringe field patterns at their edges. Understanding field distributions requires an understanding of how the guided modes develop, propagate, and interfere with each other and with the incident far-field light. Recent efforts in PEEM include the use of normal incidence excitation in addition to or in lieu of oblique incidence to alter the yield distributions. In this paper we present three cases of surface near-fields imaged in PEEM: an indium tin oxide photonic waveguide, a large gold plasmonic patch antenna, and a small gold plasmonic slot antenna. We show that the surface fields of the waveguide are those of a dual-mode waveguide and that the fields of the plasmonic antennas arise from the asymmetric surface plasmon mode excited at the perimeter of the antennas. We analyze the photoelectron yield distributions and compare and contrast the use of normal and oblique incidence for each case. Copyright © 2017 Elsevier B.V. All rights reserved.

  1. Imaging metazoan nuclear pore complexes by field emission scanning electron microscopy. (United States)

    Fichtman, Boris; Shaulov, Lihi; Harel, Amnon


    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.

  2. Microscopy (United States)

    Patricia A. Moss; Les Groom


    Microscopy is the study and interpretation of images produced by a microscope. "Interpretation" is the keyword, because the microscope enables one to see structures that are too small or too close together to be resolved by the unaided eye. (The human eye cannot separate two points or lines that are closer together than 0.1 mm.) it is important to...

  3. Electron beam confinement and image contrast enhancement in near field emission scanning electron microscopy. (United States)

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


    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.

  4. High-contrast en bloc staining of neuronal tissue for field emission scanning electron microscopy. (United States)

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


    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.

  5. Creating infinite contrast in fluorescence microscopy by using lanthanide centered emission.

    Directory of Open Access Journals (Sweden)

    Miguel R Carro-Temboury

    Full Text Available The popularity of fluorescence microscopy arises from the inherent mode of action, where the fluorescence emission from probes is used to visualize selected features on a presumed dark background. However, the background is rarely truly dark, and image processing and analysis is needed to enhance the fluorescent signal that is ascribed to the selected feature. The image acquisition is facilitated by using considerable illumination, bright probes at a relatively high concentration in order to make the fluorescent signal significantly more intense than the background signal. Here, we present two methods for completely removing the background signal in spectrally resolved fluorescence microscopy. The methodology is applicable for all probes with narrow and well-defined emission bands (Full width half-maximum < 20 nm. Here, we use two lanthanide based probes exploiting the narrow emission lines of europium(III and terbium(III ions. We used a model system with zeolites doped with lanthanides immobilized in a polymer stained with several fluorescent dyes regularly used in bioimaging. After smoothing the spectral data recorded in each pixel, they are differentiated. Method I is based on the direct sum of the gradient, while method II resolves the fluorescent signal by subtracting a background calculated via the gradient. Both methods improve signal-to-background ratio significantly and we suggest that spectral imaging of lanthanide-centered emission can be used as a tool to obtain absolute contrast in bioimaging.

  6. Creating infinite contrast in fluorescence microscopy by using lanthanide centered emission

    DEFF Research Database (Denmark)

    R. Carro-Temboury, Miguel; Arppe, Riikka Matleena; Hempel, Casper


    for completely removing the background signal in spectrally resolved fluorescence microscopy. The methodology is applicable for all probes with narrow and well-defined emission bands (Full width half-maximum lanthanide based probes exploiting the narrow emission lines of europium......(III) and terbium(III) ions. We used a model system with zeolites doped with lanthanides immobilized in a polymer stained with several fluorescent dyes regularly used in bioimaging. After smoothing the spectral data recorded in each pixel, they are differentiated. Method I is based on the direct sum of the gradient......, while method II resolves the fluorescent signal by subtracting a background calculated via the gradient. Both methods improve signal-to-background ratio significantly and we suggest that spectral imaging of lanthanide-centered emission can be used as a tool to obtain absolute contrast in bioimaging....

  7. SISGR: Room Temperature Single-Molecule Detection and Imaging by Stimulated Emission Microscopy

    Energy Technology Data Exchange (ETDEWEB)

    Xie, Xiaoliang Sunney [Harvard Univ., Cambridge, MA (United States). Dept. of Chemistry and Chemical Biology


    Single-molecule spectroscopy has made considerable impact on many disciplines including chemistry, physics, and biology. To date, most single-molecule spectroscopy work is accomplished by detecting fluorescence. On the other hand, many naturally occurring chromophores, such as retinal, hemoglobin and cytochromes, do not have detectable fluorescence. There is an emerging need for single-molecule spectroscopy techniques that do not require fluorescence. In the last proposal period, we have successfully demonstrated stimulated emission microscopy, single molecule absorption, and stimulated Raman microscopy based on a high-frequency modulation transfer technique. These first-of-a- kind new spectroscopy/microscopy methods tremendously improved our ability to observe molecules that fluorescence weakly, even to the limit of single molecule detection for absorption measurement. All of these methods employ two laser beams: one (pump beam) excites a single molecule to a real or virtual excited state, and the other (probe beam) monitors the absorption/emission property of the single. We extract the intensity change of the probe beam with high sensitivity by implementing a high-frequency phase-sensitive detection scheme, which offers orders of magnitude improvement in detection sensitivity over direct absorption/emission measurement. However, single molecule detection based on fluorescence or absorption is fundamentally limited due to their broad spectral response. It is important to explore other avenues in single molecule detection and imaging which provides higher molecular specificity for studying a wide variety of heterogeneous chemical and biological systems. This proposal aimed to achieve single-molecule detection sensitivity with near resonance stimulated Raman scattering (SRS) microscopy. SRS microscopy was developed in our lab as a powerful technique for imaging heterogeneous samples based on their intrinsic vibrational contrasts, which provides much higher molecular

  8. Field emission scanning electron microscopy of biofilm-growing bacteria involved in nosocomial infections. (United States)

    Vuotto, Claudia; Donelli, Gianfranco


    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.

  9. Microstructure-Sensitive Investigation of Fracture Using Acoustic Emission Coupled With Electron Microscopy (United States)

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


    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.

  10. Breaking Abbe's diffraction resolution limit in fluorescence microscopy with stimulated emission depletion beams of various shapes. (United States)

    Klar, T A; Engel, E; Hell, S W


    We report on the generation of various hole-centered beams in the focal region of a lens and investigate their effectiveness to break the diffraction barrier in fluorescence microscopy by stimulated emission. Patterning of the phase of the stimulating beam across the entrance pupil of the objective lens produces point-spread-functions with twofold, fourfold, and circular symmetry, which narrow down the focal spot to 65-100 nm. Comparison with high-resolution confocal images exhibits a resolution much beyond the diffraction barrier. Particles that are only 65-nm apart are resolved with focused light.

  11. Imaging the atomic orbitals of carbon atomic chains with field-emission electron microscopy (United States)

    Mikhailovskij, I. M.; Sadanov, E. V.; Mazilova, T. I.; Ksenofontov, V. A.; Velicodnaja, O. A.


    A recently developed high-field technique of atomic chains preparation has made it possible to attain the ultrahigh resolution of field-emission electron microscopy (FEEM), which can be used to direct imaging the intra-atomic electronic structure. By applying cryogenic FEEM, we are able to resolve the spatial configuration of atomic orbitals, which correspond to quantized states of the end atom in free-standing carbon atomic chains. Knowledge of the intra-atomic structure will make it possible to visualize generic aspects of quantum mechanics and also lead to approaches for a wide range of nanotechnological applications.

  12. Characterization of individual threading dislocations in GaN using ballistic electron emission microscopy. (United States)

    Im, H J; Ding, Y; Pelz, J P; Heying, B; Speck, J S


    Threading dislocations (TDs) of molecular beam epitaxy grown GaN film were studied with ultrahigh vacuum ballistic electron emission microscopy in order to quantify any fixed negative charge at identifiable TDs, with approximately 3 nm spatial and approximately 10 meV local barrier resolution. In contrast to several prior studies, we find no indication of fixed negative dislocation charge at specific TD structures, with a conservative upper limit of approximately 0.25 e(-) per c-axis unit cell. We do observe evidence of positive surface charge at TDs and at GaN step edges, which may be due to local piezoelectric fields.

  13. Parallel excitation-emission multiplexed fluorescence lifetime confocal microscopy for live cell imaging. (United States)

    Zhao, Ming; Li, Yu; Peng, Leilei


    We present a novel excitation-emission multiplexed fluorescence lifetime microscopy (FLIM) method that surpasses current FLIM techniques in multiplexing capability. The method employs Fourier multiplexing to simultaneously acquire confocal fluorescence lifetime images of multiple excitation wavelength and emission color combinations at 44,000 pixels/sec. The system is built with low-cost CW laser sources and standard PMTs with versatile spectral configuration, which can be implemented as an add-on to commercial confocal microscopes. The Fourier lifetime confocal method allows fast multiplexed FLIM imaging, which makes it possible to monitor multiple biological processes in live cells. The low cost and compatibility with commercial systems could also make multiplexed FLIM more accessible to biological research community.

  14. From the physics of secondary electron emission to image contrasts in scanning electron microscopy. (United States)

    Cazaux, Jacques


    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.

  15. Application of a grating coupler for surface plasmon polariton excitation in a photoemission electron microscopy experiment

    DEFF Research Database (Denmark)

    Leißner, Till; Jauernik, Stephan; Lemke, Christoph

    Surface plasmon polariton (SPP) excitation at a gold-vacuum interface via 800 nm light pulses mediated by a periodic array of gold ridges is probed at high lateral resolution by means of photoemission electron microscopy (PEEM). We directly monitor and quantify the coupling properties as a function...

  16. Controlling the emission of organic dyes for high sensitivity and super-resolution microscopy (United States)

    Cordes, Thorben; Stein, Ingo H.; Forthmann, Carsten; Steinhauer, Christian; Walz, Monika; Summerer, Wolfram; Person, Britta; Vogelsang, Jan; Tinnefeld, Philip


    In this paper we show that the emission of ordinary organic dyes can be controlled in order to increase photostability or to induce long off-states for superresolution microscopy. We therefore extend a recently introduced concept that utilizes triplet-state quenching via redox-reactions and recovery of the electronic ground-state by complementary redoxreactions: it is shown that different reagents in an oxidizing and reducing system (ROXS) can positively influence the fluorescence properties of organic dyes. In more detail, the effects of Trolox, a ferrocene-based compound, an oxidized quinone derivative of Trolox and nitrobenzoic acid are investigated and compared to the prototypical compounds ascorbic acid and N,N methylviologen. While the redox potential is the most important parameter for the realization of the ROXS concept it is demonstrated that also kinetic aspects have to be taken into account to explain the properties of the specific redox agents. Photostabilization and the induction of off-states are of paramount importance for fluorescence microscopy in general and especially for superresolution microscopy based on "blinking" molecules.

  17. The lateral photoemission distribution from a defined cluster/substrate system as probed by photoemission electron microscopy

    Energy Technology Data Exchange (ETDEWEB)

    Munzinger, M; Wiemann, C; Rohmer, M; Guo, L; Aeschlimann, M; Bauer, M [Department of Physics, TU Kaiserslautern, Erwin-Schroedingerstr. 46, 67663 Kaiserslautern (Germany)


    We used photoemission electron microscopy (PEEM) to investigate the lateral distribution of the photoemission yield from a defined system of silver clusters supported by a highly oriented pyrolytic graphite (HOPG) substrate. For threshold photoemission using conventional photoemission (PE) and two-photon photoemission (2PPE) we find that distinct, well-separated emitters are responsible for the measured integral photoemission yield. Complementary characterization of the surface using STM shows that the emitter density as probed by PEEM is reduced by about three orders of magnitude in comparison to the actual cluster density. Wavelength and light polarization scans in combination with two-photon-PEEM clearly show that the origin of the 2PPE signal is related to small silver particles. Furthermore, the PEEM differentiates between inhomogeneous and homogeneous broadening effects in the 2PPE signal. This observation allows one to assign the origin of the local photoemission signal to either a distinct single silver particle or a number of coherently coupled silver particles. We conclude that the 2PPE-yield is highly selective with respect to specific properties of the supported silver particles. Our results show that in future experiments, PEEM as a highly local field probe, may be a key tool in the identification of these properties.

  18. 3D fluorescence emission difference microscopy based on spatial light modulator

    Directory of Open Access Journals (Sweden)

    Guangyuan Zhao


    Full Text Available We report three-dimensional fluorescence emission difference (3D-FED microscopy using a spatial light modulator (SLM. Zero phase, 0–2π vortex phase and binary 0-pi phase are loaded on the SLM to generate the corresponding solid, doughnut and z-axis hollow excitation spot, respectively. Our technique achieves super-resolved image by subtracting three differently acquired images with proper subtractive factors. Detailed theoretical analysis and simulation tests are proceeded to testify the performance of 3D-FED. Also, the improvement of lateral and axial resolution is demonstrated by imaging 100nm fluorescent beads. The experiment yields lateral resolution of 140nm and axial resolution of approximate 380nm.

  19. Imaging plant nuclei and membrane-associated cytoskeleton by field emission scanning electron microscopy. (United States)

    Fišerová, Jindřiška; Goldberg, Martin W


    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.

  20. Improved visualization of vertebrate nuclear pore complexes by field emission scanning electron microscopy. (United States)

    Shaulov, Lihi; Harel, Amnon


    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.

  1. Compact three-dimensional super-resolution system based on fluorescence emission difference microscopy (United States)

    Zhu, Dazhao; Chen, Youhua; Fang, Yue; Hussain, Anwar; Kuang, Cuifang; Zhou, Xiaoxu; Xu, Yingke; Liu, Xu


    A compact microscope system for three-dimensional (3-D) super-resolution imaging is presented. The super-resolution capability of the system is based on a size-reduced effective 3-D point spread function generated through the fluorescence emission difference (FED) method. The appropriate polarization direction distribution and manipulation allows the panel active area of the spatial light modulator to be fully utilized. This allows simultaneous modulation of the incident light by two kinds of phase masks to be performed with a single spatial light modulator in order to generate a 3-D negative spot. The system is more compact than standard 3-D FED systems while maintaining all the advantages of 3-D FED microscopy. The experimental results demonstrated the improvement in 3-D resolution by nearly 1.7 times and 1.6 times compared to the classic confocal resolution in the lateral and axial directions, respectively.

  2. Thirty per cent contrast in secondary-electron imaging by scanning field-emission microscopy. (United States)

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


    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.

  3. The use of field emission scanning electron microscopy to assess recombinant adenovirus stability. (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


    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.

  4. Cryo-field emission scanning electron microscopy imaging of a rigid surfactant mesophase. (United States)

    Tan, Grace; Xu, Peng; John, Vijay T; He, Jibao; McPherson, Gary L; Agarwal, Vivek; Bose, Arijit


    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.

  5. Correlative microscopy of Purkinje dendritic spines: a field emission scanning and transmission electron microscopic study. (United States)

    Castejón, O J; Castellano, A; Arismendi, G; Apkarian, R


    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.

  6. Field emission scanning electron microscopy (FE-SEM) as an approach for nanoparticle detection inside cells. (United States)

    Havrdova, M; Polakova, K; Skopalik, J; Vujtek, M; Mokdad, A; Homolkova, M; Tucek, J; Nebesarova, J; Zboril, R


    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.

  7. Ballistic electron emissions microscopy (BEEM) of ferromagnet-semiconductor interfaces; Ballistische Elektronen Emissions Mikroskopie (BEEM) an Ferromagnet-Halbleitergrenzflaechen

    Energy Technology Data Exchange (ETDEWEB)

    Obernhuber, S.


    For current research on spin-transistors it is important to know the characteristics of ferromagnet semiconductor interfaces. The ballistic electron emission microscopy (BEEM) is a method to investigate such a buried interface with nanometer resolution. In this work several ferromagnet/GaAs(110) interfaces have been analysed concerning their homogeneity and mean local Schottky-barrier heights (SBH) have been determined. In Addition, the resulting integral SBH was calculated from the distribution of the local SBHs and compared with the SBH determined from voltage/current characteristics. The areas with a low SBH dominate the current conduction across the interface. Additional BEEM measurements on (AlGaAs/GaAs) heterostructures have been performed. This heterostructures consist of 50 nm AlGaAs/GaAs layers. The results of the BEEM measurements indicate, that the GaAs QWs are defined by AlGaAs barriers. The transition from AlGaAs to GaAs is done within 10 nm. (orig.)

  8. Field-emission scanning electron microscopy analysis of morphology and enzyme distribution within an industrial biocatalytic particle

    NARCIS (Netherlands)

    Roon, van J.L.; Aelst, van A.C.; Schroën, C.G.P.H.; Tramper, J.; Beeftink, H.H.


    Field-emission scanning electron microscopy (FESEM) was used in a technical feasibility study to obtain insight into the internal morphology and the intraparticle enzyme distribution of Assemblase®, an industrial biocatalytic particle containing immobilized penicillin-G acylase. The results were

  9. Profile and Morphology of Fungal Aerosols Characterized by Field Emission Scanning Electron Microscopy (FESEM). (United States)

    Afanou, Komlavi Anani; Straumfors, Anne; Skogstad, Asbjørn; Skaar, Ida; Hjeljord, Linda; Skare, Øivind; Green, Brett James; Tronsmo, Arne; Eduard, Wijnand

    Fungal aerosols consist of spores and fragments with diverse array of morphologies; however, the size, shape, and origin of the constituents require further characterization. In this study, we characterize the profile of aerosols generated from Aspergillus fumigatus, A. versicolor, and Penicillium chrysogenum grown for 8 weeks on gypsum boards. Fungal particles were aerosolized at 12 and 20 L min-1 using the Fungal Spore Source Strength Tester (FSSST) and the Stami particle generator (SPG). Collected particles were analyzed with field emission scanning electron microscopy (FESEM). We observed spore particle fraction consisting of single spores and spore aggregates in four size categories, and a fragment fraction that contained submicronic fragments and three size categories of larger fragments. Single spores dominated the aerosols from A. fumigatus (median: 53%), while the submicronic fragment fraction was the highest in the aerosols collected from A. versicolor (median: 34%) and P. chrysogenum (median: 31%). Morphological characteristics showed near spherical particles that were only single spores, oblong particles that comprise some spore aggregates and fragments (3.5 μm). Further, the near spherical particles dominated the aerosols from A. fumigatus (median: 53%), while oblong particles were dominant in the aerosols from A. versicolor (68%) and P. chrysogenum (55%). Fiber-like particles represented 21% and 24% of the aerosols from A. versicolor and P. chrysogenum, respectively. This study shows that fungal particles of various size, shape, and origin are aerosolized, and supports the need to include a broader range of particle types in fungal exposure assessment.

  10. Ion milling coupled field emission scanning electron microscopy reveals current misunderstanding of morphology of polymeric nanoparticles. (United States)

    Francis, Donny; Mouftah, Samiha; Steffen, Robert; Beduneau, Arnaud; Pellequer, Yann; Lamprecht, Alf


    Nanoparticles (NPs) are currently used as drug delivery systems for numerous therapeutic macromolecules, e.g. proteins or DNA. Based on the preparation by double emulsion solvent evaporation a sponge-like structure was postulated entrapping hydrophilic drugs inside an internal aqueous phase. However, a direct proof of this hypothesized structure is still missing today. NPs were prepared from different polymers using a double-emulsion method and characterized for their physicochemical properties. Combining ion milling with field emission scanning electron microscopy allowed to cross section single NP and to visualize their internal morphology. The imaging procedure permitted cross-sectioning of NPs and visualization of the internal structure as well as localizing drugs associated with NPs. It was observed that none of the model actives was encapsulated inside the polymeric matrix when particle diameters were below around 470 nm but predominantly adsorbed to the particle surface. Even at larger diameters only a minority of particles of a diameter below 1 μm contained an internal phase. The properties of such drug loaded NPs, i.e. drug release or the observations in cellular uptake or even drug targeting needs to be interpreted carefully since in most cases NP surface properties are potentially dominated by the 'encapsulated' drug characteristics. Copyright © 2014 Elsevier B.V. All rights reserved.

  11. Microbe repelling coated stainless steel analysed by field emission scanning electron microscopy and physicochemical methods. (United States)

    Raulio, Mari; Järn, Mikael; Ahola, Juhana; Peltonen, Jouko; Rosenholm, Jarl B; Tervakangas, Sanna; Kolehmainen, Jukka; Ruokolainen, Timo; Narko, Pekka; Salkinoja-Salonen, Mirja


    Coating of stainless steel with diamond-like carbon or certain fluoropolymers reduced or almost eliminated adhesion and biofilm growth of Staphylococcus epidermidis, Deinococcus geothermalis, Meiothermus silvanus and Pseudoxanthomonas taiwanensis. These species are known to be pertinent biofilm formers on medical implants or in the wet-end of paper machines. Field emission scanning electron microscopic analysis showed that Staph. epidermidis, D. geothermalis and M. silvanus grew on stainless steel using thread-like organelles for adhesion and biofilm formation. The adhesion threads were fewer in number on fluoropolymer-coated steel than on plain steel and absent when the same strains were grown in liquid culture. Psx. taiwanensis adhered to the same surfaces by a mechanism involving cell ghosts on which the biofilm of live cells grew. Hydrophilic (diamond-like carbon) or hydrophobic (fluoropolymer) coatings reduced the adherence of the four test bacteria on different steels. Selected topographic parameters, including root-mean-square roughness (S (q)), skewness (S (sk)) and surface kurtosis (S (ku)), were analysed by atomic force microscopy. The surfaces that best repelled microbial adhesion of the tested bacteria had higher skewness values than those only slightly repelling. Water contact angle, measured (theta (m)) or roughness corrected (theta (y)), affected the tendency for biofilm growth in a different manner for the four test bacteria.

  12. Immunogold labeling of amelogenin in developing porcine enamel revealed by field emission scanning electron microscopy. (United States)

    Du, Chang; Fan, Daming; Sun, Zhi; Fan, Yuwei; Lakshminarayanan, Rajamani; Moradian-Oldak, Janet


    The present study describes a method using immunohistochemical labeling in combination with high-resolution imaging (field emission scanning electron microscopy) to investigate the spatial localization of amelogenins on apatite crystallites in developing porcine enamel. Cross-sections of developing enamel tissue from freeze-fractured pig third molar were treated with antiserum against recombinant mouse amelogenin and immunoreactivity confirmed by Western blot analysis. The samples were then treated with the goat anti-rabbit IgG conjugated with 10-nm gold particles. The control samples were treated with the secondary antibody only. The in-lens secondary electrons detector and quadrant back-scattering detector were employed to reveal the high-resolution morphology of enamel structures and gold particle distribution. The immunolabeling showed a preference of the gold particle localization along the side faces of the ribbon-like apatite crystals. The preferential localization of amelogenin in vivo on enamel crystals strongly supports its direct function in controlling crystal morphology. Copyright 2008 S. Karger AG, Basel.

  13. Indirect Immunodetection of Fungal Fragments by Field Emission Scanning Electron Microscopy. (United States)

    Afanou, Komlavi Anani; Straumfors, Anne; Skogstad, Asbjørn; Nayak, Ajay P; Skaar, Ida; Hjeljord, Linda; Tronsmo, Arne; Eduard, Wijnand; Green, Brett James


    Submicronic fungal fragments have been observed in in vitro aerosolization experiments. The occurrence of these particles has therefore been suggested to contribute to respiratory health problems observed in mold-contaminated indoor environments. However, the role of submicronic fragments in exacerbating adverse health effects has remained unclear due to limitations associated with detection methods. In the present study, we report the development of an indirect immunodetection assay that utilizes chicken polyclonal antibodies developed against spores from Aspergillus versicolor and high-resolution field emission scanning electron microscopy (FESEM). Immunolabeling was performed with A. versicolor fragments immobilized and fixed onto poly-l-lysine-coated polycarbonate filters. Ninety percent of submicronic fragments and 1- to 2-μm fragments, compared to 100% of >2-μm fragments generated from pure freeze-dried mycelial fragments of A. versicolor, were positively labeled. In proof-of-concept experiments, air samples collected from moldy indoor environments were evaluated using the immunolabeling technique. Our results indicated that 13% of the total collected particles were derived from fungi. This fraction comprises 79% of the fragments that were detected by immunolabeling and 21% of the spore particles that were morphologically identified. The methods reported in this study enable the enumeration of fungal particles, including submicronic fragments, in a complex heterogeneous environmental sample. Copyright © 2015, American Society for Microbiology. All Rights Reserved.

  14. Toward Fourier interferometry fluorescence excitation/emission imaging of malignant cells combined with photoacoustic microscopy (United States)

    Kohen, Elli; Hirschberg, Joseph G.; Berry, John P.; Ozkutuk, Nuri; Ornek, Ceren; Monti, Marco; Leblanc, Roger M.; Schachtschabel, Dietrich O.; Haroon, Sumaira


    Dual excitation fluorescence imaging has been used as a first step towards multi-wavelength excitation/emission fluorescence spectral imaging. Target cells are transformed keratinocytes, and other osteosarcoma, human breast and color cancer cells. Mitochondrial membrane potential probes, e.g. TMRM (tetramethylrhodamine methyl ester), Mitotracker Green (Molecular Probes, Inc., Eugene OR,USA; a recently synthesized mitochondrial oxygen probe, [PRE,P1"- pyrene butyl)-2-rhodamine ester] allow dual excitation in the UV plus in teh blue-green spectral regions. Also, using the natural endogenous probe NAD(P)H, preliminary results indicate mitochondrial responses to metabolic challenges (e.g. glucose addition), plus changes in mitochonrial distribution and morphology. In terms of application to biomedicine (for diagnostiscs, prognostsics and drug trials) three parameters have been selected in addition to the natural probe NAD(P)H, i.e. vital fluorescence probing of mitochondria, lysosomes and Golgi apparatus. It is hoped that such a multiparameter approach will allow malignant cell characterization and grading. A new area being introduced is the use of similar methodology for biotechnical applications such as the study of the hydrogen-producing alga Chlamydomonas Reinhardtii, and possible agricultural applications, such as Saccharomyces yeast for oenology. Complementation by Photoacoustic Microscopy is also contemplated, to study the internal conversion component which follows the excitation by photons.

  15. An adjustable electron achromat for cathode lens microscopy

    Energy Technology Data Exchange (ETDEWEB)

    Tromp, R.M., E-mail: [IBM T.J. Watson Research Center, 1101 Kitchawan Road, Yorktown Heights, NY 10598 (United States); Leiden Institute of Physics, Kamerlingh Onnes Laboratory, Niels Bohrweg 2, 2333 CA Leiden (Netherlands)


    Chromatic aberration correction in light optics began with the invention of a two-color-corrected achromatic crown/flint lens doublet by Chester Moore Hall in 1730. Such color correction is necessary because any single glass shows dispersion (i.e. its index of refraction changes with wavelength), which can be counteracted by combining different glasses with different dispersions. In cathode lens microscopes (such as Photo Electron Emission MicroscopyPEEM) we encounter a similar situation, where the chromatic aberration coefficient of the cathode lens shows strong dispersion, i.e. depends (non-linearly) on the energy with which the electrons leave the sample. Here I show how a cathode lens in combination with an electron mirror can be configured as an adjustable electron achromat. The lens/mirror combination can be corrected at two electron energies by balancing the settings of the electron mirror against the settings of the cathode lens. The achromat can be adjusted to deliver optimum performance, depending on the requirements of a specific experiment. Going beyond the achromat, an apochromat would improve resolution and transmission by a very significant margin. I discuss the requirements and outlook for such a system, which for now remains a wish waiting for fulfilment. - Highlights: • The properties of cathode objective lens plus electron mirror are discussed. • In analogy with light-optical achromats, cathode lens plus mirror can be configured as an electron achromat. • Unlike light optics, the electron achromat can be adjusted to best fulfill experimental requirements.

  16. Photoemission Electron Microscopy as a Tool for Studying Steel Grains (United States)

    Roese, Peter; Keutner, Christoph; Berges, Ulf; Espeter, Philipp; Westphal, Carsten


    Key properties of steel like stability, weldability, or ability for absorbing deformation energy are defined by their grain structure. The knowledge about their micrometer and submicrometer structure is of particular interest for tailor-cut macroscopic steel properties. We report on photoemission electron microscopy studies which in principle yield a higher magnification than comparable optical techniques. A flat surface without any topographic features was obtained by applying a non-etching preparation procedure. PEEM images showed very tiny phase islands embedded within a steel phase matrix. Furthermore, we developed an analysis procedure for PEEM images for dual-phase steels. As a result, it is possible to identify the individual work functions of different steel phases at the surface.

  17. Measuring Exciton Migration in Conjugated Polymer Films with Ultrafast Time Resolved Stimulated Emission Depletion Microscopy (United States)

    Penwell, Samuel

    Conjugated polymers are highly tunable organic semiconductors, which can be solution processed to form thin films, making them prime candidates for organic photovoltaic devices. One of the most important parameters in a conjugated polymer solar cell is the exciton diffusion length, which depends on intermolecular couplings, and is typically on the order of 10 nm. This mean exciton migration can vary dramatically between films and within a single film due to heterogeneities in morphology on length scales of 10's to 100's nm. To study the variability of exciton diffusion and morphology within individual conjugated polymer films, we are adapting stimulated emission depletion (STED) microscopy. STED is typically used in biology with sparse well-engineered fluorescent labels or on NV-centers in diamond. I will, however, describe how we have demonstrated the extension of STED to conjugated polymer films and nanoparticles of MEH-PPV and CN-PPV, despite the presence of two photon absorption, by taking care to first understand the material's photophysical properties. We then further adapt this approach, by introducing a second ultrafast STED pulse at a variable delay. Excitons that migrate away from the initial subdiffraction excitation volume during the ps-ns time delay, are preferentially quenched by the second STED pulse, while those that remain in the initial volume survive. The resulting effect of the second STED pulse is modulated by the degree of migration over the ultrafast time delay, thus providing a new method to study exciton migration. Since this technique utilizes subdiffraction optical excitation and detection volumes with ultrafast time resolution, it provides a means of spatially and temporally resolving measurements of exciton migration on the native length and time scales. In this way, we will obtain a spatiotemporal map of exciton distributions and migration that will help to correlate the energetic landscape to film morphology at the nanoscale.

  18. Quantum well state of cubic inclusions in hexagonal silicon carbide studied with ballistic electron emission microscopy (United States)

    Ding, Yi

    SiC is a polytypic material that may crystallize in many different close-packing sequences with cubic, hexagonal, or rhombohedral Bravais lattices. All SiC polytypes have wide bandgaps ranging from 2.39 eV in cubic SiC to 3.023--3.330 eV in common hexagonal polytypes. This, as well as many other properties favorable to electrical applications, makes SiC a very promising material in electronic device fabrication. However, the many lattice stacking sequences may impair the stability of SiC devices. In the hexagonal 4H polytype, it has been found that thin cubic SiC inclusions may be formed due to stacking fault expansion, and it has been proposed that the inclusions may behave as quantum wells because of the lower bandgap of cubic SiC. We performed ultra-high vacuum ballistic electron emission microscopy (BEEM) measurements on n-type 4H-SiC samples containing double-stacking-fault cubic inclusions to characterize the electrical properties of individual inclusions. Thin Pt films are deposited in ultra-high vacuum on the sample surfaces to form Schottky contacts. A Schottky barrier height of ˜1.01 eV is observed over the inclusions in a background of normal 4H-SiC barrier height of 1.54 eV, which directly confirms the cubic inclusions support two-dimensional propagating quantum well states, and the 0.53 eV lowering of barrier height indicates the two-dimensional conduction band minimum is located at ˜0.53 eV below the conduction band minimum of bulk 4H-SIC. We also used BEEM to study the Schottky contact between Pt and p-type 4H-SiC, and observed a second transmission channel in the BEEM spectrum that suggests a split-off valence band at ˜0.11 eV below the valence band maximum. We also measured the barrier heights of p-type and n-type Schottky contacts prepared under identical conditions and the results suggest the existence of an interfacial layer. An earlier study of threading dislocations in GaN using BEEM is also described. Although threading dislocations in Ga

  19. Davisson-Germer Award Talk: Surface Electron Microscopy with Slow Electrons (United States)

    Bauer, Ernst


    Nearly 80 years ago Davisson and Germer demonstrated the diffraction of slow electrons from surfaces but it is only about 20 years that these electrons have been used for imaging of surfaces and thin films in the Low Energy Electron Microscope (LEEM). Since then several other surface imaging methods with slow electrons have emerged, in particular synchrotron radiation excited photo emission electron microscopy (XPEEM). In LEEM the high intensity of the diffracted slow electrons allows fast image acquisition. Therefore it is tempting to combine it with the other, slower complementary methods. This has been accomplished in the Spectroscopic Photo Emission and Low Energy Electron Microscope (SPELEEM) by adding an energy filter. Today the SPELEEM allows comprehensive structural, chemical, magnetic, electronic characterization of surfaces and thin films by imaging with 10 nm lateral resolution and atomic depth resolution, diffraction and spectroscopy. Recent developments are expected to push the resolution limit into the 1 nm range by aberration correction and the time resolution into and below the picosecond range by pulsed illumination and time-delayed triggered detection. The talk will first describe the general imaging principles and then illustrate with a number of examples the possibilities and limitations of some of the methods, LEEM, Spin-Polarized LEEM (SPLEEM) and X- ray Magnetic Dichroism PEEM (XMCDPEEM). A brief outlook will conclude the presentation.

  20. Precision targeted ruthenium(ii) luminophores; highly effective probes for cell imaging by stimulated emission depletion (STED) microscopy. (United States)

    Byrne, Aisling; Burke, Christopher S; Keyes, Tia E


    Fluorescence microscopy has undergone a dramatic evolution over the past two decades with development of super-resolution far-field microscopy methods that break the light diffraction limited resolution of conventional microscopy, offering unprecedented opportunity to interrogate cellular processes at the nanoscale. However, these methods make special demands of the luminescent agents used for contrast and development of probes suited to super-resolution fluorescent methods is still relatively in its infancy. In spite of their many photophysical advantages, metal complex luminophores have not yet been considered as probes in this regard, where to date, only organic fluorophores have been applied. Here, we report the first examples of metal complex luminophores applied as probes for use in stimulated emission depletion (STED) microscopy. Exemplified with endoplasmic reticulum and nuclear targeting complexes we demonstrate that luminescent Ru(ii) polypyridyl complexes can, through signal peptide targeting, be precisely and selectively delivered to key cell organelles without the need for membrane permeabilization, to give high quality STED images of these organelles. Detailed features of the tubular ER structure are revealed and in the case of the nuclear targeting probe we exploit the molecular light switch properties of a dipyrido[3,2-a:2',3'-c]phenazine containing complex which emits only on DNA/RNA binding to give outstanding STED contrast and resolution of the chromosomes within the nucleus. Comparing performance with a member of the AlexaFluor family commonly recommended for STED, we find that the performance of the ruthenium complexes is superior across both CW and gated STED microscopy methods in terms of image resolution and photostability. The large Stokes shifts of the Ru probes permit excellent matching of the stimulating depletion laser with their emission whilst avoiding anti-Stokes excitation. Their long lifetimes make them particularly amenable to

  1. Carbon Nanotube Emissions from Arc Discharge Production: Classification of Particle Types with Electron Microscopy and Comparison with Direct Reading Techniques. (United States)

    Ludvigsson, Linus; Isaxon, Christina; Nilsson, Patrik T; Tinnerberg, Hakan; Messing, Maria E; Rissler, Jenny; Skaug, Vidar; Gudmundsson, Anders; Bohgard, Mats; Hedmer, Maria; Pagels, Joakim


    An increased production and use of carbon nanotubes (CNTs) is occurring worldwide. In parallel, a growing concern is emerging on the adverse effects the unintentional inhalation of CNTs can have on humans. There is currently a debate regarding which exposure metrics and measurement strategies are the most relevant to investigate workplace exposures to CNTs. This study investigated workplace CNT emissions using a combination of time-integrated filter sampling for scanning electron microscopy (SEM) and direct reading aerosol instruments (DRIs). Field measurements were performed during small-scale manufacturing of multiwalled carbon nanotubes using the arc discharge technique. Measurements with highly time- and size-resolved DRI techniques were carried out both in the emission and background (far-field) zones. Novel classifications and counting criteria were set up for the SEM method. Three classes of CNT-containing particles were defined: type 1: particles with aspect ratio length:width >3:1 (fibrous particles); type 2: particles without fibre characteristics but with high CNT content; and type 3: particles with visible embedded CNTs. Offline sampling using SEM showed emissions of CNT-containing particles in 5 out of 11 work tasks. The particles were classified into the three classes, of which type 1, fibrous CNT particles contributed 37%. The concentration of all CNT-containing particles and the occurrence of the particle classes varied strongly between work tasks. Based on the emission measurements, it was assessed that more than 85% of the exposure originated from open handling of CNT powder during the Sieving, mechanical work-up, and packaging work task. The DRI measurements provided complementary information, which combined with SEM provided information on: (i) the background adjusted emission concentration from each work task in different particle size ranges, (ii) identification of the key procedures in each work task that lead to emission peaks, (iii

  2. Radiative decay engineering 8: Coupled emission microscopy for lens-free high-throughput fluorescence detection. (United States)

    Zhu, Liangfu; Badugu, Ramachandram; Zhang, Douguo; Wang, Ruxue; Descrovi, Emiliano; Lakowicz, Joseph R


    Fluorescence spectroscopy and imaging are now used throughout the biosciences. Fluorescence microscopes, spectrofluorometers, microwell plate readers and microarray imagers all use multiple optical components to collect, redirect and focus the emission onto single point or array imaging detectors. For almost all biological samples, except those with regular nanoscale features, emission occurs in all directions. With the exception of complex microscope objectives with large collection angles (NA ≤ 0.5), all these instruments collect only a small fraction of the total emission. Because of the increasing knowledge base on fluorophores within near-field (fluorescence or leakage radiation from nanostructures. Copyright © 2017 Elsevier Inc. All rights reserved.

  3. Microstructure of Monoplacophora (Mollusca) shell examined by low-voltage field emission scanning electron and atomic force microscopy. (United States)

    Cruz, Renato; Weissmüller, Gilberto; Farina, Marcos


    The shell of Micropilina arntzi (Mollusca: Monoplacophora), a primitive molluscan class, was examined by using field emission scanning electron microscopy (FESEM) at low voltage and atomic force microscopy (AFM). The use of these two techniques allowed the observation of fine details of Micropilina arntzi shell and contributed to bring new features concerning the study of molluscan shell microtexture. Imaging with low-voltage FESEM provided well-defined edge contours of shell structures, while analyzing the sample with AFM gave information about the step height of stacked internal structures as well as the dimension of the particles present in their surface at a nanometric level. The shell microstructure of Monoplacophora species presents different patterns and may be a taxonomic implication in the systematic studies of the group.

  4. A study of internal oxidation in carburized steels by glow discharge optical emission spectroscopy and scanning electron microscopy

    CERN Document Server

    An, X; Rainforth, W M; Chen, L


    The internal oxidation of Cr-Mn carburizing steel was studied. Internal oxidation was induced using a commercial carburizing process. Sputter erosion coupled with glow discharge optical emission spectroscopy (GDOES) was used to determine the depth profile elemental distribution within the internal oxidation layer (<10 mu m). In addition, scanning electron microscopy (SEM) equipped with energy dispersive spectrometer (EDS) studies were carried out on selected sputter eroded surfaces. Oxide type was identified primarily by transmission electron microscopy (TEM). The carburized surface was found to consist of a continuous oxide layer, followed by a complex internal oxidation layer, where Cr and Mn oxides were found to populate grain boundaries in a globular form in the near surface region. At greater depths (5-10 mu m), Si oxides formed as a grain boundary network. The internal oxides (mainly complex oxides) grew quickly during the initial stages of the carburizing process (2 h, 800 deg. C+3 h, 930 deg. C). G...

  5. The tip-sample water bridge and light emission from scanning tunnelling microscopy


    Boyle, Michael G; Mitra, J; Dawson, Paul


    Light emission spectrum from a scanning tunnelling microscope (LESTM) is investigated as a function of relative humidity and shown to be a novel and sensitive means for probing the growth and properties of a water meniscus in the nm-scale. An empirical model of the light emission process is formulated and applied successfully to replicate the decay in light intensity and spectral changes observed with increasing relative humidity. The modelling indicates a progressive water filling of the tip...

  6. Strategies and results of field emission scanning electron microscopy (FE-SEM) in the study of parasitic protozoa. (United States)

    de Souza, Wanderley; Campanati, Loraine; Attias, Marcia


    Field emission scanning electron microscopy (FE-SEM) provides a range of strategies for investigating the structural organization of biological systems, varying from isolated macromolecules to tissue organization and whole organisms. This review covers some of the results so far obtained using FE-SEM observation and various protocols of sample fixation to analyze the structural organization of parasitic protozoa and their interaction with host cells. The employment of FE-SEM can be broadened through the use of gold-labeled molecules or tracers, gradual extraction by detergents, and cleavage techniques. These analyses provide significant contributions to the characterization of these organisms concerning ultrastructure, cytoskeleton, motility and intracellular behavior.

  7. Depth-resolved soft x-ray photoelectron emission microscopy in nanostructures via standing-wave excited photoemission

    Energy Technology Data Exchange (ETDEWEB)

    Kronast, F.; Ovsyannikov, R.; Kaiser, A.; Wiemann, C.; Yang, S.-H.; Locatelli, A.; Burgler, D.E.; Schreiber, R.; Salmassi, F.; Fischer, P.; Durr, H.A.; Schneider, C.M.; Eberhardt, W.; Fadley, C.S.


    We present an extension of conventional laterally resolved soft x-ray photoelectron emission microscopy. A depth resolution along the surface normal down to a few {angstrom} can be achieved by setting up standing x-ray wave fields in a multilayer substrate. The sample is an Ag/Co/Au trilayer, whose first layer has a wedge profile, grown on a Si/MoSi2 multilayer mirror. Tuning the incident x-ray to the mirror Bragg angle we set up standing x-ray wave fields. We demonstrate the resulting depth resolution by imaging the standing wave fields as they move through the trilayer wedge structure.

  8. 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.


    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

  9. Spectrally resolved confocal microscopy using lanthanide centred near-IR emission. (United States)

    Liao, Zhiyu; Tropiano, Manuel; Mantulnikovs, Konstantins; Faulkner, Stephen; Vosch, Tom; Sørensen, Thomas Just


    The narrow, near infrared (NIR) emission from lanthanide ions has attracted great interest, particularly with regard to developing tools for bioimaging, where the long lifetimes of lanthanide excited states can be exploited to address problems arising from autofluorescence and sample transparency. Despite the promise of lanthanide-based probes for near-IR imaging, few reports on their use are present in the literature. Here, we demonstrate that images can be recorded by monitoring NIR emission from lanthanide complexes using detectors, optical elements and a microscope that were primarily designed for the visible part of the spectrum.

  10. Spectrally resolved confocal microscopy using lanthanide centred near-IR emission

    DEFF Research Database (Denmark)

    Liao, Zhiyu; Tropiano, Manuel; Mantulnikovs, Konstantins


    The narrow, near infrared (NIR) emission from lanthanide ions has attracted great interest, particularly with regard to developing tools for bioimaging, where the long lifetimes of lanthanide excited states can be exploited to address problems arising from autofluorescence and sample transparency....... Despite the promise of lanthanide-based probes for near-IR imaging, few reports on their use are present in the literature. Here, we demonstrate that images can be recorded by monitoring NIR emission from lanthanide complexes using detectors, optical elements and a microscope that were primarily designed...

  11. Field emission scanning electron microscopy (FE-SEM) as an approach for nanoparticle detection inside cells

    Czech Academy of Sciences Publication Activity Database

    Havrdová, M.; Poláková, K.; Skopalík, J.; Vůjtek, M.; Mokdad, A.; Homolková, M.; Tuček, J.; Nebesářová, Jana; Zbořil, R.


    Roč. 67, DEC 2014 (2014), s. 149-154 ISSN 0968-4328 Institutional support: RVO:60077344 Keywords : Field emission scanning electronmicroscopy (FE-SEM) * Stem cells * Iron oxide nanoparticles * Cellular morphology * Endosomes * Cell uptake Subject RIV: FD - Oncology ; Hematology Impact factor: 1.988, year: 2014

  12. Plasmonic near-electric field enhancement effects in ultrafast photoelectron emission: correlated spatial and laser polarization microscopy studies of individual Ag nanocubes. (United States)

    Grubisic, Andrej; Ringe, Emilie; Cobley, Claire M; Xia, Younan; Marks, Laurence D; Van Duyne, Richard P; Nesbitt, David J


    Electron emission from single, supported Ag nanocubes excited with ultrafast laser pulses (λ = 800 nm) is studied via spatial and polarization correlated (i) dark field scattering microscopy (DFM), (ii) scanning photoionization microscopy (SPIM), and (iii) high-resolution transmission electron microscopy (HRTEM). Laser-induced electron emission is found to peak for laser polarization aligned with cube diagonals, suggesting the critical influence of plasmonic near-field enhancement of the incident electric field on the overall electron yield. For laser pulses with photon energy below the metal work function, coherent multiphoton photoelectron emission (MPPE) is identified as the most probable mechanism responsible for electron emission from Ag nanocubes and likely metal nanoparticles/surfaces in general.

  13. X-ray photoelectron emission spectromicroscopic analysis of arborescent lycopsid cell wall composition and Carboniferous coal ball preservation

    Energy Technology Data Exchange (ETDEWEB)

    Boyce, C. Kevin [Department of the Geophysical Sciences, University of Chicago, Chicago, IL 60637 (United States); Abrecht, Mike; Zhou, Dong; Gilbert, P.U.P.A. [Department of Physics, University of Wisconsin, Madison, WI 53706 (United States)


    Two alternative processes complicate understanding of the biochemical origins and geochemical alteration of organic matter over geologic time: selective preservation of original biopolymers and in situ generation of new geopolymers. One of the best constrained potential sources of bio- and geochemical information about extinct fossil plants is frequently overlooked. Permineralized anatomically preserved plant fossils allow analysis of individual cell and tissue types that have an original biochemical composition already known from living plants. The original composition of more enigmatic fossils can be constrained by geochemical comparisons to tissues of better understood fossils from the same locality. This strategy is possible using synchrotron-based techniques for submicron-scale imaging with X-rays over a range of frequencies in order to provide information concerning the relative abundance of different organic bonds with X-ray Absorption Near Edge Spectroscopy. In this study, X-ray PhotoElectron Emission spectroMicroscopy (X-PEEM) was used to analyze the tissues of Lepidodendron, one of the lycopsid trees that were canopy dominants of many Pennsylvanian coal swamp forests. Its periderm or bark - the single greatest biomass contributor to many Late Paleozoic coals - is found to have a greater aliphatic content and an overall greater density of organic matter than lignified wood. Because X-PEEM allows simultaneous analysis of organic matter and matrix calcite in fully mineralized fossils, this technique also has great potential for analysis of fossil preservation, including documentation of significant traces of organic matter entrained in the calcite crystal fabric that fills the cell lumens. (author)

  14. A novel method for viewing heavy metal stained and embedded biological tissue by field emission scanning electron microscopy. (United States)

    Richards, R G; ap Gwynn, I


    Backscattered electron (BSE) imaging was used to display heavy metal stained biological structures of various embedded specimens. Samples were fixed, stained and embedded in resin blocks as with preparation for the transmission electron microscope (TEM). Blocks were trimmed to center the specimens in a trapezoidal face of up to 5 mm2 and their sides painted with conductive silver paint leaving the face uncovered. Blocks were sputter coated with 6-8 nm of silver, chromium or aluminum, with aluminum providing the best specimen contrast in BSE mode. Samples were examined in a field emission scanning electron microscope operated at a high emission current of 50 microA. Both the fixation protocol and microscope operating parameters were optimized to maximize the number of BSE available from the smallest probe. An accelerating voltage of 10 keV was found optimal for resolution and contrast. The technique allowed the direct visualization of embedded samples at resolutions beyond light microscopy with good contrast, without cutting sections, and avoiding grid bars obscuring areas of interest. The two dimensional images provided averaged information on the internal structures of the specimens in relation to the predicted emission depth of the BSE. The technique could be used for rapid diagnostics in pathological examinations, or for routine preselection of areas of interest within a sample face before final trimming for ultrathin sectioning for higher resolution TEM study.

  15. Stimulated-emission-depletion microscopy with a multicolor stimulated-Raman-scattering light source. (United States)

    Rankin, Brian R; Kellner, Robert R; Hell, Stefan W


    We describe a subdiffraction-resolution far-field fluorescence microscope employing stimulated emission depletion (STED) with a light source consisting of a microchip laser coupled into a standard single-mode fiber, which, via stimulated Raman scattering (SRS), yields a comb-like spectrum of seven discrete peaks extending from the fundamental wavelength at 532 nm to 620 nm. Each of the spectral peaks can be used as STED light for overcoming the diffraction barrier. This SRS light source enables the simple implementation of multicolor STED and provides a spectral output with multiple available wavelengths from green to red with potential for further expansion.

  16. Effect of metal films on the photostabilities of emissive organic layers as probed by fluorescence microscopy (United States)

    Abbas, Sikandar; Peteanu, Linda A.


    Realization of energy efficient and cost effective electroluminescence applications of conjugated polymers, like organic light emitting diodes (OLEDs), requires a complete understanding of photo-chemical processes at metal-polymer interfaces. Therefore it is useful to study the effects of metal films on the photoluminescence of emissive organic layer fabricated on it. While investigating these processes we observed an interesting and unexpected phenomenon that, when conjugated polymer is deposited on thin gold film substrates, it exhibits remarkable photo-stability relative to that deposited on glass, even in the presence of molecular oxygen. This paper addresses the photo-stability enhancement by thin Au films and explores the photochemical mechanism behind it.

  17. Visualization of the funis of Giardia lamblia by high-resolution field emission scanning electron microscopy--new insights. (United States)

    Benchimol, Marlene; Piva, Bruno; Campanati, Loraine; de Souza, Wanderley


    Giardia lamblia is a multiflagellar parasite and one of the earliest diverging eukaryotic cells. It possesses a cytoskeleton made of several microtubular structures-an adhesive disc, four pairs of flagella, median body, and funis. This protozoan displays different types of movements, including a lateral and dorso-ventral dislocation of its posterior region, which has not been completely elucidated. In the present study, high-resolution field emission scanning electron microscopy was used to analyze the funis structure of G. lamblia trophozoites. It was shown that the funis is made of short arrays of microtubules emanating from the axonemes of the caudal flagella, which are anchored to dense rods that run parallel to the posterior-lateral flagella. After emergence of the posterior-lateral flagella, funis microtubules are anchored to the epiplasm, a fibrous layer that underlies the portion of membrane that presents tail contractility. Based on these observations a model for the tail flexion of G. lamblia is proposed.

  18. Low energy electron imaging using Medipix 2 detector

    NARCIS (Netherlands)

    Sikharulidze, I.; van Gastel, Raoul; Schramm, S.; Abrahams, J.P.; Poelsema, Bene; Tromp, R.M.; van der Molen, S.J.


    Low Energy Electron Microscopy (LEEM) and Photo-Emission Electron Microscopy (PEEM) predominantly use a combination of microchannel plate (MCP), phosphor screen and optical camera to record images formed by 10–20 keV electrons. We have tested the performance of a LEEM/PEEM instrument with a Medipix2

  19. Multi-images deconvolution improves signal-to-noise ratio on gated stimulated emission depletion microscopy

    Energy Technology Data Exchange (ETDEWEB)

    Castello, Marco [Nanobiophotonics, Nanophysics, Istituto Italiano di Tecnologia, Via Morego 30, Genoa, 16163 (Italy); DIBRIS, University of Genoa, Via Opera Pia 13, Genoa 16145 (Italy); Diaspro, Alberto [Nanobiophotonics, Nanophysics, Istituto Italiano di Tecnologia, Via Morego 30, Genoa, 16163 (Italy); Nikon Imaging Center, Via Morego 30, Genoa 16163 (Italy); Vicidomini, Giuseppe, E-mail: [Nanobiophotonics, Nanophysics, Istituto Italiano di Tecnologia, Via Morego 30, Genoa, 16163 (Italy)


    Time-gated detection, namely, only collecting the fluorescence photons after a time-delay from the excitation events, reduces complexity, cost, and illumination intensity of a stimulated emission depletion (STED) microscope. In the gated continuous-wave- (CW-) STED implementation, the spatial resolution improves with increased time-delay, but the signal-to-noise ratio (SNR) reduces. Thus, in sub-optimal conditions, such as a low photon-budget regime, the SNR reduction can cancel-out the expected gain in resolution. Here, we propose a method which does not discard photons, but instead collects all the photons in different time-gates and recombines them through a multi-image deconvolution. Our results, obtained on simulated and experimental data, show that the SNR of the restored image improves relative to the gated image, thereby improving the effective resolution.

  20. Characterisation of bicontinuous cubic liquid crystalline systems of phytantriol and water using cryo field emission scanning electron microscopy (cryo FESEM). (United States)

    Rizwan, S B; Dong, Y-D; Boyd, B J; Rades, T; Hook, S


    Cubosomes are a novel lipid particulate delivery system currently being investigated for drug delivery purposes. The present study investigates bicontinuous cubic liquid crystalline systems (bulk phase and cubosomes) formed by phytantriol and water using cryo field emission scanning electron microscopy (cryo FESEM). Previously cubosomes have been characterized by cryo transmission electron microscopy (cryo TEM) with small angle X-ray diffraction (SAXS) confirming the bicontinuous liquid crystalline type. Bulk cubic phase and cubosomes were prepared from phytantriol and Pluronic F127 and analysed using cryo FESEM and SAXS. The micrographs showed the cubic phase had a tortuous, bicontinuous nature with a non-intersecting network of water channels. The cubosomes also show the same underlying tortuous structure entirely consistent with that of the bulk cubic phase and closely resemble the mathematical description of cubosomes described using nodal surface representation. The structure of both systems was confirmed using SAXS as a bicontinuous cubic liquid crystalline phase with Pn3m geometry. Cryo FESEM provides valuable insights into the morphological features of bicontinuous cubic liquid crystalline systems. The unique details shown provide strength to support the nodal surface representation of bicontinuous cubic liquid crystalline systems. Cryo FESEM provides a new technique to complement cryo TEM and SAXS for investigating their structure and function.

  1. Cold field emission gun-scanning electron microscopy: a new tool for morphological and ultrastructural analysis of liposomes. (United States)

    De Rosa, G; De Stefano, M; Ungaro, F; La Rotonda, M I


    Liposomes are lipid vesicles largely investigated in the past 30 years as pharmaceutical carriers. In the development of new liposome-based formulations, the study of liposome surface properties remains a crucial step. For this purpose, microscopy techniques can provide useful information, although each such technique suffers from some limitations. Here, we have used cold field emission gun-scanning electron microscopy (cFEG-SEM) to acquire detailed images of liposome surface. In particular, we observed PEGylated and non-PEGylated liposomes in different size ranges. In the case of nanosized liposomes (mean diameter about 200 nm), a morphological evaluation of the whole preparation was obtained. On the other hand, in the case of giant liposomes (mean diameter about 2 microm), it was possible to observe the different surface ultrastructures of the two formulations. In particular, a regular and only slightly wrinkled surface was observed in the case of non-PEGylated liposomes, while a very irregular surface ultrastructure was visible in the case of PEGylated liposomes. This study shows, for the first time, the potential of cFEG-SEM as a new and powerful tool to obtain information on liposome morphology and, at least in the case of giant liposomes, on ultrastructure of the liposome surface.

  2. Comparative analysis of Trichuris muris surface using conventional, low vacuum, environmental and field emission scanning electron microscopy. (United States)

    Lopes Torres, Eduardo José; de Souza, Wanderley; Miranda, Kildare


    The whipworm of the genus Trichuris Roederer, 1791, is a nematode of worldwide distribution and comprises species that parasitize humans and other mammals. Infections caused by Trichuris spp. in mammals can lead to various intestinal diseases of human and veterinary interest. 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 to introduce a variety of artifacts such as sample shrinking, loss of secreted products and/or hiding of small structures due to sample coating. Low vacuum (LVSEM) and environmental scanning electron microscopy (ESEM) have been applied to a variety of insulator samples, also used in the visualization of hydrated and/or live specimens in their native state. In the present work, we used LVSEM and ESEM to analyze the surface of T. muris and analyze its interaction with the host tissue using freshly fixed or unfixed hydrated samples. Analysis of hydrated samples showed a set of new features on the surface of the parasite and the host tissue, including the presence of the secretory products of the bacillary glands on the surface of the parasite, and the presence of mucous material and eggs on the intestinal surface. Field emission scanning electron microscopy (FESEM) was also applied to reveal the detailed structure of the glandular chambers in fixed, dried and metal coated samples. Taken together, the results show that analysis of hydrated samples may provide new insights in the structural organization of the surface of helminth parasites and its interaction with the infected tissue, suggesting that the application of alternative SEM techniques may open new perspectives for analysis in taxonomy, morphology and host-parasite interaction fields. Copyright © 2013 Elsevier B.V. All rights reserved.

  3. Electron transport in ultra-thin films and ballistic electron emission microscopy (United States)

    Claveau, Y.; Di Matteo, S.; de Andres, P. L.; Flores, F.


    We have developed a calculation scheme for the elastic electron current in ultra-thin epitaxial heterostructures. Our model uses a Keldysh’s non-equilibrium Green’s function formalism and a layer-by-layer construction of the epitaxial film. Such an approach is appropriate to describe the current in a ballistic electron emission microscope (BEEM) where the metal base layer is ultra-thin and generalizes a previous one based on a decimation technique appropriated for thick slabs. This formalism allows a full quantum mechanical description of the transmission across the epitaxial heterostructure interface, including multiple scattering via the Dyson equation, which is deemed a crucial ingredient to describe interfaces of ultra-thin layers properly in the future. We introduce a theoretical formulation needed for ultra-thin layers and we compare with results obtained for thick Au(1 1 1) metal layers. An interesting effect takes place for a width of about ten layers: a BEEM current can propagate via the center of the reciprocal space (\\overlineΓ ) along the Au(1 1 1) direction. We associate this current to a coherent interference finite-width effect that cannot be found using a decimation technique. Finally, we have tested the validity of the handy semiclassical formalism to describe the BEEM current.

  4. Surface and flagella morphology of the motile form of Chromera velia revealed by field-emission scanning electron microscopy. (United States)

    Weatherby, Kate; Murray, Shauna; Carter, Dee; Slapeta, Jan


    Chromera velia(Chromerida; Alveolata) is an autotrophic protist isolated from stony corals.Ch. veliapossesses a chloroplast thought to be most closely related to the apicoplasts of non-photosynthetic apicomplexa. Phylogenetic analyses placeCh. veliaas a close relative of parasitic apicomplexa and predatory colpodellids. We have used field-emission scanning electron microscopy of cells sputter-coated with gold or chromium and non-coated cells to characterise the surface ultrastructure of the motile form ofCh. velia. In overall morphology the biflagellatedCh. veliacells resemble the colpodellidColpodella edax, but with some notable differences. The ventral side of the flagellatedCh. veliacell has two grooves extending from the anterior flagella insertion point with a ridge rising towards the anterior apex of the cell. The anterior flagellum is shorter than the posterior flagellum and possesses a distinct, small curved appendage. The insertion point of the anterior flagellum is partly enclosed by a flap extending from the cell. The posterior flagellum is approximately four times the length of the cell and possesses mastigonemes. The combination of coating techniques proved superior to the commonly used gold coating to determine fine surface ultrastructure. This new ultrastructural information forCh. veliaallowed us to emend its diagnosis. 2010 Elsevier GmbH. All rights reserved.

  5. Improvement on the visualization of cytoskeletal structures of protozoan parasites using high-resolution field emission scanning electron microscopy (FESEM). (United States)

    Sant'Anna, Celso; Campanati, Loraine; Gadelha, Catarina; Lourenço, Daniela; Labati-Terra, Letícia; Bittencourt-Silvestre, Joana; Benchimol, Marlene; Cunha-e-Silva, Narcisa Leal; De Souza, Wanderley


    The association of high resolution field emission scanning electron microscopy (FESEM), with a more efficient system of secondary electron (SE) collection and in-lens specimen position, provided a great improvement in the specimen's topographical contrast and in the generation of high-resolution images. In addition, images obtained with the use of the high-resolution backscattered electrons (BSE) detector provided a powerful tool for immunocytochemical analysis of biological material. In this work, we show the contribution of the FESEM to the detailed description of cytoskeletal structures of the protozoan parasites Herpetomonas megaseliae, Trypanosoma brucei and Giardia lamblia. High-resolution images of detergent extracted H. megaseliae and T. brucei showed the profile of the cortical microtubules, also known as sub-pellicular microtubules (SPMT), and protein bridges cross-linking them. Also, it was possible to visualize fine details of the filaments that form the lattice-like structure of the paraflagellar rod (PFR) and its connection with the axoneme. In G. lamblia, it was possible to observe the intricate structure of the adhesive disk, funis (a microtubular array) and other cytoskeletal structures poorly described previously. Since most of the stable cytoskeletal structures of this protozoan rely on tubulin, we used the BSE images to accurately map immunolabeled tubulin in its cytoskeleton. Our results suggest that the observation of detergent extracted parasites using FESEM associated to backscattered analysis of immunolabeled specimens represents a new approach for the study of parasite cytoskeletal elements and their protein associations.

  6. Ultrastructure of the innermost surface of differentiating normal and compression wood tracheids as revealed by field emission scanning electron microscopy. (United States)

    Kim, Jong Sik; Awano, Tatsuya; Yoshinaga, Arata; Takabe, Keiji


    The ultrastructure of the innermost surface of Cryptomeria japonica differentiating normal wood (NW) and compression wood (CW) was comparatively investigated by field emission electron microscopy (FE-SEM) combined with enzymatic degradation of hemicelluloses. Cellulose microfibril (CMF) bundles were readily observed in NW tracheids in the early stage of secondary cell wall formation, but not in CW tracheids because of the heavy accumulation of amorphous materials composed mainly of galactans and lignin. This result suggests that the ultrastructural deposition of cell wall components in the tracheid cell wall differ between NW and CW from the early stage of secondary cell wall formation. Delignified NW and CW tracheids showed similar structural changes during differentiating stages after xylanase or β-mannanase treatment, whereas they exhibited clear differences in ultrastructure in mature stages. Although thin CMF bundles were exposed in both delignified mature NW and CW tracheids by xylanase treatment, ultrastructural changes following β-mannanase treatment were only observed in CW tracheids. CW tracheids also showed different degradation patterns between xylanase and β-mannanase. CMF bundles showed a smooth surface in delignified mature CW tracheids treated with xylanase, whereas they had an uneven surface in delignified mature CW tracheids treated with β-mannanase, indicating that the uneven surface of CMF bundles was related to xylans. The present results suggest that ultrastructural deposition and organization of lignin and hemicelluloses in CW tracheids may differ from those of NW tracheids.

  7. Inter- and intraspecific structural variations among intervascular pit membranes, as revealed by field-emission scanning electron microscopy. (United States)

    Sano, Yuzou


    The structure of the intervascular pit membranes of four dicotyledonous species (Salix sachalinensis, Betula platyphylla var. japonica, Acer mono, and Fraxinus mandshurica var. japonica) was examined by field-emission scanning electron microscopy. The intervascular pit membranes of F. mandshurica var. japonica had thin surface layers and a dense middle layer, while no similar middle layer was detectable in the other three species. In F. mandshurica var. japonica, the entire area of each pit membrane was densely covered with microfibrils. In the other three species, by contrast, openings were found in the pit membranes. In some of the intervascular pit membranes of S. sachalinensis, B. platyphylla var. japonica, and A. mono, microfibrils were sparsely interwoven in small areas of the pit membranes and openings of up to several hundred nanometers in diameter were present in such regions. These porous regions tended to be located in peripheral areas of pit membranes. In S. sachalinensis and B. platyphylla var. japonica, ethanol-soluble extracts, whose chemical nature and function remain unknown, were heavily distributed over the intervascular pit membranes. Our observations suggest that the structure of intervascular pit membranes is more complicated than has previously been acknowledged.

  8. Field-emission scanning electron microscopy analysis of morphology and enzyme distribution within an industrial biocatalytic particle. (United States)

    van Roon, J L; van Aelst, A C; Schroën, C G P H; Tramper, J; Beeftink, H H


    Field-emission scanning electron microscopy (FESEM) was used in a technical feasibility study to obtain insight into the internal morphology and the intraparticle enzyme distribution of Assemblase, an industrial biocatalytic particle containing immobilized penicillin-G acylase. The results were compared with previous studies based on light and transmission electron microscopic techniques. The integrated FESEM approach yielded the same quantitative results as the microscopic techniques used previously. Given this technical equivalence, the integrated approach offers several advantages. First, the single preparation method and detection system avoids interpretation discrepancies between corresponding areas that were examined for different properties with different detection techniques in different samples. Second, the specimen size suitable for whole particle study is virtually unlimited, which simplifies sectioning and puts less stringent demands on the embedding technique. Furthermore, the sensitivity toward enzyme presence and distribution increases because the epitopes inside thick sections become available for labeling. Quick and unambiguous analysis of the relation between particle morphology and enzyme distribution is important because this information may be used in the future for the design of enzyme distributions in which the particle morphology can be used as a control parameter.

  9. Supernumerary human hair cells-signs of regeneration or impaired development? A field emission scanning electron microscopy study. (United States)

    Rask-Andersen, Helge; Li, Hao; Löwenheim, Hubert; Müller, Marcus; Pfaller, Kristian; Schrott-Fischer, Annelies; Glueckert, Rudolf


    Current attempts to regenerate cochlear sensorineural structures motivate further inspection of the human organ of hearing. Here, we analyzed the supernumerary inner hair cell (sIHC), a possible sign of regeneration and cell replacement. Human cochleae were studied using field emission scanning electron microscopy (FESEM; maximum resolution 2 nm) obtained from individuals aged 44, 48, and 58 years with normal sensorineural pure-tone average (PTA) thresholds (PTA <20 dB). The wasted tissue was harvested during trans-cochlear approaches and immediately fixed for ultrastructural analysis. All specimens exhibited sIHCs at all turns except at the extreme lower basal turn. In one specimen, it was possible to image and count the inner hair cells (IHCs) along the cochlea representing the 0.2 kHz-8 kHz region according to the Greenwood place/frequency scale. In a region with 2,321 IHCs, there were 120 scattered one-cell losses or 'gaps' (5%). Forty-two sIHCs were present facing the modiolus. Thirty-eight percent of the sIHCs were located near a 'gap' in the IHC row (±6 IHCs). The prevalence of ectopic inner hair cells was higher than expected. The morphology and placement could reflect a certain ongoing regeneration. Further molecular studies are needed to verify if the regenerative capacity of the human auditory periphery might have been underestimated.

  10. New method for characterizing paper coating structures using argon ion beam milling and field emission scanning electron microscopy. (United States)

    Dahlström, C; Allem, R; Uesaka, T


    We have developed a new method for characterizing microstructures of paper coating using argon ion beam milling technique and field emission scanning electron microscopy. The combination of these two techniques produces extremely high-quality images with very few artefacts, which are particularly suited for quantitative analyses of coating structures. A new evaluation method has been developed by using marker-controlled watershed segmentation technique of the secondary electron images. The high-quality secondary electron images with well-defined pores makes it possible to use this semi-automatic segmentation method. One advantage of using secondary electron images instead of backscattered electron images is being able to avoid possible overestimation of the porosity because of the signal depth. A comparison was made between the new method and the conventional method using greyscale histogram thresholding of backscattered electron images. The results showed that the conventional method overestimated the pore area by 20% and detected around 5% more pores than the new method. As examples of the application of the new method, we have investigated the distributions of coating binders, and the relationship between local coating porosity and base sheet structures. The technique revealed, for the first time with direct evidence, the long-suspected coating non-uniformity, i.e. binder migration, and the correlation between coating porosity versus base sheet mass density, in a straightforward way. © 2010 The Authors Journal compilation © 2010 The Royal Microscopical Society.

  11. Formation of macromolecular lignin in ginkgo xylem cell walls as observed by field emission scanning electron microscopy. (United States)

    Terashima, Noritsugu; Awano, Tatsuya; Takabe, Keiji; Yoshida, Masato


    Formation of macromolecular lignin in ginkgo cell walls. In the lignifying process of xylem cell walls, macromolecular lignin is formed by polymerization of monolignols on the pectic substances, hemicellulose and cellulose microfibrils that have deposited prior to the start of lignification. Observation of lignifying secondary cell walls of ginkgo tracheids by field emission scanning electron microscopy suggested that lignin-hemicellulose complexes are formed as tubular bead-like modules surrounding the cellulose microfibrils (CMFs), and that the complexes finally fill up the space between CMFs. The size of one tubular bead-like module in the middle layer of the secondary wall (S2) was tentatively estimated to be about 16+/-2 nm in length, about 25+/-1 nm in outer diameter, with a wall thickness of 4+/-2 nm; the size of the modules in the outer layer of the secondary wall (S1) was larger and they were thicker-walled than that in the middle layer (S2). Aggregates of large globular modules were observed in the cell corner and compound middle lamella. It was suggested that the structure of non-cellulosic polysaccharides and mode of their association with CMFs may be important factors controlling the module formation and lignin concentration in the different morphological regions of the cell wall.

  12. Spatially resolved band alignments at Au-hexadecanethiol monolayer-GaAs(001) interfaces by ballistic electron emission microscopy

    Energy Technology Data Exchange (ETDEWEB)

    Junay, A.; Guézo, S., E-mail:; Turban, P.; Delhaye, G.; Lépine, B.; Tricot, S.; Ababou-Girard, S.; Solal, F. [Département Matériaux-Nanosciences, Institut de Physique de Rennes, UMR 6251, CNRS-Université de Rennes 1, Campus de Beaulieu, Bât 11E, 35042 Rennes Cedex (France)


    We study structural and electronic inhomogeneities in Metal—Organic Molecular monoLayer (OML)—semiconductor interfaces at the sub-nanometer scale by means of in situ Ballistic Electron Emission Microscopy (BEEM). BEEM imaging of Au/1-hexadecanethiols/GaAs(001) heterostructures reveals the evolution of pinholes density as a function of the thickness of the metallic top-contact. Using BEEM in spectroscopic mode in non-short-circuited areas, local electronic fingerprints (barrier height values and corresponding spectral weights) reveal a low-energy tunneling regime through the insulating organic monolayer. At higher energies, BEEM evidences new conduction channels, associated with hot-electron injection in the empty molecular orbitals of the OML. Corresponding band diagrams at buried interfaces can be thus locally described. The energy position of GaAs conduction band minimum in the heterostructure is observed to evolve as a function of the thickness of the deposited metal, and coherently with size-dependent electrostatic effects under the molecular patches. Such BEEM analysis provides a quantitative diagnosis on metallic top-contact formation on organic molecular monolayer and appears as a relevant characterization for its optimization.

  13. Effects of industrial noise on circumpulpar dentin - a field emission scanning electron microscopy and energy dispersive spectroscopy analysis (United States)

    Cavacas, Maria Alzira; Tavares, Vitor; Oliveira, Maria João; Oliveira, Pedro; Sezinando, Ana; Martins dos Santos, José


    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

  14. Effects of industrial noise on circumpulpar dentin--a field emission scanning electron microscopy and energy dispersive spectroscopy analysis. (United States)

    Cavacas, Maria Alzira; Tavares, Vitor; Oliveira, Maria João; Oliveira, Pedro; Sezinando, Ana; Martins dos Santos, José


    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.

  15. Direct observation of iron-induced conformational changes of mitochondrial DNA by high-resolution field-emission in-lens scanning electron microscopy.


    Yaffee, M; Walter, P; Richter, C; Müller, M


    When respiring rat liver mitochondria are incubated in the presence of Fe(III) gluconate, their DNA (mtDNA) relaxes from the supercoiled to the open circular form dependent on the iron dose. Anaerobiosis or antioxidants fail to completely inhibit the unwinding. High-resolution field-emission in-lens scanning electron microscopy imaging, in concert with backscattered electron detection, pinpoints nanometer-range iron colloids bound to mtDNA isolated from iron-exposed mitochondria. High-resolut...

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

    Directory of Open Access Journals (Sweden)

    Schneider Andreas S


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

  17. 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


    .... 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...

  18. Immunohistochemical identification of decorin and biglycan in human dentin: a correlative field emission scanning electron microscopy/transmission electron microscopy study. (United States)

    Orsini, G; Ruggeri, A; Mazzoni, A; Papa, V; Mazzotti, G; Di Lenarda, R; Breschi, L


    Decorin and biglycan, two small leucine-rich proteoglycans, have been proposed to play important roles in matrix-mediated formation of mineralized tissues, and their three-dimensional arrangement in human dentin is still not completely understood. The aim of this study was to immunohistochemically analyze the distribution of decorin and biglycan in human predentin/dentin organic matrix under a high-resolution field emission in-lens scanning electron microscope (FEI-SEM) and a transmission electron microscope (TEM). Tooth dentin specimens were submitted to either a preembedding or a postembedding immunolabeling technique using primary antibodies antidecorin and antibiglycan and gold-conjugated secondary antibodies. Correlative FEI-SEM/TEM observations showed that the two antibodies yielded a similar labeling pattern over the processes of odontoblasts and the predentin. Decorin and biglycan were mainly associated with the collagen fibers within the predentin layer, revealing a moderate immunoreaction that was significantly higher compared to the one observed on dentin. Thus, a generally weak labeling for decorin was found in dentin, which, however, was significantly higher on odontoblast processes within dentinal tubules than in intertubular dentin. On the other hand, biglycan immunolocalization on dentin revealed few gold particles rather uniformly distributed, without showing significant differences between tubular and intertubular regions. In conclusion, this study reveals distinct distribution patterns of decorin and biglycan and their relation with collagen. Decorin's and biglycan's precise roles within prematrix and mineralized matrix in human teeth should be further clarified.

  19. Mapping the directional emission of quasi-two-dimensional photonic crystals of semiconductor nanowires using Fourier microscopy

    NARCIS (Netherlands)

    Fontana, Y.; Grzela, G.; Bakkers, E.P.A.M.; Gomez Rivas, J.


    Controlling the dispersion and directionality of the emission of nanosources is one of the major goals of nanophotonics research. This control will allow the development of highly efficient nanosources even at the single-photon level. One of the ways to achieve this goal is to couple the emission to

  20. Detection of allergens from Alternaria alternata by gold-conjugated anti-human IgE and field emission scanning electron microscopy. (United States)

    Sercombe, Jason K; Eduard, Wijnand; Romeo, Tony C; Green, Brett J; Tovey, Euan R


    Fungal allergens are present in viable and non-viable conidia, hyphae and fungal fragments. It has been shown that large quantities of allergen are released from conidia during germination. We used a gold immunolabelling technique and field emission scanning electron microscopy to examine the allergen release from Alternaria alternata conidia. Immunolabelling was associated with the hyphal tip and amorphous matter associated with the emerging hyphae. Non-specific antibody controls showed no labelling associated with germinating fungi. This suggests that material released from hyphae may be an additional source of fungal allergens.

  1. Field emission scanning electron microscopy and transmission electron microscopy studies of the chorion, plasma membrane and syncytial layers of the gastrula-stage embryo of the zebrafish Brachydanio rerio : a consideration of the structural and functional relationships with respect to cryoprotectant penetration

    NARCIS (Netherlands)

    Rawson, DM; Zhang, T; Kalicharan, D; Jongebloed, WL

    The structure of the chorion and plasma membranes of gastrula-stage zebrafish Brachydanio rerio embryos were studied using field emission scanning electron microscopy (FE-SEM) and transmission electron microscopy (TEM). These studies confirm the outer chorion membrane complex to be 1.5-2.5 mu m in

  2. Functional biocompatible magnetite-cellulose nanocomposite fibrous networks: Characterization by fourier transformed infrared spectroscopy, X-ray powder diffraction and field emission scanning electron microscopy analysis. (United States)

    Habibi, Neda


    The preparation and characterization of functional biocompatible magnetite-cellulose nano-composite fibrous material is described. Magnetite-cellulose nano-composite was prepared by a combination of the solution-based formation of magnetic nano-particles and subsequent coating with amino celluloses. Characterization was accomplished using X-ray powder diffraction (XRD), fourier transformed infrared (FTIR) and field emission scanning electron microscopy (FESEM) analysis. The peaks of Fe3O4 in the XRD pattern of nanocomposite confirm existence of the nanoparticles in the amino cellulose matrix. Magnetite-cellulose particles exhibit an average diameter of roughly 33nm as demonstrated by field emission scanning electron microscopy. Magnetite nanoparticles were irregular spheres dispersed in the cellulose matrix. The vibration corresponding to the NCH3 functional group about 2850cm(-1) is assigned in the FTIR spectra. Functionalized magnetite-cellulose nano-composite polymers have a potential range of application as targeted drug delivery system in biomedical field. Copyright © 2014 Elsevier B.V. All rights reserved.


    NARCIS (Netherlands)



    The stereociliar structures of the guinea-pig cochlear organ of Corti were studied at low-voltage (1-5 kV) with field-emission scanning electron microscope (SEM) using various pre- and post-fixation methods, such as OTOTO (OsO4/thiocarbohydrazide/OsO4/thiocarbohydrazide/OsO4) and TAO (tannic

  4. 200 keV cold field emission source using carbon cone nanotip: Application to scanning transmission electron microscopy. (United States)

    Mamishin, Shuichi; Kubo, Yudai; Cours, Robin; Monthioux, Marc; Houdellier, Florent


    We report the use of a pyrolytic carbon cone nanotip as field emission cathode inside a modern 200 kV dedicated scanning transmission electron microscope. We show an unprecedented improvement in the probe current stability while maintaining all the fundamental properties of a cold field emission source such as a small angular current density together with a high brightness. We have also studied the influence of the low extraction voltage, as enabled by the nanosized apex of the cones, on the electron optics properties of the source that prevent the formation of a virtual beam cross-over of the gun. We have addressed this resolution-limiting issue by coming up with a new electron optical source design. Copyright © 2017 Elsevier B.V. All rights reserved.

  5. Reply to ``Comment on `Imaging the atomic orbitals of carbon atomic chains with field-emission electron microscopy' '' (United States)

    Mikhailovskij, I. M.; Sadanov, E. V.; Mazilova, T. I.; Ksenofontov, V. A.; Velicodnaja, O. A.


    In our recent paper [I. M. Mikhailovskij, E. V. Sadanov, T. I. Mazilova, V. A. Ksenofontov, and O. A. Velicodnaja, Phys. Rev. B 80, 165404 (2009)], we have presented evidence for field emission from individual orbitals of self-standing carbon chains, which can be used for real-space imaging of the end-atom orbitals with a field-emission electron microscope (FEEM). In this reply to the preceding Comment, we refer to the issues brought up there, which concern the viewpoint that the observed spontaneous mutual transformations of FEEM patterns have been attributed to the ligand-induced symmetry breaking by calling attention to the role of hydrogen atoms unavoidable in most nanostructured carbon materials.

  6. Investigation of porous silica nanostructures in diatoms isolated from Kurichi and Sulur lakes of Coimbatore, India using field emission scanning electron microscopy. (United States)

    N, Seethalakshmi; R, Selvakumar


    Diatoms are unicellular algae that possess cell wall made of silica. These diatoms play a pivotal role in synthesis of variety of silica nanostructures and have adorning morphology in nature. In the present study, we have used field emission scanning electron microscopy (FE-SEM) to investigate their morphological features like pore size, shape, and porous pattern in various diatoms isolated from Kurichi and Sulur fresh water lakes, Coimbatore, Tamil Nadu, India. Diatoms were identified as Nitzschia sp., Cyclotella meneghiniana, Coscinodiscus sp. and Cyclotella atomus based on their morphological features. The arrangement of porous nanostructures in these diatoms have been characterized. The change in the nanostructures present in the diatoms have been correlated to the contamination of water bodies. Copyright © 2015 Elsevier Ltd. All rights reserved.

  7. Limitations on the use of scanning probe microscopy for the measurement of field emission from copper surfaces

    Energy Technology Data Exchange (ETDEWEB)

    Mizuno, Y.


    High electric-field breakdown in cm-wave x-band radio frequency (rf) accelerating structures is an important phenomenon limiting attainable accelerating gradients. Linear collider development requires an accelerating gradient of, at least, 70MeV/m (150MV/m surface electric field). The observed breakdown sequence usually consists of field emission (FE) from an electrically-conducting surface feature which heats the point of emission, thereby releasing gas from the surface and nearby bulk. The ionized gas makes a plasma that regeneratively heats the metal and releases more gas and electrons (via secondary emission) until a flashover occurs. The FE, however, occurs at a much lower surface electric field in experiments than predicted by theory. Experimental measurements of FE from macroscopic surface areas require the use of a geometrical field-enhancement factor, {beta}, to fit best the data to a Fowler-Nordheim (F-N) emission model with reasonable physical parameters. A newer, microscopic interpretation of the results proposes the existence of quasi-filamentary electrically-conducting channels between the metal bulk and the surface. These channels connect to emitting features near or on the surface, acting as microscopically field-enhanced electron emission sources. Analysis of emitter behavior suggesting that the macroscopic emission should primarily be due to classical geometrical-enhancement is, at least in some cases, due to the presence of these conducting nano-structures in the surface region. We describe an accelerator materials-related study of high electric field breakdown from polished copper (and other) surfaces, in dry-nitrogen gas ambient, using an atomic force microscope (AFM) modified to make F-N FE measurements. In early periods of this work, the results showed that the instrument might be capable of making F-N measurements on small surface areas of mechanically-polished and natively-oxidized high-quality copper surfaces to the GV/m level. The

  8. Collagen mineralization in human aortic valve stenosis: a field emission scanning electron microscopy and energy dispersive spectroscopy analysis. (United States)

    Perrotta, Ida; Davoli, Mariano


    Abstract Calcific aortic stenosis is a slowly progressive disorder characterized by an important extracellular matrix remodeling with fibrosis and massive deposition of minerals (primarily calcium) in the valve leaflet. The main structural components of human aortic valve are the large, thick collagen bundles that withstand the diastolic loading. Collagen has been studied in a number of reports that aim to clarify the mechanisms underlying the structural deterioration of heart valve substitutes, however to date, little is known regarding the morphological interaction between collagen and mineral crystals in the calcifying tissue of native aortic valve. Here, we have analyzed a total of 12 calcified native aortic valves by using scanning electron microscopy (SEM) with Energy Dispersive X-Ray Analysis (EDX) to depict the morphological appearance of mineralized collagen and to determine the location of calcium phosphate minerals in the collagen matrix of the valve cusp. Our results demonstrate that crystals probably nucleate and grow in the interior of the collagen fibers in the absence of surface events.

  9. Scanning ultrafast electron microscopy


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


    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...

  10. A modified ‘NanoSuit®’ preserves wet samples in high vacuum: direct observations on cells and tissues in field-emission scanning electron microscopy (United States)

    Takaku, Yasuharu; Suzuki, Hiroshi; Kawasaki, Hideya; Ohta, Isao; Ishii, Daisuke; Hirakawa, Satoshi; Tsutsui, Takami; Matsumoto, Haruko; Takehara, Sayuri; Nakane, Chinatsu; Sakaida, Kana; Suzuki, Chiaki; Muranaka, Yoshinori; Kikuchi, Hirotoshi; Konno, Hiroyuki; Shimomura, Masatsugu


    Although field-emission scanning electron microscopy (FE-SEM) has proven very useful in biomedical research, the high vacuum required (10−3 to 10−7 Pa) precludes direct observations of living cells and tissues at high resolution and often produces unwanted structural changes. We have previously described a method that allows the investigator to keep a variety of insect larvae alive in the high vacuum environment of the electron microscope by encasing the organisms in a thin, vacuum-proof suit, the ‘NanoSuit®'. However, it was impossible to protect wet tissues freshly excised from intact organisms or cultured cells. Here we describe an improved ‘NanoSuit' technique to overcome this limitation. We protected the specimens with a surface shield enhancer (SSE) solution that consists of glycerine and electrolytes and found that the fine structure of the SSE-treated specimens is superior to that of conventionally prepared specimens. The SSE-based NanoSuit affords a much stronger barrier to gas and/or liquid loss than the previous NanoSuit did and, since it allows more detailed images, it could significantly help to elucidate the ‘real' organization of cells and their functions. PMID:28405375

  11. A modified 'NanoSuit®' preserves wet samples in high vacuum: direct observations on cells and tissues in field-emission scanning electron microscopy. (United States)

    Takaku, Yasuharu; Suzuki, Hiroshi; Kawasaki, Hideya; Ohta, Isao; Ishii, Daisuke; Hirakawa, Satoshi; Tsutsui, Takami; Matsumoto, Haruko; Takehara, Sayuri; Nakane, Chinatsu; Sakaida, Kana; Suzuki, Chiaki; Muranaka, Yoshinori; Kikuchi, Hirotoshi; Konno, Hiroyuki; Shimomura, Masatsugu; Hariyama, Takahiko


    Although field-emission scanning electron microscopy (FE-SEM) has proven very useful in biomedical research, the high vacuum required (10-3 to 10-7 Pa) precludes direct observations of living cells and tissues at high resolution and often produces unwanted structural changes. We have previously described a method that allows the investigator to keep a variety of insect larvae alive in the high vacuum environment of the electron microscope by encasing the organisms in a thin, vacuum-proof suit, the 'NanoSuit®'. However, it was impossible to protect wet tissues freshly excised from intact organisms or cultured cells. Here we describe an improved 'NanoSuit' technique to overcome this limitation. We protected the specimens with a surface shield enhancer (SSE) solution that consists of glycerine and electrolytes and found that the fine structure of the SSE-treated specimens is superior to that of conventionally prepared specimens. The SSE-based NanoSuit affords a much stronger barrier to gas and/or liquid loss than the previous NanoSuit did and, since it allows more detailed images, it could significantly help to elucidate the 'real' organization of cells and their functions.

  12. Silver solder "tattoo," a novel form of oral pigmentation identified with the use of field emission scanning electron microscopy and electron dispersive spectrography. (United States)

    Hussaini, H M; Waddell, J N; Girvan, L; West, L M; Kardos, T B; Rich, A M; Seymour, G J


    The aim of this study was to investigate the use of field emission scanning electron microscopy and electron dispersive spectrography (SEM-EDS) to identify silver solder "tattoo." SEM-EDS was used to analyze material present in the connective tissue of a patient who presented with bilateral pigmentation of the mandibular lingual gingiva adjacent to the first molars. No dental restorations were present. SEM-EDS analysis identified silver, with no evidence of tin, copper, or mercury. The patient was wearing an orthodontic appliance where brackets had been soldered to the archwire with silver solder. It is hypothesed that the solder underwent electrolytic corrosion with subsequent regrouping of silver ions in the submucosa leading to blue-gray discoloration. Spectrography proved to be a powerful diagnostic tool in identifying the metal within the oral mucosa. Attention is drawn to this newly described lesion, which should be included as a differential diagnosis for pigmented oral mucosal lesions. Copyright © 2011 Mosby, Inc. All rights reserved.

  13. Oocytes as an experimental system to analyze the ultrastructure of endogenous and ectopically expressed nuclear envelope components by field-emission scanning electron microscopy. (United States)

    Stick, Reimer; Goldberg, Martin W


    Xenopus oocytes provide a powerful model system for studying the structure and function of the nuclear envelope and its components. Firstly, the nuclear envelope is easily isolated by hand under gentle conditions that have little effect on its structural organization. They can then be prepared for several types of electron microscopy (EM) including field-emission scanning EM (feSEM) (described here) and cryo-EM. They can be immuno-gold labeled to determine the localization of individual proteins. There is also enough material to analyze biochemically. Secondly, they possess an efficient transcription and translation system so that proteins of interest can be ectopically expressed by injection of either mRNA into the cytoplasm or plasmids into the nucleus. Such proteins can be tagged and mutated. They are post-translationally modified and usually incorporate into the correct compartment. We describe here methods developed to analyze the structural organization of the nuclear envelope by feSEM including the structural organization of ectopically expressed nuclear envelope proteins.

  14. Structural investigations on nanoemulsions, solid lipid nanoparticles and nanostructured lipid carriers by cryo-field emission scanning electron microscopy and Raman spectroscopy. (United States)

    Saupe, Anne; Gordon, Keith C; Rades, Thomas


    Recently, colloidal dispersions based on solid lipids (solid lipid nanoparticles, SLN) and mixtures of solid and liquid lipids (nanostructured lipid carriers, NLC) were described as innovative carrier systems. A spherical particle shape is the basis of features such as a high loading capacity and controlled drug release characteristics due to smaller lipid-water interfaces and longer diffusion pathways when compared to thin platelets. The structures of SLN and the influence of oil load (NLC) on particle properties were investigated by photon correlation spectroscopy (PCS), laser diffractometry (LD), cryo-field emission scanning electron microscopy (cryo-FESEM), Raman spectroscopy and infrared spectroscopy (IR), and compared to a conventional nanoemulsion. PCS and LD data show similar size and size distribution for SLN and NLC (approximately 210 nm, polydispersity index approximately 0.15) and suggested a long term physical stability for the dispersions which had been stored for up to 12 months at different temperatures. Using cryo-FESEM droplets (for the nanoemulsion) and almost spherical particles for SLN and NLC were observed. Raman spectroscopy resulted in spectra for NLC that are weighted to the SLN spectra, suggesting an undisturbed crystal structure. Infrared spectra of the NLC are predominantly SLN in nature. Importantly the SLN bands are unshifted in the NLC spectrum indicating that the crystalline structure is unaffected by the presence of the oil.


    Mogi, Yuko; Kagami, Yayoi; Kuwano, Kazuyoshi; Miyamura, Shinichi; Nagumo, Tamotsu; Kawano, Shigeyuki


    Gametes of the marine green alga Ulva compressa L. are biflagellate and pear shaped, with one eyespot at the posterior end of the cell. The species is at an early evolutionary stage between isogamy and anisogamy. In the past, zygote formation of green algae was categorized solely by the relative sizes of gametes produced by two mating types (+ and -). Recently, however, locations of cell fusion sites and/or mating structures of gametes have been observed to differ between mating types in several green algae (asymmetry of cell fusion site and/or mating structure positions). To use this asymmetry for determining gamete mating type, we explored a new method, field emission scanning electron microscopy (FE-SEM), for visualizing the mating structure of U. compressa. When gametes were subjected to drying stress in the process of a conventional critical-point-drying method, a round structure was observed on the cell surfaces. In the mating type MGEC-1 (mt(+) ), this structure was located on the same side of the cell as the eyespot, whereas it was on the side opposite the eyespot in the mating type MGEC-2 (mt(-) ). The gametes fuse at the round structures. TEM showed an alignment of vesicles inside the cytoplasm directly below the round structures, which are indeed the mating structures. Serial sectioning and three-dimensional construction of TEM micrographs confirmed the association of the mating structure with flagellar roots. The mating structure was associated with 1d root in the MGEC-1 gamete but with 2d root in the MGEC-2 gamete. © 2008 Phycological Society of America.

  16. Group 13 allergens as environmental and immunological markers for grass pollen allergy: studies by immunogold field emission scanning and transmission electron microscopy. (United States)

    Grote, Monika; Swoboda, Ines; Valenta, Rudolf; Reichelt, Rudolf


    Polygalacturonases were recently identified as important grass pollen allergens and designated group 13 allergens. The objective of the present study was to investigate the presence of group 13 grass pollen allergens in different grass species, their release and ultrastructural location in dry and hydrated grass pollen. Nitrocellulose-blotted allergen extracts from 12 wild and cultivated grass genera were probed with a rabbit antiserum raised against purified recombinant timothy grass pollen allergen, Phl p 13. The release kinetics of Phl p 13 from timothy grass pollen hydrated for 0.5 min to 3 h were analyzed by immunoblotting. Phl p 13 was localized in dry and hydrated grass pollen grains by immunogold field emission scanning and transmission electron microscopy. Group 13 allergens were detected in all 12 wild and cultivated grass genera representing the major subfamilies of the Poaceae. Ultrastructurally, the allergen was located in the wall and in the cytoplasm of timothy grass pollen grains. In the cytoplasm, Phl p 13 was associated with polysaccharide particles and as yet undescribed stacks of microtubule-like structures. After hydration in rain water, pollen grains expel cytoplasmic particles of respirable size containing Phl p 13, which becomes detectable in aqueous supernatants already after 0.5 min. Group 13 allergens represent one set of marker allergens which specifically occur in pollen of the major grass subfamilies and are rapidly released in association with respirable particles after pollen hydration. They may be considered as environmental markers for grass pollen exposure and group 13-specific IgE antibodies as immunological markers for genuine grass pollen sensitization. Copyright (c) 2005 S. Karger AG, Basel

  17. Fluorescence microscopy. (United States)

    Sanderson, Michael J; Smith, Ian; Parker, Ian; Bootman, Martin D


    Fluorescence microscopy is a major tool with which to monitor cell physiology. Although the concepts of fluorescence and its optical separation using filters remain similar, microscope design varies with the aim of increasing image contrast and spatial resolution. The basics of wide-field microscopy are outlined to emphasize the selection, advantages, and correct use of laser scanning confocal microscopy, two-photon microscopy, scanning disk confocal microscopy, total internal reflection, and super-resolution microscopy. In addition, the principles of how these microscopes form images are reviewed to appreciate their capabilities, limitations, and constraints for operation. © 2014 Cold Spring Harbor Laboratory Press.

  18. Electron Microscopy. (United States)

    Beer, Michael


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

  19. Ballistic Electron Emission Microscopy (BEEM) of Au/Pr{sub 2}O{sub 3}/Si structures; Ballistische Elektronen Emissions Mikroskopie (BEEM) an Au/Si und Au/Pr{sub 2}O{sub 3}/Si-Strukturen

    Energy Technology Data Exchange (ETDEWEB)

    Mauch, I.


    This thesis describes Ballistic Electron Emission Microscopy (BEEM) measurements of Au/Si(111) and Au/Pr{sub 2}O{sub 3}/Si(111) structures. This technique is based on Scanning Tunnelling Microscopy (STM). It measures the ballistic transport of hot electrons through parts of the sample and across an interface, which provides a potential barrier. One part of this work was to modify the BEEM apparatus and to implement a lock in method, which modulates the tunnel current with a small frequency. In this way it is possible to study samples with very low resistance (as low as 30 k{omega}), which widely enlarges the number of samples which are appropriate for BEEM measurement at room temperature. Both types of samples studied in this thesis had low resistance and were therefore studied using the lock in method. For the classical BEEM system Au/Si(111), we observed a pronounced dependence of the sample resistance of Au/Si(111)-7 x 7 on the preparation temperature. We developed a model for the resistance of thermal prepared Au/Si(111)-7 x 7 samples. The model identifies that the low resistance is due to the surface conductivity of the reconstructed silicon surface. If the surface is prepared at a lower temperature (but still high enough that the surface is cleaned and the silicon dioxide desorbed) rough areas remain on the surface, which reduce the surface conductivity. For BEEM measurements flat areas of the sample surface are selected. The low temperature prepared samples we were able to obtain BEEM spectra as well as images at room temperature using the lock in method. The sesquioxide of praseodymium (Pr{sub 2}O{sub 3}) is currently discussed as a possible candidate for a gate oxide in semiconductor devices, since it has some of the required material properties such as a high dielectric constant, low leakage current and epitaxial growth on Si(100). We have for the first time performed BEEM measurement of praseodymium oxide. Despite a low resistance of the structures we

  20. Domain wall velocity measurement in permalloy nanowires with X-ray magnetic circular dichroism imaging and single shot Kerr microscopy

    Energy Technology Data Exchange (ETDEWEB)

    Moore, T.A., E-mail: t.a.moore@physics.or [Fachbereich Physik, Universitaet Konstanz, Universitaetsstrasse 10, 78457 Konstanz (Germany); Klaeui, M.; Heyne, L.; Moehrke, P. [Fachbereich Physik, Universitaet Konstanz, Universitaetsstrasse 10, 78457 Konstanz (Germany); Backes, D.; Rhensius, J. [Fachbereich Physik, Universitaet Konstanz, Universitaetsstrasse 10, 78457 Konstanz (Germany); Laboratory for Micro- and Nanotechnology, Paul Scherrer Institut, 5232 Villigen PSI (Switzerland); Ruediger, U. [Fachbereich Physik, Universitaet Konstanz, Universitaetsstrasse 10, 78457 Konstanz (Germany); Heyderman, L.J. [Laboratory for Micro- and Nanotechnology, Paul Scherrer Institut, 5232 Villigen PSI (Switzerland); Mentes, T.O.; Nino, M.A.; Locatelli, A. [Sincrotrone Trieste, 34012 Basovizza-Trieste (Italy); Potenza, A.; Marchetto, H.; Cavill, S.; Dhesi, S.S. [Diamond Light Source Ltd., Harwell Science and Innovation Campus, Didcot, Oxfordshire, OX11 0DE (United Kingdom)


    Domain walls (DWs) propagated along nanoscale magnetic wires by current or field pulses could potentially be used for data storage or logic applications, but the understanding of the DW dynamics, particularly under the influence of spin-polarized current, is incomplete. Measuring the velocity can give insights into the physics of the DW motion. Here we demonstrate DW velocity measurements in permalloy (Ni{sub 80}Fe{sub 20}) nanowires (1500 nm width and 20 nm thickness) using the techniques of X-ray magnetic circular dichroism photoemission electron microscopy (XMCD-PEEM) to image the magnetic contrast in the nanowires, and single shot Kerr microscopy, which allows for dynamic measurements. The magnetic imaging yields the average velocity as well as information on the DW spin structure, whereas the single shot method highlights the stochastic nature of the DW motion.

  1. Domain wall velocity measurement in permalloy nanowires with X-ray magnetic circular dichroism imaging and single shot Kerr microscopy (United States)

    Moore, T. A.; Kläui, M.; Heyne, L.; Möhrke, P.; Backes, D.; Rhensius, J.; Rüdiger, U.; Heyderman, L. J.; Mentes, T. O.; Niño, M. Á.; Locatelli, A.; Potenza, A.; Marchetto, H.; Cavill, S.; Dhesi, S. S.


    Domain walls (DWs) propagated along nanoscale magnetic wires by current or field pulses could potentially be used for data storage or logic applications, but the understanding of the DW dynamics, particularly under the influence of spin-polarized current, is incomplete. Measuring the velocity can give insights into the physics of the DW motion. Here we demonstrate DW velocity measurements in permalloy ( Ni80Fe20) nanowires (1500 nm width and 20 nm thickness) using the techniques of X-ray magnetic circular dichroism photoemission electron microscopy (XMCD-PEEM) to image the magnetic contrast in the nanowires, and single shot Kerr microscopy, which allows for dynamic measurements. The magnetic imaging yields the average velocity as well as information on the DW spin structure, whereas the single shot method highlights the stochastic nature of the DW motion.

  2. The Verwey transition observed by spin-resolved photoemission electron microscopy

    Energy Technology Data Exchange (ETDEWEB)

    Figuera, Juan de la, E-mail: [Instituto de Química Física “Rocasolano”, CSIC, Madrid E-28006 (Spain); Tusche, Christian [Max Planck Institute of Microstructure Physics, Halle D-06120 (Germany); Forschungszentrum Jülich GmbH, Peter Grünberg Institut (PGI-6), D-52425 Jülich (Germany)


    Highlights: • First observations of magnetic domains on magnetite (001) by spin-resolved PEEM. • Spin-polarization through the Verwey transitions does not change appreciably. • Shape and distribution of domains has been observed through the Verwey transition. - Abstract: We have imaged the magnetic domains on magnetite (001) through the Verwey transition by means of spin-resolved photoemission electron microscopy. A He laboratory source is used for illumination. The magnetic domains walls above the Verwey transition are aligned with 〈110〉 in-plane directions. Below the Verwey transition, the domain structure is interpreted as arising from a distribution of areas with different monoclinic c-axis, with linear 180° domain walls within each area and ragged edges when the magnetic domain boundaries coincide with structural domain walls. The domains evolve above the Verwey transition, while they are static below.

  3. Electrostatically focused addressable field emission array chips (AFEA's) for high-speed massively parallel maskless digital E-beam direct write lithography and scanning electron microscopy (United States)

    Thomas, Clarence E.; Baylor, Larry R.; Voelkl, Edgar; Simpson, Michael L.; Paulus, Michael J.; Lowndes, Douglas H.; Whealton, John H.; Whitson, John C.; Wilgen, John B.


    Systems and methods are described for addressable field emission array (AFEA) chips. A method of operating an addressable field-emission array, includes: generating a plurality of electron beams from a pluralitly of emitters that compose the addressable field-emission array; and focusing at least one of the plurality of electron beams with an on-chip electrostatic focusing stack. The systems and methods provide advantages including the avoidance of space-charge blow-up.

  4. Acoustic microscopy

    CERN Document Server

    Briggs, Andrew


    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.

  5. Insight into the spin state at the surface of LaCoO3 revealed by photoemission electron microscopy (United States)

    Yaroslavtsev, A. A.; Izquierdo, M.; Carley, R.; Dávila, M. E.; Ünal, A. A.; Kronast, F.; Lichtenstein, A.; Scherz, A.; Molodtsov, S. L.


    The evolution of the spin transition in LaCoO3 has been investigated with photoemission electron microscopy (PEEM) as a function of temperature. The investigated temperature range spanned from a predominantly low spin configuration (125 K) to the proposed percolation limit for metallization (413 K). The data show that the spin configuration exhibits an inhomogeneous spatial distribution that is very sensitive to the surface preparation method. In the region of the semiconductor-to-metal transition (300 to 450 K), the spatial contrast is continuously reduced, indicating a smooth transition without domain percolation. These observations support a new interpretation of the temperature evolution of the system that is in agreement with current theoretical understanding of the spin transition.

  6. Correlative microscopy. (United States)

    Loussert Fonta, Céline; Humbel, Bruno M


    In recent years correlative microscopy, combining the power and advantages of different imaging system, e.g., light, electrons, X-ray, NMR, etc., has become an important tool for biomedical research. Among all the possible combinations of techniques, light and electron microscopy, have made an especially big step forward and are being implemented in more and more research labs. Electron microscopy profits from the high spatial resolution, the direct recognition of the cellular ultrastructure and identification of the organelles. It, however, has two severe limitations: the restricted field of view and the fact that no live imaging can be done. On the other hand light microscopy has the advantage of live imaging, following a fluorescently tagged molecule in real time and at lower magnifications the large field of view facilitates the identification and location of sparse individual cells in a large context, e.g., tissue. The combination of these two imaging techniques appears to be a valuable approach to dissect biological events at a submicrometer level. Light microscopy can be used to follow a labelled protein of interest, or a visible organelle such as mitochondria, in time, then the sample is fixed and the exactly same region is investigated by electron microscopy. The time resolution is dependent on the speed of penetration and fixation when chemical fixatives are used and on the reaction time of the operator for cryo-fixation. Light microscopy can also be used to identify cells of interest, e.g., a special cell type in tissue or cells that have been modified by either transfections or RNAi, in a large population of non-modified cells. A further application is to find fluorescence labels in cells on a large section to reduce searching time in the electron microscope. Multiple fluorescence labelling of a series of sections can be correlated with the ultrastructure of the individual sections to get 3D information of the distribution of the marked proteins: array

  7. Visualization of the cytostome in Trypanosoma cruzi by high resolution field emission scanning electron microscopy using secondary and backscattered electron imaging. (United States)

    Vatarunakamura, Celso; Ueda-Nakamura, Tânia; de Souza, Wanderley


    High resolution scanning electron microscopy was used to analyze the surface of epimastigote, amastigote and trypomastigote forms of Trypanosoma cruzi. Significant differences were observed between these forms and in different areas of the same cell. The cytostome found in amastigote and epimastigote forms could be easily visualized in images, which resemble those obtained only using the freeze-fracture technique. In contrast to other areas of the cell surface, the region of the cytostome, localized close to the flagellar pocket, showed a rugous surface and an opening with a diameter of 90 nm. Gold-labeled concanavalin A binds to the whole cell surface. However, the extent of binding was much higher in the region of the cytostome. The results obtained show that high resolution scanning electron microscopy is a powerful technique for analyzing the surface of protozoa.

  8. Confocal microscopy

    Indian Academy of Sciences (India)

    molecular aggregates in artificial light harvesting sys- tem it is important to elucidate the exciton dynamics of individual micro-rods which can be achieved by using confocal microscopy and polarization resolved single molecule fluorescence spectroscopy.30 41 In the present work, we have studied exciton dynamics of two.

  9. Monitoring the dynamics of the surface deformation prior to the onset of plasma emission during femtosecond laser ablation of noble metals by time-resolved reflectivity microscopy (United States)

    Carrasco-García, I.; Vadillo, José M.; Javier Laserna, J.


    The generation of an expanding plasma during laser ablation is preceded at early times by the formation and evolution of a subsurface melted front. The monitoring of such transient event can't be studied by conventional spectroscopic techniques. Pump-probe femtosecond microscopy allows the following of the surface changes during femtosecond laser ablation taking advantage of the formation of a number of dynamic Newton rings that evolve with time. Measurements at different times allow the quantification of the radial expansion velocity of the molten material. For Au and Ag, expansions in the range of 7000-12,000 m/s for Au and 3000-21,000 m/s have been calculated depending on the pump energies. Such values correspond to hypersonic velocities with Mach number between 3 and 6.

  10. Quantitative backscattered electron imaging of field emission scanning electron microscopy for discrimination of nano-scale elements with nm-order spatial resolution. (United States)

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


    Discrimination of thin film elements by backscattered electron (BSE) imaging of field emission scanning electron microscope was examined. Incident electron acceleration voltage dependence on thin films' BSE intensities in five elements (Au, Ag, Ge, Cu and Fe) on a silicon substrate was experimentally measured from 3 to 30 kV. Normalization of BSE intensities using the difference between maximum and minimum brightness was proposed and allowed reproducible comparison among the elements. Measured intensities, which have correlation with electron backscattering coefficient against atomic number, indicated the existence of adequate acceleration voltage for improvement of resolution to discriminate different elements, showing the possibility of discriminating at least these six elements simultaneously by BSE imaging with nanometer-scale spatial resolution.

  11. Scanning ultrafast electron microscopy. (United States)

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


    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.

  12. Scanning electron microscopy-energy dispersive X-ray spectrometry (SEM-EDX) and aerosol time-of-flight mass spectrometry (ATOFMS) single particle analysis of metallurgy plant emissions. (United States)

    Arndt, J; Deboudt, K; Anderson, A; Blondel, A; Eliet, S; Flament, P; Fourmentin, M; Healy, R M; Savary, V; Setyan, A; Wenger, J C


    The chemical composition of single particles deposited on industrial filters located in three different chimneys of an iron-manganese (Fe-Mn) alloy manufacturing plant have been compared using aerosol time-of-flight mass spectrometry (ATOFMS) and scanning electron microscopy-energy dispersive X-ray spectrometry (SEM-EDX). Very similar types of particles were observed using both analytical techniques. Calcium-containing particles dominated in the firing area of the sintering unit, Mn and/or Al-bearing particles were observed at the cooling area of the sintering unit, while Mn-containing particles were dominant at the smelting unit. SEM-EDX analysis of particles collected downstream of the industrial filters showed that the composition of the particles emitted from the chimneys is very similar to those collected on the filters. ATOFMS analysis of ore samples was also performed to identify particulate emissions that could be generated by wind erosion and manual activities. Specific particle types have been identified for each emission source (chimneys and ore piles) and can be used as tracers for source apportionment of ambient PM measured in the vicinity of the industrial site. Copyright © 2015 Elsevier Ltd. All rights reserved.

  13. Enhanced Emission from Single Isolated Gold Quantum Dots Investigated Using Two-Photon-Excited Fluorescence Near-Field Scanning Optical Microscopy. (United States)

    Abeyasinghe, Neranga; Kumar, Santosh; Sun, Kai; Mansfield, John F; Jin, Rongchao; Goodson, Theodore


    New approaches in molecular nanoscopy are greatly desired for interrogation of biological, organic, and inorganic objects with sizes below the diffraction limit. Our current work investigates emergent monolayer-protected gold quantum dots (nanoclusters, NCs) composed of 25 Au atoms by utilizing two-photon-excited fluorescence (TPEF) near-field scanning optical microscopy (NSOM) at single NC concentrations. Here, we demonstrate an approach to synthesize and isolate single NCs on solid glass substrates. Subsequent investigation of the NCs using TPEF NSOM reveals that, even when they are separated by distances of several tens of nanometers, we can excite and interrogate single NCs individually. Interestingly, we observe an enhanced two-photon absorption (TPA) cross section for single Au25 NCs that can be attributed to few-atom local field effects and to local field-induced microscopic cascading, indicating their potential for use in ultrasensitive sensing, disease diagnostics, cancer cell therapy, and molecular computers. Finally, we report room-temperature aperture-based TPEF NSOM imaging of these NCs for the first time at 30 nm point resolution, which is a ∼5-fold improvement compared to the previous best result for the same technique. This report unveils the unique combination of an unusually large TPA cross section and the high photostability of Au NCs to (non-destructively) investigate stable isolated single NCs using TPEF NSOM. This is the first reported optical study of monolayer-protected single quantum clusters, opening some very promising opportunities in spectroscopy of nanosized objects, bioimaging, ultrasensitive sensing, molecular computers, and high-density data storage.

  14. Atom probe field ion microscopy and related topics: A bibliography 1991

    Energy Technology Data Exchange (ETDEWEB)

    Russell, K.F.; Miller, M.K.


    This report contains a bibliography for 1991 on the following topics: Atom probe field ion microscopy; field desorption mass spectrometry; field emission; field ion microscopy; and field emission theory.

  15. Field-emission scanning electron microscopy and energy-dispersive x-ray analysis to understand the role of tannin-based dyes in the degradation of historical wool textiles. (United States)

    Restivo, Annalaura; Degano, Ilaria; Ribechini, Erika; Pérez-Arantegui, Josefina; Colombini, Maria Perla


    An innovative approach, combining field-emission scanning electron microscopy (FESEM) with energy dispersive X-ray spectroscopy (EDX) analysis, is presented to investigate the degradation mechanisms affecting tannin-dyed wool. In fact, tannin-dyed textiles are more sensitive to degradation then those dyed with other dyestuffs, even in the same conservation conditions. FESEM-EDX was first used to study a set of 48 wool specimens (artificially aged) dyed with several raw materials and mordants, and prepared according to historical dyeing recipes. EDX analysis was performed on the surface of wool threads and on their cross-sections. In addition, in order to validate the model formulated by the analysis of reference materials, several samples collected from historical and archaeological textiles were subjected to FESEM-EDX analysis. FESEM-EDX investigations enabled us to reveal the correlation between elemental composition and morphological changes. In addition, aging processes were clarified by studying changes in the elemental composition of wool from the protective cuticle to the fiber core in cross-sections. Morphological and elemental analysis of wool specimens and of archaeological and historical textiles showed that the presence of tannins increases wool damage, primarily by causing a sulfur decrease and fiber oxidation.

  16. Efficient general procedure to access a diversity of gold(0) particles and gold(I) phosphine complexes from a simple HAuCl4 source. Localization of homogeneous/heterogeneous system's interface and field-emission scanning electron microscopy study. (United States)

    Zalesskiy, Sergey S; Sedykh, Alexander E; Kashin, Alexey S; Ananikov, Valentine P


    Soluble gold precatalysts, aimed for homogeneous catalysis, under certain conditions may form nanoparticles, which dramatically change the mechanism and initiate different chemistry. The present study addresses the question of designing gold catalysts, taking into account possible interconversions and contamination at the homogeneous/heterogeneous system's interface. It was revealed that accurate localization of boundary experimental conditions for formation of molecular gold complexes in solution versus nucleation and growth of gold particles opens new opportunities for well-known gold chemistry. Within the developed concept, a series of practical procedures was created for efficient synthesis of soluble gold complexes with various phosphine ligands (R3P)AuCl (90-99% yield) and for preparation of different types of gold materials. The effect of the ligand on the particles growth in solution has been observed and characterized with high-resolution field-emission scanning electron microscopy (FE-SEM) study. Two unique types of nanostructured gold materials were prepared: hierarchical agglomerates and gold mirror composed of ultrafine smoothly shaped particles.

  17. Order and orientation control of mesoporous silica films on conducting gold substrates formed by dip-coating and self-assembly: a grazing angle of incidence small-angle X-ray scattering and field emission scanning electron microscopy study. (United States)

    Tate, Michael P; Eggiman, Brian W; Kowalski, Jonathan D; Hillhouse, Hugh W


    Grazing-angle of incidence small-angle X-ray scattering (GISAXS) and high-resolution field emission scanning electron microscopy have been used to characterize the mesophase symmetry, orientation, and long-range order in PEO20-PPO70-PEO20 (Pluronic P123) templated mesoporous silica thin films on conducting gold substrates as a function of silica-to-ethylene oxide (Si/EO) block ratio and relative humidity (RH). The films are formed by dip-coating followed by evaporation-induced self-assembly under tightly controlled RH. The general evolution of the mesophase follows the trends that are expected based on shape factors due to swelling of the PEO block. However, changes in orientation of the nanostructure relative to the substrate and the degree of long-range order are found to depend on Si/EO ratio. These effects are likely due to the dynamics of evaporation and self-assembly. Generally, at Si/EO ratios lower than 3.29, the films contained regions where the nanostructure was not oriented relative to the plane of the substrate. However, for Si/EO ratios greater than 3.62, conditions were found where the nanostructure of the film was highly oriented relative to the plane of the substrate. This is true over the range of RH studied, independent of the nanostructure symmetry. For low Si/EO ratios at the highest RH levels, the films were composed of a mixture of spherical and cylindrical pores. At high Si/EO ratios and high RH levels, the films had a highly oriented R-3m nanostructure but displayed streaking perpendicular to the substrate in the Bragg spots on GISAXS patterns. This streaking is interpreted as faulting along planes parallel to the substrate.

  18. eV-TEM: Transmission electron microscopy in a low energy cathode lens instrument

    Energy Technology Data Exchange (ETDEWEB)

    Geelen, Daniël, E-mail: [Huygens-Kamerlingh Onnes Laboratory, Leiden Institute of Physics, Leiden University, P.O. Box 9504, 2300 RA Leiden (Netherlands); Thete, Aniket [Huygens-Kamerlingh Onnes Laboratory, Leiden Institute of Physics, Leiden University, P.O. Box 9504, 2300 RA Leiden (Netherlands); Schaff, Oliver; Kaiser, Alexander [SPECS GmbH, Voltastrasse 5, D-13355 Berlin (Germany); Molen, Sense Jan van der [Huygens-Kamerlingh Onnes Laboratory, Leiden Institute of Physics, Leiden University, P.O. Box 9504, 2300 RA Leiden (Netherlands); Tromp, Rudolf [IBM T.J. Watson Research Center, 1101 Kitchawan Road, P.O. Box 218, Yorktown Heights, NY 10598 (United States)


    We are developing a transmission electron microscope that operates at extremely low electron energies, 0–40 eV. We call this technique eV-TEM. Its feasibility is based on the fact that at very low electron energies the number of energy loss pathways decreases. Hence, the electron inelastic mean free path increases dramatically. eV-TEM will enable us to study elastic and inelastic interactions of electrons with thin samples. With the recent development of aberration correction in cathode lens instruments, a spatial resolution of a few nm appears within range, even for these very low electron energies. Such resolution will be highly relevant to study biological samples such as proteins and cell membranes. The low electron energies minimize adverse effects due to radiation damage. - Highlights: • We present a new way of performing low energy transmission electron microscopy in an aberration corrected LEEM/PEEM instrument. • We show a proof of principle where we measure transmitted electrons through a suspended graphene monolayer with a preliminary setup. • We present an improved setup design that provides better control of the incident electron beam.

  19. Correlated Light Microscopy and Electron Microscopy

    NARCIS (Netherlands)

    Sjollema, Klaas A.; Schnell, Ulrike; Kuipers, Jeroen; Kalicharan, Ruby; Giepmans, Ben N. G.; MullerReichert, T; Verkade, P


    Understanding where, when, and how biomolecules (inter)act is crucial to uncover fundamental mechanisms in cell biology. Recent developments in fluorescence light microscopy (FLM) allow protein imaging in living cells and at the near molecular level. However, fluorescence microscopy only reveals

  20. High-resolution electron microscopy

    CERN Document Server

    Spence, John C H


    This new fourth edition of the standard text on atomic-resolution transmission electron microscopy (TEM) retains previous material on the fundamentals of electron optics and aberration correction, linear imaging theory (including wave aberrations to fifth order) with partial coherence, and multiple-scattering theory. Also preserved are updated earlier sections on practical methods, with detailed step-by-step accounts of the procedures needed to obtain the highest quality images of atoms and molecules using a modern TEM or STEM electron microscope. Applications sections have been updated - these include the semiconductor industry, superconductor research, solid state chemistry and nanoscience, and metallurgy, mineralogy, condensed matter physics, materials science and material on cryo-electron microscopy for structural biology. New or expanded sections have been added on electron holography, aberration correction, field-emission guns, imaging filters, super-resolution methods, Ptychography, Ronchigrams, tomogr...

  1. Introduction to fluorescence microscopy. (United States)

    Ghiran, Ionita C


    This chapter is an overview of basic principles of fluorescence microscopy, including a brief history on the invention of this type of microscopy. The chapter highlights important points related to properties of fluorochromes, resolution in fluorescence microscopy, phase contrast and fluorescence, fluorescence filters, construction of a fluorescence microscope, and tips on the correct use of this equipment.

  2. Field Emission in Vacuum Microelectronics

    CERN Document Server

    Fursey, George; Schwoebel, Paul


    Field emission is a phenomenon described by quantum mechanics. Its emission capability is millions times higher than that of any other known types of electron emission. Nowadays this phenomenon is experiencing a new life due to wonderful applications in the atomic resolution microscopy, in electronic holography, and in the vacuum micro- and nanoelectronics in general. The main field emission properties, and some most remarkable experimental facts and applications, are described in this book.

  3. Semiconductor Surface Characterization by Scanning Probe Microscopies (United States)


    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

  4. Examining Thermally Sprayed Coats By Fluorescence Microscopy (United States)

    Street, Kenneth W., Jr.; Leonhardt, Todd A.


    True flaws distinquished from those induced by preparation of specimens. Fluorescence microscopy reveals debonding, porosity, cracks, and other flaws in specimens of thermally sprayed coating materials. Specimen illuminated, and dye it contains fluoresces, emitting light at different wavelength. Filters emphasize contrast between excitation light and emission light. Specimen viewed directly or photographed on color film.

  5. The 2015 super-resolution microscopy roadmap (United States)

    Hell, Stefan W.; Sahl, Steffen J.; Bates, Mark; Zhuang, Xiaowei; Heintzmann, Rainer; Booth, Martin J.; Bewersdorf, Joerg; Shtengel, Gleb; Hess, Harald; Tinnefeld, Philip; Honigmann, Alf; Jakobs, Stefan; Testa, Ilaria; Cognet, Laurent; Lounis, Brahim; Ewers, Helge; Davis, Simon J.; Eggeling, Christian; Klenerman, David; Willig, Katrin I.; Vicidomini, Giuseppe; Castello, Marco; Diaspro, Alberto; Cordes, Thorben


    Far-field optical microscopy using focused light is an important tool in a number of scientific disciplines including chemical, (bio)physical and biomedical research, particularly with respect to the study of living cells and organisms. Unfortunately, the applicability of the optical microscope is limited, since the diffraction of light imposes limitations on the spatial resolution of the image. Consequently the details of, for example, cellular protein distributions, can be visualized only to a certain extent. Fortunately, recent years have witnessed the development of ‘super-resolution’ far-field optical microscopy (nanoscopy) techniques such as stimulated emission depletion (STED), ground state depletion (GSD), reversible saturated optical (fluorescence) transitions (RESOLFT), photoactivation localization microscopy (PALM), stochastic optical reconstruction microscopy (STORM), structured illumination microscopy (SIM) or saturated structured illumination microscopy (SSIM), all in one way or another addressing the problem of the limited spatial resolution of far-field optical microscopy. While SIM achieves a two-fold improvement in spatial resolution compared to conventional optical microscopy, STED, RESOLFT, PALM/STORM, or SSIM have all gone beyond, pushing the limits of optical image resolution to the nanometer scale. Consequently, all super-resolution techniques open new avenues of biomedical research. Because the field is so young, the potential capabilities of different super-resolution microscopy approaches have yet to be fully explored, and uncertainties remain when considering the best choice of methodology. Thus, even for experts, the road to the future is sometimes shrouded in mist. The super-resolution optical microscopy roadmap of Journal of Physics D: Applied Physics addresses this need for clarity. It provides guidance to the outstanding questions through a collection of short review articles from experts in the field, giving a thorough

  6. Comparison of multi-mode parallel detection microscopy methods (United States)

    Zhu, Dazhao; Fang, Yue; Chen, Youhua; Hussain, Anwar; Kuang, Cuifang; Ding, Zhihua; Liu, Xu


    Four microscopy resolution enhancement methods based on parallel detection were investigated in this study: confocal microscopy with four pinhole sizes, fluorescence emission difference microscopy (FED) based on parallel detection, Airyscan microscopy, and virtual k-vector modulation optical microscopy (Vikmom). These methods use different algorithms to process parallel detection data and achieve resolution improvement. We investigated these methods first by performing simulations and then experimentally. In this report, the basic theories of these methods are briefly introduced. Then, analyses and comparisons of their imaging performances, especially in terms of resolution improvement, imaging speed, and signal-to-noise ratio, are presented. Finally, the results of our comparative study are summarized.

  7. New microscopy for nanoimaging

    CERN Document Server

    Kinjo, Y; Watanabe, M


    Two types of new microscopy, namely, X-ray contact microscopy (XRCM) in combination with atomic force microscopy (AFM) and X-ray projection microscopy (XRPM) using synchrotron radiation and zone plate optics were used to image the fine structures of human chromosomes. In the XRCM plus AFM system, location of X-ray images on a photoresist has become far easier than that with our previous method using transmission electron microscopy coupled with the replica method. In addition, the images obtained suggested that the conformation of chromatin fiber differs from the current textbook model regarding the architecture of a eukaryotic chromosome. X-ray images with high contrast of the specimens could be obtained with XRPM. The resolution of each microscopy was about 30 and 200-300 nm for XRCM plus AFM and XRPM, respectively. (author)

  8. Fluorescence (Multiwave) Confocal Microscopy. (United States)

    Welzel, J; Kästle, Raphaela; Sattler, Elke C


    In addition to reflectance confocal microscopy, multiwave confocal microscopes with different laser wavelengths in combination with exogenous fluorophores allow fluorescence mode confocal microscopy in vivo and ex vivo. Fluorescence mode confocal microscopy improves the contrast between the epithelium and the surrounding soft tissue and allows the depiction of certain structures, like epithelial tumors, nerves, and glands. Copyright © 2016 Elsevier Inc. All rights reserved.

  9. Electron Microscopy Center (EMC) (United States)

    Federal Laboratory Consortium — The Electron Microscopy Center (EMC) at Argonne National Laboratory develops and maintains unique capabilities for electron beam characterization and applies those...

  10. Coherent light microscopy

    CERN Document Server

    Ferraro, Pietro; Zalevsky, Zeev


    This book deals with the latest achievements in the field of optical coherent microscopy. While many other books exist on microscopy and imaging, this book provides a unique resource dedicated solely to this subject. Similarly, many books describe applications of holography, interferometry and speckle to metrology but do not focus on their use for microscopy. The coherent light microscopy reference provided here does not focus on the experimental mechanics of such techniques but instead is meant to provide a users manual to illustrate the strengths and capabilities of developing techniques. Th

  11. Bridging fluorescence microscopy and electron microscopy

    NARCIS (Netherlands)

    Giepmans, Ben N. G.

    Development of new fluorescent probes and fluorescence microscopes has led to new ways to study cell biology. With the emergence of specialized microscopy units at most universities and research centers, the use of these techniques is well within reach for a broad research community. A major

  12. Quantitative Fluorescence Microscopy of Protein Dynamics in Living Cells

    NARCIS (Netherlands)

    S.M. Ibrahim (Shehu)


    textabstractThe advent of confocal microscopy, fast microcomputers with high storage capacity and, moreover, the availability of fluorescent proteins of various excitation and emission properties have made fluorescence microscopy the method of choice in the study of protein behaviour in living

  13. Handbook of Microscopy for Nanotechnology (United States)

    Yao, Nan; Wang, Zhong L.

    This handbook highlights various key microcopic techniques and their applications in this fast-growing field. Topics to be covered include the following: scanning near field optical microscopy, confocal optical microscopy, atomic force microscopy, magnetic force microscopy, scanning turning microscopy, high-resolution scanning electron microscopy, and many more.

  14. LEDs for fluorescence microscopy

    NARCIS (Netherlands)

    Young, I.T.; Garini, Y.; Dietrich, H.R.C.; Van Oel, W.; Liqui Lung, G.


    Traditional light sources for fluorescence microscopy have been mercury lamps, xenon lamps, and lasers. These sources have been essential in the development of fluorescence microscopy but each can have serious disadvantages: lack of near monochromaticity, heat generation, cost, lifetime of the light

  15. Lasers for nonlinear microscopy. (United States)

    Wise, Frank


    Various versions of nonlinear microscopy are revolutionizing the life sciences, almost all of which are made possible because of the development of ultrafast lasers. In this article, the main properties and technical features of short-pulse lasers used in nonlinear microscopy are summarized. Recent research results on fiber lasers that will impact future instruments are also discussed.

  16. Focused ion beam and field-emission microscopy of metallic filaments in memory devices based on thin films of an ambipolar organic compound consisting of oxadiazole, carbazole, and fluorene units (United States)

    Pearson, Christopher; Bowen, Leon; Lee, Myung Won; Fisher, Alison L.; Linton, Katherine E.; Bryce, Martin R.; Petty, Michael C.


    We report on the mechanism of operation of organic thin film resistive memory architectures based on an ambipolar compound consisting of oxadiazole, carbazole, and fluorene units. Cross-sections of the devices have been imaged by electron microscopy both before and after applying a voltage. The micrographs reveal the growth of filaments, with diameters of 50 nm–100 nm, on the metal cathode. We suggest that these are formed by the drift of aluminium ions from the anode and are responsible for the observed switching and negative differential resistance phenomena in the memory devices.

  17. Confocal Raman microscopy

    CERN Document Server

    Dieing, Thomas; Hollricher, Olaf


    This second edition provides a cutting-edge overview of physical, technical and scientific aspects related to the widely used analytical method of confocal Raman microscopy. The book includes expanded background information and adds insights into how confocal Raman microscopy, especially 3D Raman imaging, can be integrated with other methods to produce a variety of correlative microscopy combinations. The benefits are then demonstrated and supported by numerous examples from the fields of materials science, 2D materials, the life sciences, pharmaceutical research and development, as well as the geosciences.

  18. International Multidisciplinary Microscopy Congress

    CERN Document Server

    Oral, Ahmet; Ozer, Mehmet; InterM; INTERM2013


    The International Multidisciplinary Microscopy Congress (INTERM2013) was organized on October 10-13, 2013. The aim of the congress was to bring together scientists from various branches to discuss the latest advances in the field of microscopy. The contents of the congress have been broadened to a more "interdisciplinary" scope, so as to allow all scientists working on related subjects to participate and present their work. These proceedings include 39 peer-reviewed technical papers, submitted by leading academic and research institutions from over 12 countries and representing some of the most cutting-edge research available. The 39 papers are grouped into the following sections: - Applications of Microscopy in the Physical Sciences - Applications of Microscopy in the Biological Sciences

  19. Clinical specular microscopy

    Energy Technology Data Exchange (ETDEWEB)

    Hirst, L.W.; Laing, R.A.


    This book provides the general ophthalmologist with a guide to the clinical applications of specular microscopy. Important material is included on laser injury, cataract surgery, corneal transplants, glaucoma, uveitis, and trauma.

  20. Light Sheet Fluorescence Microscopy (LSFM). (United States)

    Adams, Michael W; Loftus, Andrew F; Dunn, Sarah E; Joens, Matthew S; Fitzpatrick, James A J


    The development of confocal microscopy techniques introduced the ability to optically section fluorescent samples in the axial dimension, perpendicular to the image plane. These approaches, via the placement of a pinhole in the conjugate image plane, provided superior resolution in the axial (z) dimension resulting in nearly isotropic optical sections. However, increased axial resolution, via pinhole optics, comes at the cost of both speed and excitation efficiency. Light sheet fluorescent microscopy (LSFM), a century-old idea made possible with modern developments in both excitation and detection optics, provides sub-cellular resolution and optical sectioning capabilities without compromising speed or excitation efficiency. Over the past decade, several variations of LSFM have been implemented each with its own benefits and deficiencies. Here we discuss LSFM fundamentals and outline the basic principles of several major light-sheet-based imaging modalities (SPIM, inverted SPIM, multi-view SPIM, Bessel beam SPIM, and stimulated emission depletion SPIM) while considering their biological relevance in terms of intrusiveness, temporal resolution, and sample requirements. Copyright © 2015 John Wiley & Sons, Inc.

  1. Quantitative dispersion microscopy


    Fu, Dan; Choi, Wonshik; Sung, Yongjin; Yaqoob, Zahid; Ramachandra R Dasari; Feld, Michael


    Refractive index dispersion is an intrinsic optical property and a useful source of contrast in biological imaging studies. In this report, we present the first dispersion phase imaging of living eukaryotic cells. We have developed quantitative dispersion microscopy based on the principle of quantitative phase microscopy. The dual-wavelength quantitative phase microscope makes phase measurements at 310 nm and 400 nm wavelengths to quantify dispersion (refractive index increment ratio) of live...

  2. Multiphoton microscopy with near infrared contrast agents (United States)

    Yazdanfar, Siavash; Joo, Chulmin; Zhan, Chun; Berezin, Mikhail Y.; Akers, Walter J.; Achilefu, Samuel


    While multiphoton microscopy (MPM) has been performed with a wide range of excitation wavelengths, fluorescence emission has been limited to the visible spectrum. We introduce a paradigm for MPM of near-infrared (NIR) fluorescent molecular probes via nonlinear excitation at 1550 nm. This all-NIR system expands the range of available MPM fluorophores, virtually eliminates background autofluorescence, and allows for use of fiber-based, turnkey ultrafast lasers developed for telecommunications.

  3. Low copper and high manganese levels in prion protein plaques (United States)

    Johnson, Christopher J.; Gilbert, P.U.P.A.; Abrecth, Mike; Baldwin, Katherine L.; Russell, Robin E.; Pedersen, Joel A.; McKenzie, Debbie


    Accumulation of aggregates rich in an abnormally folded form of the prion protein characterize the neurodegeneration caused by transmissible spongiform encephalopathies (TSEs). The molecular triggers of plaque formation and neurodegeneration remain unknown, but analyses of TSE-infected brain homogenates and preparations enriched for abnormal prion protein suggest that reduced levels of copper and increased levels of manganese are associated with disease. The objectives of this study were to: (1) assess copper and manganese levels in healthy and TSE-infected Syrian hamster brain homogenates; (2) determine if the distribution of these metals can be mapped in TSE-infected brain tissue using X-ray photoelectron emission microscopy (X-PEEM) with synchrotron radiation; and (3) use X-PEEM to assess the relative amounts of copper and manganese in prion plaques in situ. In agreement with studies of other TSEs and species, we found reduced brain levels of copper and increased levels of manganese associated with disease in our hamster model. We also found that the in situ levels of these metals in brainstem were sufficient to image by X-PEEM. Using immunolabeled prion plaques in directly adjacent tissue sections to identify regions to image by X-PEEM, we found a statistically significant relationship of copper-manganese dysregulation in prion plaques: copper was depleted whereas manganese was enriched. These data provide evidence for prion plaques altering local transition metal distribution in the TSE-infected central nervous system.

  4. Confocal Raman Microscopy

    CERN Document Server

    Dieing, Thomas; Toporski, Jan


    Confocal Raman Microscopy is a relatively new technique that allows chemical imaging without specific sample preparation. By integrating a sensitive Raman spectrometer within a state-of-the-art microscope, Raman microscopy with a spatial resolution down to 200nm laterally and 500nm vertically can be achieved using visible light excitation. Recent developments in detector and computer technology as well as optimized instrument design have reduced integration times of Raman spectra by orders of magnitude, so that complete images consisting of tens of thousands of Raman spectra can be acquired in seconds or minutes rather than hours, which used to be standard just one decade ago. The purpose of this book is to provide the reader a comprehensive overview of the rapidly developing field of Confocal Raman Microscopy and its applications.

  5. 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...

  6. Single particle electron microscopy

    NARCIS (Netherlands)

    Boekema, Egbert J.; Folea, Mihaela; Kouril, Roman


    Electron microscopy (EM) in combination with image analysis is a powerful technique to study protein structures at low, medium, and high resolution. Since electron micrographs of biological objects are very noisy, improvement of the signal-to-noise ratio by image processing is an integral part of

  7. and transmission electron microscopy

    African Journals Online (AJOL)


    immune-electron microscopy (IEM) from patients' feces. They reported this virus particle as the causative agent of winter vomiting outbreaks in Norwalk (Kapikian et al.,. 1972). This is the remarkable landmark study of non- bacterial gastroenteritis viruses, especially for small round structured viruses (SRSVs). After that, many.

  8. Atomic Force Microscopy

    Indian Academy of Sciences (India)

    Home; Journals; Resonance – Journal of Science Education; Volume 15; Issue 7. Atomic Force Microscopy - A Tool to Unveil the Mystery of Biological Systems ... Transcription and Disease Laboratory, Molecular Biology and Genetics Unit, Jawaharlal Nehru Centre for Advanced Scientific Research, Jakkur, Bangalore 560 ...

  9. Photoacoustic computed microscopy (United States)

    Yao, Lei; Xi, Lei; Jiang, Huabei


    Photoacoustic microscopy (PAM) is emerging as a powerful technique for imaging microvasculature at depths beyond the ~1 mm depth limit associated with confocal microscopy, two-photon microscopy and optical coherence tomography. PAM, however, is currently qualitative in nature and cannot quantitatively measure important functional parameters including oxyhemoglobin (HbO2), deoxyhemoglobin (HbR), oxygen saturation (sO2), blood flow (BF) and rate of oxygen metabolism (MRO2). Here we describe a new photoacoustic microscopic method, termed photoacoustic computed microscopy (PACM) that combines current PAM technique with a model-based inverse reconstruction algorithm. We evaluate the PACM approach using tissue-mimicking phantoms and demonstrate its in vivo imaging ability of quantifying HbO2, HbR, sO2, cerebral BF and cerebral MRO2 at the small vessel level in a rodent model. This new technique provides a unique tool for neuroscience research and for visualizing microvasculature dynamics involved in tumor angiogenesis and in inflammatory joint diseases.

  10. Second harmonic generation microscopy

    DEFF Research Database (Denmark)

    Brüggemann, Dagmar Adeline; Brewer, Jonathan R.; Risbo, Jens


    -temperature endotherm peak observable in the differential scanning calorimetry (DSC) thermograms. DSC analysis of epimysium, the connective tissue layer that enfold skeletal muscles, produces one large endotherm starting at 57 °C and peaking at 59.5 °C. SHG microscopy of collagen fibers reveals a variability of thermal...

  11. Magnetic Force Microscopy

    NARCIS (Netherlands)

    Abelmann, Leon

    Principle of MFM In magnetic force microscopy (MFM), the magnetic stray field above a very flat specimen, or sample, is detected by placing a small magnetic element, the tip, mounted on a cantilever spring very close to the surface of the sample (Figure 1). Typical dimensions are a cantilever length

  12. 3D multiplexed immunoplasmonics microscopy (United States)

    Bergeron, Éric; Patskovsky, Sergiy; Rioux, David; Meunier, Michel


    Selective labelling, identification and spatial distribution of cell surface biomarkers can provide important clinical information, such as distinction between healthy and diseased cells, evolution of a disease and selection of the optimal patient-specific treatment. Immunofluorescence is the gold standard for efficient detection of biomarkers expressed by cells. However, antibodies (Abs) conjugated to fluorescent dyes remain limited by their photobleaching, high sensitivity to the environment, low light intensity, and wide absorption and emission spectra. Immunoplasmonics is a novel microscopy method based on the visualization of Abs-functionalized plasmonic nanoparticles (fNPs) targeting cell surface biomarkers. Tunable fNPs should provide higher multiplexing capacity than immunofluorescence since NPs are photostable over time, strongly scatter light at their plasmon peak wavelengths and can be easily functionalized. In this article, we experimentally demonstrate accurate multiplexed detection based on the immunoplasmonics approach. First, we achieve the selective labelling of three targeted cell surface biomarkers (cluster of differentiation 44 (CD44), epidermal growth factor receptor (EGFR) and voltage-gated K+ channel subunit KV1.1) on human cancer CD44+ EGFR+ KV1.1+ MDA-MB-231 cells and reference CD44- EGFR- KV1.1+ 661W cells. The labelling efficiency with three stable specific immunoplasmonics labels (functionalized silver nanospheres (CD44-AgNSs), gold (Au) NSs (EGFR-AuNSs) and Au nanorods (KV1.1-AuNRs)) detected by reflected light microscopy (RLM) is similar to the one with immunofluorescence. Second, we introduce an improved method for 3D localization and spectral identification of fNPs based on fast z-scanning by RLM with three spectral filters corresponding to the plasmon peak wavelengths of the immunoplasmonics labels in the cellular environment (500 nm for 80 nm AgNSs, 580 nm for 100 nm AuNSs and 700 nm for 40 nm × 92 nm AuNRs). Third, the developed

  13. High numerical aperture tabletop soft x-ray diffraction microscopy with 70-nm resolution


    Sandberg, Richard L.; Song, Changyong; Wachulak, Przemyslaw W.; Raymondson, Daisy A.; Paul, Ariel; Amirbekian, Bagrat; Lee, Edwin; Sakdinawat, Anne E.; La-O-Vorakiat, Chan; Marconi, Mario C.; Menoni, Carmen S.; Murnane, Margaret M.; Rocca, Jorge J.; Kapteyn, Henry C.; Miao, Jianwei


    Light microscopy has greatly advanced our understanding of nature. The achievable resolution, however, is limited by optical wavelengths to ≈200 nm. By using imaging and labeling technologies, resolutions beyond the diffraction limit can be achieved for specialized specimens with techniques such as near-field scanning optical microscopy, stimulated emission depletion microscopy, and photoactivated localization microscopy. Here, we report a versatile soft x-ray diffraction microscope with 70- ...

  14. Polarized Light Microscopy (United States)

    Frandsen, Athela F.


    Polarized light microscopy (PLM) is a technique which employs the use of polarizing filters to obtain substantial optical property information about the material which is being observed. This information can be combined with other microscopy techniques to confirm or elucidate the identity of an unknown material, determine whether a particular contaminant is present (as with asbestos analysis), or to provide important information that can be used to refine a manufacturing or chemical process. PLM was the major microscopy technique in use for identification of materials for nearly a century since its introduction in 1834 by William Fox Talbot, as other techniques such as SEM (Scanning Electron Microscopy), FTIR (Fourier Transform Infrared spectroscopy), XPD (X-ray Powder Diffraction), and TEM (Transmission Electron Microscopy) had not yet been developed. Today, it is still the only technique approved by the Environmental Protection Agency (EPA) for asbestos analysis, and is often the technique first applied for identification of unknown materials. PLM uses different configurations in order to determine different material properties. With each configuration additional clues can be gathered, leading to a conclusion of material identity. With no polarizing filter, the microscope can be used just as a stereo optical microscope, and view qualities such as morphology, size, and number of phases. With a single polarizing filter (single polars), additional properties can be established, such as pleochroism, individual refractive indices, and dispersion staining. With two polarizing filters (crossed polars), even more can be deduced: isotropy vs. anisotropy, extinction angle, birefringence/degree of birefringence, sign of elongation, and anomalous polarization colors, among others. With the use of PLM many of these properties can be determined in a matter of seconds, even for those who are not highly trained. McCrone, a leader in the field of polarized light microscopy, often

  15. Non-radiative excitation fluorescence microscopy (United States)

    Riachy, Lina; Vézy, Cyrille; Jaffiol, Rodolphe


    Non-radiative Excitation Fluorescence Microscopy (NEFM) constitutes a new way to observe biological samples beyond the diffraction limit. Non-radiative excitation of the samples is achieved by coating the substrate with donor species, such as quantum dots (QDs). Thus the dyes are not excited directly by the laser source, as in common fluorescence microscopy, but through a non-radiative energy transfer. To prevent dewetting of the donor film, we have recently implemented a silanization process to covalently bond the QDs on the substrate. An homogeneous monolayer of QDs was then deposited on only one side of the coverslips. Atomic force microscopy was then used to characterize the QD layer. We highlight the potential of our method through the study of Giant Unilamellar Vesicles (GUVs) labeled with DiD as acceptor, in interaction with surface functionalized with poly-L-lysine. In the presence of GUVs, we observed a quenching of QDs emission, together with an emission of DiD located in the membrane, which clearly indicated that non-radiative energy transfer from QDs to DiD occurs.

  16. Light Sheet Fluorescence Microscopy (United States)

    Santi, Peter A.


    Light sheet fluorescence microscopy (LSFM) functions as a non-destructive microtome and microscope that uses a plane of light to optically section and view tissues with subcellular resolution. This method is well suited for imaging deep within transparent tissues or within whole organisms, and because tissues are exposed to only a thin plane of light, specimen photobleaching and phototoxicity are minimized compared to wide-field fluorescence, confocal, or multiphoton microscopy. LSFMs produce well-registered serial sections that are suitable for three-dimensional reconstruction of tissue structures. Because of a lack of a commercial LSFM microscope, numerous versions of light sheet microscopes have been constructed by different investigators. This review describes development of the technology, reviews existing devices, provides details of one LSFM device, and shows examples of images and three-dimensional reconstructions of tissues that were produced by LSFM. PMID:21339178

  17. Multimodal hyperspectral optical microscopy (United States)

    Novikova, Irina V.; Smallwood, Chuck R.; Gong, Yu; Hu, Dehong; Hendricks, Leif; Evans, James E.; Bhattarai, Ashish; Hess, Wayne P.; El-Khoury, Patrick Z.


    We describe a unique approach to hyperspectral optical microscopy, herein achieved by coupling a hyperspectral imager to various optical microscopes. Hyperspectral fluorescence micrographs of isolated fluorescent beads are first employed to ensure spectral calibration of our detector and to gauge the attainable spatial resolution of our measurements. Different science applications of our instrument are then described. Spatially over-sampled absorption spectroscopy of a single lipid (18:1 Liss Rhod PE) layer reveals that optical densities on the order of 10-3 can be resolved by spatially averaging the recorded optical signatures. This is followed by three applications in the general areas of plasmonics and bioimaging. Notably, we deploy hyperspectral absorption microscopy to identify and image pigments within a simple biological system, namely, a single live Tisochrysis lutea cell. Overall, this work paves the way for multimodal spectral imaging measurements spanning the realms of several scientific disciplines.

  18. Electron microscopy of viruses. (United States)

    Laue, Michael


    Electron microscopy is widely used in virology because viruses are generally too small for a direct inspection by light microscopy. Analysis of virus morphology is necessary in many circumstances, e.g., for the diagnosis of a virus in particular clinical situations or the analysis of virus entry and assembly. Moreover, quality control of virus particle integrity is required if a virus is propagated in cell culture, particularly if the virus genome has changed. In most cases already the basic methodology for transmission electron microscopy, i.e., negative staining and ultrathin sectioning, is sufficient to give relevant information on virus ultrastructure. This chapter gives detailed information on the principles of these basic methodologies and provides simple but reliable protocols for a quick start. Moreover, the description of standard protocols for negative staining and ultrathin sectioning are supplemented by protocols on immuno-negative staining and rapid ultrathin sectioning. Finally, principles of methods for an extended ultrastructural research using more elaborate techniques, such as cryotechniques or methods to reveal the three-dimensional virus architecture, are briefly reviewed. Copyright © 2010 Elsevier Inc. All rights reserved.

  19. Deep Learning Microscopy

    KAUST Repository

    Rivenson, Yair


    We demonstrate that a deep neural network can significantly improve optical microscopy, enhancing its spatial resolution over a large field-of-view and depth-of-field. After its training, the only input to this network is an image acquired using a regular optical microscope, without any changes to its design. We blindly tested this deep learning approach using various tissue samples that are imaged with low-resolution and wide-field systems, where the network rapidly outputs an image with remarkably better resolution, matching the performance of higher numerical aperture lenses, also significantly surpassing their limited field-of-view and depth-of-field. These results are transformative for various fields that use microscopy tools, including e.g., life sciences, where optical microscopy is considered as one of the most widely used and deployed techniques. Beyond such applications, our presented approach is broadly applicable to other imaging modalities, also spanning different parts of the electromagnetic spectrum, and can be used to design computational imagers that get better and better as they continue to image specimen and establish new transformations among different modes of imaging.

  20. Super-resolution Microscopy in Plant Cell Imaging. (United States)

    Komis, George; Šamajová, Olga; Ovečka, Miroslav; Šamaj, Jozef


    Although the development of super-resolution microscopy methods dates back to 1994, relevant applications in plant cell imaging only started to emerge in 2010. Since then, the principal super-resolution methods, including structured-illumination microscopy (SIM), photoactivation localization microscopy (PALM), stochastic optical reconstruction microscopy (STORM), and stimulated emission depletion microscopy (STED), have been implemented in plant cell research. However, progress has been limited due to the challenging properties of plant material. Here we summarize the basic principles of existing super-resolution methods and provide examples of applications in plant science. The limitations imposed by the nature of plant material are reviewed and the potential for future applications in plant cell imaging is highlighted. Copyright © 2015 Elsevier Ltd. All rights reserved.

  1. STED microscopy with a supercontinuum laser source. (United States)

    Wildanger, Dominik; Rittweger, Eva; Kastrup, Lars; Hell, Stefan W


    We report on a straightforward yet powerful implementation of stimulated emission depletion (STED) fluorescence microscopy providing subdiffraction resolution in the far-field. Utilizing the same super-continuum pulsed laser source both for excitation and STED, this implementation of STED microscopy avoids elaborate preparations of laser pulses and conveniently provides multicolor imaging. Operating at pulse repetition rates around 1 MHz, it also affords reduced photobleaching rates by allowing the fluorophore to relax from excitable metastable dark states involved in photodegradation. The imaging of dense nanoparticles and of the microtubular network of mammalian cells evidences a spatial resolution of 30-50 nm in the focal plane, i.e. by a factor of 8-9 beyond the diffraction barrier.

  2. Electrochemical force microscopy (United States)

    Kalinin, Sergei V.; Jesse, Stephen; Collins, Liam F.; Rodriguez, Brian J.


    A system and method for electrochemical force microscopy are provided. The system and method are based on a multidimensional detection scheme that is sensitive to forces experienced by a biased electrode in a solution. The multidimensional approach allows separation of fast processes, such as double layer charging, and charge relaxation, and slow processes, such as diffusion and faradaic reactions, as well as capturing the bias dependence of the response. The time-resolved and bias measurements can also allow probing both linear (small bias range) and non-linear (large bias range) electrochemical regimes and potentially the de-convolution of charge dynamics and diffusion processes from steric effects and electrochemical reactivity.

  3. Reinventing Pocket Microscopy

    CERN Document Server

    Kamal, T; Lee, W M


    The key to the success of pocket microscopes stems from the convenience for anyone to magnify the fine details (tens of micrometres) of any object on-thespot. The capability with a portable microscope lets us surpass our limited vision and is commonly used in many areas of science, industry, education. The growth of imaging and computing power in smartphones is creating the possibility of converting your smartphone into a high power pocket microscope. In this article, we briefly describe the history of pocket microscopy and elucidate how mobile technologies are set to become the next platform for pocket microscopes

  4. Increasing estimation precision in localization microscopy (United States)

    Ebeling, Carl G.

    This dissertation studies detection-based methods to increase the estimation precision of single point-source emitters in the field of localization microscopy. Localization microscopy is a novel method allowing for the localization of optical point-source emitters below the Abbe diffraction limit of optical microscopy. This is accomplished by optically controlling the active, or bright, state of individual molecules within a sample. The use of time-multiplexing of the active state allows for the temporal and spatial isolation of single point-source emitters. Isolating individual sources within a sample allows for statistical analysis on their emission point-spread function profile, and the spatial coordinates of the point-source may be discerned below the optical response of the microscope system. Localization microscopy enables the identification of individual point-source emitter locations approximately an order of magnitude below standard, diffraction-limited optical techniques. The precision of localization microscopy methods is limited by the statistical uncertainty in which the location of these sources may be estimated. By utilizing a detection-based interferometer, an interference pattern may be super-imposed over the emission signal. Theoretical analysis and Monte-Carlo simulations by means of Fisher information theory demonstrate that the incorporation of a modulation structure over the emission signal allow for a more precise estimation when compared to conventional localization methods for the same number of recorded photons. These theoretical calculation and simulations are demonstrated through the use of two proof-of-concept experiments utilizing a modified Mach-Zehnder interferometer. The first methodology improves the localization precision of a single nanoparticle over the theoretical limit for an Airy-disk point-spread function by using self-interference to spatially modulate the recorded point-spread function. Experimental analysis demonstrates

  5. Two-photon excitation STED microscopy. (United States)

    Moneron, Gael; Hell, Stefan W


    We report sub-diffraction resolution in two-photon excitation (TPE) fluorescence microscopy achieved by merging this technique with stimulated-emission depletion (STED). We demonstrate an easy-to-implement and promising laser combination based on a short-pulse laser source for two-photon excitation and a continuous-wave (CW) laser source for resolution enhancement. Images of fluorescent nanoparticles and the immunostained transcription regulator NF kappaB in mammalian cell nuclei exhibit resolutions of barrier. (c) 2009 Optical Society of America

  6. STED microscopy with continuous wave beams. (United States)

    Willig, Katrin I; Harke, Benjamin; Medda, Rebecca; Hell, Stefan W


    We report stimulated emission depletion (STED) fluorescence microscopy with continuous wave (CW) laser beams. Lateral fluorescence confinement from the scanning focal spot delivered a resolution of 29-60 nm in the focal plane, corresponding to a 5-8-fold improvement over the diffraction barrier. Axial spot confinement increased the axial resolution by 3.5-fold. We observed three-dimensional (3D) subdiffraction resolution in 3D image stacks. Viable for fluorophores with low triplet yield, the use of CW light sources greatly simplifies the implementation of this concept of far-field fluorescence nanoscopy.

  7. Correlative Stochastic Optical Reconstruction Microscopy and Electron Microscopy (United States)

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


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

  8. Multimodal hyperspectral optical microscopy

    Energy Technology Data Exchange (ETDEWEB)

    Novikova, Irina V.; Smallwood, Chuck R.; Gong, Yu; Hu, Dehong; Hendricks, Leif; Evans, James E.; Bhattarai, Ashish; Hess, Wayne P.; El-Khoury, Patrick Z.


    We describe a unique and convenient approach to multimodal hyperspectral optical microscopy, herein achieved by coupling a portable and transferable hyperspectral imager to various optical microscopes. The experimental and data analysis schemes involved in recording spectrally and spatially resolved fluorescence, dark field, and optical absorption micrographs are illustrated through prototypical measurements targeting selected model systems. Namely, hyperspectral fluorescence micrographs of isolated fluorescent beads are employed to ensure spectral calibration of our detector and to gauge the attainable spatial resolution of our measurements; the recorded images are diffraction-limited. Moreover, spatially over-sampled absorption spectroscopy of a single lipid (18:1 Liss Rhod PE) layer reveals that optical densities on the order of 10-3 may be resolved by spatially averaging the recorded optical signatures. We also briefly illustrate two applications of our setup in the general areas of plasmonics and cell biology. Most notably, we deploy hyperspectral optical absorption microscopy to identify and image algal pigments within a single live Tisochrysis lutea cell. Overall, this work paves the way for multimodal multidimensional spectral imaging measurements spanning the realms of several scientific disciples.

  9. Magnetic force microscopy (United States)

    Passeri, Daniele; Dong, Chunhua; Reggente, Melania; Angeloni, Livia; Barteri, Mario; Scaramuzzo, Francesca A; De Angelis, Francesca; Marinelli, Fiorenzo; Antonelli, Flavia; Rinaldi, Federica; Marianecci, Carlotta; Carafa, Maria; Sorbo, Angela; Sordi, Daniela; Arends, Isabel WCE; Rossi, Marco


    Magnetic force microscopy (MFM) is an atomic force microscopy (AFM) based technique in which an AFM tip with a magnetic coating is used to probe local magnetic fields with the typical AFM spatial resolution, thus allowing one to acquire images reflecting the local magnetic properties of the samples at the nanoscale. Being a well established tool for the characterization of magnetic recording media, superconductors and magnetic nanomaterials, MFM is finding constantly increasing application in the study of magnetic properties of materials and systems of biological and biomedical interest. After reviewing these latter applications, three case studies are presented in which MFM is used to characterize: (i) magnetoferritin synthesized using apoferritin as molecular reactor; (ii) magnetic nanoparticles loaded niosomes to be used as nanocarriers for drug delivery; (iii) leukemic cells labeled using folic acid-coated core-shell superparamagnetic nanoparticles in order to exploit the presence of folate receptors on the cell membrane surface. In these examples, MFM data are quantitatively analyzed evidencing the limits of the simple analytical models currently used. Provided that suitable models are used to simulate the MFM response, MFM can be used to evaluate the magnetic momentum of the core of magnetoferritin, the iron entrapment efficiency in single vesicles, or the uptake of magnetic nanoparticles into cells. PMID:25050758

  10. Digital holographic microscopy (United States)

    Barkley, Solomon; Dimiduk, Thomas; Manoharan, Vinothan

    Digital holographic microscopy is a 3D optical imaging technique with high temporal ( ms) and spatial ( 10 nm) precision. However, its adoption as a characterization technique has been limited due to the inherent difficulty of recovering 3D data from the holograms. Successful analysis has traditionally required substantial knowledge about the sample being imaged (for example, the approximate positions of particles in the field of view), as well as expertise in scattering theory. To overcome the obstacles to widespread adoption of holographic microscopy, we developed HoloPy - an open source python package for analysis of holograms and scattering data. HoloPy uses Bayesian statistical methods to determine the geometry and properties of discrete scatterers from raw holograms. We demonstrate the use of HoloPy to measure the dynamics of colloidal particles at interfaces, to ascertain the structures of self-assembled colloidal particles, and to track freely swimming bacteria. The HoloPy codebase is thoroughly tested and well-documented to facilitate use by the broader experimental community. This research is supported by NSF Grant DMR-1306410 and NSERC.

  11. Advanced microscopy of microbial cells

    DEFF Research Database (Denmark)

    Haagensen, Janus Anders Juul; Regenberg, Birgitte; Sternberg, Claus


    Growing awareness of heterogeneity in cells of microbial populations has emphasized the importance of advanced microscopy for visualization and understanding of the molecular mechanisms underlying cell-to-cell variation. In this review, we highlight some of the recent advances in confocal...... microscopy, super-resolution optical microscopy (STED, SIM, PALM) as well as atomic force microscopy and Raman spectroscopy. Using examples of bistability in microbial populations as well as biofilm development and differentiation in bacterial and yeast consortia, we demonstrate the importance of microscopy...

  12. Single atom microscopy. (United States)

    Zhou, Wu; Oxley, Mark P; Lupini, Andrew R; Krivanek, Ondrej L; Pennycook, Stephen J; Idrobo, Juan-Carlos


    We show that aberration-corrected scanning transmission electron microscopy operating at low accelerating voltages is able to analyze, simultaneously and with single atom resolution and sensitivity, the local atomic configuration, chemical identities, and optical response at point defect sites in monolayer graphene. Sequential fast-scan annular dark-field (ADF) imaging provides direct visualization of point defect diffusion within the graphene lattice, with all atoms clearly resolved and identified via quantitative image analysis. Summing multiple ADF frames of stationary defects produce images with minimized statistical noise and reduced distortions of atomic positions. Electron energy-loss spectrum imaging of single atoms allows the delocalization of inelastic scattering to be quantified, and full quantum mechanical calculations are able to describe the delocalization effect with good accuracy. These capabilities open new opportunities to probe the defect structure, defect dynamics, and local optical properties in 2D materials with single atom sensitivity.

  13. Emissions Trading

    NARCIS (Netherlands)

    Woerdman, Edwin; Backhaus, Juergen


    Emissions trading is a market-based instrument to achieve environmental targets in a cost-effective way by allowing legal entities to buy and sell emission rights. The current international dissemination and intended linking of emissions trading schemes underlines the growing relevance of this

  14. Progress in the Correlative Atomic Force Microscopy and Optical Microscopy

    Directory of Open Access Journals (Sweden)

    Lulu Zhou


    Full Text Available Atomic force microscopy (AFM has evolved from the originally morphological imaging technique to a powerful and multifunctional technique for manipulating and detecting the interactions between molecules at nanometer resolution. However, AFM cannot provide the precise information of synchronized molecular groups and has many shortcomings in the aspects of determining the mechanism of the interactions and the elaborate structure due to the limitations of the technology, itself, such as non-specificity and low imaging speed. To overcome the technical limitations, it is necessary to combine AFM with other complementary techniques, such as fluorescence microscopy. The combination of several complementary techniques in one instrument has increasingly become a vital approach to investigate the details of the interactions among molecules and molecular dynamics. In this review, we reported the principles of AFM and optical microscopy, such as confocal microscopy and single-molecule localization microscopy, and focused on the development and use of correlative AFM and optical microscopy.

  15. Progress in the Correlative Atomic Force Microscopy and Optical Microscopy. (United States)

    Zhou, Lulu; Cai, Mingjun; Tong, Ti; Wang, Hongda


    Atomic force microscopy (AFM) has evolved from the originally morphological imaging technique to a powerful and multifunctional technique for manipulating and detecting the interactions between molecules at nanometer resolution. However, AFM cannot provide the precise information of synchronized molecular groups and has many shortcomings in the aspects of determining the mechanism of the interactions and the elaborate structure due to the limitations of the technology, itself, such as non-specificity and low imaging speed. To overcome the technical limitations, it is necessary to combine AFM with other complementary techniques, such as fluorescence microscopy. The combination of several complementary techniques in one instrument has increasingly become a vital approach to investigate the details of the interactions among molecules and molecular dynamics. In this review, we reported the principles of AFM and optical microscopy, such as confocal microscopy and single-molecule localization microscopy, and focused on the development and use of correlative AFM and optical microscopy.

  16. Scanning Emitter Lifetime Imaging Microscopy for Spontaneous Emission Control

    DEFF Research Database (Denmark)

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


    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...

  17. Emission Facilities - Air Emission Plants (United States)

    NSGIC Education | GIS Inventory — Represents the Primary Facility type Air Emission Plant (AEP) point features. Air Emissions Plant is a DEP primary facility type related to the Air Quality Program....

  18. Emission inventory; Inventaire des emissions

    Energy Technology Data Exchange (ETDEWEB)

    Fontelle, J.P. [CITEPA, Centre Interprofessionnel Technique d`Etudes de la Pollution Atmospherique, 75 - Paris (France)


    Statistics on air pollutant (sulfur dioxide, nitrogen oxides and ammonium) emissions, acid equivalent emissions and their evolution since 1990 in the various countries of Europe and the USA, are presented. Emission data from the industrial, agricultural, transportation and power sectors are given, and comparisons are carried out between countries based on Gnp and population, pollution import/export fluxes and compliance to the previous emission reduction objectives

  19. Aberrations and adaptive optics in super-resolution microscopy (United States)

    Booth, Martin; Andrade, Débora; Burke, Daniel; Patton, Brian; Zurauskas, Mantas


    As one of the most powerful tools in the biological investigation of cellular structures and dynamic processes, fluorescence microscopy has undergone extraordinary developments in the past decades. The advent of super-resolution techniques has enabled fluorescence microscopy – or rather nanoscopy – to achieve nanoscale resolution in living specimens and unravelled the interior of cells with unprecedented detail. The methods employed in this expanding field of microscopy, however, are especially prone to the detrimental effects of optical aberrations. In this review, we discuss how super-resolution microscopy techniques based upon single-molecule switching, stimulated emission depletion and structured illumination each suffer from aberrations in different ways that are dependent upon intrinsic technical aspects. We discuss the use of adaptive optics as an effective means to overcome this problem. PMID:26124194

  20. Dual-mode super-resolution imaging with stimulated emission depletion microscopy and fluorescence emission difference microscopy. (United States)

    Wang, Yifan; Ma, Ye; Kuang, Cuifang; Fang, Yue; Xu, Yingke; Liu, Xu; Ding, Zhihua


    Dual-mode super-resolution imaging system with two different super-resolution imaging methods, STED and FED, is presented. Electrical shutters controlled by the host computer are introduced to switch the two imaging modes. Principles of both methods are analyzed theoretically, and enhancements in the lateral resolution and SNR are demonstrated theoretically and experimentally. Results show that both imaging methods offered by the proposed system can break the diffraction barrier. Furthermore, the presented system provides a meaningful way to image fluorescent samples by a corresponding imaging mode according to the specific characteristics of samples analyzed for study. For samples that can endure high-power illumination, it is appropriate to use the STED mode to achieve a better resolution, while for samples that are vulnerable to high intensity, the FED method is a better choice because no high-power beam is needed, and the FED method can provide better resolution than STED when no high-power beam is allowed. The flexible switching of the two super-resolution imaging modes can help researchers to make most of the advantages of each imaging method. It is believed that the presented system has the potential to be widely used in future nanoscale investigations.

  1. NDE Acoustic Microscopy Research Laboratory (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...

  2. Magnetic Resonance Force Microscopy System (United States)

    Federal Laboratory Consortium — The Magnetic Resonance Force Microscopy (MRFM) system, developed by ARL, is the world's most sensitive nuclear magnetic resonance (NMR) spectroscopic analysis tool,...

  3. Local correlation of photoemission electron microscopy and STM at a defined cluster substrate system

    Energy Technology Data Exchange (ETDEWEB)

    Rohmer, M.; Wiemann, C.; Munzinger, M.; Guo, L.; Aeschlimann, M.; Bauer, M. [University of Kaiserslautern (Germany). Department of Physics


    We describe a technique that enables photoelectron spectroscopy and STM imaging of supported clusters from identical surface areas of a size of a few {mu}m{sup 2} at a lateral resolution in the low nanometer regime. In this way we are able to locally correlate properties regarding the electronic structure of the clusters and their topography. The use of a photoemission electron microscope (PEEM) allows one to probe the local distribution of the photoemission yield. An STM-tip is used to remove clusters from their position and set local, well-defined markers at the surface that are clearly visible in the PEEM images. These markers act as reference points to identify surface areas in the PEEM image that have formerly been imaged by an STM. The present accuracy of this local correlation technique is at least 300 nm. We propose a scheme to further improve this correlation so that in future experiments even selected single clusters, which have been characterized by STM, can be addressed by local photoelectron spectroscopy as well as local time-resolved photoelectron spectroscopy. (orig.)

  4. Atom probe field ion microscopy and related topics: A bibliography 1989

    Energy Technology Data Exchange (ETDEWEB)

    Miller, M.K.; Hawkins, A.R.; Russell, K.F.


    This bibliography includes references related to the following topics: atom probe field ion microscopy (APFIM), field ion spectroscopy (FIM), field emission microscopy (FEM), liquid metal ion sources (LMIS), scanning tunneling microscopy (STM), and theory. Technique-orientated studies and applications are included. This bibliography covers the period 1989. The references contained in this document were compiled from a variety of sources including computer searches and personal lists of publications.

  5. Atom probe field-ion microscopy and related topics: A bibliography, 1988

    Energy Technology Data Exchange (ETDEWEB)

    Miller, M.K.; Hawkins, A.R.


    This bibliography includes references related to the following topics: field-ion microscopy (FIM), field emission microscopy (FEM), atom probe field-ion microscopy (APFIM), and liquid metal ion sources (LMIS). Technique-orientated studies and applications are included. The references contained in this document were compiled from a variety of sources including computer searches and personal lists of publications. To reduce the length of this document, the references have been reduced to the minimum necessary to locate the articles.

  6. Advanced computing in electron microscopy

    CERN Document Server

    Kirkland, Earl J


    This book features numerical computation of electron microscopy images as well as multislice methods High resolution CTEM and STEM image interpretation are included in the text This newly updated second edition will bring the reader up to date on new developments in the field since the 1990's The only book that specifically addresses computer simulation methods in electron microscopy

  7. Electronic Blending in Virtual Microscopy (United States)

    Maybury, Terrence S.; Farah, Camile S.


    Virtual microscopy (VM) is a relatively new technology that transforms the computer into a microscope. In essence, VM allows for the scanning and transfer of glass slides from light microscopy technology to the digital environment of the computer. This transition is also a function of the change from print knowledge to electronic knowledge, or as…

  8. Vibrational phase contrast CARS microscopy

    NARCIS (Netherlands)

    Jurna, M.


    This thesis describes a new technique that improves specificity, selectivity and sensitivity in coherent anti-Stokes Raman scattering (CARS) microscopy. CARS microscopy is a nonlinear optical technique that utilizes specific bonds of molecules, sometimes referred to as the `fingerprint' of a

  9. Spatial light interference microscopy (SLIM). (United States)

    Wang, Zhuo; Millet, Larry; Mir, Mustafa; Ding, Huafeng; Unarunotai, Sakulsuk; Rogers, John; Gillette, Martha U; Popescu, Gabriel


    We present spatial light interference microscopy (SLIM) as a new optical microscopy technique, capable of measuring nanoscale structures and dynamics in live cells via interferometry. SLIM combines two classic ideas in light imaging: Zernike's phase contrast microscopy, which renders high contrast intensity images of transparent specimens, and Gabor's holography, where the phase information from the object is recorded. Thus, SLIM reveals the intrinsic contrast of cell structures and, in addition, renders quantitative optical path-length maps across the sample. The resulting topographic accuracy is comparable to that of atomic force microscopy, while the acquisition speed is 1,000 times higher. We illustrate the novel insight into cell dynamics via SLIM by experiments on primary cell cultures from the rat brain. SLIM is implemented as an add-on module to an existing phase contrast microscope, which may prove instrumental in impacting the light microscopy field at a large scale.

  10. Transmission electron microscopy physics of image formation and microanalysis

    CERN Document Server

    Reimer, Ludwig


    "Transmission Electron Microscopy" presents the theory of image and contrastformation, and the analytical modes in transmission electron microscopy Theprinciples of particle and wave optics of electrons are described Electron-specimen interactions are discussed for evaluating the theory of scattering and phase contrast Also analysed are the kinetical and dynamical theories of electron diffraction and their applications for crystal-structure determination and imaging of lattices and their defects X-ray microanalysis and electron energy-loss spectroscopy are treated as analytical methods The third edition includes a brief discussionof Schottky emission guns, some clarification of minor details, and references to the recent literature

  11. Microscopy techniques in flavivirus research. (United States)

    Chong, Mun Keat; Chua, Anthony Jin Shun; Tan, Terence Tze Tong; Tan, Suat Hoon; Ng, Mah Lee


    The Flavivirus genus is composed of many medically important viruses that cause high morbidity and mortality, which include Dengue and West Nile viruses. Various molecular and biochemical techniques have been developed in the endeavour to study flaviviruses. However, microscopy techniques still have irreplaceable roles in the identification of novel virus pathogens and characterization of morphological changes in virus-infected cells. Fluorescence microscopy contributes greatly in understanding the fundamental viral protein localizations and virus-host protein interactions during infection. Electron microscopy remains the gold standard for visualizing ultra-structural features of virus particles and infected cells. New imaging techniques and combinatory applications are continuously being developed to push the limit of resolution and extract more quantitative data. Currently, correlative live cell imaging and high resolution three-dimensional imaging have already been achieved through the tandem use of optical and electron microscopy in analyzing biological specimens. Microscopy techniques are also used to measure protein binding affinities and determine the mobility pattern of proteins in cells. This chapter will consolidate on the applications of various well-established microscopy techniques in flavivirus research, and discuss how recently developed microscopy techniques can potentially help advance our understanding in these membrane viruses. Copyright © 2013 Elsevier Ltd. All rights reserved.

  12. Fluorescence confocal microscopy for pathologists. (United States)

    Ragazzi, Moira; Piana, Simonetta; Longo, Caterina; Castagnetti, Fabio; Foroni, Monica; Ferrari, Guglielmo; Gardini, Giorgio; Pellacani, Giovanni


    Confocal microscopy is a non-invasive method of optical imaging that may provide microscopic images of untreated tissue that correspond almost perfectly to hematoxylin- and eosin-stained slides. Nowadays, following two confocal imaging systems are available: (1) reflectance confocal microscopy, based on the natural differences in refractive indices of subcellular structures within the tissues; (2) fluorescence confocal microscopy, based on the use of fluorochromes, such as acridine orange, to increase the contrast epithelium-stroma. In clinical practice to date, confocal microscopy has been used with the goal of obviating the need for excision biopsies, thereby reducing the need for pathological examination. The aim of our study was to test fluorescence confocal microscopy on different types of surgical specimens, specifically breast, lymph node, thyroid, and colon. The confocal images were correlated to the corresponding histological sections in order to provide a morphologic parallel and to highlight current limitations and possible applications of this technology for surgical pathology practice. As a result, neoplastic tissues were easily distinguishable from normal structures and reactive processes such as fibrosis; the use of fluorescence enhanced contrast and image quality in confocal microscopy without compromising final histologic evaluation. Finally, the fluorescence confocal microscopy images of the adipose tissue were as accurate as those of conventional histology and were devoid of the frozen-section-related artefacts that can compromise intraoperative evaluation. Despite some limitations mainly related to black/white images, which require training in imaging interpretation, this study confirms that fluorescence confocal microscopy may represent an alternative to frozen sections in the assessment of margin status in selected settings or when the conservation of the specimen is crucial. This is the first study to employ fluorescent confocal microscopy on

  13. Structured illumination microscopy using photoswitchable fluorescent proteins (United States)

    Hirvonen, Liisa; Mandula, Ondrej; Wicker, Kai; Heintzmann, Rainer


    In fluorescence microscopy the lateral resolution is limited to about 200 nm because of diffraction. Resolution improvement by a factor of two can be achieved using structured illumination, where a ine grating is projected onto the sample, and the final image is reconstructed from a set of images taken at different grating positions. Further resolution improvement can be achieved by saturating the transitions involved in fluorescence emission. Recently discovered photoswitchable proteins undergo transitions that are saturable at low illumination intensity. Combining this concept with structured illumination, theoretically unlimited resolution can be achieved, where the smallest resolvable distance will be determined by signal-to-noise ratio. This work focuses on the use of the photoswitchable protein Dronpa with structured illumination to achieve nanometre scale resolution in fixed cells.

  14. Mapping surface plasmon polariton propagation via counter-propagating light pulses

    DEFF Research Database (Denmark)

    Lemke, Christoph; Leißner, Till; Jauernik, Stephan


    In an interferometric time-resolved photoemission electron microscopy (ITR-PEEM) experiment, the near-field associated with surface plasmon polaritons (SPP) can be locally sensed via interference with ultrashort laser pulses. Here, we present ITR-PEEM data of SPP propagation at a gold vacuum...

  15. Holography and transmission electron microscopy


    Matteucci, G.; Pozzi, G.; Tonomura, A.


    The basic principles and methods of off-axis electron holography are presented and illustrated by means of three examples related to its application in high resolution electron microscopy and the investigation of electric and magnetic fields in thin specimens.

  16. Fluorescence Microscopy of Single Molecules (United States)

    Zimmermann, Jan; van Dorp, Arthur; Renn, Alois


    The investigation of photochemistry and photophysics of individual quantum systems is described with the help of a wide-field fluorescence microscopy approach. The fluorescence single molecules are observed in real time.

  17. Fluorescence lifetime imaging microscopy (FLIM).

    NARCIS (Netherlands)

    van Munster, E.B.; Gadella, Th.W.J.; Rietdorf, J.


    Fluorescence lifetime imaging microscopy (FLIM) is a technique to map the spatial distribution of nanosecond excited state lifetimes within microscopic images. FLIM systems have been implemented both in the frequency domain, using sinusoidally intensity-modulated excitation light and modulated

  18. Filter-Dense Multicolor Microscopy

    National Research Council Canada - National Science Library

    Kijani, Siavash; Yrlid, Ulf; Heyden, Maria; Levin, Malin; Borén, Jan; Fogelstrand, Per


    .... However, because of the risk of bleed-through signals between fluorochromes, standard multicolor microscopy is restricted to a maximum of four fluorescence channels, including one for nuclei staining...

  19. Super-resolution microscopy of single rare-earth emitters


    Kolesov, R.; Lasse, S.; Rothfuchs, C.; Wieck, A. D.; Xia, K.; Kornher, T.; Wrachtrup, J.


    We demonstrate super-resolution imaging of single rare-earth emitting centers, namely, trivalent cerium, in yttrium aluminum garnet (YAG) crystals by means of stimulated emission depletion (STED) microscopy. The achieved all-optical resolution is $\\approx$ 80nm. Similar results were obtained on H3 color centers in diamond with resolution of $\\approx$ 60nm. In both cases, STED resolution is improving slower than the inverse square-root of the depletion beam intensity. This is caused by excited...

  20. Emission detectors

    CERN Document Server

    Bolozdynya, Alexander I


    After decades of research and development, emission detectors have recently become the most successful instrumentation used in modern fundamental experiments searching for cold dark matter, and are also considered for neutrino coherent scattering and magnetic momentum neutrino measurement. This book is the first monograph exclusively dedicated to emission detectors. Properties of two-phase working media based on noble gases, saturated hydrocarbon, ion crystals and semiconductors are reviewed.

  1. Nuclear microscopy in medical research. Investigations into degenerative diseases

    Energy Technology Data Exchange (ETDEWEB)

    Makjanic, J.; Thong, P.; Watt, F. [National University of Singapore (Singapore). Dept. of Physics


    The high energy (1-4MeV) focused ion beam (nuclear microbeam) has found uses in many scientific disciplines through a wide variety of ion beam based techniques. Of the many techniques available, the powerful combination of Particle Induced X-Ray Emission (PIXE), Rutherford Backscattering Spectrometry (RBS), and Scanning Transmission Ion Microscopy (STIM) is proving to be extremely useful, particularly in the characterisation and elemental analysis of thin specimens. In this paper we briefly review these ion beam techniques, as well as the hardware required for their application. Finally, we describe the application of the PIXE, RBS and STIM techniques in conjunction with a scanning focused 2MeV proton microbeam (nuclear microscopy). The examples chosen to illustrate the potential of nuclear microscopy are recent investigations into the degenerative diseases atherosclerosis (coronary heart disease), Parkinson`s disease and Alzheimer`s disease. (author)

  2. Particles and waves in electron optics and microscopy

    CERN Document Server

    Pozzi, Giulio


    Advances in Imaging and Electron Physics merges two long-running serials, Advances in Electronics and Electron Physics and Advances in Optical and Electron Microscopy. The series features extended articles on the physics of electron devices (especially semiconductor devices), particle optics at high and low energies, microlithography, image science, digital image processing, electromagnetic wave propagation, electron microscopy, and the computing methods used in all these domains. * Contains contributions from leading authorities on the subject matter* Informs and updates all the latest developments in the field of imaging and electron physics* Provides practitioners interested in microscopy, optics, image processing, mathematical morphology, electromagnetic fields, electron, and ion emission with a valuable resource* Features extended articles on the physics of electron devices (especially semiconductor devices), particle optics at high and low energies, microlithography, image science, and digital image pro...

  3. Magnetic force microscopy of atherosclerotic plaque

    National Research Council Canada - National Science Library

    T A Alexeeva; S V Gorobets; O Yu Gorobets; I V Demianenko; O M Lazarenko


    In this work by methods of scanning probe microscopy, namely by atomic force microscopy and magnetic force microscopy the fragments of atherosclerotic plaque section of different nature were investigated...

  4. Scanning Anode Field Emission Characterisation of Carbon Nanotube emitter arrays

    NARCIS (Netherlands)

    Berhanu, S.; Gröning, O.; Chen, Z.; Merikhi, J.; Bachmann, P.K.


    Scanning anode field emission microscopy (SAFEM) was used to characterise carbon nanotube (CNT) emitter arrays produced within Philips CediX-Technotubes' activities. Four different samples were investigated and compared. The field enhancement distributions were determined and the local field

  5. Light microscopy - Methods and protocols

    Directory of Open Access Journals (Sweden)

    CarloAlberto Redi


    Full Text Available The first part of the book (six chapters is devoted to some selected applications of bright-field microscopy while the second part (eight chapters to some fluorescence microscopy studies. Both animal and plant biology investigations are presented covering multiple fields like immunology, cell signaling, cancer biology and, surprisingly to me, ecology. This chapter is titled: Light microscopy in aquatic ecology: Methods for plankton communities studies and it is due to Maria Carolina S. Soares and colleagues from the Laboratory of Aquatic Ecology, Dept. of Biology, Federal University of Juiz de Fora (Brazil. Here they present methods to quantify the different component of planktonic communities in a step-by-step manner so that virus, bacteria, algae and animals pertaining to different taxa can be recognized and the contribution they made to the plankton composition evaluated. It descends that even how the plankton composition is changing due to environmental variations can be accurately determined....

  6. Correlative fluorescence and electron microscopy. (United States)

    Schirra, Randall T; Zhang, Peijun


    Correlative fluorescence and electron microscopy (CFEM) is a multimodal technique that combines dynamic and localization information from fluorescence methods with ultrastructural data from electron microscopy, to give new information about how cellular components change relative to the spatiotemporal dynamics within their environment. In this review, we will discuss some of the basic techniques and tools of the trade for utilizing this attractive research method, which is becoming a very powerful tool for biology labs. The information obtained from correlative methods has proven to be invaluable in creating consensus between the two types of microscopy, extending the capability of each, and cutting the time and expense associated with using each method separately for comparative analysis. The realization of the advantages of these methods in cell biology has led to rapid improvement in the protocols and has ushered in a new generation of instruments to reach the next level of correlation--integration. Copyright © 2014 John Wiley & Sons, Inc.

  7. Structured illumination microscopy and correlative microscopy to study autophagy. (United States)

    Ligeon, Laure-Anne; Barois, Nicolas; Werkmeister, Elisabeth; Bongiovanni, Antonino; Lafont, Frank


    Autophagy is a predominant eukaryotic mechanism for the engulfment of "portions" of cytoplasm allowing their degradation to recycle metabolites. The autophagy is ubiquitous among the life kingdom revealing the importance of this pathway that appears more complex than previously thought. Several reviews have already addressed how to monitor this pathway and have highlighted the existence of new routes such as the LC3-associated phagocytosis (LAP) and the non-canonical autophagy. The principal difference between autophagosomes and LAP vacuoles is that the former has two limiting membranes positives for LC3 whereas the latter has one. Herein, we propose to emphasize the use of correlative light electron microscopy (CLEM) to answer some autophagy's related questions. The structured illumination microscopy (SIM) relatively easy to implement allows to better observe the Atg proteins recruitment and localization during the autophagy process. While LC3 recruitment is performed using light microscopy the ultrastructural morphological analysis of LC3-vacuoles is ascertained by electron microscopy. Hence, these combined and correlated approaches allow to tackle the LAP vs. autophagosome issue. Copyright © 2015. Published by Elsevier Inc.

  8. Multiphoton Microscopy for Ophthalmic Imaging

    Directory of Open Access Journals (Sweden)

    Emily A. Gibson


    Full Text Available We review multiphoton microscopy (MPM including two-photon autofluorescence (2PAF, second harmonic generation (SHG, third harmonic generation (THG, fluorescence lifetime (FLIM, and coherent anti-Stokes Raman Scattering (CARS with relevance to clinical applications in ophthalmology. The different imaging modalities are discussed highlighting the particular strength that each has for functional tissue imaging. MPM is compared with current clinical ophthalmological imaging techniques such as reflectance confocal microscopy, optical coherence tomography, and fluorescence imaging. In addition, we discuss the future prospects for MPM in disease detection and clinical monitoring of disease progression, understanding fundamental disease mechanisms, and real-time monitoring of drug delivery.

  9. Illuminating Electron Microscopy of Photocatalysts

    DEFF Research Database (Denmark)

    Cavalca, Filippo

    energy into chemical bonds. By means of Transmission Electron Microscopy (TEM) it is possible to gain insight in the fundamentals of their reaction mechanisms, chemical behaviour, structure and morphology before, during and after reaction using in situ investigations. In particular, the environmental TEM...... (ETEM) is the instrument of choice employed in this thesis to perform such studies. Typically, photocatalysts work in gaseous or liquid atmosphere upon light illumination. We aim at reproducing their working conditions in situ. The ETEM allows exposing specimens to a controlled gas atmosphere, thus...... the microscope that allows electron microscopy under nonconventional TEM conditions and new kinds of in situ spectroscopy....

  10. Magnetic Force Microscopy in Liquids. (United States)

    Ares, Pablo; Jaafar, Miriam; Gil, Adriana; Gómez-Herrero, Julio; Asenjo, Agustina


    In this work, the use of magnetic force microscopy (MFM) to acquire images of magnetic nanostructures in liquid environments is presented. Optimization of the MFM signal acquisition in liquid media is performed and it is applied to characterize the magnetic signal of magnetite nanoparticles. The ability for detecting magnetic nanostructures along with the well-known capabilities of atomic force microscopy in liquids suggests potential applications in fields such as nanomedicine, nanobiotechnology, or nanocatalysis. © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  11. Quantitative super-resolution microscopy

    NARCIS (Netherlands)

    Harkes, Rolf


    Super-Resolution Microscopy is an optical fluorescence technique. In this thesis we focus on single molecule super-resolution, where the position of single molecules is determined. Typically these molecules can be localized with a 10 to 30nm precision. This technique is applied in four different

  12. Mechanics in Steels through Microscopy

    NARCIS (Netherlands)

    Zandbergen, H.W.; Tirumalasetty, G.K.

    The goal of the study consolidated in this thesis is to understand the mechanics in steels using microscopy. In particular, the mechanical response of Transformation Induced Plasticity (TRIP) steels is correlated with their microstructures. Chapter 1 introduces the current state of the art of TRIP

  13. Color-televised medical microscopy (United States)

    Heath, M. A.; Peck, J. C.


    Color television microscopy used at laboratory range magnifications, reproduces a slide image with sufficient fidelity for medical laboratory and instructional use. The system is used for instant pathological reporting between operating room and remotely located pathologist viewing a biopsy through this medium.

  14. 3D -Ray Diffraction Microscopy

    DEFF Research Database (Denmark)

    Poulsen, Henning Friis; Schmidt, Søren; Juul Jensen, Dorte


    Three-dimensional X-ray diffraction (3DXRD) microscopy is a fast and non-destructive structural characterization technique aimed at the study of individual crystalline elements (grains or subgrains) within mm-sized polycrystalline specimens. It is based on two principles: the use of highly...

  15. Vacuum scanning capillary photoemission microscopy

    DEFF Research Database (Denmark)

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


    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...

  16. Advanced Microscopy of Microbial Cells

    DEFF Research Database (Denmark)

    Haagensen, Janus Anders Juul; Regenberg, Birgitte; Sternberg, Claus


    Growing awareness of heterogeneity in cells of microbial populations has emphasized the importance of advanced microscopy for visualization and understanding of the molecular mechanisms underlying cell-to-cell variation. In this review, we highlight some of the recent advances in confocal...... for visualization of variation between cells in phenotypic traits such as gene expression....

  17. High Resolution Scanning Ion Microscopy

    NARCIS (Netherlands)

    Castaldo, V.


    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

  18. Filter-Dense Multicolor Microscopy.

    Directory of Open Access Journals (Sweden)

    Siavash Kijani

    Full Text Available Immunofluorescence microscopy is a unique method to reveal the spatial location of proteins in tissues and cells. By combining antibodies that are labeled with different fluorochromes, the location of several proteins can simultaneously be visualized in one sample. However, because of the risk of bleed-through signals between fluorochromes, standard multicolor microscopy is restricted to a maximum of four fluorescence channels, including one for nuclei staining. This is not always enough to address common scientific questions. In particular, the use of a rapidly increasing number of marker proteins to classify functionally distinct cell populations and diseased tissues emphasizes the need for more complex multistainings. Hence, multicolor microscopy should ideally offer more channels to meet the current needs in biomedical science. Here we present an enhanced multi-fluorescence setup, which we call Filter-Dense Multicolor Microscopy (FDMM. FDMM is based on condensed filter sets that are more specific for each fluorochrome and allow a more economic use of the light spectrum. FDMM allows at least six independent fluorescence channels and can be applied to any standard fluorescence microscope without changing any operative procedures for the user. In the present study, we demonstrate an FDMM setup of six channels that includes the most commonly used fluorochromes for histology. We show that the FDMM setup is specific and robust, and we apply the technique on typical biological questions that require more than four fluorescence microscope channels.

  19. Mechanics in Steels through Microscopy

    NARCIS (Netherlands)

    Tirumalasetty, G.K.


    The goal of the study consolidated in this thesis is to understand the mechanics in steels using microscopy. In particular, the mechanical response of Transformation Induced Plasticity (TRIP) steels is correlated with their microstructures. Chapter 1 introduces the current state of the art of TRIP

  20. Light Microscopy at Maximal Precision (United States)

    Bierbaum, Matthew; Leahy, Brian D.; Alemi, Alexander A.; Cohen, Itai; Sethna, James P.


    Microscopy is the workhorse of the physical and life sciences, producing crisp images of everything from atoms to cells well beyond the capabilities of the human eye. However, the analysis of these images is frequently little more accurate than manual marking. Here, we revolutionize the analysis of microscopy images, extracting all the useful information theoretically contained in a complex microscope image. Using a generic, methodological approach, we extract the information by fitting experimental images with a detailed optical model of the microscope, a method we call parameter extraction from reconstructing images (PERI). As a proof of principle, we demonstrate this approach with a confocal image of colloidal spheres, improving measurements of particle positions and radii by 10-100 times over current methods and attaining the maximum possible accuracy. With this unprecedented accuracy, we measure nanometer-scale colloidal interactions in dense suspensions solely with light microscopy, a previously impossible feat. Our approach is generic and applicable to imaging methods from brightfield to electron microscopy, where we expect accuracies of 1 nm and 0.1 pm, respectively.

  1. Turning Microscopy in the Medical Curriculum Digital

    DEFF Research Database (Denmark)

    Vainer, Ben; Mortensen, Niels Werner; Poulsen, Steen Seier


    an administrative, an economic, and a teaching perspective. This fully automatic digital microscopy system has been received positively by both teachers and students, and a decision was made to convert all courses involving microscopy to the virtual microscopy format. As a result, conventional analog microscopy...

  2. Fluorescent dyes with large Stokes shifts for super-resolution optical microscopy of biological objects: A review.


    Sednev, M.; Belov, V.; Hell, S.


    The review deals with commercially available organic dyes possessing large Stokes shifts and their applications as fluorescent labels in optical microscopy based on stimulated emission depletion (STED). STED microscopy breaks Abbe’s diffraction barrier and provides optical resolution beyond the diffraction limit. STED microscopy is non-invasive and requires photostable fluorescent markers attached to biomolecules or other objects of interest. Up to now, in most biology-related STED experiment...

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

    Tomitori, Masahiko; Sasahara, Akira


    to extend a model sample prepared for the microscopies towards a microscale sample while keeping the intrinsic properties found by the microscopies.In this study we present our trial of developing microscopic combined instruments among FIM, field emission microscopy (FEM), STM, AFM and scanning electron microscopy (SEM), in which we prepared and characterized the tips for the SPM, and in addition, the sample preparation to take a correlation between nanoscale and microscale properties of functional materials. Recently, we developed a simple sample preparation method of a rutile single crystal TiO2 covered with an epitaxially-grown monolayer of SiO2 by annealing the crystals in a furnace at high temperatures in air; the crystal samples were placed into a quartz container in the furnace [1]. The vapor of SiO evaporated from the quartz container were adsorbed on the crystal while the crystal surfaces being fully oxidized in air. The SiO2-TiO2 composite systems are promising to protect catalytic TiO2 performance; the photo-catalytic activity is kept by coating with hard and stable SiO2 layers and to extend the lifetime of water super-hydrophilicity even in dark, though understanding of their properties is insufficient due to the lack of techniques to fabricate a well-characterized system on a nanoscale to conduct control experiments. The SiO2 overlayers were observed by low energy electron diffraction (LEED) in vacuum and frequency-modulation (FM) AFM in water [1,2], and water contact angles (WCA) were measured [2]. Although the WCA measurement seems a classic characterization, this method possesses a high potential to make a bridge by controlling the environmental conditions. We will discuss the details. © The Author 2014. Published by Oxford University Press on behalf of The Japanese Society of Microscopy. All rights reserved. For permissions, please e-mail:

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

    Directory of Open Access Journals (Sweden)

    Stephen A. Boppart


    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.

  5. Breaking the diffraction barrier in fluorescence microscopy at low light intensities by using reversibly photoswitchable proteins


    Hofmann, M.; Eggeling, C.; Jakobs, S.; Hell, S.


    Fluorescence microscopy is indispensable in many areas of science, but until recently, diffraction has limited the resolution of its lens-based variant. The diffraction barrier has been broken by a saturated depletion of the marker's fluorescent state by stimulated emission, but this approach requires picosecond laser pulses of GW/cm2 intensity. Here, we demonstrate the surpassing of the diffraction barrier in fluorescence microscopy with illumination intensities that are eight orders of magn...

  6. Macromolecular-scale resolution in biological fluorescence microscopy. (United States)

    Donnert, Gerald; Keller, Jan; Medda, Rebecca; Andrei, M Alexandra; Rizzoli, Silvio O; Lührmann, Reinhard; Jahn, Reinhard; Eggeling, Christian; Hell, Stefan W


    We demonstrate far-field fluorescence microscopy with a focal-plane resolution of 15-20 nm in biological samples. The 10- to 12-fold multilateral increase in resolution below the diffraction barrier has been enabled by the elimination of molecular triplet state excitation as a major source of photobleaching of a number of dyes in stimulated emission depletion microscopy. Allowing for relaxation of the triplet state between subsequent excitation-depletion cycles yields an up to 30-fold increase in total fluorescence signal as compared with reported stimulated emission depletion illumination schemes. Moreover, it enables the reduction of the effective focal spot area by up to approximately 140-fold below that given by diffraction. Triplet-state relaxation can be realized either by reducing the repetition rate of pulsed lasers or by increasing the scanning speed such that the build-up of the triplet state is effectively prevented. This resolution in immunofluorescence imaging is evidenced by revealing nanoscale protein patterns on endosomes, the punctuated structures of intermediate filaments in neurons, and nuclear protein speckles in mammalian cells with conventional optics. The reported performance of diffraction-unlimited fluorescence microscopy opens up a pathway for addressing fundamental problems in the life sciences.


    Directory of Open Access Journals (Sweden)

    Merete Krog Raarup


    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.

  8. Selective sensitivity in Kerr microscopy (United States)

    Soldatov, I. V.; Schäfer, R.


    A new technique for contrast separation in wide-field magneto-optical Kerr microscopy is introduced. Utilizing the light from eight light emitting diodes, guided to the microscope by glass fibers and being switched synchronously with the camera exposure, domain images with orthogonal in-plane sensitivity can be displayed simultaneously at real-time, and images with pure in-plane or polar contrast can be obtained. The benefit of this new method of contrast separation is demonstrated for Permalloy films, a NdFeB sinter magnet, and a cobalt crystal. Moreover, the new technique is shown to strongly enhance the sensitivity of Kerr microscopy by eliminating parasitic contrast contributions occurring in conventional setups. A doubling of the in-plane domain contrast and a sensitivity to Kerr rotations as low as 0.6 mdeg is demonstrated.

  9. Advanced Methods in Fluorescence Microscopy

    Directory of Open Access Journals (Sweden)

    Luke Fritzky


    Full Text Available It requires a good deal of will power to resist hyperbole in considering the advances that have been achieved in fluorescence microscopy in the last 25 years. Our effort has been to survey the modalities of microscopic fluorescence imaging available to cell biologists and perhaps useful for diagnostic pathologists. The gamut extends from established confocal laser scanning through multiphoton and TIRF to the emerging technologies of super-resolution microscopy that breech the Abbé limit of resolution. Also considered are the recent innovations in structured and light sheet illumination, the use of FRET and molecular beacons that exploit specific characteristics of designer fluorescent proteins, fluorescence speckles, and second harmonic generation for native anisometric structures like collagen, microtubules and sarcomeres.

  10. All-optical photoacoustic microscopy

    Directory of Open Access Journals (Sweden)

    Sung-Liang Chen


    Full Text Available Three-dimensional photoacoustic microscopy (PAM has gained considerable attention within the biomedical imaging community during the past decade. Detecting laser-induced photoacoustic waves by optical sensing techniques facilitates the idea of all-optical PAM (AOPAM, which is of particular interest as it provides unique advantages for achieving high spatial resolution using miniaturized embodiments of the imaging system. The review presents the technology aspects of optical-sensing techniques for ultrasound detection, such as those based on optical resonators, as well as system developments of all-optical photoacoustic systems including PAM, photoacoustic endoscopy, and multi-modality microscopy. The progress of different AOPAM systems and their representative applications are summarized.

  11. A history of urine microscopy. (United States)

    Cameron, J Stewart


    The naked-eye appearance of the urine must have been studied by shamans and healers since the Stone Age, and an elaborate interpretation of so-called Uroscopy began around 600 AD as a form of divination. A 1000 years later, the first primitive monocular and compound microscopes appeared in the Netherlands, and along with many other objects and liquids, urine was studied from around 1680 onwards as the enlightenment evolved. However, the crude early instruments did not permit fine study because of chromatic and linear/spherical blurring. Only after complex multi-glass lenses which avoided these problems had been made and used in the 1820s in London by Lister, and in Paris by Chevalier and Amici, could urinary microscopy become a practical, clinically useful tool in the 1830s. Clinical urinary microscopy was pioneered by Rayer and his pupils in Paris (especially Vigla), in the late 1830s, and spread to UK and Germany in the 1840s, with detailed descriptions and interpretations of cells and formed elements of the urinary sediment by Nasse, Henle, Robinson and Golding Bird. Classes were held, most notably by Donné in Paris. After another 50 years, optical microscopy had reached its apogee, with magnifications of over 1000 times obtainable free of aberration, using immersion techniques. Atlases of the urinary sediment were published in all major European countries and in the US. Polarised light and phase contrast was used also after 1900 to study urine, and by the early 20th century, photomicroscopy (pioneered by Donné and Daguerre 50 years previously, but then ignored) became usual for teaching and recording. In the 1940s electron microscopy began, followed by detection of specific proteins and cells using immunofluorescent antibodies. All this had been using handheld methodology. Around 1980, machine-assisted observations began, and have dominated progress since.

  12. Quantum state atomic force microscopy


    Passian, Ali; Siopsis, George


    New classical modalities of atomic force microscopy continue to emerge to achieve higher spatial, spectral, and temporal resolution for nanometrology of materials. Here, we introduce the concept of a quantum mechanical modality that capitalizes on squeezed states of probe displacement. We show that such squeezing is enabled nanomechanically when the probe enters the van der Waals regime of interaction with a sample. The effect is studied in the non-contact mode, where we consider the paramete...

  13. Light Sheet Fluorescence Microscopy (LSFM)


    Adams, Michael W.; Loftus, Andrew F.; Dunn, Sarah E.; Joens, Matthew S.; Fitzpatrick, James A. J.


    The development of confocal microscopy techniques introduced the ability to optically section fluorescent samples in the axial dimension, perpendicular to the image plane. These approaches, via the placement of a pinhole in the conjugate image plane, provided superior resolution in the axial (z) dimension resulting in nearly isotropic optical sections. However, increased axial resolution, via pinhole optics, comes at the cost of both speed and excitation efficiency. Light Sheet Fluorescent Mi...

  14. Low power super resolution fluorescence microscopy by lifetime modification and image reconstruction. (United States)

    Marsh, Richard J; Culley, Siân; Bain, Angus J


    We demonstrate a new method for obtaining sub-diffraction resolution in fluorescence microscopy. The technique involves the analysis of the time evolution of fluorescence images in the presence of weak and unstructured (fundamental Gaussian) continuous wave stimulated emission depletion. A reduced point spread functions (PSF) is obtained by the recombination of time segments of the evolving image. A significant reduction in the PSF for 20 nm fluorescent beads (ca. 240 nm to 125 nm) is obtained with an on-sample power of 7.5 mW (17 MW/cm2) - substantially lower than that required for spatially structured stimulated emission depletion microscopy.

  15. THz wave emission microscope (United States)

    Yuan, Tao

    Sensing and imaging using Terahertz (THz) radiation has attracted more and more interest in the last two decades thanks to the abundant material 'finger prints' in the THz frequency range. The low photon energy also makes THz radiation an attractive tool for nondestructive evaluation of materials and devices, biomedical applications, security checks and explosive screening. Due to the long wavelength, the far-field THz wave optical systems have relatively low spatial resolution. This physical limitation confines THz wave sensing and imaging to mostly macro-size samples. To investigate local material properties or micro-size structures and devices, near-field technology has to be employed. In this dissertation, the Electro-Optical THz wave emission microscope is investigated. The basic principle is to focus the femtosecond laser to a tight spot on a thin THz emitter layer to produce a THz wave source with a similar size as the focus spot. The apparatus provides a method for placing a THz source with sub-wavelength dimension in the near-field range of the investigated sample. Spatial resolution to the order of one tenth of the THz wavelength is demonstrated by this method. The properties of some widely used THz wave emission materials under tight focused pump light are studied. As an important branch of THz time domain spectroscopy (THz-TDS), THz wave emission spectroscopy has been widely used as a tool to investigate the material physics, such as energy band structure, carrier dynamics, material nonlinear properties and dynamics. As the main work of this dissertation, we propose to combine the THz wave emission spectroscopy with scanning probe microscopy (SPM) to build a tip-assisted THz wave emission microscope (TATEM), which is a valuable extension to current SPM science and technology. Illuminated by a femtosecond laser, the biased SPM tip forms a THz wave source inside the sample beneath the tip. The source size is proportional to the apex size of the tip so

  16. Use of Kelvin probe force microscopy for identification of CVD grown graphene flakes on copper foil (United States)

    Kumar, Rakesh; Mehta, B. R.; Kanjilal, D.


    Graphene flakes have been grown by chemical vapour deposition (CVD) method on Cu foils. The obtained graphene flakes have been characterized by optical microscopy, field emission scanning electron microscopy, Kelvin probe force microscopy (KPFM) and Raman spectroscopy. The graphene flakes grown on Cu foil comprise mainly single layer graphene and confirm that the nucleation for graphene growth starts very quickly. Moreover, KPFM has been found to be a valuable technique to differentiate between covered and uncovered portion of Cu foil by graphene flakes deposited for shorter duration. The results show that KPFM can be a very useful technique in understanding the mechanism of graphene growth.

  17. Breaking the diffraction barrier in fluorescence microscopy by optical shelving. (United States)

    Bretschneider, Stefan; Eggeling, Christian; Hell, Stefan W


    We report the breaking of the diffraction resolution barrier in far-field fluorescence microscopy by transiently shelving the fluorophore in a metastable dark state. Using a relatively modest light intensity of several kW/cm(2) in a focal distribution featuring a local zero, we confine the fluorescence emission to a spot whose diameter is a fraction of the wavelength of light. Nanoscale far-field optical resolution down to 50 nm is demonstrated by imaging microtubules in a mammalian cell and proteins on the plasma membrane of a neuron. The presence of dark states in virtually any fluorescent molecule opens up a new venue for far-field microscopy with resolution that is no longer limited by diffraction.

  18. Signatures of plasmoemission in two photon photoemission electron microscopy (United States)

    Meyer zu Heringdorf, Frank-J.; Kahl, Philip; Makris, Andreas; Sindermann, Simon; Podbiel, Daniel; Horn-von Hoegen, Michael


    The imaging of surface plasmon polariton waves in two photon photoemission microscopy has been intensely studied during the past years, with a focus on contrast mechanisms and light-plasmon interaction. The possibility of photoemission from the plasmonic fields alone has so far not been addressed in such experiments. This was justified, since the intensity of the plasmonic fields at the surface was comparatively weak and nonlinear plasmonic effects were not to be expected. Here we discuss the properties of grating couplers for creation of intense and short plasmon polariton pulses for which the emission of electrons purely from the plasmonic field cannot be neglected any more. Two examples for signatures of such nonlinear plasmoemission effects in experimental two photon photoemission microscopy images are discussed.

  19. Transmission electron microscopy physics of image formation and microanalysis

    CERN Document Server

    Reimer, Ludwig


    Transmission Electron Microscopy presents the theory of image and contrast formation, and the analytical modes in transmission electron microscopy. The principles of particle and wave optics of electrons are described. Electron-specimen interactions are discussed for evaluating the theory of scattering and phase contrast. Also discussed are the kinematical and dynamical theories of electron diffraction and their applications for crystal-structure analysis and imaging of lattices and their defects. X-ray micronanalysis and electron energy-loss spectroscopy are treated as analytical methods. Specimen damage and contamination by electron irradiation limits the resolution for biological and some inorganic specimens. This fourth edition includes discussion of recent progress, especially in the area of Schottky emission guns, convergent-beam electron diffraction, electron tomography, holography and the high resolution of crystal lattices.

  20. 4D electron microscopy: principles and applications. (United States)

    Flannigan, David J; Zewail, Ahmed H


    The transmission electron microscope (TEM) is a powerful tool enabling the visualization of atoms with length scales smaller than the Bohr radius at a factor of only 20 larger than the relativistic electron wavelength of 2.5 pm at 200 keV. The ability to visualize matter at these scales in a TEM is largely due to the efforts made in correcting for the imperfections in the lens systems which introduce aberrations and ultimately limit the achievable spatial resolution. In addition to the progress made in increasing the spatial resolution, the TEM has become an all-in-one characterization tool. Indeed, most of the properties of a material can be directly mapped in the TEM, including the composition, structure, bonding, morphology, and defects. The scope of applications spans essentially all of the physical sciences and includes biology. Until recently, however, high resolution visualization of structural changes occurring on sub-millisecond time scales was not possible. In order to reach the ultrashort temporal domain within which fundamental atomic motions take place, while simultaneously retaining high spatial resolution, an entirely new approach from that of millisecond-limited TEM cameras had to be conceived. As shown below, the approach is also different from that of nanosecond-limited TEM, whose resolution cannot offer the ultrafast regimes of dynamics. For this reason "ultrafast electron microscopy" is reserved for the field which is concerned with femtosecond to picosecond resolution capability of structural dynamics. In conventional TEMs, electrons are produced by heating a source or by applying a strong extraction field. Both methods result in the stochastic emission of electrons, with no control over temporal spacing or relative arrival time at the specimen. The timing issue can be overcome by exploiting the photoelectric effect and using pulsed lasers to generate precisely timed electron packets of ultrashort duration. The spatial and temporal resolutions

  1. NICHD Microscopy and Imaging Core (MIC) (United States)

    Federal Laboratory Consortium — The NICHD Microscopy and Imaging Core (MIC) is designed as a multi-user research facility providing training and instrumentation for high resolution microscopy and...

  2. Polyethyleneimine as tracer for electron microscopy

    NARCIS (Netherlands)

    Schurer, Jacob Willem


    In this thesis the development of a tracer particle for use in electron microscopy is described. Attempts were made to use this tracer particle in immuno-electron microscopy and to trace negatively charged tissue components. ... Zie: Summary

  3. Acoustic Microscopy at Cryogenic Temperatures. (United States)


    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

  4. Diffraction phase and fluorescence microscopy. (United States)

    Park, Yongkeun; Popescu, Gabriel; Badizadegan, Kamran; Dasari, Ramachandra R; Feld, Michael S


    We have developed diffraction phase and fluorescence (DPF) microscopy as a new technique for simultaneous quantitative phase imaging and epi-fluorescence investigation of live cells. The DPF instrument consists of an interference microscope, which is incorporated into a conventional inverted fluorescence microscope. The quantitative phase images are characterized by sub-nanometer optical path-length stability over periods from milliseconds to a cell lifetime. The potential of the technique for quantifying rapid nanoscale motions in live cells is demonstrated by experiments on red blood cells, while the composite phase-fluorescence imaging mode is exemplified with mitotic kidney cells.

  5. Microscopy using randomized speckle illumination (United States)

    Perinchery, Sandeep M.; Shinde, Anant; Murukeshan, V. M.


    It is well known for structured illumination microscopy (SIM) that the lateral resolution by a factor of two beyond the classical diffraction limit is achieved using spatially structured illumination in wide-field fluorescence microscope. In the state of art SIM systems, grating patterns are generally generated by physical gratings or by spatial light modulators such as digital micro mirrors (DMD), liquid crystal displays (LCD). In this study, using a combination of LCD and ground glasses, size controlled randomized speckle patterns are generated as an illumination source for the microscope. Proof of concept of using speckle illumination in SIM configuration is tested by imaging fixed BPAE cells.

  6. Visual-servoing optical microscopy (United States)

    Callahan, Daniel E.; Parvin, Bahram


    The present invention provides methods and devices for the knowledge-based discovery and optimization of differences between cell types. In particular, the present invention provides visual servoing optical microscopy, as well as analysis methods. The present invention provides means for the close monitoring of hundreds of individual, living cells over time: quantification of dynamic physiological responses in multiple channels; real-time digital image segmentation and analysis; intelligent, repetitive computer-applied cell stress and cell stimulation; and the ability to return to the same field of cells for long-term studies and observation. The present invention further provides means to optimize culture conditions for specific subpopulations of cells.

  7. Super-resolution microscopy reveals compartmentalization of peroxisomal membrane proteins

    DEFF Research Database (Denmark)

    Galiani, Silvia; Waithe, Dominic; Reglinski, Katharina


    the use of super-resolution optical stimulated emission depletion microscopy to investigate with sub-60-nm resolution the heterogeneous spatial organization of the peroxisomal proteins PEX5, PEX14, and PEX11 around actively importing peroxisomes, showing distinct differences between these peroxins......Membrane-associated events during peroxisomal protein import processes play an essential role in peroxisome functionality. Many details of these processes are not known due to missing spatial resolution of technologies capable of investigating peroxisomes directly in the cell. Here, we present...

  8. High-resolution low-dose scanning transmission electron microscopy. (United States)

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


    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.

  9. Agreement between direct fluorescent microscopy and Ziehl ...

    African Journals Online (AJOL)

    Background: The sensitivity of smear microscopy for diagnosis of tuberculosis might be improved through treatment of sputum with sodium hypochlorite and application of fluorescent microscopy. This study aimed to determine the agreement between direct Fluorescent Microscopy and Ziehl-Neelsen concentration technique ...

  10. Plasmonics Enhanced Smartphone Fluorescence Microscopy

    KAUST Repository

    Wei, Qingshan


    Smartphone fluorescence microscopy has various applications in point-of-care (POC) testing and diagnostics, ranging from e.g., quantification of immunoassays, detection of microorganisms, to sensing of viruses. An important need in smartphone-based microscopy and sensing techniques is to improve the detection sensitivity to enable quantification of extremely low concentrations of target molecules. Here, we demonstrate a general strategy to enhance the detection sensitivity of a smartphone-based fluorescence microscope by using surface-enhanced fluorescence (SEF) created by a thin metal-film. In this plasmonic design, the samples are placed on a silver-coated glass slide with a thin spacer, and excited by a laser-diode from the backside through a glass hemisphere, generating surface plasmon polaritons. We optimized this mobile SEF system by tuning the metal-film thickness, spacer distance, excitation angle and polarization, and achieved ~10-fold enhancement in fluorescence intensity compared to a bare glass substrate, which enabled us to image single fluorescent particles as small as 50 nm in diameter and single quantum-dots. Furthermore, we quantified the detection limit of this platform by using DNA origami-based brightness standards, demonstrating that ~80 fluorophores per diffraction-limited spot can be readily detected by our mobile microscope, which opens up new opportunities for POC diagnostics and sensing applications in resource-limited-settings.


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


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

  12. Plasmonics Enhanced Smartphone Fluorescence Microscopy. (United States)

    Wei, Qingshan; Acuna, Guillermo; Kim, Seungkyeum; Vietz, Carolin; Tseng, Derek; Chae, Jongjae; Shir, Daniel; Luo, Wei; Tinnefeld, Philip; Ozcan, Aydogan


    Smartphone fluorescence microscopy has various applications in point-of-care (POC) testing and diagnostics, ranging from e.g., quantification of immunoassays, detection of microorganisms, to sensing of viruses. An important need in smartphone-based microscopy and sensing techniques is to improve the detection sensitivity to enable quantification of extremely low concentrations of target molecules. Here, we demonstrate a general strategy to enhance the detection sensitivity of a smartphone-based fluorescence microscope by using surface-enhanced fluorescence (SEF) created by a thin metal-film. In this plasmonic design, the samples are placed on a silver-coated glass slide with a thin spacer, and excited by a laser-diode from the backside through a glass hemisphere, generating surface plasmon polaritons. We optimized this mobile SEF system by tuning the metal-film thickness, spacer distance, excitation angle and polarization, and achieved ~10-fold enhancement in fluorescence intensity compared to a bare glass substrate, which enabled us to image single fluorescent particles as small as 50 nm in diameter and single quantum-dots. Furthermore, we quantified the detection limit of this platform by using DNA origami-based brightness standards, demonstrating that ~80 fluorophores per diffraction-limited spot can be readily detected by our mobile microscope, which opens up new opportunities for POC diagnostics and sensing applications in resource-limited-settings.

  13. Multifunctional scanning ion conductance microscopy. (United States)

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


    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. Kelvin probe force microscopy in liquid using electrochemical force microscopy

    Directory of Open Access Journals (Sweden)

    Liam Collins


    Full Text Available Conventional closed loop-Kelvin probe force microscopy (KPFM has emerged as a powerful technique for probing electric and transport phenomena at the solid–gas interface. The extension of KPFM capabilities to probe electrostatic and electrochemical phenomena at the solid–liquid interface is of interest for a broad range of applications from energy storage to biological systems. However, the operation of KPFM implicitly relies on the presence of a linear lossless dielectric in the probe–sample gap, a condition which is violated for ionically-active liquids (e.g., when diffuse charge dynamics are present. Here, electrostatic and electrochemical measurements are demonstrated in ionically-active (polar isopropanol, milli-Q water and aqueous NaCl and ionically-inactive (non-polar decane liquids by electrochemical force microscopy (EcFM, a multidimensional (i.e., bias- and time-resolved spectroscopy method. In the absence of mobile charges (ambient and non-polar liquids, KPFM and EcFM are both feasible, yielding comparable contact potential difference (CPD values. In ionically-active liquids, KPFM is not possible and EcFM can be used to measure the dynamic CPD and a rich spectrum of information pertaining to charge screening, ion diffusion, and electrochemical processes (e.g., Faradaic reactions. EcFM measurements conducted in isopropanol and milli-Q water over Au and highly ordered pyrolytic graphite electrodes demonstrate both sample- and solvent-dependent features. Finally, the feasibility of using EcFM as a local force-based mapping technique of material-dependent electrostatic and electrochemical response is investigated. The resultant high dimensional dataset is visualized using a purely statistical approach that does not require a priori physical models, allowing for qualitative mapping of electrostatic and electrochemical material properties at the solid–liquid interface.

  15. EDITORIAL: Scanning probe microscopy: a visionary development Scanning probe microscopy: a visionary development (United States)

    Demming, Anna


    , Lezec H J, Ebbesen T W, Pellerin K M, Lewen G D, Nahata A and Linke R A 2002 Giant optical transmission of sub-wavelength apertures: physics and applications Nanotechnology 13 429-32 [8] Barrera E W, Pujol M C, Díaz F, Choi S B, Rotermund F, Park K H, Jeong M S and Cascales C 2011 Emission properties of hydrothermal Yb3+, Er3+ and Yb3+, Tm3+-codoped Lu2O3 nanorods: upconversion, cathodoluminescence and assessment of waveguide behaviour Nanotechnology 22 075205 [9] Nonnenmacher M, O'Boyle M P and Wickramasinghe H K 1991 Kelvin probe force microscopy Appl. Phys. Lett. 58 2921-3 [10] Cohen G, Halpern E, Nanayakkara S U, Luther J M, Held C, Bennewitz R, Boag A and Rosenwaks Y 2013 Reconstruction of surface potential from Kelvin probe force microscopy images Nanotechnology 24 295702 [11] 1986 The Nobel Prize in Physics prizes/physics/laureates/1986/ index.html

  16. Nanoscale spatial analysis of clay minerals containing cesium by synchrotron radiation photoemission electron microscopy (United States)

    Yoshigoe, Akitaka; Shiwaku, Hideaki; Kobayashi, Toru; Shimoyama, Iwao; Matsumura, Daiju; Tsuji, Takuya; Nishihata, Yasuo; Kogure, Toshihiro; Ohkochi, Takuo; Yasui, Akira; Yaita, Tsuyoshi


    A synchrotron radiation photoemission electron microscope (SR-PEEM) was applied to demonstrate the pinpoint analysis of micrometer-sized weathered biotite clay particles with artificially adsorbed cesium (Cs) atoms. Despite the insulating properties of the clay, we observed the spatial distributions of constituent elements (Si, Al, Cs, Mg, and Fe) without charging issues and clarified reciprocal site-correlations among these elements with nanometer resolution. We found that Cs atoms were likely to be adsorbed evenly over the entire particle; however, we identified an occupational conflict between Cs and Mg atoms, implying that Cs sorption involves ion exchange processes. Spatially resolved X-ray absorption spectra (XAS) of the Cs4,5 M-edge region showed Cs to be present in a monocation state (Cs+) as typically observed for Cs compounds. Further pinpoint XAS measurements were also performed at the Fe L2,3-edge to determine the chemical valence of the Fe atoms. The shapes of the spectra were similar to those for Fe2O3, indicating that Fe in the clay was in a 3+ oxidation state. From these observations, we infer that charge compensation facilitates Cs adsorption in the vicinity of a substitution site where Si4+ ions are replaced by Fe3+ ions in SiO4 tetrahedral sheets. Our results demonstrate the utility of SR-PEEM as a tool for spatially resolved chemical analyses of various environmental substances, which is not limited by the poor conductivity of samples.

  17. Ceria-catlyzed soot oxidation studied by environmental transmission electron microscopy

    DEFF Research Database (Denmark)

    Simonsen, S.B.; Dahl, S.; Johnson, Erik


    Environmental tranmission electron microscopy (ETEM) was used to monitor in situ ceria-catalyzed oxidation of soot in relation to diesel engine emission control.  From time-lapsed ETEM image series of soot particles in contact with CeO2. or with Al2O3 as inert reference, mechanistic and kinetic...

  18. Spectral analysis in microscopy : a study of FRET and single quantum dot luminescence

    NARCIS (Netherlands)

    Frederix, Patrick Louis Theodorus Martin


    This thesis deals with the development of new techniques and luminescent markers, to improve the quality of luminescence studies in microscopy. A sensitive spectrograph that can be used for spectrally resolved emission spectroscopy in the microscope is described, including design considerations,

  19. Magnetic microscopy of layered structures

    CERN Document Server

    Kuch, Wolfgang; Fischer, Peter; Hillebrecht, Franz Ulrich


    This book presents the important analytical technique of magnetic microscopy. This method is applied to analyze layered structures with high resolution. This book presents a number of layer-resolving magnetic imaging techniques that have evolved recently. Many exciting new developments in magnetism rely on the ability to independently control the magnetization in two or more magnetic layers in micro- or nanostructures. This in turn requires techniques with the appropriate spatial resolution and magnetic sensitivity. The book begins with an introductory overview, explains then the principles of the various techniques and gives guidance to their use. Selected examples demonstrate the specific strengths of each method. Thus the book is a valuable resource for all scientists and practitioners investigating and applying magnetic layered structures.

  20. Scanning Electrochemical Microscopy in Neuroscience (United States)

    Schulte, Albert; Nebel, Michaela; Schuhmann, Wolfgang


    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.

  1. Phase Aberrations in Diffraction Microscopy

    Energy Technology Data Exchange (ETDEWEB)

    Marchesini, S; Chapman, H N; Barty, A; Howells, M R; Spence, J H; Cui, C; Weierstall, U; Minor, A M


    In coherent X-ray diffraction microscopy the diffraction pattern generated by a sample illuminated with coherent x-rays is recorded, and a computer algorithm recovers the unmeasured phases to synthesize an image. By avoiding the use of a lens the resolution is limited, in principle, only by the largest scattering angles recorded. However, the imaging task is shifted from the experiment to the computer, and the algorithm's ability to recover meaningful images in the presence of noise and limited prior knowledge may produce aberrations in the reconstructed image. We analyze the low order aberrations produced by our phase retrieval algorithms. We present two methods to improve the accuracy and stability of reconstructions.

  2. Illuminating Electron Microscopy of Photocatalysts

    DEFF Research Database (Denmark)

    Cavalca, Filippo

    Photocatalysts are of fundamental interest for sustainable energy research because of their wide range of applications and great potential for state of the art and future usages [1]. By means of Transmission Electron Microscopy (TEM) it is possible to give a deep insight in the structure......, composition and operation of photocatalysts and to provide information on the compounds inner arrangement and a fundamental contribution for their further optimization [2]. We want to construct a novel specimen holder capable of shining light onto samples inside the TEM allowing real time in situ experiments...... an Environmental TEM (ETEM) in order to expose the specimen to a controlled gas atmosphere during illumination. The aim is to perform complete and exhaustive characterization of photocatalytic materials under simulated working environment, achieving experimental data on yet uninvestigated aspects. Analysis can...

  3. Illuminating Electron Microscopy of Photocatalysts

    DEFF Research Database (Denmark)

    Cavalca, Filippo

    Photocatalysts are of fundamental interest for sustainable energy research [1]. By means of transmission electron microscopy (TEM) it is possible to obtain deep insight in the structure, composition and reactivity of photocatalysts for their further optimization [2]. We have constructed a novel...... the device inside an environmental TEM (ETEM) in order to allow specimens to be exposed to controlled gas atmospheres during illumination. The holder is presently being used to study a variety of photoreactive materials and structures, including photocatalysts, photonic devices and solar cells. Here, we...... specimen holder capable of shining light onto samples inside the TEM. The holder contains a laser diode and an optical system that guides light onto a sample with maximum power transmission. The source can be changed and tuned, in principle spanning the whole visible and UV spectrum. It is possible to use...

  4. Lensfree microscopy on a cellphone. (United States)

    Tseng, Derek; Mudanyali, Onur; Oztoprak, Cetin; Isikman, Serhan O; Sencan, Ikbal; Yaglidere, Oguzhan; Ozcan, Aydogan


    We demonstrate lensfree digital microscopy on a cellphone. This compact and light-weight holographic microscope installed on a cellphone does not utilize any lenses, lasers or other bulky optical components and it may offer a cost-effective tool for telemedicine applications to address various global health challenges. Weighing approximately 38 grams (cellphone where the samples are loaded from the side, and are vertically illuminated by a simple light-emitting diode (LED). This incoherent LED light is then scattered from each micro-object to coherently interfere with the background light, creating the lensfree hologram of each object on the detector array of the cellphone. These holographic signatures captured by the cellphone permit reconstruction of microscopic images of the objects through rapid digital processing. We report the performance of this lensfree cellphone microscope by imaging various sized micro-particles, as well as red blood cells, white blood cells, platelets and a waterborne parasite (Giardia lamblia).

  5. Scanning Probe Microscopy of Graphene (United States)

    Tautz, Pamela


    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.

  6. Epoxy Resins in Electron Microscopy (United States)

    Finck, Henry


    A method of embedding biological specimens in araldite 502 (Ciba) has been developed for materials available in the United States. Araldite-embedded tissues are suitable for electron microscopy, but the cutting qualities of the resin necessitates more than routine attention during microtomy. The rather high viscosity of araldite 502 also seems to be an unnecessary handicap. The less viscous epoxy epon 812 (Shell) produces specimens with improved cutting qualities, and has several features—low shrinkage and absence of specimen damage during cure, minimal compression of sections, relative absence of electron beam-induced section damage, etc.—which recommends it as a routine embedding material. The hardness of the cured resin can be easily adjusted by several methods to suit the materials embedded in it. Several problems and advantages of working with sections of epoxy resins are also discussed. PMID:13822825

  7. Electronic detectors for electron microscopy. (United States)

    Faruqi, A R; McMullan, G


    Electron microscopy (EM) is an important tool for high-resolution structure determination in applications ranging from condensed matter to biology. Electronic detectors are now used in most applications in EM as they offer convenience and immediate feedback that is not possible with film or image plates. The earliest forms of electronic detector used routinely in transmission electron microscopy (TEM) were charge coupled devices (CCDs) and for many applications these remain perfectly adequate. There are however applications, such as the study of radiation-sensitive biological samples, where film is still used and improved detectors would be of great value. The emphasis in this review is therefore on detectors for use in such applications. Two of the most promising candidates for improved detection are: monolithic active pixel sensors (MAPS) and hybrid pixel detectors (of which Medipix2 was chosen for this study). From the studies described in this review, a back-thinned MAPS detector appears well suited to replace film in for the study of radiation-sensitive samples at 300 keV, while Medipix2 is suited to use at lower energies and especially in situations with very low count rates. The performance of a detector depends on the energy of electrons to be recorded, which in turn is dependent on the application it is being used for; results are described for a wide range of electron energies ranging from 40 to 300 keV. The basic properties of detectors are discussed in terms of their modulation transfer function (MTF) and detective quantum efficiency (DQE) as a function of spatial frequency.

  8. Combination of widefield fluorescence imaging and nonlinear optical microscopy of oral epithelial neoplasia (United States)

    Pal, Rahul; Edward, Kert; Brown, Tyra; Ma, Liang; Yang, Jinping; McCammon, Susan; Motamedi, Massoud; Vargas, Gracie


    Multiphoton Autofluorescence Microscopy (MPAM) and Second Harmonic Generation Microscopy (SHGM) have shown the potential for noninvasive assessment of oral precancers and cancers. We have explored a combination of these nonlinear optical microscopic imaging techniques with widefield fluorescence imaging to assess morphometry similar to that of pathologic evaluation as well as information from endogenous fluorophores, which are altered with neoplastic transformation. Widefield fluorescence revealed areas of interest corresponding to sites with precancers or early tumors, generally resulting in a decrease in green emission or increase in red emission. Subsequent microscopy revealed significant differences in morphology between normal, dysplastic/neoplastic mucosa for all layers. Combination of a widefield and a microscopic technique provides a novel approach for tissue morphometric analysis along with large area assessment of tissue autofluorescence properties.

  9. Emission Inventory for Fugitive Emissions in Denmark

    DEFF Research Database (Denmark)

    Plejdrup, Marlene Schmidt; Nielsen, Ole-Kenneth; Nielsen, Malene

    This report presents the methodology and data used in the Danish inventory of fugitive emissions from fuels for the years until 2007. The inventory of fugitive emissions includes CO2, CH4, N2O, NOx, CO, NMVOC, SO2, dioxin, PAH and particulate matter. In 2007 the total Danish emission of greenhouse...

  10. Microsphere-aided optical microscopy and its applications for super-resolution imaging (United States)

    Upputuri, Paul Kumar; Pramanik, Manojit


    The spatial resolution of a standard optical microscope (SOM) is limited by diffraction. In visible spectrum, SOM can provide ∼ 200 nm resolution. To break the diffraction limit several approaches were developed including scanning near field microscopy, metamaterial super-lenses, nanoscale solid immersion lenses, super-oscillatory lenses, confocal fluorescence microscopy, techniques that exploit non-linear response of fluorophores like stimulated emission depletion microscopy, stochastic optical reconstruction microscopy, etc. Recently, photonic nanojet generated by a dielectric microsphere was used to break the diffraction limit. The microsphere-approach is simple, cost-effective and can be implemented under a standard microscope, hence it has gained enormous attention for super-resolution imaging. In this article, we briefly review the microsphere approach and its applications for super-resolution imaging in various optical imaging modalities.

  11. Potential of ultraviolet widefield imaging and multiphoton microscopy for analysis of dehydroergosterol in cellular membranes

    DEFF Research Database (Denmark)

    Wüstner, Daniel; Brewer, Jonathan R.; Bagatolli, Luis


    ) excitation microscopy to monitor DHE in living cells. Significantly reduced photobleaching in MP microscopy of DHE enabled us to acquire three-dimensional z-stacks of DHE-stained cells and to obtain high-resolution maps of DHE in surface ruffles, nanotubes, and the apical membrane of epithelial cells. We......Dehydroergosterol (DHE) is an intrinsically fluorescent sterol with absorption/emission in the ultraviolet (UV) region and biophysical properties similar to those of cholesterol. We compared the potential of UV-sensitive low-light-level wide-field (UV-WF) imaging with that of multiphoton (MP...... be resolved by MP time-lapse microscopy. Diffusion coefficients of 0.25-μm-diameter endocytic vesicles containing DHE were determined by MP spatiotemporal image correlation spectroscopy. The requirement of extremely high laser power for visualization of DHE by MP microscopy made this method less potent...

  12. Electron microscopy of pharmaceutical systems. (United States)

    Klang, Victoria; Valenta, Claudia; Matsko, Nadejda B


    During the last decades, the focus of research in pharmaceutical technology has steadily shifted towards the development and optimisation of nano-scale drug delivery systems. As a result, electron microscopic methods are increasingly employed for the characterisation of pharmaceutical systems such as nanoparticles and microparticles, nanoemulsions, microemulsions, solid lipid nanoparticles, different types of vesicles, nanofibres and many more. Knowledge of the basic properties of these systems is essential for an adequate microscopic analysis. Classical transmission and scanning electron microscopic techniques frequently have to be adapted for an accurate analysis of formulation morphology, especially in case of hydrated colloidal systems. Specific techniques such as environmental scanning microscopy or cryo preparation are required for their investigation. Analytical electron microscopic techniques such as electron energy-loss spectroscopy or energy-dispersive X-ray spectroscopy are additional assets to determine the elemental composition of the systems, but are not yet standard tools in pharmaceutical research. This review provides an overview of pharmaceutical systems of interest in current research and strategies for their successful electron microscopic analysis. Advantages and limitations of the different methodological approaches are discussed and recent findings of interest are presented. Copyright © 2012 Elsevier Ltd. All rights reserved.

  13. Electronic detectors for electron microscopy. (United States)

    Faruqi, A R; Henderson, R


    Due to the increasing popularity of electron cryo-microscopy (cryoEM) in the structural analysis of large biological molecules and macro-molecular complexes and the need for simple, rapid and efficient readout, there is a persuasive need for improved detectors. Commercial detectors, based on phosphor/fibre optics-coupled CCDs, provide adequate performance for many applications, including electron diffraction. However, due to intrinsic light scattering within the phosphor, spatial resolution is limited. Careful measurements suggest that CCDs have superior performance at lower resolution while all agree that film is still superior at higher resolution. Consequently, new detectors are needed based on more direct detection, thus avoiding the intermediate light conversion step required for CCDs. Two types of direct detectors are discussed in this review. First, there are detectors based on hybrid technology employing a separate pixellated sensor and readout electronics connected with bump bonds-hybrid pixel detectors (HPDs). Second, there are detectors, which are monolithic in that sensor and readout are all in one plane (monolithic active pixel sensor, MAPS). Our discussion is centred on the main parameters of interest to cryoEM users, viz. detective quantum efficiency (DQE), resolution or modulation transfer function (MTF), robustness against radiation damage, speed of readout, signal-to-noise ratio (SNR) and the number of independent pixels available for a given detector.

  14. Photoacoustic microscopy of human teeth (United States)

    Rao, Bin; Cai, Xin; Favazza, Christopher; Yao, Junjie; Li, Li; Duong, Steven; Liaw, Lih-Huei; Holtzman, Jennifer; Wilder-Smith, Petra; Wang, Lihong V.


    Photoacoustic microscopy (PAM) utilizes short laser pulses to deposit energy into light absorbers and sensitively detects the ultrasonic waves the absorbers generate in response. PAM directly renders a three-dimensional spatial distribution of sub-surface optical absorbers. Unlike other optical imaging technologies, PAM features label-free optical absorption contrast and excellent imaging depths. Standard dental imaging instruments are limited to X-ray and CCD cameras. Subsurface optical dental imaging is difficult due to the highly-scattering enamel and dentin tissue. Thus, very few imaging methods can detect dental decay or diagnose dental pulp, which is the innermost part of the tooth, containing the nerves, blood vessels, and other cells. Here, we conducted a feasibility study on imaging dental decay and dental pulp with PAM. Our results showed that PAM is sensitive to the color change associated with dental decay. Although the relative PA signal distribution may be affected by surface contours and subsurface reflections from deeper dental tissue, monitoring changes in the PA signals (at the same site) over time is necessary to identify the progress of dental decay. Our results also showed that deep-imaging, near-infrared (NIR) PAM can sensitively image blood in the dental pulp of an in vitro tooth. In conclusion, PAM is a promising tool for imaging both dental decay and dental pulp.

  15. Electron microscopy and forensic practice (United States)

    Kotrlý, Marek; Turková, Ivana


    Electron microanalysis in forensic practice ranks among basic applications used in investigation of traces (latents, stains, etc.) from crime scenes. Applying electron microscope allows for rapid screening and receiving initial information for a wide range of traces. SEM with EDS/WDS makes it possible to observe topography surface and morphology samples and examination of chemical components. Physical laboratory of the Institute of Criminalistics Prague use SEM especially for examination of inorganic samples, rarely for biology and other material. Recently, possibilities of electron microscopy have been extended considerably using dual systems with focused ion beam. These systems are applied mainly in study of inner micro and nanoparticles , thin layers (intersecting lines in graphical forensic examinations, analysis of layers of functional glass, etc.), study of alloys microdefects, creating 3D particles and aggregates models, etc. Automated mineralogical analyses are a great asset to analysis of mineral phases, particularly soils, similarly it holds for cathode luminescence, predominantly colour one and precise quantitative measurement of their spectral characteristics. Among latest innovations that are becoming to appear also at ordinary laboratories are TOF - SIMS systems and micro Raman spectroscopy with a resolution comparable to EDS/WDS analysis (capable of achieving similar level as through EDS/WDS analysis).

  16. Recent developments in GSDIM microscopy (United States)

    Dyba, Marcus; Simonutti, Giulio A.; Fölling, Jonas


    In the presented study we characterized the suitability of 15 conventional fluorescence dyes for GSDIM microscopy. For all dyes involved in the screening labeled secondary antibodies for immunohistochemistry are commercially available. The dye performance was tested after staining to fixed mammalian cells. Chemical environments were chosen to be compatible with the applicative and spectroscopic demands. Investigated watery environments are suitable for TIRF based applications. To the best of our knowledge, we present for the first time systematic screening for configurations of dyes embedded in solid polymer. The polymer mounting matches well to the refractive index of oil immersion optics. This is crucial for applications at high penetration depth into the sample and suitable for long-term sample storage. We rated the final super-resolution image quality additional to quantitative characterization of important spectroscopic parameters. Therefore, this dye screening is optimized for various biological imaging applications. Control of the single molecule blinking rate by 405nm light exposure is quantified, as well. It is shown that this important effect is applicable to numerous fluorescent dyes. Thus, the controlled application of low intensities of 405nm light allows to maximize recording speed. As this option is already included in commercial GSDIM microscopes the results of our study allow optimized super-resolution imaging down to ~20nm with multiple dyes and multi-color staining.

  17. Super-resolution methods for fluorescence microscopy


    Mandula, Ondrej


    Fluorescence microscopy is an important tool for biological research. However, the resolution of a standard fluorescence microscope is limited by diffraction, which makes it difficult to observe small details of a specimen’s structure. We have developed two fluorescence microscopy methods that achieve resolution beyond the classical diffraction limit. The first method represents an extension of localisation microscopy. We used nonnegative matrix factorisation (NMF) to model ...

  18. Time-Resolved Scanning Electron Microscopy

    National Research Council Canada - National Science Library

    Weber, Peter M


    .... The pulsed electron beam is obtained by rapidly switching the electron emission of a field emission tip using the AC electric field arising from exposure to the intense electromagnetic radiation...

  19. French Society of Microscopy, 10. conference; Societe Francaise des Microscopies, 10. colloque

    Energy Technology Data Exchange (ETDEWEB)

    Thibault-Penisson, J.; Cremer, Ch.; Susini, J.; Kirklanda, A.I.; Rigneault, H.; Renault, O.; Bailly, A.; Zagonel, L.F.; Barrett, N.; Bogner, A.; Gauthier, C.; Jouneau, P.H.; Thollet, G.; Fuchs, G.; Basset, D.; Deconihout, B.; Vurpillot, F.; Vella, A.; Matthieu, G.; Cadel, E.; Bostel, A.; Blavette, D.; Baumeister, W.; Usson, Y.; Zaefferer, St.; Laffont, L.; Weyland, M.; Thomas, J.M.; Midgley, P.; Benlekbir, S.; Epicier, Th.; Diop, B.N.; Roux, St.; Ou, M.; Perriat, P.; Bausach, M.; Aouine, M.; Berhault, G.; Idrissi, H.; Cottevieille, M.; Jonic, S.; Larquet, E.; Svergun, D.; Vannoni, M.A.; Boisset, N.; Ersena, O.; Werckmann, J.; Ulhaq, C.; Hirlimann, Ch.; Tihay, F.; Cuong, Pham-Huu; Crucifix, C.; Schultz, P.; Jornsanoha, P.; Thollet, G.; Masenelli-Varlot, K.; Gauthier, C.; Ludwig, W.; King, A.; Johnson, G.; Gonzalves-Hoennicke, M.; Reischig, P.; Messaoudi, C.; Ibrahim, R.; Marco, S.; Klie, R.F.; Zhao, Y.; Yang, G.; Zhu, Y.; Hue, F.; Hytch, M.; Hartmann, J.M.; Bogumilowicz, Y.; Claverie, A.; Klein, H.; Alloyeau, D.; Ricolleau, C.; Langlois, C.; Le Bouar, Y.; Loiseau, A.; Colliex, C.; Stephan, O.; Kociak, M.; Tence, M.; Gloter, A.; Imhoff, D.; Walls, M.; Nelayah, J.; March, K.; Couillard, M.; Ailliot, C.; Bertin, F.; Cooper, D.; Rivallin, P.; Dumelie, N.; Benhayoune, H.; Balossier, G.; Cheynet, M.; Pokrant, S.; Tichelaar, F.; Rouviere, J.L.; Cooper, D.; Truche, R.; Chabli, A.; Debili, M.Y.; Houdellier, F.; Warot-Fonrose, B.; Hytch, M.J.; Snoeck, E.; Calmels, L.; Serin, V.; Schattschneider, P.; Jacob, D.; Cordier, P


    This document gathers the resumes of some of the presentations made at this conference whose aim was to present the last developments and achievements of the 3 complementary microscopies: optical microscopy, electron microscopy and X-ray microscopy. The contributions have been organized around the following 12 topics: 1) new technical developments, 2) 3-dimensional imaging, 3) quantitative microscopy, 4) technical progress in photon microscopy, 5) synchrotron radiation, 6) measurements of patterns, deformations and strains, 7) materials for energy and transports, 8) nano-structures, 9) virus: structure and infection mechanisms, 10) 3-dimensional imaging for molecules, cells and cellular tissues, 11) nano-particles and colloids, and 12) liquid crystals.

  20. Atomic Force Microscopy of Coccoliths: Implications for Biomineralisation and Diagenesis

    DEFF Research Database (Denmark)

    Henriksen, Karen; Young, Jette F.; Bown, P.R.


    geochemistry, diagenesis, coccoliths, biomineralization, biological calcite, atomic force microscopy......geochemistry, diagenesis, coccoliths, biomineralization, biological calcite, atomic force microscopy...

  1. High-resolution electron microscopy of advanced materials

    Energy Technology Data Exchange (ETDEWEB)

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


    This final report chronicles a three-year, Laboratory Directed Research and Development (LDRD) project at Los Alamos National Laboratory (LANL). The High-Resolution Electron Microscopy Facility has doubled in size and tripled in quality since the beginning of the three-year period. The facility now includes a field-emission scanning electron microscope, a 100 kV field-emission scanning transmission electron microscope (FE-STEM), a 300 kV field-emission high-resolution transmission electron microscope (FE-HRTEM), and a 300 kV analytical transmission electron microscope. A new orientation imaging microscope is being installed. X-ray energy dispersive spectrometers for chemical analysis are available on all four microscopes; parallel electron energy loss spectrometers are operational on the FE-STEM and FE-HRTEM. These systems enable evaluation of local atomic bonding, as well as chemical composition in nanometer-scale regions. The FE-HRTEM has a point-to-point resolution of 1.6 {angstrom}, but the resolution can be pushed to its information limit of 1 {angstrom} by computer reconstruction of a focal series of images. HRTEM has been used to image the atomic structure of defects such as dislocations, grain boundaries, and interfaces in a variety of materials from superconductors and ferroelectrics to structural ceramics and intermetallics.

  2. Nuclear microscopy of biological specimens

    Energy Technology Data Exchange (ETDEWEB)

    Watt, F.; Grime, G.W. (Nuclear Physics Lab., Univ. of Oxford (UK)); Brook, A.J. (Clore Lab., Univ. of Buckingham (UK)); Gadd, G.M. (Dept. of Biological Sciences, Univ. of Dundee (UK)); Perry, C.C. (Dept. of Chemistry, Brunel Univ., Uxbridge (UK)); Pearce, R.B. (Dept. of Plant Sciences, Univ. of Oxford (UK)); Turnau, K. (Dept. of Botany, Jagiellonian Univ., Krakow (Poland)); Watkinson, S.C. (Dept. of Plant Sciences, Univ. of Oxford (UK))


    Recent developments in technology have enabled the scanning proton microprobe to scan at submicron spatial resolution on a routine basis. The use of the powerful combination of techniques PIXE (proton induced X-ray emission), nuclear (or Rutherford) backscattering (RBS), and secondary electron detection operating at this resolution will open up new areas in many scientific disciplines. This paper describes some of the work carried out in the biological sciences over the last year, using the Oxford SPM facility. Collaborations with biological scientists have drawn attention to the wealth of information that can be derived when these techniques are applied to micro-organisms, cells and plant tissue. Briefly described here are investigations into the uptake of heavy metals by the alga Pandorina morum, the structure of the diatom Stephanopyxis turris, the presence of various types of crystal structures within the cells of Spirogyra, the heavy metal uptake of a mycorrhizal fungus present in the bracken (Pteridium aquilinum) root, the role of sphagnum moss in the absorption of inorganic elements, the measurement of heavy metals in environmentally-adapted cells of the yeast Saccharomyces cerevisiae, and the elemental distribution in the growing tip of a spore from the plant Equisetum arvense, with special emphasis placed on the visual interpretation of the elemental and secondary-electron maps provided by the nuclear microscopical techniques. (orig.).

  3. Helium ion microscopy for high-resolution visualization of the articular cartilage collagen network. (United States)

    Vanden Berg-Foels, W S; Scipioni, L; Huynh, C; Wen, X


    The articular cartilage collagen network is an important research focus because network disruption results in cartilage degeneration and patient disability. The recently introduced helium ion microscope (HIM), with its smaller probe size, longer depth of field and charge neutralization, has the potential to overcome the inherent limitations of electron microscopy for visualization of collagen network features, particularly at the nanoscale. In this study, we evaluated the capabilities of the helium ion microscope for high-resolution visualization of the articular cartilage collagen network. Images of rabbit knee cartilage were acquired with a helium ion microscope; comparison images were acquired with a field emission scanning electron microscope (FE-SEM) and a transmission electron microscope (TEM). Sharpness of example high-resolution helium ion microscope and field emission scanning electron microscope images was quantified using the 25-75% rise distance metric. The helium ion microscope was able to acquire high-resolution images with unprecedented clarity, with greater sharpness and three-dimensional-like detail of nanoscale fibril morphologies and fibril connections, in samples without conductive coatings. These nanoscale features could not be resolved by field emission scanning electron microscopy, and three-dimensional network structure could not be visualized with transmission electron microscopy. The nanoscale three-dimensional-like visualization capabilities of the helium ion microscope will enable new avenues of investigation in cartilage collagen network research. © 2012 The Authors Journal of Microscopy © 2012 Royal Microscopical Society.

  4. Multiphoton microscopy imaging of developing tooth germs

    Directory of Open Access Journals (Sweden)

    Pei-Yu Pan


    Conclusion: In this study, a novel multiphoton microscopy database of images from developing tooth germs in mice was set up. We confirmed that multiphoton laser microscopy is a powerful tool for investigating the development of tooth germ and is worthy for further application in the study of tooth regeneration.

  5. Magnetic force microscopy : Quantitative issues in biomaterials

    NARCIS (Netherlands)

    Passeri, D.; Dong, C.; Reggente, M.; Angeloni, L.; Barteri, M.; Scaramuzzo, F.A.; De Angelis, F.; Marinelli, F.; Antonelli, F.; Rinaldi, F.; Marianecci, C.; Carafa, M.; Sorbo, A.; Sordi, D.; Arends, I.W.C.E.; Rossi, M.


    Magnetic force microscopy (MFM) is an atomic force microscopy (AFM) based technique in which an AFM tip with a magnetic coating is used to probe local magnetic fields with the typical AFM spatial resolution, thus allowing one to acquire images reflecting the local magnetic properties of the samples

  6. CLAFEM: Correlative light atomic force electron microscopy. (United States)

    Janel, Sébastien; Werkmeister, Elisabeth; Bongiovanni, Antonino; Lafont, Frank; Barois, Nicolas


    Atomic force microscopy (AFM) is becoming increasingly used in the biology field. It can give highly accurate topography and biomechanical quantitative data, such as adhesion, elasticity, and viscosity, on living samples. Nowadays, correlative light electron microscopy is a must-have tool in the biology field that combines different microscopy techniques to spatially and temporally analyze the structure and function of a single sample. Here, we describe the combination of AFM with superresolution light microscopy and electron microscopy. We named this technique correlative light atomic force electron microscopy (CLAFEM) in which AFM can be used on fixed and living cells in association with superresolution light microscopy and further processed for transmission or scanning electron microscopy. We herein illustrate this approach to observe cellular bacterial infection and cytoskeleton. We show that CLAFEM brings complementary information at the cellular level, from on the one hand protein distribution and topography at the nanometer scale and on the other hand elasticity at the piconewton scales to fine ultrastructural details. Copyright © 2017 Elsevier Inc. All rights reserved.

  7. Scanning transmission electron microscopy imaging and analysis

    CERN Document Server

    Pennycook, Stephen J


    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

  8. Confocal Raman Microscopy; applications in tissue engineering

    NARCIS (Netherlands)

    van Apeldoorn, Aart A.


    This dissertation describes the use of confocal Raman microscopy and spectroscopy in the field of tissue engineering. Moreover, it describes the combination of two already existing technologies, namely scanning electron microscopy and confocal Raman spectroscopy in one apparatus for the enhancement

  9. Structured illumination microscopy and its new developments

    Directory of Open Access Journals (Sweden)

    Jianling Chen


    Full Text Available Optical microscopy allows us to observe the biological structures and processes within living cells. However, the spatial resolution of the optical microscopy is limited to about half of the wavelength by the light diffraction. Structured illumination microscopy (SIM, a type of new emerging super-resolution microscopy, doubles the spatial resolution by illuminating the specimen with a patterned light, and the sample and light source requirements of SIM are not as strict as the other super-resolution microscopy. In addition, SIM is easier to combine with the other imaging techniques to improve their imaging resolution, leading to the developments of diverse types of SIM. SIM has great potential to meet the various requirements of living cells imaging. Here, we review the recent developments of SIM and its combination with other imaging techniques.

  10. On slit-lamp microscopy. (United States)

    Schmidt, T A


    examination of the vitreous good brightness of the slit image is required for stereoscopic examination with as large an angle as possible between microscope and illumination. The lateral parts of the most peripheral fundus cannot be examined with the vertical slit in connection with the three-mirror lens. However, this is possible with the horizontal and tilted position of the slit and intermediate positions with an oblique slit. The slit must form an angle with the microscope in order to examine vitreous and fundus in optical section. With the indentation contact lenses ciliary processes and pars plana are now accessible to slit-lamp microscopy.

  11. Moderate emissions grandfathering


    Knight, Carl


    Emissions grandfathering holds that a history of emissions strengthens an agent’s claim for future emission entitlements. Though grandfathering appears to have been influential in actual emission control frameworks, it is rarely taken seriously by philosophers. This article presents an argument for thinking this an oversight. The core of the argument is that members of countries with higher historical emissions are typically burdened with higher costs when transitioning to a given lower level...

  12. Ion cyclotron emission by spontaneous emission

    Energy Technology Data Exchange (ETDEWEB)

    Da Costa, O. [Commission of the European Communities, Abingdon (United Kingdom). JET Joint Undertaking; Gresillon, D. [Ecole Polytechnique, 91 - Palaiseau (France). Lab. de Physique des Milieux Ionises


    The goal of the study is to examine whether the spontaneous emission can account for ICE (ion cyclotron emission) experimental results, or part of them. A straightforward approach to plasma emission is chosen, investigating the near equilibrium wave radiation by gyrating ions, and thus building from the majority and fast fusion ions the plasma fluctuations and emission on the fast magnetoacoustic or compressional Alfven wave mode in the IC frequency range. Similarities with the ICE experiments are shown: the emission temperature in the presence of fast ions (even in a very small amount), the strong fast ion emission increase with the harmonic, the fine double-line splitting of each peak, the linear but not proportional increase of the peak width with the harmonic. 3 refs., 2 figs.

  13. Atom probe field ion microscopy and related topics: A bibliography 1993

    Energy Technology Data Exchange (ETDEWEB)

    Godfrey, R.D.; Miller, M.K.; Russell, K.F.


    This bibliography, covering the period 1993, includes references related to the following topics: atom probe field ion microscopy (APFIM), field emission (FE), and field ion microscopy (FIM). Technique-oriented studies and applications are included. The references contained in this document were compiled from a variety of sources including computer searches and personal lists of publications. To reduce the length of this document, the references have been reduced to the minimum necessary to locate the articles. The references are listed alphabetically by authors, an Addendum of references missed in previous bibliographies is included.

  14. Electron microscopy of Mg/TiO{sub 2} photocatalyst morphology for deep desulfurization of diesel

    Energy Technology Data Exchange (ETDEWEB)

    Yin, Yee Cia, E-mail: [Department of Chemical Engineering, Universiti Teknologi PETRONAS, 31750 Tronoh, Perak (Malaysia); Kait, Chong Fai, E-mail:; Fatimah, Hayyiratul, E-mail:; Wilfred, Cecilia, E-mail: [Department of Fundamental and Applied Sciences, Universiti Teknologi PETRONAS, 31750 Tronoh, Perak (Malaysia)


    A series of Mg/TiO{sub 2} photocatalysts were prepared and characterized using Field Emission Scanning Electron Microscopy (FESEM) and High-Resolution Transmission Electron Microscopy (HRTEM). The average particle sizes of the photocatalysts were ranging from 25.7 to 35.8 nm. Incorporation of Mg on TiO{sub 2} did not lead to any surface lattice distortion to TiO{sub 2}. HRTEM data indicated the presence of MgO and Mg(OH){sub 2} mixture at low Mg loading while at higher Mg loading, the presence of lamellar Mg-oxyhydroxide intermediates and Mg(OH){sub 2}.

  15. Gabor domain optical coherence microscopy (United States)

    Murali, Supraja

    Time domain Optical Coherence Tomography (TD-OCT), first reported in 1991, makes use of the low temporal coherence properties of a NIR broadband laser to create depth sectioning of up to 2mm under the surface using optical interferometry and point to point scanning. Prior and ongoing work in OCT in the research community has concentrated on improving axial resolution through the development of broadband sources and speed of image acquisition through new techniques such as Spectral domain OCT (SD-OCT). In SD-OCT, an entire depth scan is acquired at once with a low numerical aperture (NA) objective lens focused at a fixed point within the sample. In this imaging geometry, a longer depth of focus is achieved at the expense of lateral resolution, which is typically limited to 10 to 20 mum. Optical Coherence Microscopy (OCM), introduced in 1994, combined the advantages of high axial resolution obtained in OCT with high lateral resolution obtained by increasing the NA of the microscope placed in the sample arm. However, OCM presented trade-offs caused by the inverse quadratic relationship between the NA and the DOF of the optics used. For applications requiring high lateral resolution, such as cancer diagnostics, several solutions have been proposed including the periodic manual re-focusing of the objective lens in the time domain as well as the spectral domain C-mode configuration in order to overcome the loss in lateral resolution outside the DOF. In this research, we report for the first time, high speed, sub-cellular imaging (lateral resolution of 2 mum) in OCM using a Gabor domain image processing algorithm with a custom designed and fabricated dynamic focus microscope interfaced to a Ti:Sa femtosecond laser centered at 800 nm within an SD-OCM configuration. It is envisioned that this technology will provide a non-invasive replacement for the current practice of multiple biopsies for skin cancer diagnosis. The research reported here presents three important advances

  16. Overview of optical microscopy and optical microspectroscopy (United States)

    Ager, Joel W.


    Optical microscopy has historically been a major tool for semiconductor inspection. As the ULSI design rule continues to decline to 0.25 μm and below, standard optical microscopy methods will arrive at their resolution limit. In the first part of this paper an overview of currently used optical microscopy techniques will be given. The resolution limit for optical imaging will be discussed, and novel methods for increasing resolution, including deep UV microscopy and confocal laser microscopy, will be presented. The second part of the paper will discuss an emerging technology for contamination analysis in semiconductor processing, microspectroscopy. Three topics in this area will be discussed with an emphasis on applications to off-line defect identification in process development: (1) micro-Raman spectroscopy, (2) micro-fluorescence or micro-photoluminescence spectroscopy, and (3) micro-reflectivity. It will be shown that these microspectroscopy methods can provide composition information for defects down to 1 μm in size that is not accessible through the more commonly used methods such as scanning electron microscopy with energy dispersive spectroscopy (SEM/EDS) and scanning Auger microscopy. Classes of defects where optical micro-spectroscopy methods are useful include ceramic particles, thin films of organic material, and dielectric films.

  17. Confocal light scattering and absorption spectroscopic microscopy (United States)

    Qiu, Le; Vitkin, Edward; Salahuddin, Saira; Zaman, Munir M.; Andersson, Charlotte; Freedman, Steven D.; Hanlon, Eugene B.; Itzkan, Irving; Perelman, Lev T.


    We have developed a novel optical method for observing submicron intracellular structures in living cells which is called confocal light absorption and scattering spectroscopic (CLASS) microscopy. It combines confocal microscopy, a well-established high-resolution microscopic technique, with light scattering spectroscopy (LSS). CLASS microscopy requires no exogenous labels and is capable of imaging and continuously monitoring individual viable cells, enabling the observation of cell and organelle functioning at scales on the order of 100 nm. In addition, it provides not only size information but also information about the biochemical and physical properties of the cell.

  18. Fluorescence spectroscopy and confocal microscopy of the mycotoxin citrinin in condensed phase and hydrogel films. (United States)

    Lauer, Milena H; Gehlen, Marcelo H; de Jesus, Karen; Berlinck, Roberto G S


    The emission spectra, quantum yields and fluorescence lifetimes of citrinin in organic solvents and hydrogel films have been determined. Citrinin shows complex fluorescence decays due to the presence of two tautomers in solution and interconversion from excited-state double proton transfer (ESDPT) process. The fluorescence decay times associated with the two tautomers have values near 1 and 5 ns depending on the medium. In hydrogel films of agarose and alginate, fluorescence imaging showed that citrinin is not homogeneously dispersed and highly emissive micrometer spots may be formed. Fluorescence spectrum and decay analysis are used to recognize the presence of citrinin in hydrogel films using confocal fluorescence microscopy and spectroscopy.

  19. Nanoscopy for nanoscience: how super-resolution microscopy extends imaging for nanotechnology. (United States)

    Johnson, Sam A


    Imaging methods have presented scientists with powerful means of investigation for centuries. The ability to resolve structures using light microscopes is though limited to around 200 nm. Fluorescence-based super-resolution light microscopy techniques of several principles and methods have emerged in recent years and offer great potential to extend the capabilities of microscopy. This resolution improvement is especially promising for nanoscience where the imaging of nanoscale structures is inherently restricted by the resolution limit of standard forms of light microscopy. Resolution can be improved by several distinct approaches including structured illumination microscopy, stimulated emission depletion, and single-molecule positioning methods such as photoactivated localization microscopy and stochastic optical reconstruction microscopy and several derivative variations of each of these. These methods involve substantial differences in the resolutions achievable in the different axes, speed of acquisition, compatibility with different labels, ease of use, hardware complexity, and compatibility with live biological samples. The field of super-resolution imaging and its application to nanotechnology is relatively new and still rapidly developing. An overview of how these methods may be used with nanomaterials is presented with some examples of pioneering uses of these approaches. © 2014 Wiley Periodicals, Inc.

  20. Tropospheric Emission Spectrometer and Airborne Emission Spectrometer (United States)

    Glavich, T.; Beer, R.


    The Tropospheric Emission Spectrometer (TES) is an instrument being developed for the NASA Earth Observing System Chemistry Platform. TES will measure the distribution of ozone and its precursors in the lower atmosphere. The Airborne Emission Spectrometer (AES) is an aircraft precursor to TES. Applicable descriptions are given of instrument design, technology challenges, implementation and operations for both.

  1. Dispersion-enhanced third-harmonic microscopy (United States)

    Stock, Christian; Zlatanov, Kaloyan; Halfmann, Thomas


    We demonstrate strong enhancements of signal yield and image contrast in third-harmonic microscopy by appropriate choice of driving laser wavelength to modulate the phase-matching conditions of the conversion process by dispersion control. Tuning the laser wavelength in the range of 1010 - 1350 nm at samples containing interfaces with water and glass, we obtained large signal enhancements up to a factor of 19, and improvements in the image contrast by an order of magnitude. The effect is most pronounced at interfaces with media of small and/or not too different nonlinear optical susceptibilities, e.g., as it is the case in typical samples in harmonic microscopy. Beyond the demonstration of this new variant of third-harmonic microscopy, our findings are also of relevance to a proper choice of laser systems for harmonic microscopy setups.

  2. Accuracy and Precision in Quantitative Fluorescence Microscopy

    National Research Council Canada - National Science Library

    Jennifer C. Waters


    .... With advances in digital cameras and the discovery and development of genetically encoded fluorophores, there has been a huge increase in the use of fluorescence microscopy to quantify spatial...

  3. Halo-free Phase Contrast Microscopy

    National Research Council Canada - National Science Library

    Tan H Nguyen; Mikhail Kandel; Haadi M Shakir; Catherine Best-popescu; Jyothi Arikkath; Minh N Do; Gabriel Popescu


    We present a new approach for retrieving halo-free phase contrast microscopy (hfPC) images by upgrading the conventional PC microscope with an external interferometric module, which generates sufficient data for reversing the halo artifact...

  4. [Atomic force microscopy involved in protein study]. (United States)

    Lu, Zhengjian; Chen, Guoping; Wang, Jianhua


    Atomic force microscopy is a rather new type of nano microscopic technology. It has some advantages, such as high resolution (sub-nano scale); avoidance of special sample preparation; real-time detection of samples under nearly physiological environment; in situ study of samples under water environment; feasibility of investigating physical and chemical properties of samples at molecular level, etc. In recent years, the application of atomic force microscopy in protein study has brought about outstanding achievements. In this paper are introduced the principle and operation modes of atomic force microscopy, also presented are its application in protein imaging, adsorption, folding-and-unfolding, assembly, and single molecular recognition. Additionally, the future application of atomic force microscopy in protein study is prospected.

  5. Multiphoton microscopy: An introduction to gastroenterologists

    Institute of Scientific and Technical Information of China (English)

    Hye Jin Cho; Hoon Jai Chun; Eun Sun Kim; Bong Rae Cho


    Multiphoton microscopy, relying on the simultaneous absorption of two or more photons by a fluorophore, has come to occupy a prominent place in modern biomedical research with its ability to allow real-time observation of a single cell and molecules in intact tissues. Multiphoton microscopy exhibits nonlinear optical contrast properties, which can make it possible to provide an exceptionally large depth penetration with less phototoxicity. This system becomes more and more an inspiring tool for a non-invasive imaging system to realize "optical biopsy" and to examine the functions of living cells. In this review, we briefly present the physical principles and properties of multiphoton microscopy as well as the current applications in biological fields. In addition, we address what we see as the future potential of multiphoton microscopy for gastroenterologic research.

  6. Multiphoton microscopy in defining liver function (United States)

    Thorling, Camilla A.; Crawford, Darrell; Burczynski, Frank J.; Liu, Xin; Liau, Ian; Roberts, Michael S.


    Multiphoton microscopy is the preferred method when in vivo deep-tissue imaging is required. This review presents the application of multiphoton microscopy in defining liver function. In particular, multiphoton microscopy is useful in imaging intracellular events, such as mitochondrial depolarization and cellular metabolism in terms of NAD(P)H changes with fluorescence lifetime imaging microscopy. The morphology of hepatocytes can be visualized without exogenously administered fluorescent dyes by utilizing their autofluorescence and second harmonic generation signal of collagen, which is useful in diagnosing liver disease. More specific imaging, such as studying drug transport in normal and diseased livers are achievable, but require exogenously administered fluorescent dyes. If these techniques can be translated into clinical use to assess liver function, it would greatly improve early diagnosis of organ viability, fibrosis, and cancer.

  7. Filter-Dense Multicolor Microscopy: e0119499

    National Research Council Canada - National Science Library

    Siavash Kijani; Ulf Yrlid; Maria Heyden; Malin Levin; Jan Borén; Per Fogelstrand


    .... However, because of the risk of bleed-through signals between fluorochromes, standard multicolor microscopy is restricted to a maximum of four fluorescence channels, including one for nuclei staining...

  8. Application of spectroscopy and super-resolution microscopy: Excited state

    Energy Technology Data Exchange (ETDEWEB)

    Bhattacharjee, Ujjal [Iowa State Univ., Ames, IA (United States)


    Photophysics of inorganic materials and organic molecules in complex systems have been extensively studied with absorption and emission spectroscopy.1-4 Steady-state and time-resolved fluorescence studies are commonly carried out to characterize excited-state properties of fluorophores. Although steady-state fluorescence measurements are widely used for analytical applications, time-resolved fluorescence measurements provide more detailed information about excited-state properties and the environment in the vicinity of the fluorophore. Many photophysical processes, such as photoinduced electron transfer (PET), rotational reorientation, solvent relaxation, and energy transfer, occur on a nanosecond (10-9 s) timescale, thus affecting the lifetime of the fluorophores. Moreover, time-resolved microscopy methods, such as lifetimeimaging, combine the benefits of the microscopic measurement and information-rich, timeresolved data. Thus, time-resolved fluorescence spectroscopy combined with microscopy can be used to quantify these processes and to obtain a deeper understanding of the chemical surroundings of the fluorophore in a small area under investigation. This thesis discusses various photophysical and super-resolution microscopic studies of organic and inorganic materials, which have been outlined below.

  9. Ultrafast Photon Counting Applied to Resonant Scanning STED Microscopy (United States)

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


    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

  10. Scanning electron microscopy physics of image formation and microanalysis

    CERN Document Server

    Reimer, Ludwig


    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...

  11. Synchrotron X-ray microscopy of marine calcifiers: how plankton record past climate change (United States)

    Redfern, S. A. T.; Branson, O.; Read, E.


    We have used STXM and PEEM to reveal the underpinning chemistry and nanoscale structure behind palaeo-climate geochemical signatures, such as trace Mg in shells- proposed proxies for palaeo-ocean temperature. This has allowed us to test the chemical assumptions and mechanisms underpinning the use of such empirical proxies. We have determined the control on driving chemical variations in biogenic carbonates using STXM at the absorption edge of Mg, B, and Na in the shells of modern plankton. The power of these observations lies in their ability to link changes in chemistry, microstructure, and growth process in biogenic carbonate to environmental influences. We have seen that such changes occur at length scales of tens of nanometres and demonstrated that STXM provides an invaluable route to understanding chemical environment and key heterogeneity at the appropriate length scale. This new understanding provides new routes for future measurements of past climate variation in the sea floor fossil record.

  12. Three-dimensional characterization of extreme ultraviolet mask blank defects by interference contrast photoemission electron microscopy. (United States)

    Lin, Jingquan; Weber, Nils; Escher, Matthias; Maul, Jochen; Han, Hak-Seung; Merkel, Michael; Wurm, Stefan; Schönhense, Gerd; Kleineberg, Ulf


    A photoemission electron microscope based on a new contrast mechanism "interference contrast" is applied to characterize extreme ultraviolet lithography mask blank defects. Inspection results show that positioning of interference destructive condition (node of standing wave field) on surface of multilayer in the local region of a phase defect is necessary to obtain best visibility of the defect on mask blank. A comparative experiment reveals superiority of the interference contrast photoemission electron microscope (Extreme UV illumination) over a topographic contrast one (UV illumination with Hg discharge lamp) in detecting extreme ultraviolet mask blank phase defects. A depth-resolved detection of a mask blank defect, either by measuring anti-node peak shift in the EUV-PEEM image under varying inspection wavelength condition or by counting interference fringes with a fixed illumination wavelength, is discussed.

  13. Tutorial on Photoacoustic Microscopy and Computed Tomography


    Wang, Lihong V.


    The field of photoacoustic tomography has experienced considerable growth in the past few years. Although several commercially available pure optical imaging modalities, including confocal microscopy, two-photon microscopy, and optical coherence tomography, have been highly successful, none of these technologies can provide penetration beyond ~1 mm into scattering biological tissues, because they are based on ballistic and quasi-ballistic photons. Heretofore, there has been a void in high-res...

  14. Concepts for nanoscale resolution in fluorescence microscopy. (United States)

    Hell, Stefan W; Dyba, Marcus; Jakobs, Stefan


    Spatio-temporal visualization of cellular structures by fluorescence microscopy has become indispensable in biology. However, the resolution of conventional fluorescence microscopy is limited by diffraction to about 180 nm in the focal plane and to about 500 nm along the optic axis. Recently, concepts have emerged that overcome the diffraction resolution barrier fundamentally. Formed on the basis of reversible saturable optical transitions, these concepts might eventually allow us to investigate hitherto inaccessible details within live cells.

  15. Direct electron detection in transmission electron microscopy


    Jin, Liang


    Since the first prototype of a transmission electron microscope was built in 1931 by Ernst Ruska and Max Knoll, Transmission Electron Microscopy (TEM) has proved to be an essential imaging tool for physicists, material scientists, and biologists. To record the TEM images for analysis, electron microscopists have used specialized electron micrograph film for a long time, until the new developments in TEM, such as electron tomography and cryo- electron microscopy, pushed for the needs of digita...

  16. Scanning Electron Microscopy in modern dentistry research


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


    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...

  17. Imaging DNA Structure by Atomic Force Microscopy. (United States)

    Pyne, Alice L B; Hoogenboom, Bart W


    Atomic force microscopy (AFM) is a microscopy technique that uses a sharp probe to trace a sample surface at nanometre resolution. For biological applications, one of its key advantages is its ability to visualize substructure of single molecules and molecular complexes in an aqueous environment. Here, we describe the application of AFM to determine superstructure and secondary structure of surface-bound DNA. The method is also readily applicable to probe DNA-DNA interactions and DNA-protein complexes.

  18. A high throughput spectral image microscopy system (United States)

    Gesley, M.; Puri, R.


    A high throughput spectral image microscopy system is configured for rapid detection of rare cells in large populations. To overcome flow cytometry rates and use of fluorophore tags, a system architecture integrates sample mechanical handling, signal processors, and optics in a non-confocal version of light absorption and scattering spectroscopic microscopy. Spectral images with native contrast do not require the use of exogeneous stain to render cells with submicron resolution. Structure may be characterized without restriction to cell clusters of differentiation.

  19. Light Sheet Fluorescence Microscopy: A Review


    Santi, Peter A.


    Light sheet fluorescence microscopy (LSFM) functions as a non-destructive microtome and microscope that uses a plane of light to optically section and view tissues with subcellular resolution. This method is well suited for imaging deep within transparent tissues or within whole organisms, and because tissues are exposed to only a thin plane of light, specimen photobleaching and phototoxicity are minimized compared to wide-field fluorescence, confocal, or multiphoton microscopy. LSFMs produce...

  20. Differential-concentration scanning ion conductance microscopy


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


    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...

  1. Imaging spin dynamics in monolayer WS2 by time-resolved Kerr rotation microscopy (United States)

    McCormick, Elizabeth J.; Newburger, Michael J.; Luo, Yunqiu Kelly; McCreary, Kathleen M.; Singh, Simranjeet; Martin, Iwan B.; Cichewicz, Edward J., Jr.; Jonker, Berend T.; Kawakami, Roland K.


    Monolayer transition metal dichalcogenides (TMD) have immense potential for future spintronic and valleytronic applications due to their 2D nature and long spin/valley lifetimes. We investigate the origin of these long-lived states in n-type WS2 using time-resolved Kerr rotation microscopy and photoluminescence microscopy with ~1 µm spatial resolution. Comparing the spatial dependence of the Kerr rotation signal and the photoluminescence reveals a correlation with neutral exciton emission, which is likely due to the transfer of angular momentum to resident conduction electrons with long spin/valley lifetimes. In addition, we observe an unexpected anticorrelation between the Kerr rotation and trion emission, which provides evidence for the presence of long-lived spin/valley-polarized dark trions. We also find that the spin/valley polarization in WS2 is robust to magnetic fields up to 700 mT, indicative of spins and valleys that are stabilized with strong spin–orbit fields.

  2. Development and testing of hyperbaric atomic force microscopy (AFM) and fluorescence microscopy for biological applications. (United States)

    D'Agostino, D P; McNally, H A; Dean, J B


    A commercially available atomic force microscopy and fluorescence microscope were installed and tested inside a custom-designed hyperbaric chamber to provide the capability to study the effects of hyperbaric gases on biological preparations, including cellular mechanism of oxidative stress. In this report, we list details of installing and testing atomic force microscopy and fluorescence microscopy inside a hyperbaric chamber. The pressure vessel was designed to accommodate a variety of imaging equipment and ensures full functionality at ambient and hyperbaric conditions (≤85 psi). Electrical, gas and fluid lines were installed to enable remote operation of instrumentation under hyperbaric conditions, and to maintain viable biological samples with gas-equilibrated superfusate and/or drugs. Systems were installed for vibration isolation and temperature regulation to maintain atomic force microscopy performance during compression and decompression. Results of atomic force microscopy testing demonstrate sub-nanometre resolution at hyperbaric pressure in dry scans and fluid scans, in both contact mode and tapping mode. Noise levels were less when measurements were taken under hyperbaric pressure with air, helium (He) and nitrogen (N(2) ). Atomic force microscopy and fluorescence microscopy measurements were made on a variety of living cell cultures exposed to hyperbaric gases (He, N(2) , O(2) , air). In summary, atomic force microscopy and fluorescence microscopy were installed and tested for use at hyperbaric pressures and enables the study of cellular and molecular effects of hyperbaric gases and pressure per se in biological preparations. © 2012 The Authors Journal of Microscopy © 2012 Royal Microscopical Society.

  3. Non-contact lateral force microscopy (United States)

    Weymouth, A. J.


    The goal of atomic force microscopy (AFM) is to measure the short-range forces that act between the tip and the surface. The signal recorded, however, includes long-range forces that are often an unwanted background. Lateral force microscopy (LFM) is a branch of AFM in which a component of force perpendicular to the surface normal is measured. If we consider the interaction between tip and sample in terms of forces, which have both direction and magnitude, then we can make a very simple yet profound observation: over a flat surface, long-range forces that do not yield topographic contrast have no lateral component. Short-range interactions, on the other hand, do. Although contact-mode is the most common LFM technique, true non-contact AFM techniques can be applied to perform LFM without the tip depressing upon the sample. Non-contact lateral force microscopy (nc-LFM) is therefore ideal to study short-range forces of interest. One of the first applications of nc-LFM was the study of non-contact friction. A similar setup is used in magnetic resonance force microscopy to detect spin flipping. More recently, nc-LFM has been used as a true microscopy technique to systems unsuitable for normal force microscopy.

  4. Backside absorbing layer microscopy: Watching graphene chemistry. (United States)

    Campidelli, Stéphane; Abou Khachfe, Refahi; Jaouen, Kevin; Monteiller, Jean; Amra, Claude; Zerrad, Myriam; Cornut, Renaud; Derycke, Vincent; Ausserré, Dominique


    The rapid rise of two-dimensional nanomaterials implies the development of new versatile, high-resolution visualization and placement techniques. For example, a single graphene layer becomes observable on Si/SiO2 substrates by reflected light under optical microscopy because of interference effects when the thickness of silicon oxide is optimized. However, differentiating monolayers from bilayers remains challenging, and advanced techniques, such as Raman mapping, atomic force microscopy (AFM), or scanning electron microscopy (SEM) are more suitable to observe graphene monolayers. The first two techniques are slow, and the third is operated in vacuum; hence, in all cases, real-time experiments including notably chemical modifications are not accessible. The development of optical microscopy techniques that combine the speed, large area, and high contrast of SEM with the topological information of AFM is therefore highly desirable. We introduce a new widefield optical microscopy technique based on the use of previously unknown antireflection and absorbing (ARA) layers that yield ultrahigh contrast reflection imaging of monolayers. The BALM (backside absorbing layer microscopy) technique can achieve the subnanometer-scale vertical resolution, large area, and real-time imaging. Moreover, the inverted optical microscope geometry allows its easy implementation and combination with other techniques. We notably demonstrate the potentiality of BALM by in operando imaging chemical modifications of graphene oxide. The technique can be applied to the deposition, observation, and modification of any nanometer-thick materials.

  5. What Is Emissions Trading? (United States)

    Learn the basics about how emissions trading uses a market-based policy tool used to control large amounts of pollution emissions from a group of sources in order to protect human health and the environment.

  6. World Emission RETRO ANTHRO (United States)

    Washington University St Louis — Anthropogenic and vegetation fire emissions data were generated monthly covering a period of 1960 to 2000. Anthropogenic emissions in the RETRO inventory are derived...

  7. National Emission Inventory (United States)

    U.S. Environmental Protection Agency — The National Emission Inventory contains measured, modeled, and estimated data for emissions of all known source categories in the US (stationary sources, fires,...

  8. Emissions Modeling Clearinghouse (United States)

    U.S. Environmental Protection Agency — The Emissions Modeling Clearinghouse (EMCH) supports and promotes emissions modeling activities both internal and external to the EPA. Through this site, the EPA...

  9. Control of Emissions (United States)

    Parrish, Clyde F. (Inventor); Chung, Landy (Inventor)


    Methods and apparatus utilizing chlorine dioxide and hydrogen peroxide are useful to reduce NOx emissions, as well as SOx and mercury (or other heavy metal) emissions, from combustion flue gas streams.

  10. Biodiesel Emissions Analysis Program (United States)

    Using existing data, the EPA's biodiesel emissions analysis program sought to quantify the air pollution emission effects of biodiesel for diesel engines that have not been specifically modified to operate on biodiesel.

  11. Measuring the Spatial Distribution of Dielectric Constants in Polymers through Quasi-Single Molecule Microscopy


    Hess, Chelsea M.; Riley, Erin A.; Palos-Chávez, Jorge; Reid, Philip J.


    The variation in dielectric constant is measured for thin films of poly(methyl methacrylate) (PMMA) and poly(vinylidene fluoride) (PVDF) using confocal fluorescence microscopy. Spatial variation in the local dielectric constant of the polymer films on the ~250 nm length scale is measured using the solvochromatic emission from incorporated nile red (NR) at “quasi-single molecule” (10−7 M) and true single molecule (SM) concentrations (10−9 M). Correlation of the NR fluorescence wavelength maxim...

  12. A novel method for enhancing the lateral resolution and image SNR in confocal microscopy (United States)

    Chen, Youhua; Zhu, Dazhao; Fang, Yue; Kuang, Cuifang; Liu, Xu


    There is always a tradeoff between the resolution and the signal-to-noise ratio (SNR) in confocal microscopy. In particular, the pinhole size is very important for maintaining a balance between them. In this paper, we propose a method for improving the lateral resolution and image SNR in confocal microscopy without making any changes to the hardware. By using the fluorescence emission difference (FED) approach, we divide the images acquired by different pinhole sizes into one image acquired by the central pinhole and several images acquired by ring-shaped pinholes. Then, they are added together with the deconvolution method. Simulation and experimental results for fluorescent particles and cells show that our method can achieve a far better resolution than a large pinhole and a higher SNR than a small pinhole. Moreover, our method can improve the performance of classic confocal laser scanning microscopy (CLSM) to a certain extent, especially CLSM with a continuously variable pinhole.

  13. High-sensitivity chemical imaging for biomedicine by SRS microscopy (Conference Presentation) (United States)

    Min, Wei


    Innovations in spectroscopy principles and microscopy technology have significantly impacted modern biology and medicine. While most of the contemporary bio-imaging modalities harness electronic transition, nuclear spin or radioactivity, vibrational spectroscopy has not been widely used yet. Here we will discuss an emerging chemical imaging platform, stimulated Raman scattering (SRS) microscopy, which can enhance the otherwise feeble spontaneous Raman eight orders of magnitude by virtue of stimulated emission. When coupled with stable isotopes (e.g., deuterium and 13C) or bioorthogonal chemical moieties (e.g., alkynes), SRS microscopy is well suited for probing in vivo metabolic dynamics of small bio-molecules which cannot be labeled by bulky fluorophores. Physical principle of the underlying optical spectroscopy and exciting biomedical applications such as imaging lipid metabolism, protein synthesis, DNA replication, protein degradation, RNA synthesis, glucose uptake, drug trafficking and tumor metabolism will be presented.

  14. Visualization of Oil Body Distribution in Jatropha curcas L. by Four-Wave Mixing Microscopy (United States)

    Ishii, Makiko; Uchiyama, Susumu; Ozeki, Yasuyuki; Kajiyama, Sin'ichiro; Itoh, Kazuyoshi; Fukui, Kiichi


    Jatropha curcas L. (jatropha) is a superior oil crop for biofuel production. To improve the oil yield of jatropha by breeding, the development of effective and reliable tools to evaluate the oil production efficiency is essential. The characteristics of the jatropha kernel, which contains a large amount of oil, are not fully understood yet. Here, we demonstrate the application of four-wave mixing (FWM) microscopy to visualize the distribution of oil bodies in a jatropha kernel without staining. FWM microscopy enables us to visualize the size and morphology of oil bodies and to determine the oil content in the kernel to be 33.2%. The signal obtained from FWM microscopy comprises both of stimulated parametric emission (SPE) and coherent anti-Stokes Raman scattering (CARS) signals. In the present situation, where a very short pump pulse is employed, the SPE signal is believed to dominate the FWM signal.

  15. Galactic Diffuse Polarized Emission

    Indian Academy of Sciences (India)

    Diffuse polarized emission by synchrotron is a key tool to investigate magnetic fields in the Milky Way, particularly the ordered component of the large scale structure. Key observables are the synchrotron emission itself and the RM is by Faraday rotation. In this paper the main properties of the radio polarized diffuse emission ...

  16. Bridging the Emissions Gap

    NARCIS (Netherlands)

    Blok, K.


    The analyses in Chapters 2 and 3 of this report concluded that the emissions gap in 2020 will likely be between 8 and 13 GtCO2e. The chapters also estimated the difference between BaU emissions in 2020 and the emissions level consistent with a “likely” chance of staying within the 2°C target to

  17. Self-labelling enzymes as universal tags for fluorescence microscopy, super-resolution microscopy and electron microscopy (United States)

    Liss, Viktoria; Barlag, Britta; Nietschke, Monika; Hensel, Michael


    Research in cell biology demands advanced microscopy techniques such as confocal fluorescence microscopy (FM), super-resolution microscopy (SRM) and transmission electron microscopy (TEM). Correlative light and electron microscopy (CLEM) is an approach to combine data on the dynamics of proteins or protein complexes in living cells with the ultrastructural details in the low nanometre scale. To correlate both data sets, markers functional in FM, SRM and TEM are required. Genetically encoded markers such as fluorescent proteins or self-labelling enzyme tags allow observations in living cells. Various genetically encoded tags are available for FM and SRM, but only few tags are suitable for CLEM. Here, we describe the red fluorescent dye tetramethylrhodamine (TMR) as a multimodal marker for CLEM. TMR is used as fluorochrome coupled to ligands of genetically encoded self-labelling enzyme tags HaloTag, SNAP-tag and CLIP-tag in FM and SRM. We demonstrate that TMR can additionally photooxidize diaminobenzidine (DAB) to an osmiophilic polymer visible on TEM sections, thus being a marker suitable for FM, SRM and TEM. We evaluated various organelle markers with enzymatic tags in mammalian cells labelled with TMR-coupled ligands and demonstrate the use as efficient and versatile DAB photooxidizer for CLEM approaches. PMID:26643905

  18. Functionalization of atomic force microscopy Akiyama tips for magnetic force microscopy measurements


    Stiller, Markus; Barzola-Quiquia, Jose; Esquinazi, Pablo D.; Sangiao, Soraya; De Teresa, Jose M.; Meijer, Jan; Abel, Bernd


    In this work we have used focused electron beam induced deposition of cobalt to functionalize atomic force microscopy Akiyama tips for application in magnetic force microscopy. The grown tips have a content of 90% Co after exposure to ambient air. The magnetic tips were characterized using energy dispersive X-ray spectroscopy and scanning electron microscopy. In order to investigate the magnetic properties, current loops were prepared by electron beam lithography. Measurements at room tempera...

  19. Distinguishing magnetic and electrostatic interactions by a Kelvin probe force microscopy-magnetic force microscopy combination

    National Research Council Canada - National Science Library

    Jaafar, Miriam; Iglesias-Freire, Oscar; Serrano-Ramón, Luis; Ibarra, Manuel Ricardo; de Teresa, Jose Maria; Asenjo, Agustina


    .... In particular, magnetic force microscopy (MFM) is used to characterize the domain configuration in ferromagnetic materials such as thin films grown by physical techniques or ferromagnetic nanostructures...

  20. Biological Applications of Near-field Optical Microscopy

    NARCIS (Netherlands)

    van Hulst, N.F.; Moers, M.H.P.; Moers, M.H.P.


    Presents several biological applications of near field optical microscopy, in combination with force microscopy. Aperture near field scanning optical microscopy (NSOM) with fluorescence detection gives (bio)chemical specificity and orientational information, in addition to the simultaneously

  1. Shipping emissions in ports


    Merk, Olaf


    Shipping emissions in ports are substantial, accounting for 18 million tonnes of CO2 emissions, 0.4 million tonnes of NOx, 0.2 million of SOx and 0.03 million tonnes of PM10 in 2011. Around 85% of emissions come from containerships and tankers. Containerships have short port stays, but high emissions during these stays. Most of CO2 emissions in ports from shipping are in Asia and Europe (58%), but this share is low compared to their share of port calls (70%). European ports have much less emi...

  2. International emissions trading

    DEFF Research Database (Denmark)

    Boom, Jan Tjeerd

    This thesis discusses the design and political acceptability of international emissions trading. It is shown that there are several designs options for emissions trading at the national level that have a different impact on output and thereby related factors such as employment and consumer prices....... The differences in impact of the design make that governments may prefer different designs of emissions trading in different situations. The thesis furthermore establishes that international emissions trading may lead to higher overall emissions, which may make it a less attractive instrument....

  3. [Synovial fluid crystal identification by electron microscopy]. (United States)

    Nero, Patrícia; Nogueira, Isabel; Vilar, Rui; Pimentão, J Bravo; Branco, Jaime C


    In clinical practice crystal identification in synovial fluid is made by polarized light microscopy and with some specific stainings. Nevertheless, sometimes we are unable to identify crystals by these means, either because they are too small or because they are widespread on the fluid. To compare the identification of crystals in synovial fluid from patients with non-infectious monoarthritis but no history of local trauma or articular disease, using polarized light and electronic microscopy. We analized synovial fluid samples from patients with non-infectious monoarthritis and no history of local trauma or articular disease. First we used a polarized light microscope and alizarin red staining. Later we used conventional transmission electron microscopy and energy dispersive spectroscopy, in order to identify and characterize crystals. Fourty-five samples from 23 synovial fluids were analyzed. Under polarized light microscopy we identified crystals on 11 samples: 3 with calcium pyrophosphate crystals, 6 with calcium basic phosphate crystals and 2 with sodium monourate crystals. On the remaining 12 samples we were unable to identify crystals. Samples were then analyzed by conventional transmission electron microscopy and energy dispersive spectroscopy confirming the presence of the previously identified crystals. On the remainig 12 samples we were able to identify calcium basic phosphate crystals. Microcrystals seem to be an universal finding in synovial fluid of patients with osteoarthritis. The prevention of their deposition in joints might contribute to stop joint damage in this disease.

  4. X-ray microscopy of human malaria

    Energy Technology Data Exchange (ETDEWEB)

    Magowan, C.; Brown, J.T.; Mohandas, N.; Meyer-Ilse, W. [Ernest Orlando Lawrence Berkeley National Lab., CA (United States)


    Associations between intracellular organisms and host cells are complex and particularly difficult to examine. X-ray microscopy provides transmission images of subcellular structures in intact cells at resolutions superior to available methodologies. The spatial resolution is 50-60nm with a 1 micron depth of focus, superior to anything achievable with light microscopy. Image contrast is generated by differences in photoelectric absorption by the atoms in different areas (i.e. subcellular structures) throughout the full thickness of the sample. Absorption due to carbon dominates among all the elements in the sample at 2.4 nm x-ray wavelength. Thus images show features or structures, in a way not usually seen by other types of microscopy. The authors used soft x-ray microscopy to investigate structural development of Plasmodium falciparum malaria parasites in normal and genetically abnormal erythrocytes, and in infected erythrocytes treated with compounds that have anti-malarial effects. X-ray microscopy showed newly elaborated structures in the cytosol of unstained, intact erythrocytes, redistribution of mass (carbon) in infected erythrocytes, and aberrant parasite morphology. Better understanding of the process of intracellular parasite maturation and the interactions between the parasite and its host erythrocyte can help define new approaches to the control of this deadly disease.

  5. Air Emissions Factors and Quantification (United States)

    Emissions factors are used in developing air emissions inventories for air quality management decisions and in developing emissions control strategies. This area provides technical information on and support for the use of emissions factors.

  6. Calcite biomineralization in coccoliths: Evidence from atomic force microscopy (AFM)

    DEFF Research Database (Denmark)

    Henriksen, Karen; Stipp, S.L.S.


    geochemistry, crystal orientation, coccolith function, biomineralization, biological calcite, atomic force microscopy......geochemistry, crystal orientation, coccolith function, biomineralization, biological calcite, atomic force microscopy...

  7. Low Emissions Aftertreatment and Diesel Emissions Reduction

    Energy Technology Data Exchange (ETDEWEB)



    Detroit Diesel Corporation (DDC) has successfully completed a five-year Low Emissions Aftertreatment and Diesel Emissions Reduction (LEADER) program under a DOE project entitled: ''Research and Development for Compression-Ignition Direct-Injection Engines (CIDI) and Aftertreatment Sub-Systems''. The objectives of the LEADER Program were to: Demonstrate technologies that will achieve future federal Tier 2 emissions targets; and Demonstrate production-viable technical targets for engine out emissions, efficiency, power density, noise, durability, production cost, aftertreatment volume and weight. These objectives were successfully met during the course of the LEADER program The most noteworthy achievements in this program are listed below: (1) Demonstrated Tier 2 Bin 3 emissions target over the FTP75 cycle on a PNGV-mule Neon passenger car, utilizing a CSF + SCR system These aggressive emissions were obtained with no ammonia (NH{sub 3}) slip and a combined fuel economy of 63 miles per gallon, integrating FTP75 and highway fuel economy transient cycle test results. Demonstrated feasibility to achieve Tier 2 Bin 8 emissions levels without active NOx aftertreatment. (2) Demonstrated Tier 2 Bin 3 emissions target over the FTP75 cycle on a light-duty truck utilizing a CSF + SCR system, synergizing efforts with the DOE-DDC DELTA program. This aggressive reduction in tailpipe out emissions was achieved with no ammonia slip and a 41% fuel economy improvement, compared to the equivalent gasoline engine-equipped vehicle. (3) Demonstrated Tier 2 near-Bin 9 emissions compliance on a light-duty truck, without active NOx aftertreatment devices, in synergy with the DOE-DDC DELTA program. (4) Developed and applied advanced combustion technologies such as ''CLEAN Combustion{copyright}'', which yields simultaneous reduction in engine out NOx and PM emissions while also improving engine and aftertreatment integration by providing favorable

  8. Laser scanning laser diode photoacoustic microscopy system. (United States)

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


    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.

  9. Optofluidic time-stretch quantitative phase microscopy. (United States)

    Guo, Baoshan; Lei, Cheng; Wu, Yi; Kobayashi, Hirofumi; Ito, Takuro; Yalikun, Yaxiaer; Lee, Sangwook; Isozaki, Akihiro; Li, Ming; Jiang, Yiyue; Yasumoto, Atsushi; Di Carlo, Dino; Tanaka, Yo; Yatomi, Yutaka; Ozeki, Yasuyuki; Goda, Keisuke


    Innovations in optical microscopy have opened new windows onto scientific research, industrial quality control, and medical practice over the last few decades. One of such innovations is optofluidic time-stretch quantitative phase microscopy - an emerging method for high-throughput quantitative phase imaging that builds on the interference between temporally stretched signal and reference pulses by using dispersive properties of light in both spatial and temporal domains in an interferometric configuration on a microfluidic platform. It achieves the continuous acquisition of both intensity and phase images with a high throughput of more than 10,000 particles or cells per second by overcoming speed limitations that exist in conventional quantitative phase imaging methods. Applications enabled by such capabilities are versatile and include characterization of cancer cells and microalgal cultures. In this paper, we review the principles and applications of optofluidic time-stretch quantitative phase microscopy and discuss its future perspective. Copyright © 2017 Elsevier Inc. All rights reserved.

  10. Transmission Electron Microscopy Physics of Image Formation

    CERN Document Server

    Kohl, Helmut


    Transmission Electron Microscopy: Physics of Image Formation presents the theory of image and contrast formation, and the analytical modes in transmission electron microscopy. The principles of particle and wave optics of electrons are described. Electron-specimen interactions are discussed for evaluating the theory of scattering and phase contrast. Also discussed are the kinematical and dynamical theories of electron diffraction and their applications for crystal-structure analysis and imaging of lattices and their defects. X-ray microanalysis and electron energy-loss spectroscopy are treated as analytical methods. Specimen damage and contamination by electron irradiation limits the resolution for biological and some inorganic specimens. This fifth edition includes discussion of recent progress, especially in the area of aberration correction and energy filtering; moreover, the topics introduced in the fourth edition have been updated. Transmission Electron Microscopy: Physics of Image Formation is written f...

  11. Confocal microscopy in the diagnosis of melanoma

    Directory of Open Access Journals (Sweden)

    Apostolović-Stojanović Milica


    Full Text Available Melanoma is the most deadly form of skin cancer of melanocytic origin. The tumor has a high malignant potential and early metastasis. Prognosis is directly linked to the stage of the disease. Diagnosing thin melanoma at an early stage offers patients their best chance for survival. The crucial innovation in the early recognition of melanoma was the development of in vivo examination of the skin in high-resolution, by confocal microscopy. Confocal microscopy and its modifications provides a “virtual biopsy“, owing to melanosomes and melanin, which are a source of endogenous contrast. Confocal scanning laser microscopy (CSLM provides visualization of microanatomical structures and cellular detail in real time (pigmented keratinocytes, melanocytes, melanosomes and melanophages in the epidermis, dermoepidermal junction and superficial dermis at a resolution equivalent to the resolution of conventional microscopes. [Projekat Ministarstva nauke Republike Srbije, br. 41002

  12. Fluorescence microscopy: A tool to study autophagy (United States)

    Rai, Shashank; Manjithaya, Ravi


    Autophagy is a cellular recycling process through which a cell degrades old and damaged cellular components such as organelles and proteins and the degradation products are reused to provide energy and building blocks. Dysfunctional autophagy is reported in several pathological situations. Hence, autophagy plays an important role in both cellular homeostasis and diseased conditions. Autophagy can be studied through various techniques including fluorescence based microscopy. With the advancements of newer technologies in fluorescence microscopy, several novel processes of autophagy have been discovered which makes it an essential tool for autophagy research. Moreover, ability to tag fluorescent proteins with sub cellular targets has enabled us to evaluate autophagy processes in real time under fluorescent microscope. In this article, we demonstrate different aspects of autophagy in two different model organisms i.e. yeast and mammalian cells, with the help of fluorescence microscopy.

  13. Spectroscopy and atomic force microscopy of biomass. (United States)

    Tetard, L; Passian, A; Farahi, R H; Kalluri, U C; Davison, B H; Thundat, T


    Scanning probe microscopy has emerged as a powerful approach to a broader understanding of the molecular architecture of cell walls, which may shed light on the challenge of efficient cellulosic ethanol production. We have obtained preliminary images of both Populus and switchgrass samples using atomic force microscopy (AFM). The results show distinctive features that are shared by switchgrass and Populus. These features may be attributable to the lignocellulosic cell wall composition, as the collected images exhibit the characteristic macromolecular globule structures attributable to the lignocellulosic systems. Using both AFM and a single case of mode synthesizing atomic force microscopy (MSAFM) to characterize Populus, we obtained images that clearly show the cell wall structure. The results are of importance in providing a better understanding of the characteristic features of both mature cells as well as developing plant cells. In addition, we present spectroscopic investigation of the same samples.

  14. Aberration corrected Lorentz scanning transmission electron microscopy

    Energy Technology Data Exchange (ETDEWEB)

    McVitie, S., E-mail:; McGrouther, D.; McFadzean, S.; MacLaren, D.A.; O’Shea, K.J.; Benitez, M.J.


    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.

  15. [Watching dance of the molecules - CARS microscopy]. (United States)

    Korczyński, Jaroslaw; Kubiak, Katarzyna; Węgłowska, Edyta


    CARS (Coherent Anti-Stokes Raman Scattering) microscopy is an imaging method for living cells visualization as well as for food or cosmetics material analysis without the need for staining. The near infrared laser source generates the CARS signal - the characteristic intrinsic vibrational contrast of the molecules in a sample which is no longer caused by staining, but by the molecules themselves. It provides the benefit of a non-toxic, non-destructive and almost noninvasive method for sample imaging. CARS can easily be combined with fluorescence confocal microscopy so it is an excellent complementary imaging method. In this article we showed some of the applications for this technology: imaging of lipid droplets inside human HaCaT cells and analysis of the composition of cosmetic products. Moreover we believe, that soon new fields of application become accessible for this rapidly developing branch of microscopy.

  16. Ultrafast Science Opportunities with Electron Microscopy

    Energy Technology Data Exchange (ETDEWEB)

    DURR, HERMANN; Wang, X.J., ed.


    X-rays and electrons are two of the most fundamental probes of matter. When the Linac Coherent Light Source (LCLS), the world’s first x-ray free electron laser, began operation in 2009, it transformed ultrafast science with the ability to generate laser-like x-ray pulses from the manipulation of relativistic electron beams. This document describes a similar future transformation. In Transmission Electron Microscopy, ultrafast relativistic (MeV energy) electron pulses can achieve unsurpassed spatial and temporal resolution. Ultrafast temporal resolution will be the next frontier in electron microscopy and can ideally complement ultrafast x-ray science done with free electron lasers. This document describes the Grand Challenge science opportunities in chemistry, material science, physics and biology that arise from an MeV ultrafast electron diffraction & microscopy facility, especially when coupled with linac-based intense THz and X-ray pump capabilities.

  17. Total internal reflection fluorescence (TIRF) microscopy. (United States)

    Trache, Andreea; Meininger, Gerald A


    Total internal reflection fluorescence (TIRF) microscopy represents a method of exciting and visualizing fluorophores present in the near-membrane region of live or fixed cells grown on coverslips. TIRF microscopy is based on the total internal reflection phenomenon that occurs when light passes from a high-refractive medium (e.g., glass) into a low-refractive medium (e.g., cell, water). The evanescent field produced by total internally reflected light excites the fluorescent molecules at the cell-substrate interface and is accompanied by minimal exposure of the remaining cell volume. This technique provides high-contrast fluorescence images, with very low background and virtually no out-of-focus light, ideal for visualization and spectroscopy of single-molecule fluorescence near a surface. This unit presents, in a concise manner, the principle of operation, instrument diversity, and TIRF microscopy applications for the study of biological samples. Copyright 2008 by John Wiley & Sons, Inc.

  18. Digital image inpainting and microscopy imaging. (United States)

    Stanciu, Stefan G; Hristu, Radu; Stanciu, George A


    A considerable amount of image processing techniques known as inpainting techniques have been recently developed aiming to provide solutions for filling in missing or damaged regions in a digital image. Typical such techniques reconstruct a defined area by using information from its neighborhood, for example, by completing inside the missing region the isophote lines arriving at its boundaries. In this article, we show that inpainting techniques have considerable potential usefulness in microscopy imaging, even though experimenting and using them in this domain has been almost entirely neglected up until now. In this purpose, we experiment the "curvature-preserving" partial differential equations as a solution to inpainting regions in images collected by several optical and scanning probe microscopy techniques. The results achieved are presented along with a discussion on typical problematic scenarios of microscopy imaging for which this type of techniques can provide a viable solution. Copyright © 2011 Wiley Periodicals, Inc.

  19. Ultrafast imaging of surface plasmons propagating on a gold surface. (United States)

    Gong, Yu; Joly, Alan G; Hu, Dehong; El-Khoury, Patrick Z; Hess, Wayne P


    We record time-resolved nonlinear photoemission electron microscopy (tr-PEEM) images of propagating surface plasmons (PSPs) launched from a lithographically patterned rectangular trench on a flat gold surface. Our tr-PEEM scheme involves a pair of identical, spatially separated, and interferometrically locked femtosecond laser pulses. Power-dependent PEEM images provide experimental evidence for a sequential coherent nonlinear photoemission process, in which one laser source launches a PSP through a linear interaction, and the second subsequently probes the PSP via two-photon photoemission. The recorded time-resolved movies of a PSP allow us to directly measure various properties of the surface-bound wave packet, including its carrier wavelength (783 nm) and group velocity (0.95c). In addition, tr-PEEM images reveal that the launched PSP may be detected at least 250 μm away from the coupling trench structure.

  20. Applications of microscopy in Salmonella research. (United States)

    Malt, Layla M; Perrett, Charlotte A; Humphrey, Suzanne; Jepson, Mark A


    Salmonella enterica is a Gram-negative enteropathogen that can cause localized infections, typically resulting in gastroenteritis, or systemic infection, e.g., typhoid fever, in humans and many other animals. Understanding the mechanisms by which Salmonella induces disease has been the focus of intensive research. This has revealed that Salmonella invasion requires dynamic cross-talk between the microbe and host cells, in which bacterial adherence rapidly leads to a complex sequence of cellular responses initiated by proteins translocated into the host cell by a type 3 secretion system. Once these Salmonella-induced responses have resulted in bacterial invasion, proteins translocated by a second type 3 secretion system initiate further modulation of cellular activities to enable survival and replication of the invading pathogen. Elucidation of the complex and highly dynamic pathogen-host interactions ultimately requires analysis at the level of single cells and single infection events. To achieve this goal, researchers have applied a diverse range of microscopy techniques to analyze Salmonella infection in models ranging from whole animal to isolated cells and simple eukaryotic organisms. For example, electron microscopy and high-resolution light microscopy techniques such as confocal microscopy can reveal the precise location of Salmonella and its relationship to cellular components. Widefield light microscopy is a simpler approach with which to study the interaction of bacteria with host cells and often has advantages for live cell imaging, enabling detailed analysis of the dynamics of infection and cellular responses. Here we review the use of imaging techniques in Salmonella research and compare the capabilities of different classes of microscope to address specific types of research question. We also provide protocols and notes on some microscopy techniques used routinely in our own research.

  1. Structural examination of lithium niobate ferroelectric crystals by combining scanning electron microscopy and atomic force microscopy (United States)

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


    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

  2. Observation of DNA Molecules Using Fluorescence Microscopy and Atomic Force Microscopy (United States)

    Ito, Takashi


    This article describes experiments for an undergraduate instrumental analysis laboratory that aim to observe individual double-stranded DNA (dsDNA) molecules using fluorescence microscopy and atomic force microscopy (AFM). dsDNA molecules are observed under several different conditions to discuss their chemical and physical properties. In…

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

    NARCIS (Netherlands)

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


    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

  4. Visualization of mobility by atomic force microscopy. (United States)

    Ando, Toshio; Kodera, Noriyuki


    Intrinsically disordered regions (IDRs) of proteins are very thin and hence hard to be visualized by electron microscopy. Thus far, only high-speed atomic force microscopy (HS-AFM) can visualize them. The molecular movies identify the alignment of IDRs and ordered regions in an intrinsically disordered protein (IDP) and show undulation motion of the IDRs. The visualized tail-like structures contain the information of mechanical properties of the IDRs. Here, we describe methods of HS-AFM visualization of IDPs and methods of analyzing the obtained images to characterize IDRs.

  5. Light sheet microscopy in cell biology. (United States)

    Tomer, Raju; Khairy, Khaled; Keller, Philipp J


    Light sheet-based fluorescence microscopy (LSFM) is emerging as a powerful imaging technique for the life sciences. LSFM provides an exceptionally high imaging speed, high signal-to-noise ratio, low level of photo-bleaching, and good optical penetration depth. This unique combination of capabilities makes light sheet-based microscopes highly suitable for live imaging applications. Here, we provide an overview of light sheet-based microscopy assays for in vitro and in vivo imaging of biological samples, including cell extracts, soft gels, and large multicellular organisms. We furthermore describe computational tools for basic image processing and data inspection.

  6. Multimodal CARS microscopy of structured carbohydrate biopolymers (United States)

    Slepkov, Aaron D.; Ridsdale, Andrew; Pegoraro, Adrian F.; Moffatt, Douglas J.; Stolow, Albert


    We demonstrate the utility of multimodal coherent anti-Stokes Raman scattering (CARS) microscopy for the study of structured condensed carbohydrate systems. Simultaneous second-harmonic generation (SHG) and spectrally-scanned CARS microscopy was used to elucidate structure, alignment, and density in cellulose cotton fibers and in starch grains undergoing rapid heat-moisture swelling. Our results suggest that CARS response of the O-H stretch region (3000 cm−1–3400 cm−1), together with the commonly-measured C-H stretch (2750 cm−1–2970 cm−1) and SHG provide potentially important structural information and contrast in these materials. PMID:21258555

  7. IR microscopy utilizing intense supercontinuum light source

    DEFF Research Database (Denmark)

    Dupont, Sune; Petersen, Christian; Thøgersen, Jan


    . The supercontinuum light source has a high brightness and spans the infrared region from 1400 nm to 4000 nm. This combination allows contact free high resolution hyper spectral infrared microscopy. The microscope is demonstrated by imaging an oil/water sample with 20 μm resolution.......Combining the molecular specificity of the infrared spectral region with high resolution microscopy has been pursued by researchers for decades. Here we demonstrate infrared supercontinuum radiated from an optical fiber as a promising new light source for infrared microspectroscopy...

  8. Resonant Scanning with Large Field of View Reduces Photobleaching and Enhances Fluorescence Yield in STED Microscopy. (United States)

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


    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.

  9. Transmission electron microscopy characterization of nanomaterials

    CERN Document Server


    Third volume of a 40volume series on nanoscience and nanotechnology, edited by the renowned scientist Challa S.S.R. Kumar. This handbook gives a comprehensive overview about Transmission electron microscopy characterization of nanomaterials. Modern applications and state-of-the-art techniques are covered and make this volume an essential reading for research scientists in academia and industry.

  10. Fluorescence confocal polarizing microscopy: Three-dimensional ...

    Indian Academy of Sciences (India)

    Three-dimensional imaging of the director. O D LAVRENTOVICH. Chemical Physics ... cholesteric LCs. Keywords. 3D imaging; confocal microscopy; liquid crystals; dislocations. PACS Nos 07.60. ... magnetic resonance, x-ray diffraction, optical phase retardation, etc., suffer from the same deficiency: they produce only an ...

  11. Multiphoton microscopy imaging of developing tooth germs. (United States)

    Pan, Pei-Yu; Chen, Rung-Shu; Ting, Chih-Liang; Chen, Wei-Liang; Dong, Chen-Yuan; Chen, Min-Huey


    Traditionally, tooth germ is observed by histological investigation with hematoxylin and eosin stain and information may loss during the process. The purpose of this study is to use multiphoton laser fluorescence microscopy to observe the developing tooth germs of mice for building up the database of the images of tooth germs and compare with those from conventional histological analysis. Tooth germs were isolated from embryonic and newborn mice with age of Embryonic Day 14.5 and Postnatal Days 1, 3, 5, and 7. Comparison of the images of tooth germ sections in multiphoton microscopy with the images of histology was performed for investigating the molar tooth germs. It was found that various signals arose from different structures of tooth germs. Pre-dentin and dentin have strong second-harmonic generation signals, while ameloblasts and enamel tissues were shown with strong autofluorescence signals. In this study, a novel multiphoton microscopy database of images from developing tooth germs in mice was set up. We confirmed that multiphoton laser microscopy is a powerful tool for investigating the development of tooth germ and is worthy for further application in the study of tooth regeneration. Copyright © 2012. Published by Elsevier B.V.

  12. Reconstruction of Undersampled Atomic Force Microscopy Images

    DEFF Research Database (Denmark)

    Jensen, Tobias Lindstrøm; Arildsen, Thomas; Østergaard, Jan


    Atomic force microscopy (AFM) is one of the most advanced tools for high-resolution imaging and manipulation of nanoscale matter. Unfortunately, standard AFM imaging requires a timescale on the order of seconds to minutes to acquire an image which makes it complicated to observe dynamic processes...

  13. Dark Field Microscopy for Analytical Laboratory Courses (United States)

    Augspurger, Ashley E.; Stender, Anthony S.; Marchuk, Kyle; Greenbowe, Thomas J.; Fang, Ning


    An innovative and inexpensive optical microscopy experiment for a quantitative analysis or an instrumental analysis chemistry course is described. The students have hands-on experience with a dark field microscope and investigate the wavelength dependence of localized surface plasmon resonance in gold and silver nanoparticles. Students also…

  14. Contrast in coherent raman scattering microscopy

    NARCIS (Netherlands)

    Garbacik, E.T.


    Coherent anti-Stokes Raman scattering (CARS) microscopy is becoming a widely used technique for sub-micron, chemically-selective imaging at high rates of speed In this thesis I discuss three methods for increasing the specificity and selectivity of coherent Raman experiments. The first method is the

  15. Cathodoluminescence Microscopy of Nanostructures on Transparent Substrates

    NARCIS (Netherlands)

    Narváez, A.C.


    Cathodoluminescence (CL), the excitation of light by an electron beam, has gained attention as an analysis tool for investigating the optical response of a structure, at a resolution that approaches that in electron microscopy, in the nanometer range. However, the application possibilities are

  16. Cathodoluminescence Microscopy of nanostructures on glass substrates

    NARCIS (Netherlands)

    Narvaez, A.C.; Weppelman, I.G.C.; Moerland, R.J.; Liv, N.; Zonnevylle, A.C.; Kruit, P.; Hoogenboom, J.P.


    Cathodoluminescence (CL) microscopy is an emerging analysis technique in the fields of biology and photonics, where it is used for the characterization of nanometer sized structures. For these applications, the use of transparent substrates might be highly preferred, but the detection of CL from

  17. The 2015 super-resolution microscopy roadmap

    NARCIS (Netherlands)

    Hell, Stefan W.; Sahl, Steffen J.; Bates, Mark; Zhuang, Xiaowei; Heintzmann, Rainer; Booth, Martin J.; Bewersdorf, Joerg; Shtengel, Gleb; Hess, Harald; Tinnefeld, Philip; Honigmann, Alf; Jakobs, Stefan; Testa, Ilaria; Cognet, Laurent; Lounis, Brahim; Ewers, Helge; Davis, Simon J.; Eggeling, Christian; Klenerman, David; Willig, Katrin I.; Vicidomini, Giuseppe; Castello, Marco; Diaspro, Alberto; Cordes, Thorben


    Far-field optical microscopy using focused light is an important tool in a number of scientific disciplines including chemical, (bio) physical and biomedical research, particularly with respect to the study of living cells and organisms. Unfortunately, the applicability of the optical microscope is

  18. Cryo-electron microscopy of vitreous sections. (United States)

    Chlanda, Petr; Sachse, Martin


    More than 30 years ago two groups independently reported the vitrification of pure water, which was until then regarded as impossible without a cryoprotectant [1, 2]. This opened the opportunity to cryo-electron microscopy (cryo-EM) to observe biological samples at nanometer scale, close to their native state. However, poor electron penetration through biological samples sets the limit for sample thickness to less than the average size of the mammalian cell. In order to image bulky specimens at the cell or tissue level in transmission electron microscopy (TEM), a sample has to be either thinned by focused ion beam or mechanically sectioned. The latter technique, Cryo-Electron Microscopy of Vitreous Section (CEMOVIS), employs cryo-ultramicrotomy to produce sections with thicknesses of 40-100 μm of vitreous biological material suitable for cryo-EM. CEMOVIS consists of trimming and sectioning a sample with a diamond knife, placing and attaching the section onto an electron microscopy grid, transferring the grid to the cryo-electron microscope and imaging. All steps must be carried on below devitrification temperature to obtain successful results. In this chapter we provide a step-by-step guide to produce and image vitreous sections of a biological sample.

  19. Atomic Resolution Microscopy of Nitrides in Steel

    DEFF Research Database (Denmark)

    Danielsen, Hilmar Kjartansson


    MN and CrMN type nitride precipitates in 12%Cr steels have been investigated using atomic resolution microscopy. The MN type nitrides were observed to transform into CrMN both by composition and crystallography as Cr diffuses from the matrix into the MN precipitates. Thus a change from one...

  20. Tapping mode atomic force microscopy in liquid

    NARCIS (Netherlands)

    Putman, Constant A.J.; Putman, C.A.J.; van der Werf, Kees; de Grooth, B.G.; van Hulst, N.F.; Greve, Jan


    We show that standard silicon nitride cantilevers can be used for tapping mode atomic force microscopy (AFM) in air, provided that the energy of the oscillating cantilever is sufficiently high to overcome the adhesion of the water layer. The same cantilevers are successfully used for tapping mode

  1. Confocal microscopy imaging of the biofilm matrix

    DEFF Research Database (Denmark)

    Schlafer, Sebastian; Meyer, Rikke L


    The extracellular matrix is an integral part of microbial biofilms and an important field of research. Confocal laser scanning microscopy is a valuable tool for the study of biofilms, and in particular of the biofilm matrix, as it allows real-time visualization of fully hydrated, living specimens...

  2. Towards high-speed scanning tunneling microscopy

    NARCIS (Netherlands)

    Tabak, Femke Chantal


    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. Scanning electron microscopy study of Trichomonas gallinae. (United States)

    Tasca, Tiana; De Carli, Geraldo A


    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.

  4. Light Microscopy Module (LMM)-Emulator (United States)

    Levine, Howard G.; Smith, Trent M.; Richards, Stephanie E.


    The Light Microscopy Module (LMM) is a microscope facility developed at Glenn Research Center (GRC) that provides researchers with powerful imaging capability onboard the International Space Station (ISS). LMM has the ability to have its hardware recongured on-orbit to accommodate a wide variety of investigations, with the capability of remotely acquiring and downloading digital images across multiple levels of magnication.

  5. Fluorescence confocal polarizing microscopy: Three-dimensional ...

    Indian Academy of Sciences (India)

    A PM image bears only two-dimensional (2D) information, integrating the 3D pattern of optical birefringence over the path of light. Recently, we proposed a technique to image 3D director patterns by fluorescence confocal polarizing microscopy (FCPM). The technique employs the property of LC to orient the fluorescent dye ...

  6. Global Seabird Ammonia Emissions (United States)

    Riddick, S. N.; Blackall, T. D.; Dragosits, U.; Daunt, F. H.; Braban, C. F.; Tang, Y. S.; Trathan, P.; Wanless, S.; Sutton, M. A.


    Seabird colonies represent a major source of atmospheric ammonia (NH3) in remote coastal and marine systems in temperate, tropical and polar regions. Previous studies have shown that NH3 emissions from Scottish seabird colonies were substantial - of similar magnitude to the most intensive agricultural point source emissions. The UK data were used to model global seabird NH3 emissions and suggested that penguins are a major source of emissions on and around the Antarctic continent. The largest seabird colonies are in the order of millions of seabirds. Due to the isolation of these colonies from anthropogenic nitrogen sources, they may play a major role in the nitrogen cycle within these ecosystems. A global seabird database was constructed and used in conjunction with a species-specific seabird bioenergetics model to map the locations of NH3 emissions from seabird colonies. The accuracy of the modelled emissions was validated with field data of NH3 emissions measured at key seabird colonies in different climatic regions of the world: temperate (Isle of May, Scotland), tropical (Ascension Island) and polar (Signy Island, South Georgia). The field data indicated good agreement between modelled and measured NH3 emissions. The measured NH3 emissions also showed the variability of emission with climate. Climate dependence of seabird NH3 emissions may have further implications under a changing global climate. Seabird colonies represent NH3 emission ‘hotspots’, often far from anthropogenic sources, and are likely to be the major source of nitrogen input to these remote coastal ecosystems. The direct manuring by seabirds at colony locations may strongly influence species richness and biodiversity. The subsequent volatilisation and deposition of NH3 increases the spatial extent of seabird influence on nitrogen cycling in their local ecosystem. As many seabird populations are fluctuating due to changing food supply, climate change or anthropogenic pressures, these factors

  7. Scanning Probe Microscopy of Organic Solar Cells (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

  8. VOC emissions chambers (United States)

    Federal Laboratory Consortium — In order to support the development of test methods and reference materials for volatile organic compounds (VOC) emissions from building materials and furnishings,...

  9. Ammonia emissions in Europe

    DEFF Research Database (Denmark)

    Jacobsen, Brian H.


    The NEC (National Emission Ceiling) directive has set targets for the 2010 ammonia emissions from a number of European countries. The target will be reached by most EU-countries and the total emission for EU-27 has been reduced by 22% from 1990 to 2007. Denmark is one of the countries...... technology is adopted quicker and that the farm has the right location. It is concluded that the new application process so far has not lived up to the high expectations at the outset. Despite this, the paper concludes that Denmark is likely to reduce emission by 50% from 1990 to 2020 and reach the likely...

  10. Bridging the Emissions Gap


    Blok, K.


    The analyses in Chapters 2 and 3 of this report concluded that the emissions gap in 2020 will likely be between 8 and 13 GtCO2e. The chapters also estimated the difference between BaU emissions in 2020 and the emissions level consistent with a “likely” chance of staying within the 2°C target to be 14 GtCO2e. This chapter explores the potential for bridging this gap using a sector policy approach. Firstly, the chapter provides a summary and update of the estimated emission reduction potential ...

  11. Leica solution: CARS microscopy at video rates (United States)

    Lurquin, V.


    Confocal and multiphoton microscopy are powerful techniques to study morphology and dynamics in cells and tissue, if fluorescent labeling is possible or autofluorescence is strong. For non-fluorescent molecules, Coherent anti-Stokes Raman scattering (CARS) microscopy provides chemical contrast based on intrinsic and highly specific vibrational properties of molecules eliminating the need for labeling. Just as other multiphoton techniques, CARS microscopy possesses three-dimensional sectioning capabilities. Leica Microsystems has combined the CARS imaging technology with its TCS SP5 confocal microscope to provide several advantages for CARS imaging. For CARS microscopy, two picosecond near-infrared lasers are overlapped spatially and temporally and sent into the scanhead of the confocal system. The software allows programmed, automatic switching between these light sources for multi-modal imaging. Furthermore the Leica TCS SP5 can be equipped with a non-descanned detector which will significantly enhance the signal. The Leica TCS SP5 scanhead combines two technologies in one system: a conventional scanner for maximum resolution and a resonant scanner for high time resolution. The fast scanner allows imaging speeds as high as 25 images/per second at a resolution of 512×512 pixel. This corresponds to true video-rate allowing to follow processes at these time-scales as well as the acquisition of three-dimensional stacks in a few seconds. This time resolution is critical to study live animals or human patients for which heart beat and muscle movements lead to a blurring of the image if the acquisition time is high. Furthermore with the resonant scanhead the sectioning is truly confocal and does not suffer from spatial leakage. In summary, CARS microscopy combined with the tandem scanner makes the Leica TCS SP5 a powerful tool for three-dimensional, label-free imaging of chemical and biological samples in vitro and in vivo.

  12. Further observations on cerebellar climbing fibers. A study by means of light microscopy, confocal laser scanning microscopy and scanning and transmission electron microscopy. (United States)

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


    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.

  13. Outsourcing CO2 Emissions (United States)

    Davis, S. J.; Caldeira, K. G.


    CO2 emissions from the burning of fossil fuels are the primary cause of global warming. Much attention has been focused on the CO2 directly emitted by each country, but relatively little attention has been paid to the amount of emissions associated with consumption of goods and services in each country. This consumption-based emissions inventory differs from the production-based inventory because of imports and exports of goods and services that, either directly or indirectly, involved CO2 emissions. Using the latest available data and reasonable assumptions regarding trans-shipment of embodied carbon through third-party countries, we developed a global consumption-based CO2 emissions inventory and have calculated associated consumption-based energy and carbon intensities. We find that, in 2004, 24% of CO2 emissions are effectively outsourced to other countries, with much of the developed world outsourcing CO2 emissions to emerging markets, principally China. Some wealthy countries, including Switzerland and Sweden, outsource over half of their consumption-based emissions, with many northern Europeans outsourcing more than three tons of emissions per person per year. The United States is both a big importer and exporter of emissions embodied in trade, outsourcing >2.6 tons of CO2 per person and at the same time as >2.0 tons of CO2 per person are outsourced to the United States. These large flows indicate that CO2 emissions embodied in trade must be taken into consideration when considering responsibility for increasing atmospheric greenhouse gas concentrations.

  14. Air Emission Inventory for the INEEL -- 1999 Emission Report

    Energy Technology Data Exchange (ETDEWEB)

    Zohner, Steven K


    This report presents the 1999 calendar year update of the Air Emission Inventory for the Idaho National Engineering and Environmental Laboratory (INEEL). The INEEL Air Emission Inventory documents sources and emissions of nonradionuclide pollutants from operations at the INEEL. The report describes the emission inventory process and all of the sources at the INEEL, and provides nonradionuclide emissions estimates for stationary sources.

  15. Correlative atomic force microscopy and localization-based super-resolution microscopy: revealing labelling and image reconstruction artefacts. (United States)

    Monserrate, Aitor; Casado, Santiago; Flors, Cristina


    Hybrid microscopy: A correlative microscopy tool that combines in situ super-resolution fluorescence microscopy based on single-molecule localization and atomic force microscopy is presented. Direct comparison with high- resolution topography allows the authors to improve fluorescence labeling and image analysis in super-resolution imaging. © 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  16. 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:; 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)


    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.

  17. Study of Thermal-Field Emission Properties and Investigation of Temperature dependent Noise in the Emission Current form vertical Carbon nanotube emitters

    KAUST Repository

    Kolekar, Sadhu


    We have investigated temperature dependent field electron emission characteristics of vertical carbon nanotubes (CNTs). The generalized expression for electron emission from well defined cathode surface is given by Millikan and Lauritsen [1] for the combination of temperature and electric field effect. The same expression has been used to explain the electron emission characteristics from vertical CNT emitters. Furthermore, this has been applied to explain the electron emission for different temperatures ranging from room temperature to 1500 K. The real-time field electron emission images at room temperature and 1500 K are recorded by using Charge Coupled Device (CCD), in order to understand the effect of temperature on electron emission spots in image morphology (as indicated by ring like structures) and electron emission spot intensity of the emitters. Moreover, the field electron emission images can be used to calculate the total number of emitters per cm2 for electron emission. The calculated number of emitters per cm2 is 4.5x107 and, the actual number emitters per cm2 present for electron emission calculated from Atomic Force Microscopy (AFM) data is 1.2x1012. The measured Current-Voltage (I-V) characteristics obey the Folwer-Nordheim (F-N) type behavior. The fluctuations in the emission current are recorded at different temperatures and, temperature dependence of power spectral density obeys power law relation s(f)=I2/f2 with that of emission current and frequency.

  18. Database of emission lines (United States)

    Binette, L.; Ortiz, P.; Joguet, B.; Rola, C.


    A widely accessible data bank (available through Netscape) and consiting of all (or most) of the emission lines reported in the litterature is being built. It will comprise objects as diverse as HII regions, PN, AGN, HHO. One of its use will be to define/refine existing diagnostic emission line diagrams.

  19. Uncertainties in emission inventories

    NARCIS (Netherlands)

    Aardenne, van J.A.


    Emission inventories provide information about the amount of a pollutant that is emitted to the atmosphere as a result of a specific anthropogenic or natural process at a given time or place. Emission inventories can be used for either policy or scientific purposes. For

  20. Diesel Emissions Quantifier (DEQ) (United States)

    .The Diesel Emissions Quantifier (Quantifier) is an interactive tool to estimate emission reductions and cost effectiveness. Publications EPA-420-F-13-008a (420f13008a), EPA-420-B-10-035 (420b10023), EPA-420-B-10-034 (420b10034)

  1. Controlling spontaneous emission

    DEFF Research Database (Denmark)

    Lodahl, Peter

    Control over spontaneous emission of light is of great importance in quantum optics. It is essential for diverse applications such as miniature lasers, light-emitting diodes, and single-photon sources for quantum information. We present experimental studies on spontaneous emission of CdSe quantum...

  2. Photo- and thermionic emission of MWPECVD nanocrystalline diamond films

    Energy Technology Data Exchange (ETDEWEB)

    Cicala, G., E-mail: [CNR-IMIP, Via Amendola 122/D, 70126 Bari (Italy); Magaletti, V. [ALTA S.p.A., Via Gherardesca 5, 56121 Ospedaletto, Pisa (Italy); Valentini, A.; Nitti, M.A. [Department of Physics, University of Bari “A. Moro”, Via Orabona 4, 70126 Bari (Italy); Bellucci, A.; Trucchi, D.M. [CNR-ISM UOS Montelibretti, Via Salaria km 29.300, 00015 Monterotondo Scalo, RM (Italy)


    Highlights: • Photo- and thermionic emission of MWPECVD NCD films was compared. • The amount of incorporated hydrogen in the film was tuned by changing the grain size. • Higher emission was found in NCD film with BL grown at higher deposition temperature. - Abstract: Nanocrystalline diamond (NCD) films with and without a diamond buffer layer (BL) have been grown on p-type silicon substrates by microwave plasma enhanced chemical vapor deposition technique at different values of deposition temperature (652–884 °C). The photo- and thermionic electron emission properties of NCD films have been investigated, illustrated and explained by analyzing the surface morphology and the grain shape determined by atomic force microscopy, the chemical-structural properties by Raman spectroscopy and nanocrystallites size by X-ray diffraction. The NCD films with BL grown at the highest deposition temperature have shown the highest photo- and thermionic emission currents.

  3. Concepts in Imaging and Microscopy: Exploring Biological Structure and Function with Confocal Microscopy

    National Research Council Canada - National Science Library

    Michael Dailey; Glen Marrs; Jakob Satz; Marc Waite


    .... The utility of confocal microscopy relies on its fundamental capacity to reject out-of-focus light, thus providing sharp, high-contrast images of cells and subcellular structures within thick samples...

  4. Observed Barium Emission Rates (United States)

    Stenbaek-Nielsen, H. C.; Wescott, E. M.; Hallinan, T. J.


    The barium releases from the CRRES satellite have provided an opportunity for verifying theoretically calculated barium ion and neutral emission rates. Spectra of the five Caribbean releases in the summer of 1991 were taken with a spectrograph on board a U.S. Air Force jet aircraft. Because the line of sight release densities are not known, only relative rates could be obtained. The observed relative rates agree well with the theoretically calculated rates and, together with other observations, confirm the earlier detailed theoretical emission rates. The calculated emission rates can thus with good accuracy be used with photometric observations. It has been postulated that charge exchange between neutral barium and oxygen ions represents a significant source for ionization. If so. it should be associated with emissions at 4957.15 A and 5013.00 A, but these emissions were not detected.

  5. High-speed atomic force microscopy combined with inverted optical microscopy for studying cellular events


    Suzuki, Yuki; Sakai, Nobuaki; Yoshida, Aiko; Uekusa, Yoshitsugu; Yagi, Akira; Imaoka, Yuka; Ito, Shuichi; Karaki, Koichi; Takeyasu, Kunio


    A hybrid atomic force microscopy (AFM)-optical fluorescence microscopy is a powerful tool for investigating cellular morphologies and events. However, the slow data acquisition rates of the conventional AFM unit of the hybrid system limit the visualization of structural changes during cellular events. Therefore, high-speed AFM units equipped with an optical/fluorescence detection device have been a long-standing wish. Here we describe the implementation of high-speed AFM coupled with an optic...

  6. Multiscale photoacoustic microscopy and computed tomography (United States)

    Wang, Lihong V.


    Photoacoustic tomography (PAT) is probably the fastest growing biomedical imaging technology owing to its capability of high-resolution sensing of rich optical contrast in vivo at depths beyond the optical transport mean free path (~1 mm in the skin). Existing high-resolution optical imaging technologies, such as confocal microscopy and two-photon microscopy, have fundamentally impacted biomedicine but cannot reach such depths. Taking advantage of low ultrasonic scattering, PAT indirectly improves tissue transparency by 100 to 1000 fold and consequently enables deeply penetrating functional and molecular imaging at high spatial resolution. Further, PAT holds the promise of in vivo imaging at multiple length scales ranging from subcellular organelles to organs with the same contrast origin, an important application in multiscale systems biology research. PMID:20161535

  7. Fundamentals of fluorescence and fluorescence microscopy. (United States)

    Wolf, David E


    This chapter discusses the fundamental physics of fluorescence. The application of fluorescence to microscopy represents an important transition in the development of microscopy, particularly as it applies to biology. It enables quantitating the amounts of specific molecules within a cell, determining whether molecules are complexing on a molecular level, measuring changes in ionic concentrations within cells and organelles, and measuring molecular dynamics. This chapter also discusses the issues important to quantitative measurement of fluorescence and focuses on four of quantitative measurements of fluorescence--boxcar-gated detection, streak cameras, photon correlation, and phase modulation. Although quantitative measurement presents many pitfalls to the beginner, it also presents significant opportunities to one skilled in the art. This chapter also examines how fluorescence is measured in the steady state and time domain and how fluorescence is applied in the modern epifluorescence microscope. Copyright © 2007 Elsevier Inc. All rights reserved.

  8. Hollow-tip scanning photoelectron microscopy (United States)

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


    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).

  9. Reflectance Confocal Microscopy in Lentigo Maligna. (United States)

    Gamo, R; Pampín, A; Floristán, U


    Lentigo maligna is the most common type of facial melanoma. Diagnosis is complicated, however, as it shares clinical and dermoscopic characteristics with other cutaneous lesions of the face. Reflectance confocal microscopy is an imaging technique that permits the visualization of characteristic features of lentigo maligna. These include a disrupted honeycomb pattern and pagetoid cells with a tendency to show folliculotropism. These cells typically have a dendritic morphology, although they may also appear as round cells measuring over 20μm with atypical nuclei. Poorly defined dermal papillae and atypical cells may be seen at the dermal-epidermal junction and can form bridges resembling mitochondrial structures. Other characteristic findings include junctional swelling with atypical cells located around the follicles, resembling caput medusae. Reflectance confocal microscopy is a very useful tool for diagnosing lentigo maligna. Copyright © 2016 AEDV. Publicado por Elsevier España, S.L.U. All rights reserved.

  10. Diffractive elements performance in chromatic confocal microscopy

    Energy Technology Data Exchange (ETDEWEB)

    Garzon, J; Duque, D; Alean, A; Toledo, M [Grupo de Optica y EspectroscopIa, Centro de Ciencia Basica, Universidad Pontificia Bolivariana. Medellin (Colombia); Meneses, J [Laboratorio de Optica y Tratamiento de Senales, Instituto de Fisica, Universidad Industrial de Santander, Bucaramanga (Colombia); Gharbi, T, E-mail: [Laboratoire d' Optique P. M. Duffieux, UMR-6603 CNR/Universite de Franche-Comte. 16 route de Gray, 25030 Besancon Cedex (France)


    The Confocal Laser Scanning Microscopy (CLSM) has been widely used in the semiconductor industry and biomedicine because of its depth discrimination capability. Subsequent to this technique has been developed in recent years Chromatic Confocal Microscopy. This method retains the same principle of confocal and offers the added advantage of removing the axial movement of the moving system. This advantage is usually accomplished with an optical element that generates a longitudinal chromatic aberration and a coding system that relates the axial position of each point of the sample with the wavelength that is focused on each. The present paper shows the performance of compact chromatic confocal microscope when some different diffractive elements are used for generation of longitudinal chromatic aberration. Diffractive elements, according to the process and manufacturing parameters, may have different diffraction efficiency and focus a specific wavelength in a specific focal position. The performance assessment is carried out with various light sources which exhibit an incoherent behaviour and a broad spectral width.

  11. Pump-probe optical coherence microscopy (United States)

    Wan, Qiujie; Applegate, Brian E.


    High-resolution optical molecular imaging has become a vital tool for understanding and measuring physiologically important biometrics on the cellular and subcellular level. In spite of significant recent advances in microscopy, molecular imaging of most endogenous biomolecular species remains elusive. Directly imaging endogenous biomolecules without the aid of exogenous tags is highly desirable. We developed pump-probe optical coherence microscopy (PPOCM) based on our previous success in integrating pump-probe absorption spectroscopy with optical coherence tomography. A fixed human skin tissue with melanoma was imaged by the PPOCM system. The preliminary results show that PPOCM can provide better can clear contrast between normal tissue and melanoma than OCM. This system also can be used to image other chromophores.

  12. Light sheet fluorescence microscopy: a review. (United States)

    Santi, Peter A


    Light sheet fluorescence microscopy (LSFM) functions as a non-destructive microtome and microscope that uses a plane of light to optically section and view tissues with subcellular resolution. This method is well suited for imaging deep within transparent tissues or within whole organisms, and because tissues are exposed to only a thin plane of light, specimen photobleaching and phototoxicity are minimized compared to wide-field fluorescence, confocal, or multiphoton microscopy. LSFMs produce well-registered serial sections that are suitable for three-dimensional reconstruction of tissue structures. Because of a lack of a commercial LSFM microscope, numerous versions of light sheet microscopes have been constructed by different investigators. This review describes development of the technology, reviews existing devices, provides details of one LSFM device, and shows examples of images and three-dimensional reconstructions of tissues that were produced by LSFM.

  13. Nuclear microscopy of sperm cell elemental structure

    Energy Technology Data Exchange (ETDEWEB)

    Bench, G.S.; Balhorn, R.; Friz, A.M.; Freeman, S.P.H.T.


    Theories suggest there is a link between protamine concentrations in individual sperm and male fertility. Previously, biochemical analyses have used pooled samples containing millions of sperm to determine protamine concentrations. These methods have not been able to determine what percentage of morphologically normal sperm are biochemically defective and potentially infertile. Nuclear microscopy has been utilized to measure elemental profiles at the single sperm level. By measuring the amount of phosphorus and sulfur, the total DNA and protamine content in individual sperm from fertile bull and mouse semen have been determined. These values agree with results obtained from other biochemical analyses. Nuclear microscopy shows promise for measuring elemental profiles in the chromatin of individual sperm. The technique may be able to resolve theories regarding the importance of protamines to male fertility and identify biochemical defects responsible for certain types of male infertility.

  14. Nanostructural analysis by atomic force microscopy followed by light microscopy on the same archival slide. (United States)

    Wagner, Mathias; Kaehler, Dirk; Anhenn, Olaf; Betz, Thomas; Awad, Sally; Shamaa, Ali; Theegarten, Dirk; Linder, Roland


    Integrated information on ultrastructural surface texture and chemistry increasingly plays a role in the biomedical sciences. Light microscopy provides access to biochemical data by the application of dyes. Ultrastructural representation of the surface structure of tissues, cells, or macromolecules can be obtained by scanning electron microscopy (SEM). However, SEM often requires gold or coal coating of biological samples, which makes a combined examination by light microscopy and SEM difficult. Conventional histochemical staining methods are not easily applicable to biological material subsequent to such treatment. Atomic force microscopy (AFM) gives access to surface textures down to ultrastructural dimensions without previous coating of the sample. A combination of AFM with conventional histochemical staining protocols for light microscopy on a single slide is therefore presented. Unstained cores were examined using AFM (tapping mode) and subsequently stained histochemically. The images obtained by AFM were compared with the results of histochemistry. AFM technology did not interfere with any of the histochemical staining protocols. Ultrastructurally analyzed regions could be identified in light microscopy and histochemical properties of ultrastructurally determined regions could be seen. AFM-generated ultrastructural information with subsequent staining gives way to novel findings in the biomedical sciences. (c) 2009 Wiley-Liss, Inc.

  15. Introduction to Conventional Transmission Electron Microscopy (United States)

    de Graef, Marc


    This book covers the fundamentals of conventional transmission electron microscopy (CTEM) as applied to crystalline solids. In addition to including a large selection of worked examples and homework problems, the volume is accompanied by a supplementary website ( containing interactive modules and over 30,000 lines of free Fortran 90 source code. The work is based on a lecture course given by Marc De Graef in the Department of Materials Science and Engineering at Carnegie Mellon University.

  16. Confocal reference free traction force microscopy


    Bergert, Martin; Lendenmann, Tobias; Z?ndel, Manuel; Ehret, Alexander E.; Panozzo, Daniele; Richner, Patrizia; Kim, David K.; Kress, Stephan J. P.; Norris, David J.; Sorkine-Hornung, Olga; Mazza, Edoardo; Poulikakos, Dimos; Ferrari, Aldo


    The mechanical wiring between cells and their surroundings is fundamental to the regulation of complex biological processes during tissue development, repair or pathology. Traction force microscopy (TFM) enables determination of the actuating forces. Despite progress, important limitations with intrusion effects in low resolution 2D pillar-based methods or disruptive intermediate steps of cell removal and substrate relaxation in high-resolution continuum TFM methods need to be overcome. Here ...

  17. Photoacoustic microscopy of blood pulse wave


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


    Blood pulse wave velocity (PWV) is an important physiological parameter that characterizes vascular stiffness. In this letter, we present electrocardiogram-synchronized, photoacoustic microscopy for noninvasive quantification of the PWV in the peripheral vessels of living mice. Interestingly, blood pulse wave-induced fluctuations in blood flow speed were clearly observed in arteries and arterioles, but not in veins or venules. Simultaneously recorded electrocardiograms served as references to...

  18. Biological cryo‐electron microscopy in China (United States)


    Abstract Cryo‐electron microscopy (cryo‐EM) plays an increasingly more important role in structural biology. With the construction of an arm of the Chinese National Protein Science Facility at Tsinghua University, biological cryo‐EM has entered a phase of rapid development in China. This article briefly reviews the history of biological cryo‐EM in China, describes its current status, comments on its impact on the various biological research fields, and presents future outlook. PMID:27534377

  19. Biological cryo?electron microscopy in China


    Wang, Hong Wei; Lei, Jianlin; Shi, Yigong


    Abstract Cryo?electron microscopy (cryo?EM) plays an increasingly more important role in structural biology. With the construction of an arm of the Chinese National Protein Science Facility at Tsinghua University, biological cryo?EM has entered a phase of rapid development in China. This article briefly reviews the history of biological cryo?EM in China, describes its current status, comments on its impact on the various biological research fields, and presents future outlook.

  20. Atomic Force Microscopy of Biological Membranes


    Frederix, Patrick L.T.M.; Bosshart, Patrick D.; Engel, Andreas


    Atomic force microscopy (AFM) is an ideal method to study the surface topography of biological membranes. It allows membranes that are adsorbed to flat solid supports to be raster-scanned in physiological solutions with an atomically sharp tip. Therefore, AFM is capable of observing biological molecular machines at work. In addition, the tip can be tethered to the end of a single membrane protein, and forces acting on the tip upon its retraction indicate barriers that occur during the process...

  1. Scanning electron microscopy of superficial white onychomycosis* (United States)

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


    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

  2. Scanning electron microscopy of molluscum contagiosum*


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


    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...

  3. High-energy electron diffraction and microscopy

    CERN Document Server

    Peng, L M; Whelan, M J


    This book provides a comprehensive introduction to high energy electron diffraction and elastic and inelastic scattering of high energy electrons, with particular emphasis on applications to modern electron microscopy. Starting from a survey of fundamental phenomena, the authors introduce the most important concepts underlying modern understanding of high energy electron diffraction. Dynamical diffraction in transmission (THEED) and reflection (RHEED) geometries is treated using ageneral matrix theory, where computer programs and worked examples are provided to illustrate the concepts and to f

  4. Scanning electron microscopy of cold gases (United States)

    Santra, Bodhaditya; Ott, Herwig


    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.

  5. Laser-based dual-space microscopy (United States)

    Alotaibi, Maged; Gedies, Robert; Alzayed, Abdullah; Aldawsari, Fahad; Dominguez, Daniel; Grave de Peralta, Luis


    We show using two-dimensional simulations that the dual-space microscopy (DSM) phase-recovery algorithm converges to the correct result for arbitrary samples. We present experimental results obtained by implementing the DSM technique using a laser. We demonstrate that DSM produces synthetic images with a large field of view when a laser is used as the illumination source. However, speckles affect the quality of the obtained images.

  6. Quantitative pupil analysis in stimulated emission depletion microscopy using phase retrieval

    DEFF Research Database (Denmark)

    Kromann, Emil B; Gould, Travis J; Juette, Manuel F


    no instrument modifications, for obtaining an equivalent to the complex pupil function at the back aperture of the objective and show that it provides quantitative information about aberration sources (including aberrations induced by the objective or sample). We show the accuracy of this field representation...

  7. Proton induced X-ray emission and electron microscopy analysis of induced mutants of sorghum

    CSIR Research Space (South Africa)

    Mbambo, Z


    Full Text Available referred to as trace elements (Ager et al., 2003). In higher concentrations some trace elements (for example iron and copper) are toxic to cells. Many organisms therefore have evolved mechanisms to regulate the uptake of trace elements, their excretion... and intracellular translocation and localisation within cells. These regulatory mechanisms are intended to assist in balancing out the need for protection against toxic reactivity of some elements in cells, as well as the need to ensure adequate availability when...

  8. Non-descanned multifocal multiphoton microscopy with a multianode photomultiplier tube

    Directory of Open Access Journals (Sweden)

    Jae Won Cha


    Full Text Available Multifocal multiphoton microscopy (MMM improves imaging speed over a point scanning approach by parallelizing the excitation process. Early versions of MMM relied on imaging detectors to record emission signals from multiple foci simultaneously. For many turbid biological specimens, the scattering of emission photons results in blurred images and degrades the signal-to-noise ratio (SNR. We have recently demonstrated that a multianode photomultiplier tube (MAPMT placed in a descanned configuration can effectively collect scattered emission photons from each focus into their corresponding anodes significantly improving image SNR for highly scattering specimens. Unfortunately, a descanned MMM has a longer detection path resulting in substantial emission photon loss. Optical design constraints in a descanned geometry further results in significant optical aberrations especially for large field-of-view (FOV, high NA objectives. Here, we introduce a non-descanned MMM based on MAPMT that substantially overcomes most of these drawbacks. We show that we improve signal efficiency up to fourfold with limited image SNR degradation due to scattered emission photons. The excitation foci can also be spaced wider to cover the full FOV of the objective with minimal aberrations. The performance of this system is demonstrated by imaging interneuron morphological structures deep in the brains of living mice.

  9. Non-descanned multifocal multiphoton microscopy with a multianode photomultiplier tube

    Energy Technology Data Exchange (ETDEWEB)

    Cha, Jae Won; Yew, Elijah Y. S. [Department of Mechanical Engineering, Massachusetts Institute of Technology, Cambridge, MA (United States); Kim, Daekeun [Department of Mechanical Engineering, Dankook University (Korea, Republic of); Subramanian, Jaichandar [Picower Institute for Learning and Memory, Massachusetts Institute of Technology, Cambridge, MA (United States); Nedivi, Elly [Picower Institute for Learning and Memory, Massachusetts Institute of Technology, Cambridge, MA (United States); Departments of Brain and Cognitive Sciences, and Biology, Massachusetts Institute of Technology, Cambridge, MA (United States); So, Peter T. C. [Department of Mechanical Engineering, Massachusetts Institute of Technology, Cambridge, MA (United States); Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, MA (United States)


    Multifocal multiphoton microscopy (MMM) improves imaging speed over a point scanning approach by parallelizing the excitation process. Early versions of MMM relied on imaging detectors to record emission signals from multiple foci simultaneously. For many turbid biological specimens, the scattering of emission photons results in blurred images and degrades the signal-to-noise ratio (SNR). We have recently demonstrated that a multianode photomultiplier tube (MAPMT) placed in a descanned configuration can effectively collect scattered emission photons from each focus into their corresponding anodes significantly improving image SNR for highly scattering specimens. Unfortunately, a descanned MMM has a longer detection path resulting in substantial emission photon loss. Optical design constraints in a descanned geometry further results in significant optical aberrations especially for large field-of-view (FOV), high NA objectives. Here, we introduce a non-descanned MMM based on MAPMT that substantially overcomes most of these drawbacks. We show that we improve signal efficiency up to fourfold with limited image SNR degradation due to scattered emission photons. The excitation foci can also be spaced wider to cover the full FOV of the objective with minimal aberrations. The performance of this system is demonstrated by imaging interneuron morphological structures deep in the brains of living mice.

  10. Extended full-field optical coherence microscopy (United States)

    Dubois, Arnaud


    Full-field optical coherence microscopy (FF-OCM) is a recent optical technology based on low-coherence interference microscopy for semi-transparent sample imaging with ˜ 1 μm spatial resolution. FF-OCM has been successfully applied to three-dimensional imaging of various biological tissues at cellular-level resolution. The contrast of FF-OCM images results from the intensity of light backscattered by the sample microstructures. This contrast mechanism, based on refractive index changes, provides information on the internal architectural morphology of the sample. In this paper, we present a multimodal FF-OCM system, capable of measuring simultaneously the intensity, the power spectrum and the phase-retardation of light backscattered by the sample being imaged. Tomographic fluorescence-based images can also be produced by coupling to the FF-OCM set-up a fluorescence microscopy system with structured illumination. Fluorescence targeted probes can be used to identify molecular components of subcellular scattering structures. Compared to conventional FF-OCM, this multimodal system provides enhanced imaging contrasts at the price of a moderate increase in experimental complexity and cost.

  11. Differential dynamic microscopy of bidisperse colloidal suspensions. (United States)

    Safari, Mohammad S; Poling-Skutvik, Ryan; Vekilov, Peter G; Conrad, Jacinta C


    Research tasks in microgravity include monitoring the dynamics of constituents of varying size and mobility in processes such as aggregation, phase separation, or self-assembly. We use differential dynamic microscopy, a method readily implemented with equipment available on the International Space Station, to simultaneously resolve the dynamics of particles of radius 50 nm and 1 μm in bidisperse aqueous suspensions. Whereas traditional dynamic light scattering fails to detect a signal from the larger particles at low concentrations, differential dynamic microscopy exhibits enhanced sensitivity in these conditions by accessing smaller wavevectors where scattering from the large particles is stronger. Interference patterns due to scattering from the large particles induce non-monotonic decay of the amplitude of the dynamic correlation function with the wavevector. We show that the position of the resulting minimum contains information on the vertical position of the particles. Together with the simple instrumental requirements, the enhanced sensitivity of differential dynamic microscopy makes it an appealing alternative to dynamic light scattering to characterize samples with complex dynamics.

  12. Invited Review Article: Pump-probe microscopy (United States)

    Fischer, Martin C.; Wilson, Jesse W.; Robles, Francisco E.; Warren, Warren S.


    Multiphoton microscopy has rapidly gained popularity in biomedical imaging and materials science because of its ability to provide three-dimensional images at high spatial and temporal resolution even in optically scattering environments. Currently the majority of commercial and home-built devices are based on two-photon fluorescence and harmonic generation contrast. These two contrast mechanisms are relatively easy to measure but can access only a limited range of endogenous targets. Recent developments in fast laser pulse generation, pulse shaping, and detection technology have made accessible a wide range of optical contrasts that utilize multiple pulses of different colors. Molecular excitation with multiple pulses offers a large number of adjustable parameters. For example, in two-pulse pump-probe microscopy, one can vary the wavelength of each excitation pulse, the detection wavelength, the timing between the excitation pulses, and the detection gating window after excitation. Such a large parameter space can provide much greater molecular specificity than existing single-color techniques and allow for structural and functional imaging without the need for exogenous dyes and labels, which might interfere with the system under study. In this review, we provide a tutorial overview, covering principles of pump-probe microscopy and experimental setup, challenges associated with signal detection and data processing, and an overview of applications.

  13. Invited Review Article: Pump-probe microscopy (United States)

    Wilson, Jesse W.; Robles, Francisco E.; Warren, Warren S.


    Multiphoton microscopy has rapidly gained popularity in biomedical imaging and materials science because of its ability to provide three-dimensional images at high spatial and temporal resolution even in optically scattering environments. Currently the majority of commercial and home-built devices are based on two-photon fluorescence and harmonic generation contrast. These two contrast mechanisms are relatively easy to measure but can access only a limited range of endogenous targets. Recent developments in fast laser pulse generation, pulse shaping, and detection technology have made accessible a wide range of optical contrasts that utilize multiple pulses of different colors. Molecular excitation with multiple pulses offers a large number of adjustable parameters. For example, in two-pulse pump-probe microscopy, one can vary the wavelength of each excitation pulse, the detection wavelength, the timing between the excitation pulses, and the detection gating window after excitation. Such a large parameter space can provide much greater molecular specificity than existing single-color techniques and allow for structural and functional imaging without the need for exogenous dyes and labels, which might interfere with the system under study. In this review, we provide a tutorial overview, covering principles of pump-probe microscopy and experimental setup, challenges associated with signal detection and data processing, and an overview of applications. PMID:27036751

  14. Phase-contrast scanning transmission electron microscopy. (United States)

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


    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:

  15. Gradient field microscopy of unstained specimens (United States)

    Kim, Taewoo; Sridharan, Shamira; Popescu, Gabriel


    We present a phase derivative microscopy technique referred to as gradient field microscopy (GFM), which provides the first-order derivatives of the phase associated with an optical field passing through a transparent specimen. GFM utilizes spatial light modulation at the Fourier plane of a bright field microscope to optically obtain the derivatives of the phase and increase the contrast of the final image. The controllable spatial modulation pattern allows us to obtain both one component of the field gradient (derivative along one direction) and the gradient intensity, which offers some advantages over the regular differential interference contrast (DIC) microscopy. Most importantly, unlike DIC, GFM does not use polarizing optics and, thus, it is applicable to birefringent samples. We demonstrate these features of GFM with studies of static and dynamic biological cells (HeLa cells and red blood cells). We show that GFM is capable of qualitatively providing information about cell membrane fluctuations. Specifically, we captured the disappearance of the bending mode of fluctuations in osmotically swollen red blood cells. PMID:22418558

  16. Fluctuation microscopy analysis of amorphous silicon models

    Energy Technology Data Exchange (ETDEWEB)

    Gibson, J.M., E-mail: [Northeastern University, Department of Physics, Boston MA 02115 (United States); FAMU/FSU Joint College of Engineering, 225 Pottsdamer Street, Tallahassee, FL 32310 (United States); Treacy, M.M.J. [Arizona State University, Department of Physics, Tempe AZ 85287 (United States)


    Highlights: • Studied competing computer models for amorphous silicon and simulated fluctuation microscopy data. • Show that only paracrystalline/random network composite can fit published data. • Specifically show that pure random network or random network with void models do not fit available data. • Identify a new means to measure volume fraction of ordered material. • Identify unreported limitations of the Debye model for simulating fluctuation microscopy data. - Abstract: Using computer-generated models we discuss the use of fluctuation electron microscopy (FEM) to identify the structure of amorphous silicon. We show that a combination of variable resolution FEM to measure the correlation length, with correlograph analysis to obtain the structural motif, can pin down structural correlations. We introduce the method of correlograph variance as a promising means of independently measuring the volume fraction of a paracrystalline composite. From comparisons with published data, we affirm that only a composite material of paracrystalline and continuous random network that is substantially paracrystalline could explain the existing experimental data, and point the way to more precise measurements on amorphous semiconductors. The results are of general interest for other classes of disordered materials.

  17. Scanning Ion Conductance Microscopy of Live Keratinocytes (United States)

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


    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.

  18. Clinical applications of corneal confocal microscopy

    Directory of Open Access Journals (Sweden)

    Mitra Tavakoli


    Full Text Available Mitra Tavakoli1, Parwez Hossain2, Rayaz A Malik11Division of Cardiovascular Medicine, University of Manchester and Manchester Royal Infirmary, Manchester, UK; 2University of Southampton, Southampton Eye Unit, Southampton General Hospital, Southampton, UKAbstract: Corneal confocal microscopy is a novel clinical technique for the study of corneal cellular structure. It provides images which are comparable to in-vitro histochemical techniques delineating corneal epithelium, Bowman’s layer, stroma, Descemet’s membrane and the corneal endothelium. Because, corneal confocal microscopy is a non invasive technique for in vivo imaging of the living cornea it has huge clinical potential to investigate numerous corneal diseases. Thus far it has been used in the detection and management of pathologic and infectious conditions, corneal dystrophies and ecstasies, monitoring contact lens induced corneal changes and for pre and post surgical evaluation (PRK, LASIK and LASEK, flap evaluations and Radial Keratotomy, and penetrating keratoplasty. Most recently it has been used as a surrogate for peripheral nerve damage in a variety of peripheral neuropathies and may have potential in acting as a surrogate marker for endothelial abnormalities.Keywords: corneal confocal microscopy, cornea, infective keratitis, corneal dystrophy, neuropathy

  19. Axial tomography in live cell laser microscopy (United States)

    Richter, Verena; Bruns, Sarah; Bruns, Thomas; Weber, Petra; Wagner, Michael; Cremer, Christoph; Schneckenburger, Herbert


    Single cell microscopy in a three-dimensional (3-D) environment is reported. Cells are grown in an agarose culture gel, located within microcapillaries and observed from different sides after adaptation of an innovative device for sample rotation. Thus, z-stacks can be recorded by confocal microscopy in different directions and used for illustration in 3-D. This gives additional information, since cells or organelles that appear superimposed in one direction, may be well resolved in another one. The method is tested and validated with single cells expressing a membrane or a mitochondrially associated green fluorescent protein, or cells accumulating fluorescent quantum dots. In addition, axial tomography supports measurements of cellular uptake and distribution of the anticancer drug doxorubicin in the nucleus (2 to 6 h after incubation) or the cytoplasm (24 h). This paper discusses that upon cell rotation an enhanced optical resolution in lateral direction compared to axial direction can be utilized to obtain an improved effective 3-D resolution, which represents an important step toward super-resolution microscopy of living cells.

  20. Assessment of the contribution of electron microscopy to nanoparticle characterization sampled with two cascade impactors. (United States)

    Noël, Alexandra; L'Espérance, Gilles; Cloutier, Yves; Plamondon, Philippe; Boucher, Julie; Philippe, Suzanne; Dion, Chantal; Truchon, Ginette; Zayed, Joseph


    This study assessed the contribution of electron microscopy to the characterization of nanoparticles and compared the degree of variability in sizes observed within each stage when sampled by two cascade impactors: an Electrical Low Pressure Impactor (ELPI) and a Micro-Orifice Uniform Deposit Impactor (MOUDI). A TiO(2) nanoparticle (5 nm) suspension was aerosolized in an inhalation chamber. Nanoparticles sampled by the impactors were collected on aluminum substrates or TEM carbon-coated copper grids using templates, specifically designed in our laboratories, for scanning and transmission electron microscopy (SEM, TEM) analysis, respectively. Nanoparticles were characterized using both SEM and TEM. Three different types of diameters (inner, outer, and circular) were measured by image analysis based on count and volume, for each impactor stage. Electron microscopy, especially TEM, is well suited for the characterization of nanoparticles. The MOUDI, probably because of the rotation of its collection stages, which can minimize the resuspension of particles, gave more stable results and smaller geometric standard deviations per stage. Our data suggest that the best approach to estimate particle size by electron microscopy would rely on geometric means of measured circular diameters. Overall, the most reliable data were provided by the MOUDI and the TEM sampling technique on carbon-coated copper grids for this specific experiment. This study indicates interesting findings related to the assessment of impactors combined with electron microscopy for nanoparticle characterization. For future research, since cascade impactors are extensively used to characterize nano-aerosol exposure scenarios, high-performance field emission scanning electron microscopy (FESEM) should also be considered.

  1. Resolution of multiple GFP color variants and dyes using two-photon microscopy and imaging spectroscopy (United States)

    Lansford, Rusty; Bearman, Gregory H.; Fraser, Scott E.


    The imaging of living cells and tissues using laser-scanning microscopy is offering dramatic insights into the spatial and temporal controls of biological processes. The availability of genetically encoded labels such as green fluorescent protein (GFP) offers unique opportunities by which to trace cell movements, cell signaling or gene expression dynamically in developing embryos. Two-photon laser scanning microscopy (TPLSM) is ideally suited to imaging cells in vivo due to its deeper tissue penetration and reduced phototoxicity; however, in TPLSM the excitation and emission spectra of GFP and its color variants [e.g., CyanFP (CFP); yellowFP (YFP)] are insufficiently distinct to be uniquely imaged by conventional means. To surmount such difficulties, we have combined the technologies of TPLSM and imaging spectroscopy to unambiguously identify CFP, GFP, YFP, and redFP (RFP) as well as conventional dyes, and have tested the approach in cell lines. In our approach, a liquid crystal tunable filter was used to collect the emission spectrum of each pixel within the TPLSM image. Based on the fluorescent emission spectra, supervised classification and linear unmixing analysis algorithms were used to identify the nature and relative amounts of the fluorescent proteins expressed in the cells. In a most extreme case, we have used the approach to separate GFP and fluorescein, separated by only 7 nm, and appear somewhat indistinguishable by conventional techniques. This approach offers the needed ability to concurrently image multiple colored, spectrally overlapping marker proteins within living cells.

  2. Testing and Comparison of Imaging Detectors for Electrons in the Energy Range 10–20 keV (United States)

    Matheson, J.; Moldovan, G.; Kirkland, A.; Allinson, N.; Abrahams, J. P.


    Interest in direct detectors for low-energy electrons has increased markedly in recent years. Detection of electrons in the energy range up to low tens of keV is important in techniques such as photoelectron emission microscopy (PEEM) and electron backscatter diffraction (EBSD) on scanning electron microscopes (SEMs). The PEEM technique is used both in the laboratory and on synchrotron light sources worldwide. The ubiquity of SEMs means that there is a very large market for EBSD detectors for materials studies. Currently, the most widely used detectors in these applications are based on indirect detection of incident electrons. Examples include scintillators or microchannel plates (MCPs), coupled to CCD cameras. Such approaches result in blurring in scintillators/phosphors, distortions in optical systems, and inefficiencies due the limited active area of MCPs. In principle, these difficulties can be overcome using direct detection in a semiconductor device. Growing out of a feasibility study into the use of a direct detector for use on an XPEEM, we have built at Rutherford Appleton Laboratory a system to illuminate detectors with an electron beam of energy up to 20 keV . We describe this system in detail. It has been used to measure the performance of a custom back-thinned monolithic active pixel sensor (MAPS), a detector based on the Medipix2 chip, and a commercial detector based on MCPs. We present a selection of the results from these measurements and compare and contrast different detector types.

  3. Phase transitions in biogenic amorphous calcium carbonate (United States)

    Gong, Yutao

    Geological calcium carbonate exists in both crystalline phases and amorphous phases. Compared with crystalline calcium carbonate, such as calcite, aragonite and vaterite, the amorphous calcium carbonate (ACC) is unstable. Unlike geological calcium carbonate crystals, crystalline sea urchin spicules (99.9 wt % calcium carbonate and 0.1 wt % proteins) do not present facets. To explain this property, crystal formation via amorphous precursors was proposed in theory. And previous research reported experimental evidence of ACC on the surface of forming sea urchin spicules. By using X-ray absorption near-edge structure (XANES) spectroscopy and photoelectron emission microscopy (PEEM), we studied cross-sections of fresh sea urchin spicules at different stages (36h, 48h and 72h after fertilization) and observed the transition sequence of three mineral phases: hydrated ACC → dehydrated ACC → biogenic calcite. In addition, we unexpectedly found hydrated ACC nanoparticles that are surrounded by biogenic calcite. This observation indicates the dehydration from hydrated ACC to dehydrated ACC is inhibited, resulting in stabilization of hydrated ACC nanoparticles. We thought that the dehydration was inhibited by protein matrix components occluded within the biomineral, and we designed an in vitro assay to test the hypothesis. By utilizing XANES-PEEM, we found that SM50, the most abundant occluded matrix protein in sea urchin spicules, has the function to stabilize hydrated ACC in vitro.

  4. Polarization-dependent Imaging Contrast (PIC) mapping reveals nanocrystal orientation patterns in carbonate biominerals

    Energy Technology Data Exchange (ETDEWEB)

    Gilbert, Pupa U.P.A., E-mail: [University of Wisconsin-Madison, Departments of Physics and Chemistry, Madison, WI 53706 (United States)


    Highlights: Black-Right-Pointing-Pointer Nanocrystal orientation shown by Polarization-dependent Imaging Contrast (PIC) maps. Black-Right-Pointing-Pointer PIC-mapping of carbonate biominerals reveals their ultrastructure at the nanoscale. Black-Right-Pointing-Pointer The formation mechanisms of biominerals is discovered by PIC-mapping using PEEM. -- Abstract: Carbonate biominerals are one of the most interesting systems a physicist can study. They play a major role in the CO{sub 2} cycle, they master templation, self-assembly, nanofabrication, phase transitions, space filling, crystal nucleation and growth mechanisms. A new imaging modality was introduced in the last 5 years that enables direct observation of the orientation of carbonate single crystals, at the nano- and micro-scale. This is Polarization-dependent Imaging Contrast (PIC) mapping, which is based on X-ray linear dichroism, and uses PhotoElectron Emission spectroMicroscopy (PEEM). Here we present PIC-mapping results from biominerals, including the nacre and prismatic layers of mollusk shells, and sea urchin teeth. We describe various PIC-mapping approaches, and show that these lead to fundamental discoveries on the formation mechanisms of biominerals.

  5. Design of an ultrahigh-energy-resolution and wide-energy-range soft X-ray beamline. (United States)

    Xue, L; Reininger, R; Wu, Y-Q; Zou, Y; Xu, Z-M; Shi, Y-B; Dong, J; Ding, H; Sun, J-L; Guo, F-Z; Wang, Y; Tai, R-Z


    A new ultrahigh-energy-resolution and wide-energy-range soft X-ray beamline has been designed and is under construction at the Shanghai Synchrotron Radiation Facility. The beamline has two branches: one dedicated to angle-resolved photoemission spectroscopy (ARPES) and the other to photoelectron emission microscopy (PEEM). The two branches share the same plane-grating monochromator, which is equipped with four variable-line-spacing gratings and covers the 20-2000 eV energy range. Two elliptically polarized undulators are employed to provide photons with variable polarization, linear in every inclination and circular. The expected energy resolution is approximately 10 meV at 1000 eV with a flux of more than 3 × 10(10) photons s(-1) at the ARPES sample positions. The refocusing of both branches is based on Kirkpatrick-Baez pairs. The expected spot sizes when using a 10 µm exit slit are 15 µm × 5 µm (horizontal × vertical FWHM) at the ARPES station and 10 µm × 5 µm (horizontal × vertical FWHM) at the PEEM station. The use of plane optical elements upstream of the exit slit, a variable-line-spacing grating and a pre-mirror in the monochromator that allows the influence of the thermal deformation to be eliminated are essential for achieving the ultrahigh-energy resolution.

  6. Carbon emissions in China

    Energy Technology Data Exchange (ETDEWEB)

    Liu, Zhu [Harvard Univ., Cambridge, MA (United States). Sustainability Science Program


    This study analyzes the spatial-temporal pattern and processes of China's energy-related carbon emissions. Based on extensive quantitative analysis, it outlines the character and trajectory of China's energy-related carbon emissions during the period 1995-2010, examining the distribution pattern of China's carbon emissions from regional and sectoral perspectives and revealing the driving factors of China's soaring emission increase. Further, the book investigates the supply chain carbon emissions (the carbon footprints) of China's industrial sectors. Anthropogenic climate change is one of the most serious challenges currently facing humankind. China is the world's largest developing country, top primary energy consumer and carbon emitter. Achieving both economic growth and environmental conservation is the country's twofold challenge. Understanding the status, features and driving forces of China's energy-related carbon emissions is a critical aspect of attaining global sustainability. This work, for the first time, presents both key findings on and a systematic evaluation of China's carbon emissions from energy consumption. The results have important implications for global carbon budgets and burden-sharing with regard to climate change mitigation. The book will be of great interest to readers around the world, as it addresses a topic of truly global significance.

  7. Quantitative high dynamic range beam profiling for fluorescence microscopy

    Energy Technology Data Exchange (ETDEWEB)

    Mitchell, T. J., E-mail:; Saunter, C. D.; O’Nions, W.; Girkin, J. M.; Love, G. D. [Centre for Advanced Instrumentation and Biophysical Sciences Institute, Department of Physics, Durham University, Durham DH1 3LE (United Kingdom)


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

  8. Single-molecule super-resolution microscopy reveals how light couples to a plasmonic nanoantenna on the nanometer scale. (United States)

    Wertz, Esther; Isaacoff, Benjamin P; Flynn, Jessica D; Biteen, Julie S


    The greatly enhanced fields near metal nanoparticles have demonstrated remarkable optical properties and are promising for applications from solar energy to biosensing. However, direct experimental study of these light-matter interactions at the nanoscale has remained difficult due to the limitations of optical microscopy. Here, we use single-molecule fluorescence imaging to probe how a plasmonic nanoantenna modifies the fluorescence emission from a dipole emitter. We show that the apparent fluorophore emission position is strongly shifted upon coupling to an antenna and that the emission of dyes located up to 90 nm away is affected by this coupling. To predict this long-ranged effect, we present a framework based on a distance-dependent partial coupling of the dye emission to the antenna. Our direct interpretation of these light-matter interactions will enable more predictably optimized, designed, and controlled plasmonic devices and will permit reliable plasmon-enhanced single-molecule nanoscopy.

  9. National Greenhouse Gas Emission Inventory (United States)

    U.S. Environmental Protection Agency — The National Greenhouse Gas Emission Inventory contains information on direct emissions of greenhouse gases as well as indirect or potential emissions of greenhouse...

  10. Managing Air Quality - Emissions Inventories (United States)

    This page describes the role of emission inventories in the air quality management process, a description of how emission inventories are developed, and where U.S. emission inventory information can be found.

  11. Visualization of early influenza A virus trafficking in human dendritic cells using STED microscopy.

    Directory of Open Access Journals (Sweden)

    Faezzah Baharom

    Full Text Available Influenza A viruses (IAV primarily target respiratory epithelial cells, but can also replicate in immune cells, including human dendritic cells (DCs. Super-resolution microscopy provides a novel method of visualizing viral trafficking by overcoming the resolution limit imposed by conventional light microscopy, without the laborious sample preparation of electron microscopy. Using three-color Stimulated Emission Depletion (STED microscopy, we visualized input IAV nucleoprotein (NP, early and late endosomal compartments (EEA1 and LAMP1 respectively, and HLA-DR (DC membrane/cytosol by immunofluorescence in human DCs. Surface bound IAV were internalized within 5 min of infection. The association of virus particles with early endosomes peaked at 5 min when 50% of NP+ signals were also EEA1+. Peak association with late endosomes occurred at 15 min when 60% of NP+ signals were LAMP1+. At 30 min of infection, the majority of NP signals were in the nucleus. Our findings illustrate that early IAV trafficking in human DCs proceeds via the classical endocytic pathway.

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

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


    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.

  13. Enhancement of field emission characteristics of carbon nanotubes on oxidation. (United States)

    Mathur, Ashish; Roy, Susanta Sinha; Ray, Sekhar Chandra; Hazra, Kiran Shankar; Hamilton, Jeremy; Dickinson, Calum; McLaughlin, James; Misra, Devi Shankar


    Vertically aligned multi-walled carbon nanotubes (CNTs) were grown on p-type silicon wafer using thermal chemical vapor deposition process and subsequently treated with oxygen plasma for oxidation. It was observed that the electron field emission (EFE) characteristics are enhanced. It showed that the turn-on electric field (E(TOE)) of CNTs decreased from 0.67 (untreated) to 0.26 V/microm (oxygen treated). Raman spectra showed that the numbers of defects are increased, which are generated by oxygen-treatment, and absorbed molecules on the CNTs are responsible for the enhancement of EFE. Scanning electron microscopy and Transmission electron microscopy images were used to identify the quality and physical changes of the nanotube morphology and surfaces; revealing the evidence of enhancement in the field emission properties after oxygen-plasma treatment.

  14. Schottky barrier measurements on individual GaAs nanowires by X-ray photoemission microscopy

    Energy Technology Data Exchange (ETDEWEB)

    Di Mario, Lorenzo [IMM-CNR, via del Fosso del Cavaliere 100, 00133 Rome (Italy); Turchini, Stefano, E-mail: [ISM-CNR, via del Fosso del Cavaliere 100, 00133 Rome (Italy); Zamborlini, Giovanni; Feyer, Vitaly [Peter Grünberg Institute (PGI-6) and JARA-FIT, Research Center Jülich, 52425 Jülich (Germany); Tian, Lin [IMM-CNR, via del Fosso del Cavaliere 100, 00133 Rome (Italy); Schneider, Claus M. [Peter Grünberg Institute (PGI-6) and JARA-FIT, Research Center Jülich, 52425 Jülich (Germany); Fakultät für Physik and Center for Nanointegration Duisburg-Essen (CENIDE), Universität Duisburg-Essen, D-47048 Duisburg (Germany); Rubini, Silvia [IOM-CNR, TASC Laboratory, Basovizza 34149, Trieste (Italy); Martelli, Faustino, E-mail: [IMM-CNR, via del Fosso del Cavaliere 100, 00133 Rome (Italy)


    Highlights: • The Schottky barrier at the interface between Cu and GaAs nanowires was measured. • Individual nanowires were investigated by X-ray Photoemission Microscopy. • The Schottky barrier at different positions along the nanowire was evaluated. - Abstract: We present measurements of the Schottky barrier height on individual GaAs nanowires by means of x-ray photoelectron emission microscopy (XPEEM). Values of 0.73 and 0.51 eV, averaged over the entire wires, were measured on Cu-covered n-doped and p-doped GaAs nanowires, respectively, in agreement with results obtained on bulk material. Our measurements show that XPEEM can become a feasible and reliable investigation tool of interface formation at the nanoscale and pave the way towards the study of size-dependent effects on semiconductor-based structures.

  15. Spatial distribution of Na+-K+-ATPase in dendritic spines dissected by nanoscale superresolution STED microscopy

    Directory of Open Access Journals (Sweden)

    Bondar Alexander


    Full Text Available Abstract Background The Na+,K+-ATPase plays an important role for ion homeostasis in virtually all mammalian cells, including neurons. Despite this, there is as yet little known about the isoform specific distribution in neurons. Results With help of superresolving stimulated emission depletion microscopy the spatial distribution of Na+,K+-ATPase in dendritic spines of cultured striatum neurons have been dissected. The found compartmentalized distribution provides a strong evidence for the confinement of neuronal Na+,K+-ATPase (α3 isoform in the postsynaptic region of the spine. Conclusions A compartmentalized distribution may have implications for the generation of local sodium gradients within the spine and for the structural and functional interaction between the sodium pump and other synaptic proteins. Superresolution microscopy has thus opened up a new perspective to elucidate the nature of the physiological function, regulation and signaling role of Na+,K+-ATPase from its topological distribution in dendritic spines.

  16. Emission control of InGaN nanocolumns grown by molecular-beam epitaxy on Si(111) substrates

    Energy Technology Data Exchange (ETDEWEB)

    Albert, S.; Bengoechea-Encabo, A.; Sanchez-Garcia, M. A.; Calleja, E. [ISOM and Departamento de Ingenieria Electronica, ETSI Telecomunicacion, Universidad Politecnica de Madrid, Ciudad Universitaria s/n, 28040 Madrid (Spain); Lefebvre, P. [ISOM and Departamento de Ingenieria Electronica, ETSI Telecomunicacion, Universidad Politecnica de Madrid, Ciudad Universitaria s/n, 28040 Madrid (Spain); Universite Montpellier 2, F-34095 Montpellier, Cedex 5 (France); Jahn, U.; Trampert, A. [Paul-Drude-Institut fuer Festkoeperelektronik, Hausvogteiplatz 5-7, 10117 Berlin (Germany)


    This work studies the effect of the growth temperature on the morphology and emission characteristics of self-assembled InGaN nanocolumns grown by plasma assisted molecular beam epitaxy. Morphology changes are assessed by scanning electron microscopy, while emission is measured by photoluminescence. Within the growth temperature range of 750 to 650 deg. C, an increase in In incorporation for decreasing temperature is observed. This effect allows tailoring the InGaN nanocolumns emission line shape by using temperature gradients during growth. Depending on the gradient rate, span, and sign, broad emission line shapes are obtained, covering the yellow to green range, even yielding white emission.

  17. Field emission properties of single crystal chromium disilicide nanowires

    Energy Technology Data Exchange (ETDEWEB)

    Valentin, L. A.; Carpena-Nunez, J.; Yang, D.; Fonseca, L. F. [Department of Physics, University of Puerto Rico, Rio Piedras Campus, P.O. Box 70377, San Juan, 00931 (Puerto Rico)


    The composition, crystal structure, and field emission properties of high-crystallinity chromium disilicide (CrSi{sub 2}) nanowires synthesized by a vapor deposition method have been studied. High resolution transmission electron microscopy, energy dispersive spectroscopy, and selected area electron diffraction studies confirm the single-crystalline structure and composition of the CrSi{sub 2} nanowires. Field emission measurements show that an emission current density of 0.1 {mu}A/cm{sup 2} was obtained at a turn-on electric field intensity of 2.80 V/{mu}m. The maximum emission current measured was 1.86 mA/cm{sup 2} at 3.6 V/{mu}m. The relation between the emission current density and the electric field obtained follows the Fowler-Nordheim equation, with an enhancement coefficient of 1140. The electrical conductivity of single nanowires was measured by using four-point-probe specialized microdevices at different temperatures, and the calculated values are close to those reported in previous studies for highly conductive single crystal bulk CrSi{sub 2}. The thermal tolerance of the nanowires was studied up to a temperature of 1100 Degree-Sign C. The stability of the field emission current, the I-E values, their thermal tolerance, and high electrical conductivity make CrSi{sub 2} nanowires a promising material for field emission applications.

  18. Field emission properties of single crystal chromium disilicide nanowires (United States)

    Valentín, L. A.; Carpena-Nuñez, J.; Yang, D.; Fonseca, L. F.


    The composition, crystal structure, and field emission properties of high-crystallinity chromium disilicide (CrSi2) nanowires synthesized by a vapor deposition method have been studied. High resolution transmission electron microscopy, energy dispersive spectroscopy, and selected area electron diffraction studies confirm the single-crystalline structure and composition of the CrSi2 nanowires. Field emission measurements show that an emission current density of 0.1 μA/cm2 was obtained at a turn-on electric field intensity of 2.80 V/μm. The maximum emission current measured was 1.86 mA/cm2 at 3.6 V/μm. The relation between the emission current density and the electric field obtained follows the Fowler-Nordheim equation, with an enhancement coefficient of 1140. The electrical conductivity of single nanowires was measured by using four-point-probe specialized microdevices at different temperatures, and the calculated values are close to those reported in previous studies for highly conductive single crystal bulk CrSi2. The thermal tolerance of the nanowires was studied up to a temperature of 1100 °C. The stability of the field emission current, the I-E values, their thermal tolerance, and high electrical conductivity make CrSi2 nanowires a promising material for field emission applications.

  19. 2011 NATA - Emissions Sources (United States)

    U.S. Environmental Protection Agency — This dataset includes all emissions sources that were modeled in the 2011 National Air Toxics Assessment (NATA), inlcluding point, nonpoint, and mobile sources, and...

  20. National Emission Inventory (NEI) (United States)

    U.S. Environmental Protection Agency — This data exchange allows states to submit data to the US Environmental Protection Agency's National Emissions Inventory (NEI). NEI is a national database of air...